Thermo Scientific Chromatography Columns and Consumables 2014-2015 Your world of chromatography predictability Thermo Scientific Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories 4 As a leader in LC column technology including silica, polymer and porous graphitic carbon manufacturing, bonded phase production and column packing for over 35 years, you can rely on the quality of Thermo Scientific HPLC products. Here we showcase our latest and most comprehensive range of innovative columns, accessories and equipment for fast LC, analytical HPLC and biomolecule separations. Introduction . . . . . . . . . . . . . . . . . . 4-001 Column Selection . . . . . . . . . . . . . . 4-002 Column Protection . . . . . . . . . . . . . 4-016 Columns for Fast and Analytical LC . . 4-022 Columns for Biomolecules . . . . . . . 4-110 LC Accessories . . . . . . . . . . . . . . . . 4-158 LC Equipment . . . . . . . . . . . . . . . . . 4-186 LC Reagents . . . . . . . . . . . . . . . . . . 4-193 LC and LC-MS Instrument Parts . . 4-209 Technical Information. . . . . . . . . . . 4-231 Featured Products Accucore Columns Syncronis Columns Hypersil GOLD Columns Ultimate core performance speed and selectivity combined Consistent, reproducible separations, column after column, time after time Outstanding peak shape for your separations PAGE 4-042 PAGE 4-074 PAGE 4-024 MAbPac Columns Acclaim PepMap Columns Viper Connectors Unrivalled resolution and efficiency in the analysis of protein variants The standard for peptide separations in proteomics Simple, dead volume free plumbing of HPLC and UHPLC systems PAGE 4-126 PAGE 4-149 LC Columns and Accessories Section Contents PAGE 4-158 www.thermoscientific.com/chromatography 4-001 LC Columns and Accessories LC Column Selection Information in the following section will help you make an informed decision on the appropriate HPLC column for your application, based on stationary phase use, analyte properties, LC-MS requirements or USP specifications. You will also find a useful table of Thermo Scientific phases with specifications, as well as recommended Thermo Scientific alternatives for other popular columns. Refer to the Advanced User Graphic (AUG) on the corresponding product page (illustrated to the right) for more help and information on general purpose column selection. The AUG will show you Hydrophobicity which gives the relative retention on the column. Generally, the higher the hydrophobicity, the greater the retention of neutral compounds and the higher the organic content in the mobile phase. A lower value indicates a need for higher aqueous mobile phases to achieve comparable retention and resolution. The recommended pH Range for the column is illustrated, outside of which column lifetimes will diminish. The Pore Size is shown, with larger pore size columns being more applicable to larger analytes such as proteins or peptides. The percentage Carbon Load is related to the hydrophobicity. Below the icon, you will see the particle sizes available, as well as the USP code. These graphics are designed to allow you to quickly compare the main characteristics of multiple stationary phases, allowing you to choose quickly the most appropriate stationary phase for your analysis. For additional help in column selection, please see the back cover to contact our expert Technical Support and tap into our expertise to help make the best choice for your application. Hydrophobicity Low High pH Range 0 2 to 8 14 Pore Size 0 80Å 300 Carbon Load (%) 0 5 Particle Size 2.6µm 25 USP L11 Advanced User Graphic (AUG) Common HPLC Phases and Their Uses Thermo Scientific Chromatography Columns and Consumables 2014-2015 Common Name 4-002 Alternative Name Functional Group Normal Phase Reverse Ion HILIC Phase Exchange • • Application Silica Silica -OH C1 SAS -(CH3) 3 • Least retentive of all alkyl group bonded phases for non-polar solutes. Typically used for moderately polar and multi-functional compounds. C4 Butyl -C 4H9 • Shorter retention than C8, C18. Separation of peptides and proteins. C8 MOS -C 8H17 • Less retentive than C18; normally used for small peptides and proteins, pharmaceuticals, steroids, environmental samples. C18 ODS -C18H37 • Most retentive of the alkyl-bonded phases. Used widely for pharmaceuticals, steroids, fatty acids, phthalates, environmental etc. Cyano CPS, CN -(CH2) 3CN • • Unique selectivity for polar compounds, more suitable than base silica for normal phase gradient separations. When used in reversed phase, the selectivity is different to that of the C8 and C18 phases. Useful for a wide range of pharmaceutical applications and for mixtures of very different solutes. • • Amino HILIC: Carbohydrate analysis and other polar compounds. Weak anion exchange: anions and organic acids. Normal Phase: Alternative selectivity to silica. Good for aromatics. APS -(CH2) 3NH2 -(CH3)C 6H5 • Aromatic compounds and moderately polar compounds. PFP -C 6F 5 • Extra selectivity and retention for halogenated, polar compounds and isomers. Phenyl Pentafluorophenyl Non-polar and moderately polar organic compounds. Diol -(CH 2) 2O CH 2 (CH 2 OH) 2 • • • • SCX Strong Cation Exchanger -RSO3H- • SAX Strong Anion Exchanger -RN+(CH3) 3 • AX Anion Exchanger Polyethyleneimine (PEI) -(CH2CH2NH-)n Porous graphitic carbon PGC 100% carbon • • • • Reversed Phase: Proteins, peptides. Normal Phase: Similar selectivity to silica, but less polar. Organic bases. Organic acids, nucleotides and nucleosides. Organic acids, nucleotides and oligonucleotides. Particularly useful for the separation of highly polar compounds that are difficult to retain using conventional silica based columns; separation of structurally similar compounds (e.g., isomers, diastereoisomers). Before beginning a new analysis, consider the physical and chemical properties of the analyte(s), the mode of analysis and how the analyte(s) will interact with the surface of the chromatographic phase. To aid column selection, the following guide may be useful. Non-polar Analyte Solubilty Analyte Chemical Properties Acidic pKa > 2 Non-polar Mode of Separation Recommended Phase Anion exchange / Reversed-phase mixed mode Reversed phase Polar solvents Moderately-polar Reversed phase Reversed phase Basic pKa < 10 Cation exchange / Reversed-phase mixed mode 4-069 Acclaim Trinity P1 4-091 Accucore C18 4-045 Accucore RP-MS 4-044 Accucore C8 4-046 Accucore C30 4-052 Accucore Phenyl-Hexyl 4-049 Acclaim 120 C18 4-061 Acclaim 120 C8 4-062 Acclaim C30 4-064 Acclaim Phenyl-1 4-063 Hypersil GOLD 4-026 Hypersil GOLD C8 4-027 Hypersil GOLD C4 4-028 Syncronis C18 4-075 Syncronis C8 4-076 Accucore aQ 4-047 Accucore Polar Premium 4-048 Accucore PFP 4-051 Accucore Phenyl-X 4-050 Acclaim PolarAdvantage 4-065 Acclaim PolarAdvantage II 4-066 Hypersil GOLD aQ 4-029 Hypersil GOLD PFP 4-030 Hypersil GOLD Phenyl 4-031 Syncronis aQ 4-077 Syncronis Phenyl 4-078 Accucore C18 4-045 Accucore RP-MS 4-044 Accucore C8 4-046 Accucore C30 4-052 Accucore Phenyl-Hexyl 4-049 Acclaim 120 C18 4-061 Acclaim 120 C8 4-062 Acclaim C30 4-064 Acclaim Phenyl-1 4-063 Hypersil GOLD 4-026 Hypersil GOLD C8 4-027 Hypersil GOLD C4 4-028 Syncronis C18 4-075 Syncronis C8 4-076 Acclaim Mixed-Mode WCX-1 4-070 Acclaim Trinity P1 4-091 www.thermoscientific.com/chromatography Column Selection Neutral Acclaim Mixed-Mode WAX-1 LC Columns and Accessories >> HPLC Stationary Phase Column Selection 4-003 LC Columns and Accessories HPLC Stationary Phase Column Selection continued Polar Analyte Solubilty Analyte Chemical Properties Mode of Separation Polar retention effect on graphite Anion exchange Acidic pKa < 2 HILIC Polar retention effect on graphite Polar solvents Neutral HILIC Polar retention effect on graphite Cation exchange Basic pKa > 10 HILIC Thermo Scientific Chromatography Columns and Consumables 2014-2015 Non-polar solvents 4-004 Normal phase Recommended Phase Hypercarb 4-084 Hypersil GOLD AX 4-034 Hypersil GOLD SAX 4-035 Accucore Urea-HILIC 4-054 Accucore 150-Amide-HILIC 4-146 Acclaim HILIC-10 4-067 Syncronis HILIC 4-081 Hypercarb 4-084 Accucore HILIC 4-053 Acclaim Mixed-Mode HILIC-1 4-068 Acclaim Mixed-Mode WAX-1 4-069 Acclaim Mixed-Mode WCX-1 4-070 Acclaim Trinity P1 4-091 Hypersil GOLD Amino 4-033 Hypersil GOLD HILIC 4-037 Syncronis Amino 4-079 Syncronis HILIC 4-081 Hypercarb 4-084 Acclaim Mixed-Mode WCX-1 4-070 Acclaim Trinity P1 4-091 Accucore Urea-HILIC 4-054 Accucore 150-Amide-HILIC 4-146 Acclaim HILIC-10 4-067 Syncronis HILIC 4-081 Hypersil GOLD Amino 4-033 Hypersil GOLD CN 4-032 Hypersil GOLD Silica 4-036 Syncronis Amino 4-079 Syncronis Silica 4-080 The above table recommends columns for the separation of small molecule analytes. Column recommendations for the separation of biomolecules can be found in the table on page 4-111 The Thermo Scientific range offers a broad array of column designs and stationary phases optimized for LC-MS applications. Use the following table to help you choose your column format to best meet your application needs. A variety of HPLC column hardware configurations are available, designed to give superior results for high speed, high sensitivity, high efficiency and convenience. A wide range of particle and monolithic stationary phases allows choices for optimized selectivity. Column Hardware Selection for LC-MS LC-MS Application High throughput analysis Column Hardware Design Javelin HTS columns High sensitivity analysis Acclaim PepMap nano, capillary and micro columns EASY-Spray columns Proteomics analysis Acclaim PepMap nano columns, Accucore nano columns, nanoViper PepSwift monolithic columns Various HPLC columns, throughout this LC section, can also be used for LC-MS application. Typical flow rates and MS source compatibility for these columns are shown in the table on the next page www.thermoscientific.com/chromatography Column Selection Acclaim PepMap nano columns, classic EASY-Column Description Direct-connection columns Slim design, 20mm length, 1mm to 4.6mm ID Nano, capillary and micro columns 0.075mm to 1mm ID; 50 to 250mm length nanoViper format offers fingertight dead volume free connection to 1000 bar Combined column/emitter design with nanoViper connection. Heated flexible silica columns 50µm and 75µm ID; 150 to 500mm length nanoViper offers fingertight dead volume free connection to 1000 bar Flexible silica columns 50μm and 75μm ID; 50 to 500mm length Trap column 20mm x 100μm ID Flexible silica columns 50μm and 75μm ID; 50 to 500mm length Trap column 20mm x 100μm ID Flexible silica columns 100mm x 75μm Trap column 20mm x 100μm ID Flexible fused silica columns, nanoViper connections 100µm to 500µm ID, 50 to 250mm length Trap columns 5mm x 200µm ID LC Columns and Accessories >> Column Format Selection for LC-MS 4-005 LC Columns and Accessories Column Selection for LC-MS continued Packed column selection for LC-MS Analyte Molecular Weight Sample Polarity Interface Relative Ionization Sensitivity Low Column ID (mm) Flow Rate (µL/min) Low 4.6, 4.0, 3.0 2000 – 200 High 2.1, 1.0 200 – 50 Low 4.6, 4.0, 3.0 2000 – 200 High 2.1, 1.0 200 – 50 Low 2.1, 1.0 200 – 50 1.0 – 0.3 50 – 5 Acclaim PepMap (RSLC) capillary and micro 0.2 – 0.05 2 – 0.2 EASY-Spray column, EASY-Column, Acclaim PepMap (RSLC) nano, Accucore nano 2.1, 1.0 200 – 50 1.0 – 0.3 50 – 5 Acclaim PepMap (RSLC) capillary and micro 0.2 – 0.05 2 – 0.2 EASY-Spray column, EASY-Column, Acclaim PepMap (RSLC) nano, Accucore nano 2.1, 1.0 200 – 50 1.0 – 0.3 50 – 5 Acclaim PepMap (RSLC) capillary and micro 0.2 – 0.05 2 – 0.2 EASY-Spray column, EASY-Column, Acclaim PepMap (RSLC) nano, Accucore nano APCI APCI < 1000 Da Medium ESI High High (or ionizable) Low ESI High Low > 1000 Da Column Hardware ESI High Javelin HTS, Analytical Analytical, Javelin HTS Javelin HTS, Analytical Analytical, Javelin HTS Analytical, Javelin HTS Analytical, Javelin HTS Analytical, Javelin HTS Monolith columns for LC-MS Analyte Molecular Weight Column ID (mm) Flow Rate (µL/min) < 1000 Da 0.1, 0.2, 0.5 0.7 – 25 PepSwift Monolith > 1000 Da 1.0 40 – 200 ProSwift Monolith Thermo Scientific Chromatography Columns and Consumables 2014-2015 ProSwift is also available in larger IDs for high throughput applications. 4-006 Column Hardware Description Octadecyl silane (C18) chemically bonded to porous or ceramic microparticles, 1.5 to 10μm in diameter, or a monolithic rod L3 Porous silica particles, 1.5 to 10μm in diameter, or a monolithic rod L7 Octyl silane chemically bonded to totally or superficially porous silica particles, 1.5 to 10μm in diameter, or a monolithic rod L8 An essentially monomolecular layer of aminopropylsilane chemically bonded to totally porous silica gel support, 1.5 to 10μm in diameter Nitrile groups chemically bonded to porous silica particles, 1.5 to 10μm in diameter Phenyl groups chemically bonded to porous silica particles, 1.5 to 10μm in diameter L10 L11 L13 Trimethylsilane chemically bonded to porous silica particles, 3 to 10μm in diameter L14 Silica gel having a chemically bonded, strongly basic quaternary ammonium anion exchange coating, 5 to 10μm in diameter Hexylsilane (C6) chemically bonded to totally porous silica particles, 3 to 10μm in diameter L15 Recommended Phase Acclaim 120 C18 Acclaim 300 C18 Accucore C18 Accucore aQ Accucore 150-C18 Accucore XL C18 BioBasic 18 Hypersil GOLD Hypersil GOLD aQ Acclaim PepMap 100 C18 Syncronis C18 Syncronis aQ Accucore HILIC Hypersil GOLD Silica Syncronis Silica Acclaim 120 C8 Accucore C8 Accucore XL C8 BioBasic 8 Hypersil GOLD C8 Acclaim PepMap 100 C8 Syncronis C8 Hypersil GOLD Amino Syncronis Amino Page 4-061 4-142 4-045 4-047 4-144 4-056 4-140 4-026 4-029 4-149 4-075 4-077 4-053 4-036 4-080 4-061 4-046 4-057 4-140 4-027 4-150 4-076 4-033 4-079 Hypersil GOLD CN 4-032 Accucore Phenyl-Hexyl Hypersil GOLD Phenyl Syncronis Phenyl BETASIL C1 Hypersil SAS Hypersil GOLD SAX Hypersil SAX 4-049 4-031 4-078 Inquire 4-107 4-035 4-109 BETASIL C6 Inquire www.thermoscientific.com/chromatography Column Selection USP Code L1 LC Columns and Accessories >> HPLC Column Selection by U.S. Pharmacopeia Specifications* 4-007 LC Columns and Accessories HPLC Column Selection by U.S. Pharmacopeia Specifications* continued USP Code L17 L19 L20 L26 L33 L34 L43 L46 L50 L52 Thermo Scientific Chromatography Columns and Consumables 2014-2015 L58 4-008 L59 L60 L62 L78 Description Strong cation exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the hydrogen form, 6 to 12μm in diameter Strong cation exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the calcium form, about 9μm in diameter Recommended Phase HyperREZ XP Carbohydrate H + HyperREZ XP Organic Acids Page 4-156 4-156 HyperREZ XP Carbohydrate Ca2+ HyperREZ XP Sugar Alcohols BETASIL Diol 4-156 Dihydroxypropane groups chemically bonded to porous silica or hybrid particles, 1.5 to 10μm in diameter Butyl silane chemically bonded to totally porous silica Accucore 150-C4 particles, 1.5 to 10μm in diameter BioBasic 4 Hypersil GOLD C4 Acclaim PepMap 300 C4 Packing having the capacity to separate dextrans BioBasic SEC 120 by molecular size over a range of 4,000 to 500,000 BioBasic SEC 300 daltons. It is spherical, silica-based, and processed BioBasic SEC 1000 to provide pH stability MAbPac SEC-1 Strong cation exchange resin consisting of sulfonated HyperREZ XP Carbohydrate Pb2+ cross-linked styrene-divinylbenzene copolymer in the lead form, 7 to 9μm in diameter Pentafluorophenyl groups chemically bonded to silica Accucore PFP particles by a propyl spacer, 1.5 to 10μm in diameter Hypersil GOLD PFP Polystyrene/divinylbenzene substrate agglomerated OmniPac PAX-100 with quaternary amine functionalized latex beads about 9 to 11um in diameter Multifunction resin with reverse-phase retention and OmniPac PAX-500 strong anion-exchange functionalities. The resin consists of ethylvinylbenzene, 55% cross-linked with divinylbenzene copolymer, 3 to 15µm in diameter, and a surface area of not less than 350m2 per g. Substrate is coated with quaternary ammonium functionalized latex particles consisting of styrene cross-linked with divinylbenzene A strong cation exchange resin made of porous silica BioBasic SCX with sulfopropyl groups 5 to 10μm in diameter Strong cation exchange resin consisting of sulfonated HyperREZ XP Carbohydrate Na+ cross-linked styrene-divinylbenzene copolymer in the sodium form, about 6 to 30μm in diameter Packing for the size exclusion separation of proteins BioBasic SEC 300 (5μm) (separation by molecular weight) over the range of MAbPac SEC-1 5 to 7,000 kDa. The packing is spherical, 1.5 to 10um, silica or hybrid packing with a hydrophilic coating Spherical, porous silica gel, 10μm or less in diameter, Acclaim PolarAdvantage the surface of which has been covalently modified Acclaim PolarAdvantage II with alkyl amide groups and endcapped Accucore Polar Premium C30 silane bonded phase on a fully porous spherical Acclaim C30 silica, 3 to 15µm in diameter Accucore C30 A silane ligand that consists of both reversed-phase Acclaim Mixed-Mode WAX-1 (an alkyl chain longer than C8) and anion-exchange (primary, secondary, tertiary or quaternary amino groups) functional groups chemically bonded to porous or non-porous or ceramic micro-particles, 1.0 to 50μm in diameter or a monolithic rod 4-156 Inquire 4-145 4-140 4-028 4-149 4-139 4-139 4-139 4-129 4-156 4-051 4-030 4-103 4-103 4-141 4-156 4-139 4-129 4-065 4-066 4-048 4-064 4-052 4-069 *These are the recommended Thermo Scientific HPLC columns for various USP categories although other columns for each category are also available. The tables below list Thermo Scientific HPLC sorbents offered for small molecules separations. For Thermo Scientific phases for the separations of biomolecules see page 4-112. Please also refer to the Advanced User Graphic (AUG) for each HPLC phase on the pages indicated. Phase Particle Type Pore Nominal Surface % Endcapping Carbon Size (Å) Area (m2/g) USP Code Phase Code Page 2.2, 3, 5 3 2.2, 3, 5 3 3, 5 2.2, 3, 5 2.2, 3, 5 3 3 3 3 3 3, 5 3, 5 3, 5 3, 5 3, 5 3, 5 3, 5 2.2 5, 7 120 300 120 120 200 120 120 120 300 300 300 120 120 120 120 120 120 120 120 120 300 300 100 300 300 200 300 300 300 100 100 300 300 300 300 300 300 300 300 300 300 1000 18 8 11 13 13 16 16 8 – – – – – – – – – – – – – Yes Yes Yes Yes Proprietary Yes Yes Yes Proprietary Proprietary Proprietary Proprietary Proprietary Proprietary Yes Yes Yes Yes Yes Yes – L1 L1 L7 L11 L62 L60 L60 – – – – – – L78 – – – – – – – – – – – – – – – – – – – – – – – – – – – – 4-061 4-142 4-062 4-063 4-064 4-065 4-066 4-067 4-091 4-092 4-099 4-068 4-069 4-070 4-093 4-094 4-095 4-097 4-100 4-101 4-088 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 80 80 80 80 150 80 80 80 150 80 80 130 130 130 130 90 130 130 130 90 130 130 7 9 5 9 8 5 6 5 5 – – Yes Yes Yes Polar Yes Yes Yes Yes Yes – – – L1 L7 L1 L60 L11 – L43 L62 L3 – 176 171 172 173 280 179 279 174 278 175 277 4-044 4-045 4-046 4-047 4-048 4-049 4-050 4-051 4-052 4-053 4-054 4 4 80 80 90 90 7 4 Yes Yes L1 L7 741 742 4-056 4-057 www.thermoscientific.com/chromatography Column Selection Acclaim Phases 120 C18 Spherical, fully porous silica 300 C18 Spherical, fully porous silica 120 C8 Spherical, fully porous silica Phenyl-1 Spherical, fully porous silica C30 Spherical, fully porous silica PA Spherical, fully porous silica PA II Spherical, fully porous silica HILIC-10 Spherical, fully porous silica Trinity P1 Nano polymer silica hybrid Trinity P2 Nano polymer silica hybrid Trinity Q1 Nano polymer silica hybrid Mixed Mode HILIC-1 Spherical, fully porous silica Mixed Mode WAX-1 Spherical, fully porous silica Mixed Mode WCX-1 Spherical, fully porous silica Organic Acid Spherical, fully porous silica Surfactant Spherical, fully porous silica Surfactant Plus Spherical, fully porous silica Explosives Spherical, fully porous silica Carbamate Spherical, fully porous silica Carbonyl C18 Spherical, fully porous silica SEC Spherical, resin Accucore Phases RP-MS Spherical, solid core silica C18 Spherical, solid core silica C8 Spherical, solid core silica aQ Spherical, solid core silica Polar Premium Spherical, solid core silica Phenyl-Hexyl Spherical, solid core silica Phenyl-X Spherical, solid core silica PFP Spherical, solid core silica C30 Spherical, solid core silica HILIC Spherical, solid core silica Urea-HILIC Spherical, solid core silica Accucore XL Phases C18 Spherical, solid core silica C8 Spherical, solid core silica Particle Size (µm) LC Columns and Accessories >> Thermo Scientific HPLC Phases 4-009 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-010 Thermo Scientific HPLC Phases continued Phase Particle Type Hypercarb Phase Hypercarb Spherical, porous graphitic carbon Hypersil Phases ODS (C18) Spherical, fully porous silica ODS-2 (C18) Spherical, fully porous silica MOS (C8) Spherical, fully porous silica MOS-2 (C8) Spherical, fully porous silica SAS (C1) Spherical, fully porous silica Phenyl Spherical, fully porous silica Phenyl-2 Spherical, fully porous silica CPS Spherical, fully porous silica CPS-2 Spherical, fully porous silica APS-2 Spherical, fully porous silica Silica Spherical, fully porous silica SAX Spherical, fully porous silica Hypersil BDS Phases C18 Spherical, fully porous silica C8 Spherical, fully porous silica Phenyl Spherical, fully porous silica Cyano Spherical, fully porous silica Hypersil GOLD Phases C18 selectivity Spherical, fully porous silica C8 Spherical, fully porous silica C4 Spherical, fully porous silica aQ Spherical, fully porous silica PFP Spherical, fully porous silica Phenyl Spherical, fully porous silica CN (Cyano) Spherical, fully porous silica Amino Spherical, fully porous silica AX Spherical, fully porous silica SAX Spherical, fully porous silica Silica Spherical, fully porous silica HILIC Spherical, fully porous silica Hypersil Green Phase PAH Spherical, fully porous silica Syncronis Phases C18 Spherical, fully porous silica C8 Spherical, fully porous silica aQ Spherical, fully porous silica Phenyl Spherical, fully porous silica Amino Spherical, fully porous silica Silica Spherical, fully porous silica HILIC Spherical, fully porous silica Particle Size (µm) Pore Nominal Surface % Endcapping Carbon Size (Å) Area (m2/g) USP Code Phase Code Page 3, 5 250 120 100 – – 350 4-084 3, 5 3, 5 3, 5 5 5 5 5 3, 5 5 3, 5 3, 5 5 120 80 120 120 120 120 120 120 120 120 120 120 170 220 170 170 170 170 170 170 170 170 170 170 10 11 6.5 6.5 2.5 5 5 4 4 1.9 – 2.5 Yes Yes No Yes Yes No Yes No Yes No – Yes L1 L1 L7 L7 L13 L11 L11 L10 L10 L8 L3 L14 301 316 302 303 305 309 319 308 318 307 300 341 4-106 4-107 4-107 4-107 4-107 4-108 4-108 4-108 4-108 4-108 4-109 4-109 2.4, 3, 5 2.4, 3, 5 3, 5 3, 5 130 130 130 130 170 170 170 170 11 7 5 4 Yes Yes Yes Yes L1 L7 L11 L10 281 282 289 288 4-105 4-105 4-105 4-105 1.9, 3, 5, 12 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 1.9, 3, 5 175 175 175 175 175 175 175 175 175 175 175 175 220 220 220 220 220 220 220 220 220 220 220 220 10 8 5 12 8 8.5 4 2 6 2.5 – 6 Yes Yes Yes Polar Yes Yes Yes Yes No Yes – No L1 L7 L26 L1 L43 L11 L10 L8 – L14 L3 – 250 252 255 253 254 259 258 257 261 263 251 265 4-026 4-027 4-028 4-029 4-030 4-031 4-032 4-033 4-034 4-035 4-036 4-037 3, 5 120 170 13.5 Yes – 311 4-102 1.7, 3, 5 1.7, 3, 5 1.7, 3, 5 1.7, 3, 5 1.7, 3, 5 1.7, 3, 5 1.7, 3, 5 100 100 100 100 100 100 100 320 320 320 320 320 320 320 16 10 19 11 4 – 5 Yes Yes Polar Yes Yes – – L1 L7 L1 L11 L8 L3 – 971 972 973 979 977 970 975 4-075 4-076 4-077 4-078 4-079 4-080 4-081 To find a suitable Thermo Scientific alternative to another manufacturer’s columns, refer to the selection guide below. The Thermo Scientific alternative phases are selected based on a combination of physical and chemical similarities as well as mode of retention. These alternatives are not guaranteed to The following table is not complete in terms of manufacturer or products offered. Although every effort is made to ensure that the product information provided is as accurate as possible, some errors may occur in collation and transcription. We cannot accept any responsibility for the use of the following information. Manufacturer ACT ACT ACT ACT ACT ACT ACT Waters Waters Waters Waters Waters Waters Phenomenex Phenomenex Phenomenex Phenomenex Grace Grace Grace Grace Grace Grace Bio-Rad Bio-Rad Bio-Rad Bio-Rad Bio-Rad Bio-Rad Phenomenex Supelco Supelco Supelco Supelco Supelco Supelco Supelco Pore Size (Å) 100 100 100 100 300 300 300 130 100 130 130 130 100 100 200 200 200 190 100 100 190 190 190 – – – – – – 125 100 100 90 90 90 90 90 Area (m2/g) 300 300 300 300 100 100 100 185 230 185 185 185 230 200 25 25 25 200 450 450 200 200 200 – – – – – – 320 450 450 150 150 150 150 150 % Carbon 15.5 9 9.5 14 9 5 2.6 – 15 18 13 15 11 – – – – 12 20 24 8 4 – – – – – – – 15 25 15 – – – – – Recommended Thermo Scientific Alternative Syncronis C18 Syncronis C8 Syncronis Phenyl Syncronis aQ BioBasic 18 BioBasic 8 BioBasic 4 Hypersil GOLD Silica (1.9µm) Hypersil GOLD (1.9µm) Hypersil GOLD (1.9µm) Hypersil GOLD C8 (1.9µm) Hypersil GOLD Phenyl (1.9µm) Hypersil GOLD aQ (1.9µm) Accucore 150-C18 Accucore 150-C18 Accuccore 150-C4 Accuccore 150-C4 Hypersil GOLD Hypersil GOLD aQ Syncronis C18 Hypersil GOLD C8 Hypersil GOLD CN Hypersil GOLD Silica HyperREZ XP Carbohydrate Ca2+ HyperREZ XP Carbohydrate H+ HyperREZ XP Carbohydrate Ca2+ HyperREZ XP Carbohydrate H+ HyperREZ XP Carbohydrate Na+ HyperREZ XP Carbohydrate Pb2+ Hypersil GOLD aQ Syncronis C18 Syncronis C8 Accucore C18 Accucore C8 Accucore PFP Accucore HILIC Accucore Phenyl-Hexyl Page 4-075 4-076 4-078 4-077 4-140 4-140 4-140 4-036 4-026 4-026 4-027 4-031 4-029 4-144 4-144 4-145 4-145 4-026 4-029 4-075 4-027 4-032 4-036 4-156 4-156 4-156 4-156 4-156 4-156 4-029 4-075 4-076 4-045 4-046 4-051 4-053 4-049 Supelco 90 150 – Accucore Polar Premium 4-048 Supelco 160 80 – Accucore 150-C18 4-144 Supelco 100 450 19 Syncronis Phenyl 4-078 www.thermoscientific.com/chromatography Column Selection Phase ACE C18 ACE C8 ACE Phenyl ACE AQ ACE C18-300 ACE C8-300 ACE C4-300 ACQUITY UPLC ™ BEH HILIC ACQUITY UPLC HSS C18 ACQUITY UPLC BEH C18 ACQUITY UPLC BEH C8 ACQUITY UPLC BEH Phenyl ACQUITY UPLC HSS T3 Aeris PEPTIDE XB-C18 Aeris WIDEPORE XB-C18 Aeris WIDEPORE XB-C8 Aeris WIDEPORE XB-C4 Alltima™ HP C18 Alltima HP C18 AQ Alltima HP C18 HiLoad Alltima HP C8 Alltima HP CN Alltima HP Silica Aminex™ HPX42C Aminex HPX72S Aminex HPX87C Aminex HPX87H Aminex HPX87N Aminex HPX87P AQUA™ C18 Ascentis™ C18 Ascentis C8 Ascentis Express C18 Ascentis Express C8 Ascentis Express F5 Ascentis Express HILIC Ascentis Express Phenyl-Hexyl Ascentis Express RP-Amide Ascentis Express Peptide ES-C18 Ascentis Phenyl provide the same retention or selectivity, but should be suitably similar in character to allow a similar or improved separation to be achieved with some method optimization. The user should refer to the individual phase information to ensure that the characteristics of the alternative match the requirements of their separation. LC Columns and Accessories >> HPLC Column Selection by Manufacturer 4-011 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-012 HPLC Column Selection by Manufacturer continued Phase Atlantis™ dC18 Atlantis T3 Atlantis HILIC Silica Atlantis dC18 Capcell Core C18 Capcell Pak C18 AQ Cortecs C18 Cortecs C18+ Cortecs HILIC Discovery™ BIO Wide Pore C18 Discovery BIO Wide Pore C8 Discovery C18 Discovery C8 Discovery Cyano Gemini ™ C18 Halo C18 Halo C8 Halo HILIC Halo PFP Halo Phenyl-Hexyl Halo RP-Amide HALO Peptide ES-C18 Inertsil™ C4 Inertsil C8 Inertsil ODS3V Inertsil Phenyl Inertsil Silica J’Sphere M80 Jupiter™ C18 Jupiter C4 Kinetex C18 Kinetex C8 Kinetex HILIC Kinetex PFP Kinetex Phenyl-Hexyl Kinetex XB-C18 Kromasil C18 Kromasil C4 Kromasil Silica LiChrospher™ CN LiChrospher Diol LiChrospher NH 2 LiChrospher RP 18 LiChrospher RP-18e LiChrospher RP-8 LiChrospher RP-8e Luna™ C18 (2) Luna C8 (2) Luna CN Luna HILIC Luna NH2 Luna PFP (2) Manufacturer Waters Waters Waters Waters Shiseido Shiseido Waters Waters Waters Supelco Supelco Supelco Supelco Supelco Phenomenex AMT AMT AMT AMT AMT AMT AMT GL Sciences GL Sciences GL Sciences GL Sciences GL Sciences YMC Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Akzo-Nobel Akzo-Nobel Akzo-Nobel Merck Merck Merck Merck Merck Merck Merck Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Pore Size (Å) 100 100 100 100 90 120 90 90 90 300 300 180 180 180 110 90 90 90 90 90 90 160 150 150 100 150 150 80 300 300 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 200 100 100 Area (m2/g) 330 300 300 330 150 300 100 100 100 – – 200 200 200 375 150 150 150 150 150 150 80 320 320 450 320 320 – 170 170 – – – – – – 340 340 340 350 350 350 350 350 350 350 400 400 400 200 400 400 % Carbon 12 14 – 12 7 11 6.6 5.7 – – – 14 – – 14 – – – – – – – 8 11 15 10 – 14 13 5 12 10 – 9 – 12 19 8 – 7 – 5 21 22 13 13 18 14 – 5.7 10 5.7 Recommended Thermo Scientific Alternative Acclaim Polar Advantage II Hypersil GOLD Hypersil GOLD Silica Hypersil GOLD aQ Accucore C18 Acclaim Polar Advantage II Accucore C18 Accucore Polar Premium Accucore HILIC BioBasic 18 BioBasic 8 Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD CN Hypersil GOLD Accucore C18 Accucore C8 Accucore HILIC Accucore PFP Accucore Phenyl-Hexyl Accucore Polar Premium Accucore 150-C18 Hypersil GOLD C4 Syncronis C8 Syncronis C18 Syncronis Phenyl Syncronis Silica Acclaim PolarAdvantage II BioBasic 18 BioBasic C4 Accucore C18 Accucore C8 Accucore HILIC Accucore PFP Accucore Phenyl-Hexyl Accucore C18 Syncronis C18 Hypersil GOLD C4 Syncronis Silica Hypersil GOLD CN BETASIL Diol Syncronis Amino Syncronis C18 Syncronis C18 Syncronis C8 Syncronis C8 Syncronis C18 Syncronis C8 Hypersil GOLD CN BETASIL Diol Syncronis Amino Hypersil GOLD PFP Page 4-066 4-026 4-036 4-029 4-045 4-066 4-045 4-048 4-053 4-140 4-140 4-026 4-027 4-032 4-026 4-045 4-046 4-053 4-051 4-049 4-048 4-144 4-028 4-076 4-075 4-078 4-080 4-066 4-140 4-140 4-045 4-046 4-053 4-051 4-049 4-045 4-075 4-028 4-080 4-032 Inquire 4-079 4-075 4-075 4-076 4-076 4-075 4-076 4-032 Inquire 4-079 4-030 Pore Size (Å) 100 100 125 125 125 125 60 60 60 60 60 110 110 110 110 90 90 90 100 100 100 100 100 100 100 100 100 100 300 300 300 300 300 120 120 120 120 140 140 140 140 140 120 120 120 120 120 100 300 120 120 120 120 Area (m2/g) 400 400 330 330 330 330 120 120 120 120 120 340 340 340 340 130 130 130 350 350 350 350 350 350 350 350 350 350 100 100 100 100 100 170 170 170 170 – – – – – 170 170 170 170 170 – – 120 120 120 130 % Carbon – – 10 – 4 – 7 – – 5 – 18 18 7 14 7.5 4.5 3 17 24 – 9 – – 4 – 7 10 7 – – – – 2 10 4 5 11 11 6 4 5 – 2 5 3 – 20 2 8 5 7.5 – Recommended Thermo Scientific Alternative BioBasic SCX Syncronis Silica Hypersil GOLD Hypersil GOLD CN Hypersil APS-2 Hypersil GOLD Phenyl Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD CN Hypersil GOLD Phenyl Hypersil GOLD Silica Syncronis C18 Syncronis C18 Hypersil GOLD CN Syncronis aQ Accucore C18 Accucore Phenyl-Hexyl Accucore PFP Syncronis C18 Syncronis C18 Syncronis Phenyl Syncronis C8 Hypersil GOLD CN Hypersil SAX Syncronis Amino BETASIL Diol BioBasic SCX Hypersil GOLD SAX BioBasic 18 BioBasic 4 BioBasic Phenyl BioBasic 8 BioBasic CN Hypersil SAS Hypersil GOLD Hypersil GOLD C4 Hypersil GOLD CN Hypersil GOLD Hypersil GOLD (1.9µm) Hypersil GOLD C8 Hypersil GOLD CN Hypersil GOLD Phenyl Hypersil GOLD Hypersil GOLD Amino Hypersil GOLD Phenyl Hypersil GOLD SAX Hypersil GOLD Silica Syncronis C18 BioBasic 4 Accucore C18 Accucore C8 Accucore C18 Accucore aQ Page 4-141 4-080 4-026 4-032 4-108 4-031 4-026 4-027 4-032 4-031 4-036 4-075 4-075 4-032 4-077 4-045 4-049 4-051 4-075 4-075 4-078 4-076 4-032 4-109 4-079 Inquire 4-141 4-035 4-140 4-140 Inquire 4-140 Inquire 4-107 4-026 4-028 4-032 4-026 4-026 4-027 4-032 4-031 4-026 4-033 4-031 4-035 4-036 4-075 4-140 4-045 4-046 4-045 4-047 www.thermoscientific.com/chromatography Column Selection Manufacturer Phenomenex Phenomenex Waters Waters Waters Waters Waters Waters Waters Waters Waters Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Macherey-Nagel Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Restek Agilent Agilent Agilent Agilent LC Columns and Accessories >> Phase Luna SCX Luna Silica (2) µBondapak™ C18 µBondapak CN µBondapak NH2 µBondapak Phenyl Nova-Pak™ (HR) C18 Nova-Pak C8 Nova-Pak CN Nova-Pak Phenyl Nova-Pak Silica NUCLEODUR ™ C18 EC NUCLEODUR C18 Gravity NUCLEODUR CN NUCLEODUR Pyramid Nucleoshell™ RP 18 Nucleoshell Phenyl-Hexyl Nucleoshell PFP Nucleosil™ 100 C18 Nucleosil 100 C18 AB Nucleosil 100 C 6 H 5 Nucleosil 100 C8 Nucleosil 100 CN Nucleosil 100 N(CH 3) 2 Nucleosil 100 NH2 Nucleosil 100 OH Nucleosil 100 SA Nucleosil 100 SB Nucleosil 300 C18 Nucleosil 300 C4 Nucleosil 300 C 6 H 5 Nucleosil 300 C8 Nucleosil 300 CN Pinnacle™ C1 Pinnacle C18 Pinnacle C4 Pinnacle CN Pinnacle DB C18 Pinnacle DB C18 1.9µm Pinnacle DB C8 Pinnacle DB Cyano Pinnacle DB Phenyl Pinnacle IBD Pinnacle NH 2 Pinnacle Phenyl Pinnacle SAX Pinnacle Silica Pinnacle Ultra C18 Pinnacle Wide Pore C4 Poroshell™ 120 EC-C18 Poroshell 120 EC-C8 Poroshell 120 SB-C18 Poroshell 120 SB-Aq 4-013 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-014 HPLC Column Selection by Manufacturer continued Phase Poroshell 120 Phenyl-Hexyl Poroshell 120 Bonus-RP Poroshell SB-C18 Poroshell Extend-C18 Poroshell 300 SB-C8 Poroshell 300 SB-C3 Primesep™ Prodigy™ C8 Prodigy ODS2 Prodigy ODS-3 Prodigy ODS-3V Prodigy Phenyl-3 Purospher™ RP-18 Purospher STAR-8e Purospher STAR RP-18e Waters™ Spherisorb™ C1 Waters Spherisorb C6 Waters Spherisorb C8 Waters Spherisorb CN Waters Spherisorb NH2 Waters Spherisorb ODS1 Waters Spherisorb ODS2 Waters Spherisorb ODSB Waters Spherisorb Phenyl Waters Spherisorb SAX Waters Spherisorb SCX Waters Spherisorb W (silica) SunFire™ C18 SunFire C8 SunShell™ C18 SunShell C8 SunShell PFP Supelcosil™ LC-1 Supelcosil LC-18 Supelcosil LC-18DB Supelcosil LC-8 Supelcosil LC-CN Supelcosil LC-NH2 Supelcosil LC-Si Symmetry™ C18 Symmetry C8 Synergi™ Hydro-RP TSKgel™ G2000SW (incl XL) TSKgel Octyl-80TS TSKgel ODS-120A TSKgel ODS-120T TSKgel ODS-80TM TSKgel Super Octyl TSKgel Super ODS TSKgel Super Phenyl TSKgel SuperSW3000 Manufacturer Agilent Agilent Agilent Agilent Agilent Agilent SieLC Phenomenex Phenomenex Phenomenex Phenomenex Phenomenex Merck Merck Merck Waters Waters Waters Waters Waters Waters Waters Waters Waters Waters Waters Waters Waters Waters ChromaNik ChromaNik ChromaNik Supelco Supelco Supelco Supelco Supelco Supelco Supelco Waters Waters Phenomenex Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Tosoh Pore Size (Å) 120 120 300 300 300 300 – 150 150 100 100 100 60 120 120 80 80 80 80 80 80 80 80 80 80 80 80 90 90 90 90 90 120 120 120 120 120 120 120 100 100 80 125 80 120 120 80 110 110 110 250 Area (m2/g) 130 130 45 45 45 45 – 310 310 450 450 450 500 300 300 200 200 200 200 200 200 200 200 200 200 200 200 340 340 150 150 150 170 170 170 170 170 170 170 335 335 475 – 200 200 200 200 – – – – % Carbon 9 9.5 2.8 4 1.5 1.1 – 13 18 16 16 10 – – – 2 5 6 3 2 6 12 12 3 – – – 16 16 7 4.5 4.5 – 11 11 – – – – 19 12 19 – 11 22 22 15 5 8 3 – Recommended Thermo Scientific Alternative Accucore Phenyl-Hexyl Accucore Polar Premium Accucore 150-C18 Accucore 150-C18 Accucore 150-C4 Accucore 150-C4 Acclaim Mixed-Mode Columns Syncronis C8 Syncronis C18 Syncronis C18 Hypersil GOLD Syncronis Phenyl Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD Hypersil SAS BETASIL C6 Hypersil GOLD C8 Hypersil GOLD CN Hypersil APS-2 Hypersil GOLD Hypersil GOLD Hypersil GOLD Hypersil GOLD Phenyl Hypersil SAX BioBasic SCX Hypersil GOLD Silica Syncronis C18 Syncronis C8 Accucore C18 Accucore C8 Accucore PFP Hypersil SAS Hypersil GOLD Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD CN Hypersil GOLD Amino Hypersil GOLD Silica Syncronis C18 Syncronis C8 Syncronis aQ BioBasic SEC 120 Hypersil GOLD C8 Hypersil GOLD Syncronis C18 Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD Hypersil GOLD Phenyl BioBasic SEC 300 Page 4-049 4-048 4-144 4-144 4-145 4-145 4-068 4-076 4-075 4-075 4-026 4-078 4-026 4-027 4-026 4-107 Inquire 4-027 4-032 4-108 4-026 4-026 4-026 4-031 4-109 4-141 4-036 4-075 4-076 4-045 4-046 4-051 4-107 4-026 4-026 4-027 4-032 4-033 4-036 4-075 4-076 4-077 4-139 4-027 4-026 4-075 4-026 4-027 4-026 4-031 4-139 Pore Size (Å) 90 90 Area (m2/g) – 250 % Carbon – – Recommended Thermo Scientific Alternative Page Hypersil GOLD 4-026 Hypersil GOLD 4-026 Grace Grace Grace Grace Grace Grace 300 300 300 300 300 300 – – – – – 70 – – – – – – BioBasic 18 BioBasic 18 BioBasic 8 BioBasic 4 BioBasic 18 BioBasic 18 4-140 4-140 4-140 4-140 4-140 4-140 Waters Waters Waters Waters Waters Waters YMC YMC YMC YMC YMC YMC YMC YMC YMC YMC YMC YMC YMC Agilent Agilent Agilent Agilent Agilent Agilent Agilent Agilent – – 130 – 125 125 – 120 120 120 120 120 120 120 300 120 120 120 120 80 80 80 95 95 95 95 80 – – 185 – 180 180 – 300 300 300 300 – 300 300 150 300 350 – 300 180 180 180 160 160 160 160 180 – – – – 16 12 – 7 10 7 – – 16 17 6 9 16 – 4 10 8 8 8 6 8 6 10 Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD Silica Hypersil GOLD Phenyl Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD C8 HyPURITY C4 Acclaim C8 Hypersil GOLD CN BETASIL Diol Hypersil GOLD Amino Syncronis aQ Syncronis C18 BioBasic 18 Syncronis Phenyl Syncronis C18 Syncronis Silica BETASIL C1 Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD Phenyl Hypersil GOLD Hypersil GOLD C8 Hypersil GOLD (1.9µm) Hypersil GOLD C8 (1.9µm) Hypersil GOLD (1.9µm) 4-026 4-027 4-036 4-031 4-026 4-027 4-027 4-109 4-062 4-032 Inquire 4-033 4-077 4-075 4-140 4-078 4-075 4-080 Inquire 4-026 4-027 4-031 4-026 4-027 4-026 4-027 4-026 Agilent 80 180 7.5 Hypersil GOLD C8 (1.9µm) 4-027 Agilent Agilent Agilent Agilent Agilent Agilent Agilent Agilent 80 80 80 300 80 300 80 80 180 180 180 45 180 45 180 180 4 – 10 3 6 2 4 6 Hypersil GOLD CN (1.9µm) Hypersil GOLD aQ Hypersil GOLD BioBasic 18 Hypersil GOLD C8 BioBasic 8 Hypersil GOLD CN Hypersil GOLD Phenyl 4-032 4-029 4-026 4-140 4-027 4-140 4-032 4-031 www.thermoscientific.com/chromatography Column Selection Manufacturer Grace Grace LC Columns and Accessories >> Phase Vydac™ 201SP C18 Vydac 201SP Selectapore 90M C18 Vydac 201TP C18 Vydac 202TP C18 Vydac 208TP C8 Vydac 214TP Vydac 218TP Vydac 218WP Selectapore 300M C18 XBridge™ C18 XBridge C8 XBridge HILIC XBridge Phenyl XTerra™ MS C18 XTerra MS C8 YMCbasic™ YMC-Pack™ C4 YMC-Pack C8 YMC-Pack CN YMC-Pack Diol YMC-Pack NH 2 YMC-Pack ODS AQ YMC-Pack ODS-A YMC-Pack ODS-A YMC-Pack Phenyl YMC-Pack Pro C18 YMC-Pack Silica YMC-Pack TMS (C1) Zorbax™ Eclipse XDB C18 Zorbax Eclipse XDB C8 Zorbax Eclipse XDB Phenyl Zorbax Eclipse Plus C18 Zorbax Eclipse Plus C8 Zorbax RRHT Eclipse Plus C18 Zorbax RRHT Eclipse Plus C8 Zorbax RRHT Eclipse XDB-C18 Zorbax RRHT Eclipse XDB-C8 Zorbax RRHT SB-CN Zorbax SB Aq Zorbax SB C18 Zorbax SB C18 Zorbax SB C8 Zorbax SB C8 Zorbax SB CN Zorbax SB Phenyl 4-015 LC Columns and Accessories Thermo Scientific LC Columns Column Protection Extend column lifetime and improve performance • Guards and filters to protect your analytical column • Economical extension of column lifetime • Multiple formats for optimum performance and efficiency • Drop-in designs for quick and easy guard and filter replacement • UHPLC Filter cartridges and holder to protect Hypersil GOLD 1.9µm columns To extend the lifetime and performance of your analytical and preparative columns, we recommend that they be protected from contamination by sample and solvent debris and interferences from the sample matrix. The most cost-effective and efficient way of trapping these unwanted system components is by use of a filter or packed guards. Column performance should not be affected by adding a guard or filter unit to the HPLC system. The chromatogram shown demonstrates how the column’s chromatographic performance is unaffected by the addition of a guard unit during the analysis of procainamides. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Guard and Filter Selection 4-016 Guard columns are positioned between the injector and the analytical column, removing strongly adsorbed sample components before the sample reaches the analytical column. The simple rule of thumb in guard selection is to choose one that matches your analytical column. The internal diameters should match as closely as possible, and the packing material should Interferences be the same particle size and chemistry as the analytical column. If a guard cartridge system is used, the replacement of the packed cartridges should be simple and fast. Analytes: Procainamides Without a guard With C18 guard Pre-column filters are positioned between the solvent inlet filter and the column inlet. They are designed to trap particulate matter from the fluid path. They do not remove sample interferences or contaminants that are dissolved in the fluid path. These units are designed to be wholly disposable or have replaceable filters in a re-useable holder. Replaceable 0.2µm Thermo Scientific UHLPC filter cartridges protect Hypersil GOLD 1.9µm and Syncronis 1.7µm columns against particles, enhancing column lifetime. Its low dead volume design maintains chromatographic performance without degrading peak shape and causes minimal efficiency loss through dispersion. The UHPLC filter adds a minimal increase in backpressure, so can be fitted to any length column. Peak resolution and shape are unaffected by the addition of a guard column Use a pre-column guard to trap any remaining interferences Isolate interferences or analytes using SPE and/or Sample Successful, Reproducible Analysis Particulates and Interferences and/or Particulates Choosing a guard or filter based on sample make-up Filter Sample through a 0.45 µm filter Use a pre-column filter to trap particulates Drop-in guard cartridges offer convenience, economy and effective protection for extending column lifetime • The 10mm design offers maximum protection with minimal increase in retention • Fit Thermo Scientific™ UNIGUARD™ direct connection and stand alone holders Drop-in Guard Cartridges Ordering Guide 1.0mm ID 2.1mm ID 3.0mm ID 4.0/4.6mm ID Quantity 25003-011001 25005-011001 25003-012101 25005-012101 25003-013001 25005-013001 25003-014001 25005-014001 4 Pack 4 Pack 25203-011001 25205-011001 25203-012101 25205-012101 25203-013001 25205-013001 25203-014001 25205-014001 4 Pack 4 Pack 25303-011001 25305-011001 25303-012101 25305-012101 25303-013001 25305-013001 25303-014001 25305-014001 4 Pack 4 Pack 25403-011001 25405-011001 25403-012101 25405-012101 25403-013001 25405-013001 25403-014001 25405-014001 4 Pack 4 Pack 25803-011001 25805-011001 25803-012101 25805-012101 25803-013001 25805-013001 25803-014001 25805-014001 4 Pack 4 Pack 25903-011001 25905-011001 25903-012101 25905-012101 25903-013001 25905-013001 25903-014001 25905-014001 4 Pack 4 Pack – 74104-012101 74104-013001 74104-014001 4 Pack – 74204-012101 74204-013001 74204-014001 4 Pack – 97105-012101 97105-013001 97105-014001 4 Pack – 97205-012101 97205-013001 97205-014001 4 Pack – 97305-012101 97305-013001 97305-014001 4 Pack – 97905-012101 97905-013001 97905-014001 4 Pack 35003-011001 35005-011001 35003-012101 35005-012101 35003-013001 35005-013001 35003-014001 35005-014001 2 Pack 2 Pack Column Protection Particle Size (µm) Length (mm) Hypersil GOLD 3 10 5 10 Hypersil GOLD C8 3 10 5 10 Hypersil GOLD aQ 3 10 5 10 Hypersil GOLD PFP 3 10 5 10 Hypersil GOLD CN 3 10 5 10 Hypersil GOLD Phenyl 3 10 5 10 Accucore XL C18 4 10 Accucore XL C8 4 10 Syncronis C18 5 10 Syncronis C8 5 10 Syncronis aQ 5 10 Syncronis Phenyl 5 10 Hypercarb 3 10 5 10 LC Columns and Accessories >> Drop-in Guard Cartridges This table provides a sample of the guard cartridges for the most popular Thermo Scientific HPLC stationary phases. Drop-in guard cartridges are available in other Thermo Scientific phases. For information on guard cartridges for other Thermo Scientific phases, please consult the appropriate section of the catalogue or contact Customer Services for more information. www.thermoscientific.com/chromatography 4-017 LC Columns and Accessories Defender Guard Cartridges Thermo Scientific™ Defender™ Guard Cartridges have been designed specifically to work with high speed, high efficiency separations. Accucore Defender Guard Cartridges (4/pk) Ordering Guide Description Accucore RP-MS Accucore C18 Accucore C8 Accucore aQ Accucore Polar Premium Accucore Phenyl-Hexyl Accucore Phenyl-X Particle Size (µm) 2.6 2.6 2.6 2.6 2.6 2.6 2.6 Length (mm) 10 10 10 10 10 10 10 2.1mm ID 17626-012105 17126-012105 17226-012105 17326-012105 28026-012105 17926-012105 27926-012105 Particle Size (µm) Accucore PFP 2.6 Accucore C30 2.6 Accucore HILIC 2.6 Accucore Urea-HILIC 2.6 Accucore 150-C18 2.6 Accucore 150-C4 2.6 Accucore 150-Amide-HILIC 2.6 Description Length (mm) 10 10 10 10 10 10 10 2.1mm ID 17426-012105 27826-012105 17526-012105 27726-012105 16126-012105 16526-012105 16726-012105 Other guard column dimension are available in some Accucore phases. Please call your local Customer Service for more information. UNIGUARD Direct-Connection Guard Cartridge Holders Reusable, stainless steel guard cartridge holders that match directly to the analytical column inlet – eliminating requirement for extra fittings • With PEEK 1/16in male outlet that fits most columns • 1/16in female inlet can be used with various standard fittings UNIGUARD Direct-Connection Guard Cartridge Holders Ordering Guide Thermo Scientific Chromatography Columns and Consumables 2014-2015 Description UNIGUARD Drop-In /Defender Guard Cartridge Holder Standard Replacement Tip Waters Columns Replacement Tip 4-018 1.0mm ID 851-00 2.1mm ID 852-00 3.0mm ID 852-00 4.0/4.6mm ID 850-00 Quantity 1 Each 850-RT 850-WT 850-RT 850-WT 850-RT 850-WT 850-RT 850-WT 1 Each 1 Each Thermo Scientific™ Acclaim™ Guard Cartridges are available in the same packing and internal diameters as the Acclaim analytical column chemistries. A re-usable holder allows for easy cartridge replacement; extending the life of the analytical column. Acclaim Guard Cartridges Ordering Guide Acclaim Guard Cartridge Ordering Guide (2/pk) Particle Size (µm) 5 Length (mm) 10 ID (mm) 2.1 3.0 4.6 120 C18 120 C8 069689 071981 069695 069688 071979 069696 Polar Advantage 069691 071983 069698 Polar Advantage II 069692 071985 069699 C30 075722 075721 075720 Mixed-Mode HILIC-1 069694 071913 069706 Mixed-Mode WAX-1 069686 071909 069704 Mixed-Mode WCX-1 085455 071911 069705 Acclaim Guard Cartridge Ordering Guide (2/pk) continued Description Length (mm) 10 10 2.1mm ID 3.0mm ID 4.6mm ID 079934 074263 071974 074261 071973 074262 Acclaim Specialty Column Guard Cartridge Ordering Guide (2/pk) Description Acclaim Trinity P1 Acclaim Trinity P2 Acclaim Trinity Q1 Acclaim Organic Acid Acclaim Surfactant Plus Acclaim Surfactant Acclaim Explosives E1 Acclaim Explosives E2 Acclaim Carbamate Acclaim Carbonyl C18 Particle Size (µm) 3 3 5 5 5 5 5 5 5 5 Length (mm) 10 10 10 10 10 10 10 10 10 10 2.1mm ID 3.0mm ID 4.6mm ID 071391 085435 079719 – 082773 069693 – – 072930 079012 071390 085436 079720 071987 078959 071991 – 071989 072929 079013 – – – 069700 078960 069701 069702 069703 072928 079014 Column Protection Phenyl-1 HILIC-10 Particle Size (µm) 3 5 LC Columns and Accessories >> Acclaim Guards Cartridges Acclaim Guard Holder Ordering Guide Format Acclaim Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Cat. No. 069580 069707 074188 www.thermoscientific.com/chromatography 4-019 LC Columns and Accessories Javelin Direct-Connection Column Filters One-piece filter protects HPLC systems • Direct-connection design for maximum efficiency • Replace entire disposable filter unit for easy changes • Recommended for use as dedicated filters for a column rather than the HPLC system • 1/16in CPI tip attaches directly to HPLC column inlet without tubing or wrenches • 0.5µm porosity Javelin Direct-Connection Column Filter Ordering Guide Description Javelin Column Filter 2.1mm ID 88200 3.0mm ID 88700 4.0/4.6mm ID 88400 Quantity 4 Pack Cat. No 60140-412 Quantity 10 Pack ColumnSaver Precolumn Filters Filter mesh size 2μm ColumnSaver Precolumn Filters Ordering Guide Filter Mesh Size (µm) 2 UNIFILTER Direct-Connection HPLC Filter Systems Quickly replaced for minimal down time • Replaceable 0.5µm drop-in filter enhances column lifetime and improved performance • Holder attached directly to the inlet of your analytical system for maximum convenience Thermo Scientific Chromatography Columns and Consumables 2014-2015 UNIFILTER Direct-Connection HPLC Filter Systems Ordering Guide 4-020 Description UNIFILTER Direct Connection Holder Replacement Filter, 0.5µm Replacement Filter, 0.5µm Replacement Tip, CPI, Standard Replacement Tip, Waters End-fitting 2.1/3.0mm ID 27002 22016 22017 850-RT 850-WT 4.0/4.6mm ID 27000 22150 22155 850-RT 850-WT Quantity 1 Each 1 Each 5 Pack 1 Each 1 Each UHPLC Filter Column protection for Hypersil GOLD 1.9µm and Syncronis 1.7µm columns without compromising performance • Low volume filter cartridge design • Maintain peak shape • Minimal efficiency loss through dispersion UHPLC Filter Ordering Guide Description UHPLC Direct Connect Filter Holder 2.1mm ID Replacement Filter Cartridge, 0.2μm 1.0mm ID Replacement Filter Cartridge, 0.2μm Cat. No. 27006 22180 22185 Quantity 1 Each 5 Pack 5 Pack LC Columns and Accessories >> Notes Column Protection www.thermoscientific.com/chromatography 4-021 LC Columns and Accessories Thermo Scientific Columns for Fast and Analytical HPLC The Thermo Scientific LC column portfolio offers a comprehensive range of high quality options for fast and analytical HPLC. The table below shows our major column families with details of relative hydrophobicity, key attributes and how the attributes are achieved. Use this table and the information in the subsequent catalogue pages to find the best Thermo Scientific LC columns for your separation needs. Hydrophobicity Key Attribute Accucore Columns Page 4-042 Achieved through • 2.6 and 4µm solid core Ultimate Performance silica particles • Core Enhanced Technology Low Hypersil GOLD Columns Page 4-024 • 1.9, 3, 5 and 12µm Selectivity & Peak Shape Acclaim Columns Page 4-060 Medium/High Traditional, Speciality & Application Specific Thermo Scientific Chromatography Columns and Consumables 2014-2015 Syncronis Columns Page 4-074 4-022 Hypercarb Columns Page 4-084 Very High • 2.2, 3 and 5µm ultra-pure silica particles • Innovative and unique bonded phases • 1.7, 3 and 5µm high Ruggedness & Reproducibility High ultra-high purity silica particles • Wide range of conventional phases Extended Separation Capabilities surface area silica particles • Dense bonding, double endcapping, rigorous testing • 3 and 5µm 100% porous graphitic carbon particles • Polar retention effect on graphite (PREG) Increasing Impedan 5µm Accucore 2.6µm Effect of particle size and type An alternative to small diameter particles is the Core Enhanced Technology used in Accucore HPLC columns. As shown in the impedance plot below, Accucore columns 45,000 generate higher efficiencies in shorter times than columns packed with 5µm or 3µm particles Increasing Efficiency and does so at low backpressures. 15,000 15 3µm u opt 5 1.9µm u opt 1 2 3 u (mm/s) 4 N2 Ε Impedance ΔP Backpressure 4,500 t0 Retention time of unretained peak Ν Efficiency η Mobile phase viscosity Impedance Plot 5µm Accucore 2.6µm E= 1,500 6 5 3µm 45,000 Pt0 N2 4,500 Increasing Efficiency Increasing Flow Rate Speed and Resolution Peak Capacity Miniaturization The general chromatographic resolution equation shows that resolution is directly proportional to efficiency. High efficiencies across a wider range of linear velocities mean that shorter columns and/or faster flow rates can be used to increase the speed of separations without sacrificing resolution. As an alternative to speeding up analysis the excellent resolution offered by high efficiency columns can also be used to improve complex separations through an increase in peak capacity – the number of peaks that can be separated in a given gradient time. The sensitivity formula also shows that peak concentration can be increased through the use of shorter columns and more importantly, with narrower column internal diameters. The resolution equation also shows that a wide range of different bonded phases, each offering a different selectivity, is a useful way to improve resolution. RS = 1 (α − 1) N k' 1 + k' 4 α α Selectivity N Efficiency k’ Capacity factor Poor Resolution nc = 1 + nc tg W (t ) g W Peak capacity Gradient time When transferring a method to a different column geometry adjustments must be made to the following parameters: • Flow Rate • Injection Volume • Gradient Profile A convenient method transfer tool is available at the Chromatography Resource Center (www.thermoscientific.com/crc). Peak width (10% height) Sensitivity Optimization According to the formula shown below, sensitivity is increased in high efficiency separations by increasing the concentration of the peak and thus the detector signal to noise ratio. In order to preserve high efficiency, and therefore resolution and sensitivity, the HPLC system in use should be optimized to reduce any potential causes of peak broadening. See page 4-231 for details of this optimization. cmax∝ N Vi L dc2 (1 + k' ) Columns for Analytical LC 0 Pt0 5µm u opt 10 E= Increasing Impedance Advantage of Small Particles H (µm) Increasing Column Efficiency N) It is now well established that columns packed with smaller diameter particles generate higher 1,500 efficiencies over a wider range of flow rates than larger particle columns – as shown in the plot below. LC Columns and Accessories >> Principles of Fast LC cmax Concentration at peak apex N Efficiency Vi Injection volume L Column length dc Column internal diameter k’ Capacity factor Good Resolution www.thermoscientific.com/chromatography 4-023 LC Columns and Accessories Thermo Scientific Hypersil GOLD HPLC Columns Outstanding peak shape for your separations • Excellent peak symmetry • Narrow peaks for outstanding efficiency • Increased sensitivity and improved resolution • Variety of chemistries • 1.9 to 12µm particles Thermo Scientific™ Hypersil GOLD™ columns are exceptionally reproducible for reliable chromatography, column after column. This allows the user to be confident that assays developed with Hypersil GOLD columns will be robust and stable for the life of the assay, making them an ideal choice for new method development. Built on more than 35 years of experience in product development and manufacturing of HPLC media and columns, we successfully continue to extend the capabilities of this state-of-the-art family of columns, designed for improved chromatography. Hypersil GOLD columns are manufactured in ISO 9001:2008 accredited laboratories under strict protocols using a robust manufacturing procedure and extensive quality control testing. Improved Selectivity, Resolution and Productivity Thermo Scientific Chromatography Columns and Consumables 2014-2015 Hypersil GOLD columns are available in an array of chemistries to optimize separations and maximize productivity: 4-024 • Hypersil GOLD offers outstanding peak shape using generic gradients with C18 selectivity • Hypersil GOLD C8 offers similar selectivity but with less retention • Hypersil GOLD aQ can be used for challenging reverse phase separations employing highly aqueous mobile phases • Hypersil GOLD PFP can offer alternative selectivity in reverse phase applications • Hypersil GOLD Phenyl offers alternative selectivity and is particularly suitable for aromatic and moderately polar compounds • Hypersil GOLD CN can be used for both reversed and normal phase separations • Hypersil GOLD C4 has short alkyl chain length, low hydrophobicity column for less retention • Hypersil GOLD Amino demonstrates excellent chromatographic properties in three modes: weak anion exchange, reversed phase and normal phase. • Hypersil GOLD AX can be used to separate proteins, peptides, other anionic species and polar molecules • Hypersil GOLD SAX is a highly stable silica-based quarternary amine strong anion exchange column, designed for aqueous mobile phase • Hypersil GOLD Silica is a powerful and efficient tool in the chromatography of non-polar and moderately polar organic compounds by normal phase • Hypersil GOLD HILIC columns retain polar analytes that are problematic using reversed phase columns These chemistries offer alternative selectivities in the same column family, providing enhanced retention or changes in elution order for flexibility in method development. Each phase is made with the same care and attention to quality that defines all Thermo Scientific columns. Hypersil GOLD, 5µm, 150 x 4.6mm Mobile Phase: Gradient: Flow Rate: Detection: Temperature: Sample: A: H2O + 0.1% Formic acid B: MeOH + 0.1% Formic acid 20 to 50% B in 15 min 1mL/min UV at 280nm 25°C 1. Catechin 2. Epigallocatechin Gallate 3. Epicatechin 4. Gallocatechin Gallate 5. Epicatechin Gallate 6. Catechin Gallate 11 0 7 5 8 300 9 10 Competitor 18 column 5 10 15 20 7 3 2 1 0 5 8 6 5 10 9 100 10 Hypersil GOLD column 15 Competitor 1 200 25 11 4 Competitor 2 20 25 The improved peak symmetry provides additional peak height to increase sensitivity of analysis of trace components. Resolution of analytes is improved using a Hypersil GOLD column. Data courtesy of M. Euerby, AstraZeneca, Charnwood, UK. Solutions for High Throughput Screening, Capillary to Preparative Analysis Improved Sensitivity Good peak shape means greater sensitivity. When peaks exhibit tailing, peak height is reduced causing the sensitivity of the analysis to be compromised. The more symmetrical the chromatographic peaks, the more confidence you derive from your data. Using Hypersil GOLD columns, peak height is enhanced and peak integration calculations are optimized. 2 3 H250-001 4 5 1 7 Robust assay development requires a clear definition of resolution expectations. Narrow symmetrical chromatographic peaks ensure that optimum resolution is achieved. Obtaining narrow peak widths is especially challenging for basic pharmaceutical compounds. The figure above shows how Hypersil GOLD columns provide excellent resolution between critical pairs, aiding in separation of closely related species. Excellent reproducibility is illustrated with the percent carbon on the Hypersil GOLD media pH Stability Hypersil GOLD columns are well suited to extended pH applications. Hypersil GOLD columns have been shown to produce robust assays at high pH. At low pH, excellent column stability and reproducibility are illustrated. Dimensions: 5µm, 150 x 4.6mm Part Number: Mobile Phase: 25005-154630 A: 0.1% ammonia pH 10.6 B: MeOH + 0.1% ammonia 5 – 100% B in 15 min 1.0mL/min 10µL UV at 254nm 30°C Gradient: Flow: Injection: Detection: Temperature: 6 Enhanced Resolution Columns for Analytical LC Hypersil GOLD columns are available in particle sizes and column designs to meet all separation needs, including improved resolution, enhanced sensitivity and faster analyses. From 1.9µm to 12µm particles, Hypersil GOLD columns offer chromatographic solutions with consistent separations and performance. Specialized hardware includes Thermo Scientific™ KAPPA™ capillary columns, PicoFrit™ and IntegraFrit nanobore columns, Thermo Scientific™ Javelin™ HTS direct-connection columns and Thermo Scientific™ DASH™ HTS columns, designed for high throughput screening. Enhanced peak height can be particularly critical when low concentrations of an analyte are present, for example in an impurity assay. The increase in sensitivity gained with the Hypersil GOLD columns over competitor C18 columns is illustrated above. LC Columns and Accessories >> 6 3 2 1 Hypersil GOLD Signal 4 Stability of Hypersil GOLD columns at low pH. No loss of retention after 28L of mobile phase in 19.5 days of analysis. 1. Uracil 2. Acetaminophen 3. N-Acetyl procainamide 4. n-Propionyl procainamide 5. Oxazepam 6. Diazepam 7. Diisopropylphthalate 8. Di-n-propylphthalate 8 High pH stability assay (pH 10.6) of Hypersil GOLD columns www.thermoscientific.com/chromatography 4-025 Hydrophobicity Endcapped, ultrapure, silica-based columns with exceptional peak shape and resolution for HPLC and LC/MS pH Range • Significant reduction in peak tailing while retaining C18 selectivity • Excellent resolution, efficiency and sensitivity 300 Particle Size 1.9μm, 3μm, 5μm, 12μm USP L1 H2O / MeCN β-blockers +0.1%foric grad 15 to 100% 1min; 0.5ml/min; 30C; 1.9um Gold 20x2.1mm 550C; 3.5kV; 20V Seven in 1inminute 100 SM: 7G 6 NL: 1.02E5 Base Peak MS Blockers24 90 80 70 60 7 1 Gradient: Flow rate: Temperature: Detection: System: Analytes: 5 50 4 40 2 10 0 0.0 0.1 Column: 1.9µm Hypersil GOLD 20 x 2.1mm Part Number: 25002-022130 Mobile phase: 3 0.2 0.3 0.4 0.5 Minutes 0.6 0.7 0.8 0.9 1.0 H250-101 14 175 0 RT: 0.00 - 1.00 1 to 11 Pore Size Carbon Load (%) • Confidence in the accuracy and quality of analytical data 20 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High 0 30 4-026 Low 0 Relative Abundance LC Columns and Accessories Hypersil GOLD A – H 2O+0.1%formic acid B – MeCN+0.1%formic acid 15 to 100% B in 1min 0.5ml/min 30°C +ESI Thermo Scientific™Surveyor™ MSQ 1. Atenolol 2. Nadolol 3. Pindolol 4. Timolol 5. Metoprolol 6. Oxprenolol 7. Propanolol 10 25 Hydrophobicity Recommended for analytes with medium hydrophobicity or when a less hydrophobic phase is required to obtain optimum retention pH Range Low High 0 2 to 9 14 Pore Size • Similar selectivity to C18 columns but with reduced retention • Lower hydrophobicity, allowing compounds to elute quicker 0 175 300 Carbon Load (%) • Faster separations • Excellent peak shape 0 • High efficiency Particle Size 1.9μm, 3μm, 5μm 8 25 USP L7 • Outstanding sensitivity β-carotene 1 600 Column: Hypersil GOLD C8, 5µm, 150 x 4.6mm Part Number: 25205-154630 400 mAU 300 MeOH 1.5mL/min 25°C UV @ 450 nm 1. Lutein 2. Lycopene 3. β-Carotene 200 2 3 100 0 0 1 2 3 Minutes 4 5 6 7 H252-009 www.thermoscientific.com/chromatography Columns for Analytical LC Mobile phase: Flow rate: Temperature: Detection: Analytes: 500 LC Columns and Accessories >> Hypersil GOLD C8 4-027 LC Columns and Accessories Hypersil GOLD C4 Hydrophobicity Lower hydrophobicity than C18 or C8 recommended for very hydrophobic analytes pH Range 2 to 8 • Lower hydrophobicity • Faster separations 0 175 • High efficiency 0 • Outstanding sensitivity Particle Size 1.9μm, 3μm, 5μm 5 Fatty acids Column: Hypersil GOLD C4 1.9µm, 100 x 2.1mm Part Number: 25502-102130 1 Mobile phase: Flow rate: Temperature: Detection: Injection volume: Sample: 160 mAU 2 40 0 0.0 0.2 0.4 0.6 300 Carbon Load (%) • Excellent peak shape 200 14 Pore Size 80 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High 0 120 4-028 Low 0.8 1.0 Minutes 1.2 1.4 1.6 1.8 2.0 H255-002 H 2O / MeCN (20:80) 0.55mL/min 30°C 200 nm 1µL 1. Linolenic acid 2. Linoleic acid 25 USP L26 Hydrophobicity Hypersil GOLD aQ polar endcapped C18 columns provide a controlled interaction mechanism by which polar analytes can be retained and resolved Low High pH Range 0 2 to 9 14 Pore Size • Polar endcapped C18 phase for alternative selectivity 0 175 300 Carbon Load (%) • Retention and resolution of polar analytes • Excellent peak shape 0 • Stable in 100% aqueous mobile phases Particle Size 1.9μm, 3μm, 5μm 12 25 USP L1 Water soluble vitamins 1200 2 Column: Hypersil GOLD aQ, 5µm, 150 x 4.6mm Part Number: 25305-154630 3 8 6 1000 Mobile phase: Gradient: Flow Rate: Detection: Sample: 800 1 mAU 400 4 200 7 5 0 0 1 2 3 4 5 6 7 8 Minutes 9 10 11 12 13 14 H253-003 www.thermoscientific.com/chromatography Columns for Analytical LC 600 A: 50 mM KH 2 PO 4, pH 3.5 B: MeOH 0 – 100% B in 15 min 1mL/min UV @ 205nm 1. Vitamin B1 (thiamine) 2. Vitamin B6 (pyridoxine) 3. Vitamin B3 (nicotinamide) 4. Vitamin B5 (pantothenic acid) 5. Folic Acid 6. Vitamin B12 (cyanocobalamin) 7. Vitamin H (biotin) 8. Vitamin B2 (riboflavin) LC Columns and Accessories >> Hypersil GOLD aQ 4-029 LC Columns and Accessories Hypersil GOLD PFP Hydrophobicity Introduction of a fluorine group into the stationary phase causes significant changes in solute-stationary phase interaction pH Range • The fluorine atoms around the phenyl ring enhance pi-pi interactions • Alternative selectivity to C18 Carbon Load (%) with aromatic molecules 0 • Selectivity for non-halogenated polar compounds Polyphenols uAU 2 NL: 6.87E4 Channel A UV HGOLDPF PPP3 Column: Hypersil GOLD PFP 1.9µm, 50 x 2.1mm Part Number: 25402-052130 Mobile phase: Flow rate: Temperature: Detection: Injection Volume: Sample: 3 40000 30000 20000 4 1 10000 0 0.0 0.5 1.0 1.5 2.0 Minutes 2.5 3.0 3.5 H254-012 0.1% Acetic Acid 0.5mL/min 25°C UV @ 280nm (2µL Flow Cell) 0.5µL 1. Pyrogallol 2. Hydroquinone 3. Resorcinol 4. Phenol 14 175 8 Particle Size 1.9μm, 3μm, 5μm • Excellent peak shape and sensitivity 70000 2 to 8 0 • Extra retention for halogenated species 50000 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High 0 Pore Size 60000 4-030 Low 300 25 USP L43 Hydrophobicity Contains a C4 linker which allows for superior alignment of the phenyl ring with aromatic molecules pH Range Low High 0 2 to 8 14 Pore Size • Enhanced pi-pi interactions with aromatics • Moderate hydrophobicity 0 175 300 Carbon Load (%) • Outstanding peak shape and sensitivity 0 8 Particle Size 1.9μm, 3μm, 5μm 25 USP L11 Antidepressants 100 1 3 80 mAU Column: Hypersil GOLD Phenyl 1.9µm, 50 x 2.1mm Part Number: 25902-052130 Mobile phase: 5 60 6 2 40 imp 0 0.0 1.0 2.0 Minutes 3.0 4.0 H259-006 www.thermoscientific.com/chromatography Columns for Analytical LC 7 8 4 20 A – 0.1% Formic acid B – 0.1% Formic acid in MeCN Gradient: 10 – 60% B in 3.4mins, 60 - 90% B in 0.24 min Flow rate: 0.5mL/min Temperature: 60°C Injection Volume: 0.7µL Detection: UV @ 225 and 254 nm Analytes: 1. Uracil 2. Acetaminophen 3. p-Hydroxybenzoic acid 4. o-Hydroxybenzoic acid 5. Oxazepam 6. Diazepam 7. Di-isopropyl phthalate 8. Di-n-propyl phthalate LC Columns and Accessories >> Hypersil GOLD Phenyl 4-031 LC Columns and Accessories Hypersil GOLD CN Hydrophobicity Hypersil GOLD CN columns can be used for both normal phase and reversed phase separations pH Range 2 to 8 • Excellent peak shape 0 175 • Less retention for faster analysis 0 4 Particle Size 1.9μm, 3μm, 5μm Organic acids Column: Hypersil GOLD CN, 5µm, 150 x 4.6mm Part Number: 25805-154630 5 70 Mobile Phase: mVolts Isocratic: Flow Rate: Temperature: Detection: Sample: 4 3 2 55 50 0.0 2.5 5.0 300 Carbon Load (%) • Outstanding senstivity 60 14 Pore Size • Provide alternative selectivity with lower hydrophobicity 1 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High 0 65 4-032 Low 7.5 10.0 Minutes 12.5 15.0 17.5 H258-002 A: 25 mM KH 2 PO 4 pH2 B: MeOH 95% A: 5% B 1.5mL/min 25°C UV @ 230nm 1. 4-Fluorobenzoic 2. o-Toluic Acid 3. p-Toluic Acid 4. 2,4,6-Trimethylbenzoic Acid 5. 2,5-Dimethylbenzoic Acid 25 USP L10 Hydrophobicity A high performance aminopropyl phase that gives excellent chromatographic properties in three modes: weak anion exchange, reversed phase and normal phase Low High pH Range 0 2 to 8 14 Pore Size • Retains anions and organic acids in weak anion exchange 0 175 300 Carbon Load (%) • Excellent for carbohydrate analysis in reversed phase • Alternative selectivity to silica columns in normal phase chromatography 0 • Outstanding peak shape and sensitivity Particle Size 1.9μm, 3μm, 5μm 2 25 USP L8 Sugars 14 Column: Hypersil GOLD Amino, 5µm, 150 x 4.6mm Part Number: 25705-154630 1 12 uRIU 8 3 6 2 4 4 2 5 MeCN/Water (80:20) 1.2mL/min 35°C RI 20 µL 1. Fructose 2. Glucose 3. Sucrose 4. Maltose 5. Lactose 0 0 2 4 6 8 10 Minutes 12 14 16 18 20 H257-001 www.thermoscientific.com/chromatography Columns for Analytical LC Mobile Phase: Flow Rate: Temperature: Detection: Injection volume: Sample: 10 LC Columns and Accessories >> Hypersil GOLD Amino 4-033 LC Columns and Accessories Hypersil GOLD AX Hydrophobicity A novel polymeric amine ligand bonded to highly pure base deactivated silica pH Range 2 to 8 • Suitable for HILIC retention and separation of highly polar molecules • Higher efficiency than polymer based ion exchange columns • Outstanding peak shape and selectivity 0 175 0 6 Vitamin C Column: Hypersil GOLD AX, 5µm, 100 x 4.6mm Part Number: 26105-104630 Mobile Phase: 80 Flow Rate: Temperature: Detection: Injection volume: mAU 20 0 0 5 10 15 Minutes 20 25 H261-002 300 Carbon Load (%) Particle Size 1.9μm, 3μm, 5μm 100 14 Pore Size • Weak anion exchange phase for multiple charged species 40 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High 0 60 4-034 Low 100 mM Ammonium Acetate pH 6.8/ MeCN (30:70) 0.5mL/min 30°C UV @ 240nm 50µL 25 Hydrophobicity A highly stable quaternary amine strong anion exchange column for aqueous and low pH mobile phases pH Range Low High 0 2 to 8 14 Pore Size • High stability to aqueous and low pH mobile phases • Ideally suited to the analysis of smaller organic molecules including nucleotides and organic acids • Outstanding peak shape and sensitivity 0 175 300 Carbon Load (%) 0 2.5 Particle Size 1.9μm, 3μm, 5μm 25 USP L14 Monophosphates Column: Hypersil GOLD SAX, 5µm, 150 x 4.6mm Part Number: 26105-154630 200 175 Mobile Phase: Flow Rate: Temperature: Detection: Injection volume: 150 125 Columns for Analytical LC 100 Aqueous KH 2 PO 4 (50 mM, pH 3) 1.0mL min-1 40°C UV @ 254nm 10µL mAU 75 50 25 0 0 1 2 3 4 Minutes 5 6 7 LC Columns and Accessories >> Hypersil GOLD SAX 8 H263-001 www.thermoscientific.com/chromatography 4-035 Hydrophobicity Unbonded, highly pure base deactivated silica media that is the backbone of the Hypersil GOLD range of columns pH Range Low 0 High 2 to 8 • Outstanding peak shape and senstivity 0 Particle Size 1.9μm, 3μm, 5μm Steroids Column: Hypersil GOLD Silica, 5µm, 150 x 4.6mm Part Number: 25105-154630 90 80 Mobile Phase: Flow Rate: Temperature: Detection: Injection volume: Sample: 70 60 50 40 30 20 10 19:1 (v/v) n-C6H14/EtOH 1.5mL min-1 30°C UV @ 254nm 5µL 1. Progesterone 2. 21-Hydroxyprogesterone-21-acetate 3. 17-a-Hydroxyprogesterone 4. Cortisone 5. 11-a-Hydroxyprogesterone 6. Corticosterone 7. Hydrocortisone 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0 4-036 1 2 3 4 5 6 7 8 Minutes 9 10 14 Pore Size • Highly pure base deactivated silica media mVolts LC Columns and Accessories Hypersil GOLD Silica 11 12 13 14 15 H251-002 The Hypersil GOLD web page contains the latest news, applications and downloads for the Hypersil GOLD HPLC column range. Visit: www.thermoscientific.com/hypersilgold 175 300 USP L3 Hydrophobicity Hypersil GOLD HILIC retains and separates polar analytes that are problematic using reversed phase columns pH Range Low High 0 2 to 8 14 Pore Size • Alternative selectivity to C18 • Improved sensitivity for MS detection 0 175 300 Carbon Load (%) • Alternative to ion-pair or derivatisation • Outstanding peak shape and selectivity 0 25 6 Particle Size 1.9μm, 3μm, 5μm USP L14 Urea Column: Hypersil GOLD HILIC, 5µm, 150 x 4.6mm Part Number: 26505-154630 100 Mobile Phase: Flow Rate: Temperature: Detection: Injection volume: Sample: 60 40 H 2 O/MeCN (10:90) + 0.1% formic adic 0.6mL/min 30°C +ESI 1µL (made up in mobile phase) 1. Urea Columns for Analytical LC Relative Abundance 80 20 0 0 1 2 3 4 5 Minutes 6 7 8 LC Columns and Accessories >> Hypersil GOLD HILIC 9 10 H265-005 www.thermoscientific.com/chromatography 4-037 LC Columns and Accessories Hypersil GOLD Ordering Guide Particle Size (µm) 1.9 Format UHPLC Column Length (mm) 20 30 50 100 3 Drop-in Guard (4/pk) 150 200 10 HPLC Column 30 50 100 150 Thermo Scientific Chromatography Columns and Consumables 2014-2015 5 4-038 Drop-in Guard (4/pk) 10 HPLC Column 30 50 100 150 250 ID (mm) 2.1 1.0 2.1 1.0 2.1 3.0 4.6 1.0 2.1 3.0 2.1 2.1 1.0 2.1 3.0 4.0/4.6 2.1 3.0 4.6 2.1 3.0 4.0 4.6 1.0 2.1 3.0 4.0 4.6 1.0 2.1 3.0 4.0 4.6 2.1 3.0 4.0/4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 GOLD 25002-022130 25002-031030 25002-032130 25002-051030 25002-052130 25002-053030 25002-054630 25002-101030 25002-102130 25002-103030 25002-152130 25002-202130 25003-011001 25003-012101 25003-013001 25003-014001 25003-032130 25003-033030 25003-034630 25003-052130 25003-053030 25003-054030 25003-054630 25003-101030 25003-102130 25003-103030 25003-104030 25003-104630 25003-151030 25003-152130 25003-153030 25003-154030 25003-154630 25005-012101 25005-013001 25005-014001 25005-032130 25005-033030 25005-034630 25005-052130 25005-053030 25005-054630 25005-102130 25005-103030 25005-104630 25005-152130 25005-153030 25005-154030 25005-154630 25005-252130 25005-253030 25005-254030 25005-254630 C8 25202-022130 – 25202-032130 25202-051030 25202-052130 25202-053030 25202-054630 25202-101030 25202-102130 25202-103030 25202-152130 25202-202130 25203-011001 25203-012101 25203-013001 25203-014001 25203-032130 25203-033030 25203-034630 25203-052130 25203-053030 25203-054030 25203-054630 25203-101030 25203-102130 25203-103030 25203-104030 25203-104630 25203-151030 25203-152130 25203-153030 25203-154030 25203-154630 25205-012101 25205-013001 25205-014001 – – – 25205-052130 25205-053030 25205-054630 25205-102130 25205-103030 25205-104630 25205-152130 25205-153030 25205-154030 25205-154630 25205-252130 25205-253030 25205-254030 25205-254630 aQ 25302-022130 – 25302-032130 25302-051030 25302-052130 25302-053030 25302-054630 25302-101030 25302-102130 25302-103030 25302-152130 25302-202130 25303-011001 25303-012101 25303-013001 25303-014001 25303-032130 25303-033030 25303-034630 25303-052130 25303-053030 25303-054030 25303-054630 25303-101030 25303-102130 25303-103030 25303-104030 25303-104630 25303-151030 25303-152130 25303-153030 25303-154030 25303-154630 25305-012101 25305-013001 25305-014001 – – – 25305-052130 25305-053030 25305-054630 25305-102130 25305-103030 25305-104630 25305-152130 25305-153030 25305-154030 25305-154630 25305-252130 25305-253030 25305-254030 25305-254630 PFP 25402-022130 – 25402-032130 25402-051030 25402-052130 25402-053030 25402-054630 25402-101030 25402-102130 25402-103030 25402-152130 25402-202130 25403-011001 25403-012101 25403-013001 25403-014001 25403-032130 25403-033030 25403-034630 25403-052130 25403-053030 25403-054030 25403-054630 25403-101030 25403-102130 25403-103030 25403-104030 25403-104630 25403-151030 25403-152130 25403-153030 25403-154030 25403-154630 25405-012101 25405-013001 25405-014001 – – – 25405-052130 25405-053030 25405-054630 25405-102130 25405-103030 25405-104630 25405-152130 25405-153030 25405-154030 25405-154630 25405-252130 25405-253030 25405-254030 25405-254630 CN – – – – 25802-052130 – – – 25802-102130 – 25802-152130 25802-202130 25803-011001 25803-012101 25803-013001 25803-014001 25803-032130 – 25803-034630 25803-052130 – – – – 25803-102130 25803-103030 – 25803-104630 25803-151030 25803-152130 25803-153030 – 25803-154630 25805-012101 25805-013001 25805-014001 – – – 25805-052130 – 25805-054630 25805-102130 25805-103030 25805-104630 25805-152130 – – 25805-154630 25805-252130 25805-253030 25805-254030 25805-254630 Amino – – – – 25702-052130 – – – 25702-102130 – 25702-152130 25702-202130 25703-011001 25703-012101 25703-013001 25703-014001 25703-032130 – 25703-034630 25703-052130 – – – – 25703-102130 25703-103030 – 25703-104630 25703-151030 25703-152130 25703-153030 – 25703-154630 25705-012101 25705-013001 25705-014001 – – – 25705-052130 – 25705-054630 25705-102130 25705-103030 25705-104630 25705-152130 – – 25705-154630 25705-252130 25705-253030 25705-254030 25705-254630 AX – – – – 26102-052130 – – – 26102-102130 – 26102-152130 26102-202130 26103-011001 26103-012101 26103-013001 26103-014001 26103-032130 – 26103-034630 26103-052130 – – – – 26103-102130 26103-103030 – 26103-104630 26103-151030 26103-152130 26103-153030 – 26103-154630 26105-012101 26105-013001 26105-014001 – – – 26105-052130 – 26105-054630 26105-102130 26105-103030 26105-104630 26105-152130 – – 26105-154630 26105-252130 26105-253030 26105-254030 26105-254630 SAX – – – – 26302-052130 – – – 26302-102130 – 26302-152130 26302-202130 26303-011001 26303-012101 26303-013001 26303-014001 26303-032130 – 26303-034630 26303-052130 – – – – 26303-102130 26303-103030 – 26303-104630 26303-151030 26303-152130 26303-153030 – 26303-154630 26305-012101 26305-013001 26305-014001 – – – 26305-052130 – 26305-054630 26305-102130 26305-103030 26305-104630 26305-152130 – – 26305-154630 26305-252130 26305-253030 26305-254030 26305-254630 Silica – – – – 25102-052130 – – – 25102-102130 – 25102-152130 25102-202130 25103-011001 25103-012101 25103-013001 25103-014001 25103-032130 – 25103-034630 25103-052130 – – – – 25103-102130 25103-103030 – 25103-104630 25103-151030 25103-152130 25103-153030 – 25103-154630 25105-012101 25105-013001 25105-014001 – – – 25105-052130 – 25105-054630 25105-102130 25105-103030 25105-104630 25105-152130 – – 25105-154630 25105-252130 25105-253030 25105-254030 25105-254630 HILIC – – – – 26502-052130 – – – 26502-102130 – 26502-152130 26502-202130 26503-011001 26503-012101 26503-013001 26503-014001 26503-032130 – 26503-034630 26503-052130 – – – – 26503-102130 26503-103030 – 26503-104630 26503-151030 26503-152130 26503-153030 – 26503-154630 26505-012101 26505-013001 26505-014001 – – – 26505-052130 – 26505-054630 26505-102130 26505-103030 26505-104630 26505-152130 – – 26505-154630 26505-252130 26505-253030 26505-254030 26505-254630 www.thermoscientific.com/chromatography Columns for Analytical LC Phenyl – – – – 25902-052130 – – – 25902-102130 – 25902-152130 25902-202130 25903-011001 25903-012101 25903-013001 25903-014001 25903-032130 – 25903-034630 25903-052130 – – – – 25903-102130 25903-103030 – 25903-104630 25903-151030 25903-152130 25903-153030 – 25903-154630 25905-012101 25905-013001 25905-014001 – – – 25905-052130 – 25905-054630 25905-102130 25905-103030 25905-104630 25905-152130 – – 25905-154630 25905-252130 25905-253030 25905-254030 25905-254630 LC Columns and Accessories >> C4 – – – – 25502-052130 – – – 25502-102130 – 25502-152130 25502-202130 25503-011001 25503-012101 25503-013001 25503-014001 25503-032130 – 25503-034630 25503-052130 – – – – 25503-102130 25503-103030 – 25503-104630 25503-151030 25503-152130 25503-153030 – 25503-154630 25505-012101 25505-013001 25505-014001 – – – 25505-052130 – 25505-054630 25505-102130 25505-103030 25505-104630 25505-152130 – – 25505-154630 25505-252130 25505-253030 25505-254030 25505-254630 4-039 LC Columns and Accessories Hypersil GOLD Ordering Guide continued Format UNIGUARD Guard Cartridge Holder Length (mm) 10 ID (mm) 1.0 2.1 3.0 4.0/4.6 Cat. No. 851-00 852-00 852-00 850-00 Particle Size (µm) 1.9 Format Javelin HTS Column (3/pk) Javelin HTS Column (3/pk) Length (mm) 10 ID (mm) 2.1 GOLD 25002-012135 20 4.0 25005-024035 Format Length (mm) 10 ID (mm) 10 20 10 21 30 50 10 21 30 50 10 21 30 50 10 21 30 50 10 20 10 21 30 50 10 21 30 50 10 21 30 50 10 21 30 50 GOLD 25005-019023 25005-019223 25005-059070 25005-059270 25005-059370 25005-059570 25005-109070 25005-109270 25005-109370 25005-109570 25005-159070 25005-159270 25005-159370 25005-159570 25005-259070 25005-259270 25005-259370 25005-259570 25012-019023 25012-019223 25012-059070 25012-059270 25012-059370 25012-059570 25012-109070 25012-109270 25012-109370 25012-109570 25012-159070 25012-159270 25012-159370 25012-159570 25012-259070 25012-259270 25012-259370 25012-259570 ID (mm) 10 20 Cat. No. C-1000 854-00 5 Particle Size (µm) 5 Preparative Guard Cartridge (3/pk) Preparative HPLC Column 50 100 150 250 Thermo Scientific Chromatography Columns and Consumables 2014-2015 12 4-040 Preparative Guard Cartridge (3/pk) 10 Preparative HPLC Column 50 100 150 250 Format Preparative Guard Cartridge Holder Length (mm) 10 aQ 25305-019023 25305-019223 25305-059070 25305-059270 25305-059370 25305-059570 25305-109070 25305-109270 25305-109370 25305-109570 25305-159070 25305-159270 25305-159370 25305-159570 25305-259070 25305-259270 25305-259370 25305-259570 – – – – – – – – – – – – – – – – – – PFP 25405-019023 25405-019223 25405-059070 25405-059270 25405-059370 25405-059570 25405-109070 25405-109270 25405-109370 25405-109570 25405-159070 25405-159270 25405-159370 25405-159570 25405-259070 25405-259270 25405-259370 25405-259570 – – – – – – – – – – – – – – – – – – LC Columns and Accessories >> Notes Columns for Analytical LC 4-041 www.thermoscientific.com/chromatography LC Columns and Accessories Thermo Scientific Accucore HPLC Columns Ultimate Core Performance – Speed and Selectivity Combined Founded on state-of-the-art Core Enhanced Technology™ and utilizing vast experience in phase bonding and packing, Thermo Scientific™ Accucore™ HPLC columns provide a unique chromatography solution to enhance laboratory workflow and efficiency. Available in a wide range of stationary phase selectivities and compatible with almost any instrument, these columns provide an excellent return on investment. Containing solid core particles, which are engineered to a diameter of 2.6μm and a very narrow particle size distribution; Accucore HPLC columns allows high speed, high resolution separation, with back pressures significantly lower than those associated with UHPLC. Eleven different stationary phases bonded using advanced technology and packed with highly controlled automated processes result in highly reproducible, rugged columns that offer a wide range of selectivities to meet all your separation needs. Thermo Scientific Chromatography Columns and Consumables 2014-2015 The key components of Core Enhanced Technology 4-042 Solid Core Particles With a solid central core and porous outer layer, these particles generate high speed, high resolution separations without excessive backpressure Tight Control of Particle Diameter Enhanced selection process keeps particle size distribution to a minimum and produces high efficiency columns Automated Packing Process Enhanced automated procedures ensure that all columns are packed with the highest quality Advanced Bonding Technology Optimized phase bonding creates a series of high coverage, robust phases View product information and application notes The Accucore web page contains the latest news, applications and downloads for the Accucore HPLC column range. Visit it at: www.thermoscientific.com/accucore Pressure Pressure(bar) (bar) HH Decreasing Efficiency fully porous, porous, 5µm 5µm fully fully porous, porous, 3µm 3µm fully fully porous, porous, <2µm <2µm fully Accucore RP-MS RP-MS 2.6µm 2.6µm Accucore 2.0 2.0 4.0 4.0 1000 1000 900 900 800 800 700 700 600 600 500 500 Typical HPLC Pressure Limit Typical HPLC Pressure Limit 400 400 300 300 200 200 100 100 0 0 0 200 0 200 (mm/s) µµ (mm/s) 6.0 6.0 8.0 8.0 10.0 10.0 Accucore HPLC columns are more efficient than 5μm and 3μm columns. fully porous, porous, 5µm 5µm fully fully porous, porous, 3µm 3µm fully fully porous, porous, <2µm <2µm fully Accucore RP-MS RP-MS 2.6µm 2.6µm Accucore Column dimensions: dimensions: 100 100 xx 2.1mn 2.1mn Column 400 400 600 600 Flow Flow Rate Rate (µL/min) (µL/min) 800 800 1000 1000 Accucore HPLC columns achieve this efficiency without excessive backpressure Columns for Analytical LC Increasing Pressure LC Columns and Accessories >> 18.0 18.0 16.0 16.0 14.0 14.0 12.0 12.0 10.0 10.0 8.0 8.0 6.0 6.0 4.0 4.0 2.0 2.0 0.0 0.0 0.0 0.0 Accucore 2.6µm HPLC Columns Optimum Conditions and Ratings Column ID (mm) 2.1 3.0 4.6 Optimum Flow Rate 400µL/min 800µL/min 1800µL/min Maximum Inj. Volume 1µL 3µL 5µL Backpressure Rating 1000 bar 1000 bar 1000 bar Temperature Rating 70°C 70°C 70°C www.thermoscientific.com/chromatography 4-043 LC Columns and Accessories Accucore RP-MS Hydrophobicity • Optimized for MS detection Low High pH Range • Excellent peak shapes • Excellent combination of speed and efficiency 0 2 to 9 Accucore RP-MS uses an optimized alkyl chain length for more effective coverage of the silica surface. This coverage results in a significant reduction in non-hydrophobic interactions and thus highly efficient peaks with very low tailing. 0 0 Particle Size 2.6µm The selectivity offered by Accucore RP-MS matches that of C18 columns. Bases Accucore RP-MS 2.6µm, 50mm x 2.1mm 4 Mobile phase: 2 200 Flow rate: Temperature: Detection: Injection volume: Back pressure: Analytes: mAU 160 1 120 3 5 80 6 65% Methanol / 35% 25mM Potassium Phosphate pH7.0 500µL/min 30°C UV at 215nm 1µL 232 bar 1.Uracil 2. Propranolol 3. Butylparaben 4. Naphthalene 5. Acenaphthene 6. Amitriptyline 40 Tf = 1.27 0 0.0 0.5 1.0 1.5 Minutes 2.0 2.5 3.0 Testosterones 2 600 Accucore RP-MS 2.6µm, 100mm x 2.1mm Mobile phase: Flow rate: Temperature: Detection: Injection volume: Analytes: 1 3 500 4-044 mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 400 300 200 100 0 0 0.5 1 1.5 Minutes 2 2.5 300 80Å Carbon Load (%) RP-MS offers slightly lower retention than C18 and this combined with high efficiencies and low peak tailing make this the phase of choice for use with MS detection. 240 14 Pore Size 60:40 (v/v) Water / Acetonitrile 0.6mL/min 40°C UV at 254nm 1µL 1. 11-Ketotestosterone 2. 19-Nortestosterone (Nandrolone) 3. Epitestosterone 7 25 Hydrophobicity • Optimum retention of non-polar compounds Low High pH Range • Hydrophobic interaction mechanism • Separates a broad range of analytes 0 1 to 11 14 Pore Size The carbon loading of Accucore C18 phase provides high retention of non-polar analytes via a predominantly hydrophobic interaction mechanism. The highly retentive nature of Accucore C18 phase means that it can be used to separate a broad range of analytes. 0 80Å 300 Carbon Load (%) 0 Particle Size 2.6µm 9 25 USP L1 Triazines 100 Accucore C18 2.6µm, 50mm x 2.1mm Mobile phase: Gradient: 3 mAU 80 2 4 60 40 5 20 0 0.0 0.5 1.0 1.5 2.0 2.5 3.0 Minutes www.thermoscientific.com/chromatography Columns for Analytical LC Flow rate: Temperature: Detection: Injection volume: Backpressure: Analytes: 6 1 A – Water ; B – Acetonitrile Time (min) %B 1.0 35 2.5 70 600µL/min 25°C UV at 280nm 2µL 298 bar 1. Simazine 2. Simetryn 3. Atrazine 4. Ametryn 5. Propazine 6. Prometryn LC Columns and Accessories >> Accucore C18 4-045 LC Columns and Accessories Hydrophobicity Accucore C8 Hydrophobicity Low High • Lower retention pHhydrophobic Range Low • Complementary steric selectivity to C18 9 0 • Low levels of secondary2 tointeractions Pore Size • Recommended for moderately polar analytes 0 14 0 300 80 Å 14 Particle Size 2.6µm 300 80 Å Carbon Load (%) Accucore C8 HPLC offer lower hydrophobic retention than columns packed Carbon Loadcolumns (%) with longer alkyl chain length material, such as C18, and are therefore recommended for analytes with medium hydrophobicity or when25a less hydrophobic 5 0 phase provides optimum retention. 0 5 25 Particle Size 2.6µm USP L7 USP L7 The low levels of secondary interactions demonstrated in the phase characterization are the result of excellent bonded phase coverage and allow users of Accucore C8 HPLC columns to benefit from excellent peak shapes. Testosterone Testosterone Column: 100 Colum 90 Mobil 80 Mobil Accucore C8 C8 2.6 µm, 5050x 2.1 mm Accucore 2.6μm, x 2.1mm 40 Injection: 30 Detection: Gradie Relative Abundance 50 80 Relative Abundance 100 70 Mobile phase A: Mobile waterphase + 0.1% acid+ 0.1% formic acid A:formic water 60 phase B: acetonitrile + 0.1% formic acid Mobile phase B: Mobile acetonitrile + 0.1% formic acid Gradient: 5–95 % B in 0.8 minutes 50 Gradient: 5–95 % B in 0.8 minutes Flow: 1500 μL/min 40 Temperature: 60 °C Flow: 1500 µL/min 30 Injection: 5μL Temperature: 60 °C 20 Detection: ESI-MS/MS%RSD Peak area 90 70 60 20 Retention time (tR /min) 0 ESI-MS/MS 0.0 %RSD tR0.2 0.4 %RSD Area 0 0.0 0.2 0.4 0.6 0.8 1.0 1.2 Minutes Tempe Injecti Detec 0.8 1.0 1.2 1.4 3.01 Minutes 0.73 %RSD tR 0.22 %RSD Area 3.01 Data from six injections. 22 22 0.73 0.6 0.22 Retention time (tR /min) Data from six injections. 1.4 Flow: 5 µL 10 10 Thermo Scientific Chromatography Columns and Consumables 2014-2015 2 to 9 0 Pore Size Testosterone 4-046 High pH Range Data Hydrophobicity • Retention and resolution of polar analytes Low • Polar endcapped C18 stationary phase for alternative selectivity • Ideal for highly aqueous mobile phases High pH Range 0 2 to 9 14 Pore Size The polar functional group used to endcap Accucore aQ phase provides an additional controlled interaction mechanism by which polar compounds can be retained and resolved, making Accucore aQ phase ideal for the quantitative analysis of trace levels of polar analytes. The wettability of reversed phase media can be increased by the introduction of polar functional groups. The polar endcapping of Accucore aQ media also makes it usable in 100% aqueous mobile phases without the risk of loss of performance or poor stability. 0 80Å 300 Carbon Load (%) 0 Particle Size 2.6µm 9 25 USP L1 Lamivudine (USP) 14050 x 2.1 mm Accucore C8 2.6 µm, Accucore aQ 2.6µm, 50mm x 2.1mm le phase A: water + 0.1% formic acid Mobile phase: le phase B: acetonitrile + 0.1% formic acid ent: 100 5–95 % B in 0.8 minutes 95:5 (v/v) Ammonium Acetate, pH 3.80 / Methanol Flow rate: 200µL/min Temperature: 35°C Detection: UV at 277nm Injection volume: 1µL Analytes: Lamivudine Asymmetry 1.36 0.00 %RSD t r %RSD Peak area 1.72 (%RSD calculated from 6 replicate injections) USP acceptance criteria: % RSD (t r, Peak Area) <2.0 mAU 120 1500 µL/min erature: 60 °C tion: 5 µL ction: ESI-MS/MS Retention time (tR /min) %RSD tR %RSD Area a from six injections. 80 60 40 20 0.73 0.22 0 0.0 3.01 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 Minutes www.thermoscientific.com/chromatography Columns for Analytical LC mn: LC Columns and Accessories >> Accucore aQ 4-047 Pore Size 0 150 Å 300 Carbon Load (%) Accucore Polar Premium Particle Size 2.6µm 8 LC Columns and Accessories 0 25 Hydrophobicity USP L60 • Rugged amide-embedded C18 phase Low High pH Range • Selectivity complementary to conventional C18 phases • Stable over a wide pH range and compatible with 100% aqueous 1.5 to 10 0 14 Pore Size mobile phase 0 150 Å 300 Carbon Load (%) Accucore Polar Premium is an exceptionally rugged polar embedded reverse phase material that offers high efficiency, wider operating pH range and unique selectivity complementary to standard C18 phases. 8 25 Particle Size 2.6µm USP L60 0 The specially designed bonded phase is stable from pH 1.5 to 10.5 and will not undergo phase collapse in 100% aqueous mobile phase. Curcuminoids (Tumeric) 600 1, 2, 3 1,2,3 Accucore Polar Premium 2.6μm, 100 x 3.0mm Fused Core C18, 100 x 3.0 mm 500 Mobile phase: Flow: Temperature: Injection: Detection: Analytes: 400 mAU Fused core C18 300 Fused Core C18 methanol : 10mM phosphoric acid, 80 : 20 800 μL/min 40 °C 6μL UV at 428nm 1. Curcumin 2. Desmethoxycurcumin 3. Bis-desmethoxycurcumin 200 1 100 2 3 Accucore Polar Premium The Accucore Polar Premium HPLC column provides desirable 600 selectivity that resolves the major and minor component 1, 2, 3 1,2,3 under simple isocratic conditions in less than three minutes, 500 while the C18 columns fail to separate these components. -50 0.00 1.00 2.00 400 3.00 Fused core C18 mAU Minutes 300 Fused Core C18 200 1 100 Thermo Scientific Chromatography Columns and Consumables 2014-2015 2 4-048 3 Accucore Polar Premium -50 0.00 1.00 2.00 Minutes 3.00 Hydrophobicity • Mixed-mode selectivity for aromatic and moderately polar analytes Low High pH Range • Enhanced Pi-pi interactions with aromatics • Moderate hydrophobicity 0 2 to 8 14 Pore Size The C6 chain in Accucore Phenyl-Hexyl phase exhibits classical RP retention and selectivity, while the phenyl ring can add special selectivity by interacting with polar groups within the solutes. This results in a mixed-mode separation mechanism. The reduced hydrophobicity of this phase makes it ideal for the separation of very non-polar compounds. 0 80Å 300 Carbon Load (%) 0 5 Particle Size 2.6µm 25 USP L11 The Phenyl-Hexyl phase should be selected for complex samples where some peaks are well resolved on a conventional alkyl phases, but are not well resolved on a conventional phenyl phase, or when other peaks are well resolved on a phenyl phase, but not well resolved on a conventional alkyl phase. Beta-agonists 100 Accucore Phenyl-Hexyl 2.6µm, 100mm x 2.1mm Mobile phase: Terbutaline 0 100 Gradient: NL: 6.51E5 m/z = 239.60-240.60 MS 3.04 Cimaterol 0 100 NL: 3.52E6 m/z = 219.60-220.60 Flow rate: Temperature: Detection: 3.16 Salbutamol 0 100 NL: 2.23E6 m/z = 301.60-302.60 MS Injection volume: Backpressure: 4.72 A – Ammonium acetate 5mM, pH 4 B – Acetonitrile Time (min) %B 0 5 1 5 10 100 0.25mL/min 40°C +ESI-MS (45ºC, 4.5kV, 60V, scan 150 – 350) 1µL 120 bar (at t0) Ractopamine 0 100 NL: 2.51E6 m/z = 213.60-214.60 MS 4.80 Columns for Analytical LC NL: 9.22E5 m/z = 225.60-226.60 2.89 LC Columns and Accessories >> Accucore Phenyl-Hexyl Clorprenaline 0 100 NL: 1.71E6 m/z = 276.60-277.60 MS 5.13 Clenbuterol 0 0 1 2 3 Minutes 4 5 6 7 www.thermoscientific.com/chromatography 4-049 0 80 Å 300 Carbon Load (%) 6 0 25 LC Columns and Accessories Particle Size 2.6µm Accucore Phenyl-X Hydrophobicity • Unique reversed-phase shape selectivity Low High pH Range • Enhanced selectivity for aromatic compounds • Compatible with highly aqueous mobile phases 2 to 8 0 • Robust, high-efficiency, low column bleed Pore Size The proprietary Accucore Phenyl-X alkyl aromatic bonded phase provides a unique selectivity when compared to other reversed phase materials such as C18 or Phenyl. Carbon Load (%) 0 80 Å 300 6 0 Phenyl-X exhibits particularly high aromatic selectivity. 14 25 Particle Size 2.6µm The advanced design of the bonded phase makes it compatible with highly aqueous mobile phases and robust, demonstrating very low bleed. Estrogens Accucore Phenyl-X 2.6μm, 100 x 2.1mm Fused Core C18, 100 x 2.1mm 1 120 Mobile phase: Flow: Temperature: Injection: Detection: Wash solvent: 100 2 mAU 80 3 4 60 40 15:40:45 (v/v) acetonitrile: methanol : water 400 μL/min 40 °C 1μL UV at 220nm Same as mobile phase 20 Fused Core C18 0 0.0 0.5 1.0 2.5 2.0 2.5 3.0 3.5 4.0 5.5 6.0 1. Estriol (E3) 2. Estradiol (E2) Minutes 150 H3C 1 H 2 75 H 3. Estrone (E1) 50 4. Ethynylestradiol 100 H3C H 1.0 2.5 2.0 2.5 3.0 3.5 4.0 5.5 6.0 OH 80 H3C HO mAU 0 0.5 OH H Accucore Phenyl-X 0.0 1 120 4 25 H HO 2.04 3 OH H H HO 100 H3 C OH H 125 mAU OH H 60H 2 3 CH 4 H40 H HO 20 Fused Core C18 Minutes 0 0.0 0.5 1.0 2.5 2.0 2.5 3.0 3.5 4.0 5.5 6.0 1. Estriol (E3) Minutes 4-050 1 125 H HO 100 mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 150 2 75 2.04 3 4 3. Estrone (E1 50 25 Accucore Phenyl-X 0 0.0 0.5 1.0 2.5 2.0 2.5 Minutes 3.0 3.5 4.0 5.5 6.0 HO Hydrophobicity • Alternative selectivity to C18 Low High pH Range • Extra retention for halogenated species • Unique selectivity for non-halogenated polar compounds 0 2 to 8 14 Pore Size The introduction of fluorine groups into the Accucore PFP (pentafluorophenyl) stationary phase causes significant changes in solute-stationary phase interactions. This can lead to extra retention and selectivity for positional isomers of halogenated compounds. PFP Columns are also well suited to the selective analysis of non-halogenated compounds, in particular polar compounds containing hydroxyl, carboxyl, nitro, or other polar groups. High selectivity is often most apparent when the functional groups are located on an aromatic or other rigid ring system. 0 80Å 300 Carbon Load (%) 0 5 Particle Size 2.6µm 25 USP L43 Positional isomers Accucore PFP 2.6µm, 50mm x 2.1mm 50 40 7 mAU 30 1 89 20 2 11 34 10 10 12 13 6 14 0 0 1 2 3 4 Minutes 5 6 7 8 A – 0.1% Formic Acid in Water A – 0.1% Formic Acid in Acetonitrile 15-30%B in 7 minutes 600µL/min 50°C UV at 270nm 2µL 1. 3,4 – Dimethoxyphenol 2. 2,6 – Dimethoxyphenol 3. 2,6 – Difluorophenol 4. 3,5 – Dimethoxyphenol 5. 2,4 – Difluorophenol 6. 2,3 – Difluorophenol 7. 3,4 – Difluorophenol 8. 3,5 – Dimethylphenol 9. 2,6 – Dimethylphenol 10. 2,6 – Dichlorophenol 11. 4 – Chloro-3-Methylphenol 12. 4 – Chloro-2-Methylphenol 13. 3,4 – Dichlorophenol 14. 3,5 – Dichlorophenol www.thermoscientific.com/chromatography Columns for Analytical LC Mobile phase: Mobile phase: Gradient: Flow rate: Temperature: Detection: Injection volume: 5 LC Columns and Accessories >> Accucore PFP 4-051 0 300 150 Å Carbon Load (%) LC Columns and Accessories 25 5 0 Accucore C30 Particle Size 2.6µm USP L62 Hydrophobicity • Ideal for separation of hydrophobic, long alkyl chain compounds Low High pH Range • High shape selectivity for structurally related isomers • Excellent aqueous-compatibility 2 to 8 0 14 Pore Size Accucore C30 offers high shape selectivity for hydrophobic, long chain, structurally related isomers, for example carotenoids and steroids. This is a different form of shape selectivity from that measured in the SS phase characterisation test. 0 300 150 Å Carbon Load (%) 25 5 0 It is also an excellent alternative to normal-phase columns for lipid analysis. The optimized bonding density of the long alkyl chains facilitated by a wider pore diameter particle result in a phase that is stable even in highly aqueous mobile phases. Particle Size 2.6µm USP L62 Vitamin K isomers 250 1 Fused Core C18 200 2+2’ mAU 150 100 Accucore C30 2.6μm, 100 x 3.0mm Fused Core C18, 100 x 3.0mm Mobile phase: methanol: 2mM ammonium acetate, 98:2 Flow: 650μL/min Temperature: 20 °C Injection: 5μL Detection: UV at 250nm Accucore C30 shows better separation for vitamin K1 isomers than the C18 column. Accucore C30 1 2 50 2’ 0 250 -50 1 Fused Core C18 200 0.00 1.00 2.00 3.00 Minutes 4.00 5.00 6.00 2+2’ Chromatogram showing the separation of Vitamin K compounds Minutes 1-Vitamin K2, 2-Vitamin K1 (trans isomer), 2’-Vitamin K1 (cis isomer) mAU 150 100 Accucore C30 1 2 Thermo Scientific Chromatography Columns and Consumables 2014-2015 50 4-052 2’ 0 -50 0.00 1.00 2.00 3.00 Minutes 28 4.00 5.00 6.00 Hydrophobicity • Enhanced retention of polar and hydrophilic analytes Low Very Low (5%) High pH Range • Alternative selectivity to C18 without ion-pair or derivatization 0 In HILIC mode the separation occurs through two mechanisms. The primary mechanism is a partitioning effect due to the enriched water layer around the polar or charged substrate material. The secondary mechanism involves interaction between the analyte and the active surface moiety. 2 to 8 14 Pore Size 0 80Å Particle Size 2.6µm 300 USP L3 Analyte properties that govern retention with HILIC phases are acidity/basicity, which determines hydrogen bonding, and polarizability which determines dipoledipole interactions. The highly organic mobile phases used with Accucore HILIC phase ensure efficient desolvation in ESI MS detection, which in turn leads to improved sensitivity. Catecholamines 2 Accucore HILIC 2.6µm, 50mm x 2.1mm Mobile phase: 120 Flow rate: Temperature: Detection: Injection volume: Backpressure: Analytes: 1 4 mAU 100 80 3 60 40 5 6 20 85:15 Acetonitrile:100mM Ammonium Formate, pH 3.2 2mL/min 40°C UV at 280nm 5µL 157 bar 1. Catechol 2. 5-HIAA 3. DOPAC 4. Serotonin 5. L-tyrosine 6. Dopamine 7. L-DOPA 7 0 0 0.5 1 1.5 2 2.5 3 3.5 Columns for Analytical LC 140 LC Columns and Accessories >> Accucore HILIC 4 Minutes www.thermoscientific.com/chromatography 4-053 2 to 8 0 14 Pore Size 0 300 80 Å LC Columns and Accessories Particle Size 2.6µm Accucore Urea-HILIC Hydrophobicity • Bonded hydrophilic stationary phase Low High pH Range • Unique selectivity compared to other HILIC phases • Low ion exchange activity 2 to 8 0 14 Pore Size Accucore Urea-HILIC has an alternative selectivity and lower ion exchange activity than other HILIC phases. 0 300 80 Å Particle Size 2.6µm The bonded hydrophilic stationary phase provides retention of broad range of polar analytes using up to 20% aqueous mobile phase. Analgesic compounds 600 Accucore Urea-HILIC 2.6μm, 100 x 2.1mm 2 Mobile phase: 1 mAU 400 3 200 0 0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 Flow: Run Time: Temperature: Injection: Injection wash solvent: Detection: Backpressure: composition 10:80:10, A : B : C A: water B: acetonitrile C: 100 mM ammonium acetate adjusted to pH 4.9 300 μL/min 2 minutes 35 °C Analgesic 2 μL into 10 μL partial loop mode.compounds water:acetonitrile 20:80 UV at 230 nm 71 bar 600 2 Minutes Acetaminophen As Mean 0.760 1.474 CV % 0.00 1.17 400Salicylic acid Aspirin 1 tR As Rs tR As Rs 0.908 1.303 2.359 1.100 1.318 3.264 0.48 200 0.92 0.49 0.00 0.63 0.48 mAU tR 3 Data from eight replicate analyses of a mixture of acetaminophen, salicylic acid and aspirin Retention time (tR), peak asymmetry (As), peak resolution (Rs) 0 0 0.2 0.4 0.6 0.8 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Minutes 4-054 1.0 1.2 1.4 Based on Core Enhanced Technology using 4μm solid core particles, Accucore XL HPLC columns allow users of conventional HPLC methods to enjoy performance far beyond that of columns packed with 5μm, 4μm or even 3μm fully porous particles. Very high separation efficiencies using standard HPLC instruments and conditions provide increased peak resolution and lower limits of detection. An ultra-stable packed bed results in exceptionally robust columns that demonstrate excellent retention and response reproducibility. 4µm Solid Core Particles for all Users The 4µm solid core particles used in Accucore XL HPLC columns have been specifically designed to get the optimum chromatographic performance from conventional HPLC instruments. • Very high efficiencies • Little decrease in efficiency as flow rate is increased • Moderate backpressures 18.0 Fully porous C18, 5μm 16.0 Fully porous C18, 3μm 14.0 Accucore XL C18, 4μm 12.0 10.0 8.0 6.0 4.0 Identical instrument and method conditions for all columns Increasing Efficiency Plate Height (H, µm) Accucore XL HPLC columns generate higher efficiencies than columns packed with 5µm and 3µm fully porous material – as shown in the van Deemter curve below. Columns for Analytical LC Efficiency LC Columns and Accessories >> Accucore XL HPLC Columns 0.0 Dimensions: Mobile Phase: Temperature: Injection Volume: Detection: Sample: 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 150 x 4.6mm 50% water: 50% acetonitrile 30°C 1µL UV at 254nm (0.1s rise time, 20 Hz) o-xylene 4.5 Linear velocity (u, mm/s) • 75% higher efficiency than 5µm fully porous • 50% higher efficiency than 3µm fully porous www.thermoscientific.com/chromatography 4-055 Accucore XL HPLC columns generate reasonable backpressures, moderately higher than fully porous 5µm and lower than fully porous 3µm, that are compatible with conventional HPLC instruments. Fully porous C18, 5μm 600 Fully porous C18, 3μm 500 400 bar conventional HPLC pressure Pressure (bar) LC Columns and Accessories Backpressure Accucore XL C18, 4μm 400 300 200 Identical instrument and method conditions for all columns 100 Dimensions: Mobile Phase: Temperature: 0 0.0 0.5 1.0 1.5 2.0 150 x 4.6mm 50% water : 50% acetonitrile 30°C 2.5 Flow rate (mL/min) • Backpressures between those generated by 3µm and 5µm fully porous particles • Within conventional HPLC instrumentation pressure limit – even at high flow rates Accucore XL C18 Hydrophobicity • Optimum retention of non-polar compounds Low • Hydrophobic interaction mechanism • Separates a broad range of analytes 30 0 80Å 0 7 Particle Size 4µm Accucore XL C18 4µm, 150 x 4.6mm Fully porous C18 5µm, 150 x 4.6mm 25 Mobile phase: 20 mAU 15 2 1 10 Flow rate: Temperature: Detection: Injection volume: Analytes: N = 10538 plates S/N = 534 5 0 -5 0 2.5 5.0 7.5 10.0 12.5 30 15.0 17.5 Minutes 25 2 1 20 20.0 22.5 25.0 27.5 66.3:33.7 (v/v) water with phosphoric acid, pH 2.5:methanol 2mL/min 30°C UV at 214nm 5µL 1. Valerophenone 2. Ibuprofen 30.0 N = 18274 plates S/N = 1202 • 73% higher efficiency 15 10 • 125% higher sensitivity 5 0 -5 0 2.5 5.0 7.5 10.0 12.5 14 15.0 17.5 Minutes 20.0 22.5 25.0 27.5 30.0 300 Carbon Load (%) Ibuprofen and Valerophenone (USP) mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 1 to 11 0 Pore Size The carbon loading of Accucore XL C18 provides high retention of non-polar analytes via a predominantly hydrophobic interaction mechanism. The highly retentive nature of the phase means that it can be used to separate a broad range of analytes. 4-056 High pH Range 25 USP L1 Hydrophobicity • Similar selectivity to C18 with lower retention Low High pH Range • Recommended for analytes with moderate hydrophobicity 2 to 9 0 Accucore XL C8 offers lower hydrophobic retention than columns packed with longer alkyl chain length material, such as C18. It is then therefore recommended for analytes with moderate hydrophobicity, or when a less hydrophobic phase provides optimum retention. 14 Pore Size 0 80Å 300 Carbon Load (%) 0 4 Particle Size 4µm 25 USP L7 Endocrine Disruptors Accucore XL C8 4µm, 150 x 4.6mm Fully porous C8 5µm, 150 x 4.6mm 400 Mobile phase A: Mobile phase B: Gradient: mAU 300 1 200 Rs = 4.12 3 2 100 5 4 0 1 2 3 4 5 Minutes 7 8 9 10 8 9 10 1 400 300 mAU 6 3 2 200 Rs = 5.14 4 100 5 Flow rate: Temperature: Detection: Injection volume: Analytes 0 0 1 2 3 4 5 Minutes 6 7 Columns for Analytical LC 0 water acetonitrile Time (min) % B 0.0 25 20.0 70 20.1 75 25.0 25 1.5mL/min 25°C UV at 220nm 5µL 1. Desethyl Atrazine 2. Simazine 3. Atrazine 4. Diuron 5. Bisphenol A LC Columns and Accessories >> Accucore XL C8 • 31% better resolution of critical pair • 37% narrower peaks • 226% higher sensitivity www.thermoscientific.com/chromatography 4-057 LC Columns and Accessories Accucore Ordering Guide Particle Size (µm) 2.6 Format Defender Guard (4/pk) Length (mm) 10 HPLC Column 30 50 100 150 250 Particle Size (µm) 4 Format Drop-in Guard (4/pk) Length (mm) 10 HPLC Column 50 100 150 Thermo Scientific Chromatography Columns and Consumables 2014-2015 250 4-058 ID (mm) 2.1 3.0 4.6 2.1 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 RP-MS 17626-012105 17626-013005 17626-014005 17626-032130 17626-052130 17626-053030 17626-054630 17626-102130 17626-103030 17626-104630 17626-152130 17626-153030 17626-154630 – C18 17126-012105 17126-013005 17126-014005 17126-032130 17126-052130 17126-053030 17126-054630 17126-102130 17126-103030 17126-104630 17126-152130 17126-153030 17126-154630 – ID (mm) 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 C18 74104-012101 74104-013001 74104-014001 74104-052130 74104-053030 74104-054630 74104-102130 74104-103030 74104-104630 74104-152130 74104-153030 74104-154630 74104-252130 74104-253030 74104-254630 C8 74204-012101 74204-013001 74204-014001 74204-052130 74204-053030 74204-054630 74204-102130 74204-103030 74204-104630 74204-152130 74204-153030 74204-154630 74204-252130 74204-253030 74204-254630 C8 17226-012105 17226-013005 17226-014005 17226-032130 17226-052130 17226-053030 17226-054630 17226-102130 17226-103030 17226-104630 17226-152130 17226-153030 17226-154630 – aQ 17326-012105 17326-013005 17326-014005 17326-032130 17326-052130 17326-053030 17326-054630 17326-102130 17326-103030 17326-104630 17326-152130 17326-153030 17326-154630 – Phenyl-Hexyl 17926-012105 17926-013005 17926-014005 17926-032130 17926-052130 17926-053030 17926-054630 17926-102130 17926-103030 17926-104630 17926-152130 17926-153030 17926-154630 – PFP 17426-012105 17426-013005 17426-014005 17426-032130 17426-052130 17426-053030 17426-054630 17426-102130 17426-103030 17426-104630 17426-152130 17426-153030 17426-154630 – C30 27826-012105 – – – 27826-052130 27826-053030 27826-054630 27826-102130 27826-103030 27826-104630 27826-152130 27826-153030 27826-154630 27826-252130 Length (mm) 10 ID (mm) 2.1 3.0 4.6 Cat. No. 852-00 852-00 850-00 UNIGUARD Guard Cartridge Holder Particle Size (µm) 2.6 Length (mm) 50 100 150 ID (mm) 2.1 HILIC 17526-012105 17526-013005 17526-014005 17526-032130 17526-052130 17526-053030 17526-054630 17526-102130 17526-103030 17526-104630 17526-152130 17526-153030 17526-154630 – Urea-HILIC 27726-012105 – – – 27726-052130 27726-053030 27726-054630 27726-102130 27726-103030 27726-104630 27726-152130 27726-153030 27726-154630 27726-252130 Format Validation Narrow Selectivity Wide Selectivity Polar Selectivity 3-column Kit 17126-052130-3V 17126-102130-3V 17126-152130-3V C18 C18 C18 17X26-052130-3VA 17X26-102130-3VA 17X26-152130-3VA C18 RP-MS aQ 17X26-052130-3VB 17X26-102130-3VB 17X26-152130-3VB C18 Phenyl-Hexyl PFP 17X26-052130-3VC 17X26-102130-3VC 17X26-152130-3VC aQ PFP HILIC Kit contains www.thermoscientific.com/chromatography Columns for Analytical LC Format Phenyl-X 27926-012105 – – – 27926-052130 27926-053030 27926-054630 27926-102130 27926-103030 27926-104630 27926-152130 27926-153030 27926-154630 27926-252130 LC Columns and Accessories >> Polar Premium 28026-012105 – – – 28026-052130 28026-053030 28026-054630 28026-102130 28026-103030 28026-104630 28026-152130 28026-153030 28026-154630 28026-252130 4-059 LC Columns and Accessories Thermo Scientific Acclaim HPLC Columns Optimal selectivity through innovative chemistry • Novel and proprietary surface chemistries for diversified selectivities • Ultrapure, porous, spherical silica • High efficiencies • Low silanol activity for good basic analyte peak shapes • Reproducible and reliable manufacturing process Thermo Scientific™ Acclaim™ columns are based on high-purity, porous silica particles, with advanced and innovative column bonding technologies. This provides complementary selectivity, high column efficiencies, and symmetrical peaks. Acclaim columns meet the high standards set by modern HPLC and LC-MS methods and are used in applications such as pharmaceutical, environmental, food and beverage, chemical, and consumer products. General reversed-phase, HILIC and Specialty phases are available. Acclaim RSLC columns, with a particle size of 2.2μm, are designed for use with UHPLC systems. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Reliability and Durability 4-060 Quality and reliability are essential to a successful analysis. The Acclaim columns are thoroughly tested individually, so that chromatographers can have full confidence in them. Manufacturing starts with an ultrapure silica substrate, using only carefully selected lots with narrow ranges of physical parameters. By design, the bonding processes are clean and repeatable with no unexpected changes in performance. Each batch of bonded silica receives a full suite of validation tests appropriate to its intended use. The bonded silica is packed in precision-polished 316 stainless steel hardware using highly reliable processes. Each packed column is tested to ensure the same great performance every time. The quality assurance reports for silica lot validation and column performance explain the test protocols, list the specifications, and show the actual chromatograms. Performance Indicators Size-exclusion Columns Acclaim columns have been designed to meet the high quality standard needed in laboratories today. The innovative surface chemistries deliver exceptional peak efficiencies for a broad range of analytes. To ensure optimal performance, all Acclaim products are thoroughly characterized using a number of performance indicators, including surface coverage of the bonded phase, metal contamination, steric selectivity, column polarity, column hydrophobicity, and low silanol activity for bases. The specialty columns are also application-tested for their specific analysis, to ensure that each lot of bonded silica provides high-performance separations. Acclaim SEC Columns: Polymeric columns designed for separating water soluble polymers and oligomers in the MW range of 100 to 1,000,000 Daltons. Reversed-phase Columns Mixed-Mode Columns Acclaim 120 C18: High-density, monolayer C18 reversed-phase columns for exceptional resolution in a variety of applications. Hydrophilic Interaction Columns Acclaim HILIC-10: Designed for separating hydrophilic compounds Acclaim Trinity P1 and P2 columns: Unique trimodal surface chemistry provides simultaneous reversed-phase, anion and cation exchange capability for unparalleled chromatographic performance and maximum flexibility in adjusting selectivity (see Specialty Columns). Acclaim 120 C8: High-density monolayer C8 reversed-phase column. Mixed-mode columns provide a unique, adjustable selectivity tool, using variation in pH, ionic strength, or organic modifier to influence the separation selectivity of acids, bases, zwitterions and neutral molecules. Acclaim Phenyl-1: A unique reversed-phase column for the superior separation of aromatic compounds with enhanced hydrolytic stability. Acclaim Mixed-Mode HILIC-1: Combines both reversed-phase and hydrophilic interaction liquid chromatography (HILIC) properties. Acclaim C30: Designed to provide high shape selectivity for separation of hydrophobic structurally related isomers. Acclaim Mixed-Mode WAX-1: High-density monolayer that incorporates both reversedphase and weak anion exchange properties. Acclaim PolarAdvantage: Sulfonamideembedded column for separating a wide variety of analytes. Acclaim Mixed-Mode WCX-1: Reversed-phase and cation exchange combined in a single column. Acclaim PolarAdvantage II: Amideembedded reversed-phase columns with enhanced hydrolytic stability. Hydrophobicity High performance reversed-phase columns for reproducible results Low High pH Range 0 • High hydrophobic retention 14 2 to 8 Pore Size • Excellent efficiencies for maximum resolution 0 • Low silanol activity for excellent peak shapes for basic analytes 300 120 Carbon Load (%) • Reproducible manufacturing practices for reproducible column-to-column performance 0 • Extremely low bleed, fully compatible with MS 25 18 Particle Size 2.2μm, 3μm, 5μm USP L1 • Available in 2.2, 3 and 5μm particle size The Acclaim 120 columns are for high resolution reversed-phase separations. The very high surface coverage and very low metal content together result in columns with excellent efficiencies. These columns provide exceptional performance for a variety of applications in the pharmaceutical, chemical, environmental, and food separations areas. A 250 mAU 4 3 1 B Standard Column 2 220 4 1 2 3 Column: 4 C RSLC Column 6× Acceleration 1 3 2 Double Velocity 12× Acceleration 0 0 0 0 10 20 30 40 50 60 0 2 4 6 Minutes 8 10 0 1 2 3 4 5 25400 A: Acclaim Acclaim 120 120 C18, A: C18,5µm, 5 µm, 4.6 × 150mm 4.6 × 150 mm B, C: Acclaim RSLC C18, .2µm, B,2.1C:× Acclaim 50mm RSLC C18, 2 µm, 2.1 × 50HOAc mm Mobile Phase: 200mM in 10% (v/v)HOAc MeOH Mobile Phase: 200 mM Flow: A: 1.00mL/min inB: 10% (v/v) 0.41mL/min MeOH Flow: A: 1.00 mL/min C: 0.82mL/min Temperature: B:20°C 0.41 mL/min Injection: A: 10µL C:B: 0.82 1.2µL mL/min Temperature: 20 °C C: 1.2µL Detector: UV,10 254µLnm, Injection: A: A: 1 Hz data rate B:B: 1.2 5 HzµL data rate C:C: 1.2 10 HzµL data rate Peaks: Sample: Detector: UV A: 1 B: C: 1 Peaks: 1. p 2. p 3. V 4. V Sample: Com 40% Reference: AO 1. p-Hydroxybenzoic acid 2. p-Hydroxybenzaldehyde 3. Vanillic acid 4. Vanillin Commercial vanilla extract in 40% ethanol, filtered AOAC Official Method 990.25 mAU Reference: Columns for Analytical LC 160 LC Columns and Accessories >> Acclaim 120 C18 0 0.0 www.thermoscientific.com/chromatography 4-061 LC Columns and Accessories Acclaim 120 C8 Hydrophobicity High performance reversed-phase columns with intermediate hydrophobic retention pH Range Low 0 High 14 2 to 8 Pore Size • Low silanol activity for excellent peak shapes for basic analytes • Excellent column efficiencies 0 300 120 Carbon Load (%) • LC-MS compatible • Reproducible manufacturing practices for reproducible column-to-column performance 0 • Available in 2.2, 3 and 5μm particle size Particle Size 2.2μm, 3μm, 5μm 11 Acclaim 120 C8 reversed-phase columns feature densely bonded monolayer C8 ligands on a high-purity, spherical porous silica substrate. The columns are a well-characterized line of LC/MS compatible C8 phases with very high surface coverage and extremely low silanol activity. These columns provide exceptional performance for a variety of applications in the pharmaceutical, environmental, food and many other industrial sectors. Triclosan Separation in Toothpaste Using an Acclaim RSLC C8 Column Triclosan in toothpaste Column: 80 Column: 2 Dimension: Dimension: HPLC System: HPLC System: Buffer: Buffer:Phase: Mobile 281.1 200 nm 400 mAU 1 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0.0 4-062 1.0 Minutes 2.0 3.0 27999 Acclaim RSLC C8, 2.2µm ® Acclaim RSLC C8, 2.2 µm 2.1 × 100mm 2.1 × 100 mm UltiMate 3000 RSLC® 3000 UltiMate 2mM Ammonium acetate pHRSLC 5 2 mM Ammonium acetate pH 5 Isocratic, 15% buffer, (v/v) 15% buffer, 85% methanol (v/v) Mobile Phase: 85% methanol Isocratic, Flow 0.200mL/min FlowRate: Rate: 0.200 mL/min Inj. 1.0µL Inj.Volume: Volume: 1.0 µL Temperature: 50°C Temperature: 50 °C Detection: Diode array detector, 281nm, Detection: Diode array detector, 281 nm, 10 Hz, 10Hz, 0.1 s resp. time and spectra 0.1 s resp. time and spectra 200–400 nm 200–400 nm Samples: containing Samples: ToothpasteToothpaste containing 0.3% triclosan0.3% triclosan Preparation: 1.0 g Toothpaste + 1.0 mL of 7.5 mg/mL Preparation: 1.0g Toothpaste + 1.0mL of 7.5mg/ mL magnesium sulfate + methanol to make 25mL. magnesium sulfate + methanol to make Sonicate and filter. 25 mL. Sonicate and filter. Peaks: 1. Saccharin Peaks: 2. Triclosan 1. Saccharin 2. Triclosan 25 USP L7 Hydrophobicity A unique reversed-phase column with high aromatic selectivity Low High pH Range 0 • High aromatic selectivity 14 2 to 8 Pore Size • High hydrophobic retention • Unique and complementary selectivity compared to any other phenyl type column • Compatibility with highly aqueous mobile phase • High efficiency and rugged packing 0 300 120 Carbon Load (%) 0 Particle Size 3µm, 5µm 13 25 USP L11 Acclaim Phenyl-1 columns provide unique selectivity of aromatic compounds for superior chromatographic performance. This column has a higher π-π interaction than other phenyl phases and provides unique selectivity for aromatic compounds while maintaining sufficient hydrophobic interaction and aqueous compatibility for superior chromatographic performance. Separation of fat-soluble vitamins 60 Column: Acclaim Phenyl-1, 3μm 2 3 mAU 4 Dimension: Mobile Phase: Temperature: Flow Rate: Inj. Volume: Detection: Peaks: 5 6 1 9 8 10 7 0 0 6 12 Minutes 18 24 30 27628 3 × 150mm Methanol/water v/v 90/10 30°C 0.5mL/min 2μL UV at 220nm (100 ppm each) 1. Retinol acetate (vitamin A acetate) 2. Vitamin D2 3. Vitamin D3 4. delta-Tocopherol 5. gamma-Tocopherol 6. alpha-Tocopherol (vitamin E) 7. Impurity (unknown) 8. Vitamin E acetate 9. Vitamin K2 10. Vitamin K1 www.thermoscientific.com/chromatography Columns for Analytical LC The Acclaim Phenyl-1 column can be used in a wide range of applications in pharmaceutical, environmental, food testing and product-quality testing. This column is ideally suited for the analysis of aromatic analytes; some examples include glucocorticosteroids, estrogens, fat-soluble vitamins and phospholipids. LC Columns and Accessories >> Acclaim Phenyl-1 4-063 LC Columns and Accessories Acclaim C30 Hydrophobicity Columns for separating structurally related isomers Low 0 • High shape selectivity 0 • Compatibility with highly aqueous mobile phase 200 300 Carbon Load (%) • High-quality: low column bleed, high efficiency and rugged packing 0 The Acclaim C30 is designed to provide high shape selectivity for separating hydrophobic structural related isomers and unique selectivity complementary to other reversed-phase columns (e.g. C18). Omega fatty acids LLA ng 100 0 14 0 Area 11 9 8 2 pA A 1 34 6 7 10 9 13 12 5 B 0 10 12 14 16 Minutes 18 20 22 24 28381 Particle Size 3µm, 5µm Column: Acclaim C30, 5 µm (4.6 × 150 mm) LC System: UltiMate 3000 RS, Dual Gradient Mobile Phases: A. Water:formic acid:mobile phase B 900:3.6:100 (v/v) B. Acetone:acetonitrile:THF:formic acid Column: Acclaim C30, 5µm (4.6 × 150mm) 675:225:100:4(v/v) LC System: UltiMate 3000 RS, Dual Gradient Gradient: Time (min) %A %B Mobile Phases: A. Water:formic acid:mobile phase B 0 100 0 900:3.6:100 (v/v) 1 40 60 B. Acetone:acetonitrile:THF:formic acid 675:225:100:4(v/v) 13 30 70 Gradient: Time (min) %A %B 22 5 95 0 24 100 5 0 95 1 40 60 29 100 70 0 13 30 0 2232 5 100 95 Flow Rate: 1.00 24 mL/min5 95 100 0 Temperature: 3029°C 100 0 Inj. Volume: 2 32 µL Flow Rate: 1.00mL/min Detection: Corona ultra, nebulizer 15 °C, filter high Temperature: 30°C Samples: A.2µL Standards in isopropanol Inj. Volume: B.Corona Saponified chicken 15°C, fat filter high Detection: ultra, nebulizer Samples: Peaks: Peaks: Standards in8. isopropanol DPA 1.A.SDA Saponified chicken fat 9. 9E, 14Z-CLA 2.B.EPA 1. SDA 10. ETA 3.2.ALA EPA 11. Adrenic 4.3.GLA ALA 12. Oleic GLA 5.4.DHA DHA 13. EDA 6.5.Arach. 6. Arach. 14. Erucic 7. LLA 7. LLA Thermo Scientific Chromatography Columns and Consumables 2014-2015 14 2 to 8 Pore Size • Unique selectivity complementary to other reversed-phase columns 6000 4-064 High pH Range 13 25 USP L62 Hydrophobicity Novel polar-embedded reversed-phase columns with unique selectivity Low 0 • Selectivity complementary to the C18 column High pH Range 14 2 to 8 Pore Size • Low silanol activity for excellent peak shape with basic compounds • Compatible with 100% aqueous mobile phase 0 300 120 Carbon Load (%) • High selectivity for hydrophobic aromatic molecules 0 • Wide range of applications Particle Size 2.2μm, 3μm, 5μm • Available in 2.2, 3 and 5μm 16 25 USP L60 Acclaim PolarAdvantage (PA) columns feature a patented bonding column chemistry that incorporates a polar sulfonamide group with an ether linkage near the silica surface. This unique chemistry provides low silanol activity, compatibility with 100% aqueous mobile phase. The Acclaim PA column offers great separation power to resolve a wide variety of polar and nonpolar analytes and supports LC/MS analysis. EPA 604 phenols 60 Column: Acclaim RSLC PolarAdvantage, 2.2μm 7 5 mAU 2 3 6 4 1 8 9 10 11 0 0 0.5 1.0 1.5 Minutes 2.0 2.5 3.0 27652 Dimensions: 3 × 50mm System: UltiMate 3000 RSLC Mobile Phases: A: 10mM formic acid + 10mM ammonium formate, pH 3.75 ± 0.05 B: Acetonitrile Gradient Time (min): -1.5 0.0 0.3 2.6 3.0 %A 70 70 70 10 10 %B 30 30 30 90 90 Flow Rate: 1.25mL/min Temperature: 30°C Injection Volume: 0.5μL Detection: UV at 280nm, 10Hz, 0.5s resp. time Sample: Calibration mix, 50μg/mL in water Peaks: 1. Phenol 2. 2,4-Dinitrophenol 3. 4-Nitrophenol 4. 2-Chlorophenol 5. 2-Nitrophenol 6. 2,4-Dimethylphenol 7. 4,6-Dinitro-2-methylphenol 8. 4-Chloro-3-methylphenol 9. 2,4-Dichlorophenol 10. 2,4,6-Trichlorophenol 11. Pentachlorophenol www.thermoscientific.com/chromatography Columns for Analytical LC Acclaim PA columns provide unique selectivity, good peak shape for acidic, basic, and neutral analytes, and full compatibility with 100% aqueous conditions. Applications include pharmaceutical, environmental, life science, food testing, and product-quality testing. LC Columns and Accessories >> Acclaim PolarAdvantage 4-065 LC Columns and Accessories Acclaim PolarAdvantage II Hydrophobicity Complementary selectivity and enhanced hydrolytic stability Low 0 • Unique selectivity complementary to the C18 column • Novel polar-embedded column chemistry for hydrolytic stability 0 • Compatible with 100% aqueous mobile phase 300 120 Carbon Load (%) • Low bleed for MS compatibility 0 • Wide range of applications Particle Size 2.2μm, 3μm, 5μm • Available in 2.2, 3 and 5μm Acclaim PolarAdvantage II (PA2) columns feature a patented surface chemistry that incorporates an amide-embedded polar group and multi-point attachment between the ligands and the silica surface. This unique chemistry provides enhanced hydrolytic stability from pH 1.5-10 with 100% aqueous mobile phases and exhibits high reversed-phase capacity, with selectivity complementary to conventional C18 columns. Turmeric 200 1, 2, 3 Area: 9.7 mAU*Min. mAU C18 1 2 PA2 0 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 14 1.5 – 10.5 Pore Size The Acclaim PA2 column is specifically designed to withstand high pH conditions, making it a good choice for the separation of both basic and acidic analytes. 4-066 High pH Range 1 2 Minutes Total area: 9.9 mAU*Min. 3 3 4 25402 Columns: Acclaim RSLC 120 C18 Acclaim RSLC PA2 Dimensions: Mobile Phase: 2.2μm, 2.1 × 100mm A: 15mM H 3PO 4 B: Methanol C18: 70% B (v/v) PA2: 80% B (v/v) 0.41mL/min 30°C UV, 428nm Turmeric extract 1. Curcumin 2. Demethoxycurcumin 3. Bis-demethoxycurcumin Isocratic: Flow: Temperature: Detector: Sample: Peaks: 16 25 USP L60 Hydrophobicity Designed with unique selectivity for hydrophilic molecules Low High pH Range 0 2 to 8 • Retains highly polar molecules that are not retained by Pore Size • Unique selectivity, complementary to reversed-phase columns 0 • Hydrolytically stable Particle Size 3µm, 5µm reversed-phase chromatography 120 14 300 • Rugged column packing • Broad application range The Acclaim HILIC-10 column is designed for separating highly hydrophilic molecules by Hydrophilic Interaction Liquid Chromatography (HILIC). This column is based on high-purity spherical porous silica covalently modified with a proprietary hydrophilic layer. 200 Column: Acclaim HILIC-10, 3μm Dimensions: Mobile Phase: 4 2 3 pA 1 7 8 5 6 0 0 2 4 6 Minutes 8 10 27129 3 × 150mm A: Heptane B: 2-Propanol/acetic acid 99.5:0.5 Step Elution (min): –6.0 0.0 0.5 4.0 10.0 %A 99 99 96 87 87 %B 1 1 4 13 13 Flow Rate: 0.50mL/min Temperature: 25°C Inj. Volume: 4μL Detector: Corona ultra, nebulizer 15°C Peaks: 1. Tristearin 2. Trilaurin 3. Distearin isomer 1 4. Dilaurin isomer 1 5. Distearin isomer 2 6. Dilaurin isomer 2 7. Monostearin 8. Monolaurin www.thermoscientific.com/chromatography Columns for Analytical LC Glycerides LC Columns and Accessories >> Acclaim HILIC-10 4-067 LC Columns and Accessories Acclaim Mixed-Mode HILIC-1 Hydrophobicity Uniquely designed for both reversed-phase and HILIC operations Low 0 • Can operate in both RP and HILIC modes 14 • Retains highly polar molecules 0 • Unique selectivity complementary to RP columns • Broader application range compared with conventional diol-based columns The Acclaim Mixed-Mode HILIC-1 column features a unique, high-efficiency, silica-based HPLC mixed-mode stationary phase that combines both reversed-phase (RP) and hydrophilic interaction liquid chromatography (HILIC) properties. This combination allows both hydrophobic interaction and hydrophilic interaction to be utilized to optimize separations. The Acclaim Mixed-Mode HILIC-1 stationary phase consists of a hydrophobic alkyl chain with a diol group at the terminus. The hydrophobic moiety provides reversed-phase retention and the terminal diol group facilitates hydrophilic interactions. This unique combination results in the adjustable selectivity, making Acclaim Mixed-Mode HILIC-1 separate mixtures that would be impossible for a C18 column. This column is suitable for a broad range of applications, including non-ionic ethoxylated surfactants, drug metabolites, lipids, polyethylene glycols (PEGs), ethoxylated surfactants, and more. Column: Acclaim Mixed-Mode HILIC-1, 5μm Reversed-Phase Mode Dimensions: Mobile Phase: AU 2 Temperature: Flow Rate: Inj. Volume: Detection: Peaks: 2 1 HILIC Mode 0 2 4 Minutes 6 8 10 24254 120 Particle Size 3µm, 5µm • High-efficiency column for high-resolution separations 1 Thermo Scientific Chromatography Columns and Consumables 2014-2015 2.5 to 7.5 Pore Size Cytosine and naphthalene 4-068 High pH Range 4.6 × 150mm CH 3CN/0.1 M NH 4OAc, pH 5.2 v/v 52/48 for RP mode v/v 92/8 for HILIC mode 30°C 1mL/min 10μL UV at 254nm 1. Cytosine (100 ppm) 2. Naphthalene (100 ppm) 300 pH Range Designed for separating anionic molecules; with powerful adjustable selectivity control Pore Size 0 0 2.5 to 7.5 120 Particle Size 3µm, 5µm • Adjustable selectivity 14 300 USP L78 • Selectivity orthogonal to reversed-phase (RP) columns • Ideal selectivity for anionic molecules • Excellent column efficiency and peak asymmetry • Multimode retention mechanisms: reversed-phase, weak anion exchange, and HILIC modes The Acclaim Mixed-Mode WAX-1 is a novel, high-efficiency silica HPLC column that combines hydrophobic and weak anion exchange characteristics. Its unique chemistry results in a multimode separation mechanism that includes reversed-phase, anion exchange, and HILIC interactions. Selectivity can be adjusted by changing ionic strength, pH, or organic solvent content. Pain relief medicine Column: Acclaim Mixed-Mode WAX-1, 5μm Dimensions: Mobile Phase: 3 Temperature: Flow Rate: Inj. Volume: Detection: Peaks: 2 AU 1 4 O OH OH 4.6 × 150mm 40/60 v/v Acetonitrile/buffer (6.8 g potassium monophosphate and 0.5 g pyrophosphate in 1000 g D.I. H2 O, pH is adjusted to 6.0 with NaOH) 30°C 1mL/min 1μL UV, 220nm 1. Caffeine 2. Acetaminophen 3. Salicylamide 4. Acetyl salicylic acid (Aspirin) 5. Salicylic acid Columns for Analytical LC The Acclaim Mixed-Mode WAX-1 surface consists of a hydrophobic alkyl chain with a tertiary amine group at the terminus. The hydrophobic moiety provides reversed-phase retention and the terminal amino group facilitates electrostatic interactions. LC Columns and Accessories >> Acclaim Mixed-Mode WAX-1 5 0 10 Minutes 20 www.thermoscientific.com/chromatography 4-069 LC Columns and Accessories Acclaim Mixed-Mode WCX-1 pH Range Designed for separating cationic molecules with adjustable selectivity control Pore Size 0 0 • Ideal selectivity for separating basic molecules • Selectivity complementary to C18 RP columns • Multimode separation mechanism: reversed-phase, weak cation exchange, anion-exclusion and HILIC The Acclaim Mixed-Mode WCX-1 is a novel, high-efficiency, silica-based column, manufactured by bonding a specially designed proprietary ligand with both hydrophobic and weak cation exchange properties. Selectivity of ionizable and neutral compounds can be controlled independently or simultaneously by tuning mobile phase ionic strength, pH or organic modifier. This column therefore can separate using multiple separation modes: reversed-phase, cation exchange, and normal-phase/ HILIC. Basic compounds are important in a variety of industrial applications, including pharmaceutical, chemical, consumer products, foods and beverages, and more. The Acclaim Mixed-Mode WCX-1 not only retains basic molecules (from highly hydrophilic to highly hydrophobic), but also separates them with symmetrical peak shapes and excellent efficiency. Pharmaceutical counterions Column: Acclaim Mixed-Mode WCX-1, 5μm Dimension: 4.6 × 150mm 1 Mobile Phase: 3 Temperature: Flow Rate: Inj. Volume: Detection: Peaks: 7 2 5 4 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0 4-070 6 4 Minutes 120 Particle Size 3µm, 5µm • Adjustable selectivity AU 2.5 to 7.5 8 40/60 v/v CH 3 CN/NH 4 OAc, pH 5.2 (20 mM total) 30°C 1mL/min 5µL UV (225 nm) 1. Maleate 50μg/mL 2. Ketoprofen 30μg/mL 3. Naproxen 30μg/mL 4. Hydrocortisone 60μg/mL 5. Dexamethasone 60μg/mL 6. Oxprenolol 300μg/mL 7. Timolol 250μg/mL 14 300 Particle Size (µm) 2.2 Format RSLC Column Length (mm) 30 50 75 100 3 HPLC Column 150 250 33 50 75 100 250 5 Guard Cartridge 10 HPLC Column 50 100 150 250 Format Acclaim Guard Cartridge Holder Acclaim Guard Cartridge-Column Coupler Acclaim Guard Kit (Holder and Coupler) 120 C18 071400 071606 068981 071605 075697 068982 071604 071399 074812 066272 059128 068971 059131 066273 059129 076186 059132 059130 063691 059133 076187 070077 069689 071981 069695 059142 059146 059143 059147 059144 059148 059145 059149 120 C8 072614 072618 072615 072619 075696 072616 072620 072617 074811 – 059122 – 059125 – 059123 076184 059126 059124 068970 059127 076185 070078 069688 071979 069696 059134 059138 059135 059139 059136 059140 059137 059141 PolarAdvantage 072621 072625 072622 072626 075698 072623 072627 072624 074813 066274 063174 068972 – 066275 061316 076214 076216 061317 063693 061318 076215 070079 069691 071983 069698 – 061319 – – – 061320 – 061321 PolarAdvantage II 071402 071609 068989 071608 075699 068990 071607 071401 074814 066276 077999 068973 063189 066277 077998 078000 078001 063187 063705 063191 077997 070080 069692 071985 069699 – – – – – 063197 – 063199 Columns for Analytical LC 150 ID (mm) 2.1 3.0 2.1 3.0 3.0 2.1 3.0 2.1 2.1 3.0 2.1 3.0 4.6 3.0 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 2.1 3.0 4.6 2.1 4.6 2.1 4.6 2.1 4.6 2.1 4.6 LC Columns and Accessories >> Acclaim Ordering Guide Cat. No. 069580 074188 069707 www.thermoscientific.com/chromatography 4-071 LC Columns and Accessories Acclaim Ordering Guide continued Acclaim Phenyl-1 Ordering Guide Particle Size (µm) 3 Format Guard Cartridge Length (mm) 10 HPLC Column 50 100 150 250 5 HPLC Column 150 250 ID (mm) 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 2.1 4.6 4.6 Phenyl-1 079934 071974 071973 078016 071972 078018 078015 074693 078017 071971 071970 071969 078014 074694 079698 088016 079697 ID (mm) 2.1 3.0 4.6 2.1 4.6 2.1 3.0 4.6 2.1 3.0 2.1 3.0 4.6 4.6 4.6 C30 078666 078663 078661 078665 078660 075725 075724 075723 078664 075726 075722 075721 075720 075719 075718 ID (mm) 2.1 3.0 4.6 2.1 3.0 4.6 HILIC-10 074259 074258 074257 074263 074261 074262 ID (mm) 3.0 2.1 3.0 2.1 3.0 4.6 2.1 4.6 4.6 HILIC-1 071912 070091 070090 069694 071913 069706 066847 066843 066844 Acclaim C30 Ordering Guide Particle Size (µm) 3 Format HPLC Column Length (mm) 50 100 150 250 Thermo Scientific Chromatography Columns and Consumables 2014-2015 5 4-072 Guard Cartridge 10 HPLC Column 150 250 Acclaim HILIC-10 Ordering Guide Particle Size (µm) 3 Format HPLC Column Length (mm) 150 5 Guard Cartridge 10 Acclaim Mixed-Mode Ordering Guide Particle Size (µm) 3 Format HPLC Column Length (mm) 50 150 5 Guard Cartridge 10 HPLC Column 150 250 WAX-1 071908 070089 070088 069686 071909 069704 067084 064984 064985 WCX-1 071910 070093 070092 085455 071911 069705 068371 068353 068352 LC Columns and Accessories >> Notes Columns for Analytical LC 4-073 www.thermoscientific.com/chromatography LC Columns and Accessories Thermo Scientific Syncronis HPLC Columns Consistent Reproducible Separations, Column after Column, Time after Time. Extensive testing and strong quality control procedures ensure the consistency of Syncronis HPLC columns – column after column. • • Thermo Scientific™ Syncronis™ HPLC columns are manufactured, packed and tested in ISO9000 accredited facilities. Each lot of silica is tested for the physical properties of the silica support and only released for production if it meets the stringent test specifications. • Available in three particle sizes: 1.7μm for rapid UHPLC separations plus 3µm and 5μm for the more traditional HPLC analysis. • Syncronis reversed phase columns are densely bonded and double endcapped to minimize the number of residual silanols available to interact with basic analytes. Syncronis columns are based on highly pure 100Å silica, with a surface area of 320m2/g, compared to 200m2/g for typical silica based material. This greater surface area ensures good retention of analytes having a range of hydrophobicity, away from the solvent front. • Each batch of chromatographic media packed into Syncronis columns is put through a series of diagnostic chromatographic tests, based on those developed by Tanaka1 to ensure consistent, predictable separations. Thermo Scientific Chromatography Columns and Consumables 2014-2015 View product information and application notes 4-074 These tests rigorously probe interactions between analytes and the stationary phase, measuring hydrophobicity, shape selectivity and secondary interactions with bases, acids and chelators. • Enhanced, automated packing methods drive consistency even further and every column is individually tested to ensure that it meets the required quality. For full details on the Syncronis column range, please request or view a copy of our Syncronis technical guide www.thermoscientific.com/syncronis Hydrophobicity Syncronis C18 columns deliver consistent predictable separations, column after column, time after time pH Range Low 0 High 2 to 9 14 Pore Size • Highly pure, high surface area silica • High carbon load for increased retention 0 300 100 Carbon Load (%) • Double endcapped for extra surface coverage • Highly inert towards basic compounds 0 • Rigorously tested to ensure quality Particle Size 1.7μm, 3μm, 5μm 16 25 USP L1 When developing a new method, one of the most important goals for the chromatographer is to achieve a consistent, reproducible separation. The selection of a highly reproducible HPLC column is essential to attaining this goal. Ziovudine Column: Syncronis C18, 5μm, 150mm x 4.6mm 2400 2000 Mobile phase: Flow rate: Temperature: Detection: Injection volume: 1. Zidovudine mAU 1600 1200 800 Water:Methanol (4:1) 1.0mL min-1 25°C 265nm 10μL Columns for Analytical LC Syncronis C18 columns show excellent column to column reproducibility, as illustrated here by the analysis of zidovudine using five separate columns. The reproducibility in terms of retention time and peak area is less than or equal to 0.5%, column to column. LC Columns and Accessories >> Syncronis C18 400 0 0 4 8 12 16 Minutes 20 24 28 32 www.thermoscientific.com/chromatography 4-075 LC Columns and Accessories Syncronis C8 Hydrophobicity Reduces hydrophobic interactions allowing compounds to elute quicker from the column. Recommended for analytes with medium hydrophobicity or when a less hydrophobic phase is required to obtain optimum retention 0 2 to 8 14 Pore Size 0 • Highly pure, high surface area silica 300 100 • Less hydrophobic than Syncronis C18 0 • Double endcapped for extra surface coverage Particle Size 1.7µm, 3µm, 5µm • Rigorously tested to ensure quality Uron herbicides 1000 Column: Syncronis C8, 5μm, 150mm x 4.6mm Mobile phase: 800 A: Water B: Acetonitrile Gradient: 35 to 60% B in 10 minutes Flow rate: 1.0mL/min Temperature: 30°C Detection: 240nm Injection volume.: 20μL 1. Tebuthiuron 2. Metoxuron 3. Monuron 4. Chlorotoluron 5. Diuron 6. Linuron 4 2 600 mAU 5 400 6 1 200 0 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High Carbon Load (%) 3 4-076 Low pH Range 2 4 6 8 Minutes 10 12 14 16 11 25 USP L7 Hydrophobicity Polar endcapped Syncronis aQ columns provide a controlled interaction mechanism that retains and resolves polar analytes. Stable in 100% aqueous mobile phase Low High pH Range 0 2 to 8 14 Pore Size • Stable in 100% aqueous mobile phase 0 300 100 Carbon Load (%) • Enhanced retention of polar compounds • Rigorously tested to ensure quality 0 Particle Size 1.7μm, 3μm, 5μm 19 25 USP L1 In comparison to a conventionally endcapped C18, the Syncronis aQ polar end-capped C18 stationary phase exhibits superior stability towards aqueous mobile phase. Syncronis aQ shows no degradation in performance after 100 injections in a buffered 100% aqueous eluent. DAD1 A, Sig=260,4 Ref=360,100 (110809B\R2000001.D) Stability of Syncronis aQ in 100% aqueous mobile phase 80 Column: Syncronis aQ, 5μm, 100mm x 4.6mm mAU 40 20 0 Ref=360,100 (110809B\R2000085.D) 0DAD1 A, Sig=260,4 1 2 3 4 Minutes 5 6 7 8 100% aqueous inj2 0 5 6 7 8 100% aqueous inj100 80 Columns for Analytical LC Mobile phase: 50mM Aqueous K 2HPO 4 (pH 6) Flow rate: 0.7mL/min Temperature: 30°C Detection: 260nm Injection volume: 2μL 1. Cytidine-5’-diphosphate 2. Adenosine-5’-triphosphate 3. Adenosine-5’-diphosphate 4. Adenosine-5’-monophosphate 60 LC Columns and Accessories >> Syncronis aQ mAU 60 40 20 0 1 2 3 Minutes 4 www.thermoscientific.com/chromatography 4-077 Hydrophobicity Provides an alternative to Syncronis C18 and is particularly useful for retention of aromatic compounds pH Range • Alternative selectivity to Syncronis C18 • Double endcapped for extra surface coverage 2 to 8 14 0 300 100 Carbon Load (%) • Highly inert towards basic compounds • Rigorously tested to ensure quality 0 11 Particle Size 1.7µm, 3µm, 5µm Oxacillin sodium (USP) Column: Syncronis Phenyl, 5μm, 300mm x 4.0mm 140 Mobile phase: Phosphate Buffer: MeCN:MeOH (70:30:10) Flow rate: 1.0mL/min (2mL/min in USP method) Temperature: 25°C Detection: 225nm Injection volume: 10μL 1. Oxacillin Sodium (0.11mg/mL) 120 100 80 60 20 0 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High Pore Size 40 4-078 Low 0 mAU LC Columns and Accessories Syncronis Phenyl 1 2 3 4 5 6 7 8 Minutes 9 10 11 12 13 14 15 25 USP L11 Hydrophobicity Provides a versatile aminopropyl phase that gives excellent chromatographic properties in four modes: weak anion exchange, reversed phase, normal phase and HILIC Low High pH Range 0 2 to 8 14 Pore Size • Highly pure, high surface area silica 0 175 300 Carbon Load (%) • Double endcapped for extra surface coverage • Rigorously tested to ensure quality 0 4 Particle Size 1.7μm, 3μm, 5μm 25 USP L8 Lactulose Column: Syncronis Amino 5μm, 150mm x 4.6mm Mobile phase: Water: MeCN (30:70) Flow rate: 1.0mL/min Temperature: 35°C Detection: RI Injection volume: 5μL 1. Lactulose 2.00 uRIU 1.50 1.00 Columns for Analytical LC 0.50 0.00 -0.25 0 1 2 3 4 5 Minutes 6 7 8 9 LC Columns and Accessories >> Syncronis Amino 10 www.thermoscientific.com/chromatography 4-079 Hydrophobicity Serves as a powerful and efficient tool for the chromatography of moderately polar organic compounds by normal phase chromatography pH Range • Highly pure, high surface area silica 2 to 8 • Excellent reproducibility for normal phase chromatography 0 • Rigorously tested to ensure quality Particle Size 1.7µm, 3µm, 5µm Tocopherols 12 Column: Syncronis Silica 5µm, 150 x 4.6mm Part Number: 97005-154630 10 Mobile phase: Hexane +0.2% propan-2-ol (IPA) Flow rate: 2.0mL/min Temperature: 40°C Detection: 254nm Injection volume: 10µL Sample: Mixture of standards (200-1000 ug/ml) of the following: α-tocopherol β -tocopherol γ -tocopherol δ-tocopherol 8 6 2 0 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 High 5 10 15 Minutes 14 Pore Size 4 4-080 Low 0 mAU LC Columns and Accessories Syncronis Silica 20 25 100 300 USP L3 Hydrophobicity Provides enhanced retention of polar and hydrophilic analytes Low High pH Range 0 • Alternative selectivity to Syncronis C18 2 to 8 14 100 300 Pore Size • Improved sensitivity with MS detection 0 • No need for ion-pair or derivatisation Carbon Load (%) • Outstanding peak shape and sensitivity 0 • Highly pure, high surface area silica particles Catecholamines Column: Syncronis HILIC, 5µm, 250 x 4.6mm Part Number: 97505-254630 500 mAU 300 2 4 3 5 6 7 0 0 5 10 Minutes 15 20 www.thermoscientific.com/chromatography Columns for Analytical LC Mobile phase: water : acetonitrile : 200mM ammonium formate (10.5 : 84.5 : 5) Flow rate: 1.0mL/min Temperature: 40°C Detection: 280nm Injection volume: 5µL Sample: 1. catechol 2. 5-HIAA 3. DOPAC 4. serotonin 5. tyrosine 6. dopamine 7. L-DOPA 1 400 100 25 Particle Size 1.7μm, 3μm, 5μm • Neutral (uncharged), highly polar surface 200 5 LC Columns and Accessories >> Syncronis HILIC 4-081 LC Columns and Accessories Syncronis Ordering Guide Particle Size (µm) 1.7 Format UHPLC Column Length (mm) 30 50 100 3 HPLC Column 30 50 100 150 5 Drop-in Guard (4/pk) 10 HPLC Column 30 50 100 150 Thermo Scientific Chromatography Columns and Consumables 2014-2015 250 4-082 Format UNIGUARD Guard Cartridge Holder Length (mm) 10 ID (mm) 2.1 2.1 3.0 4.6 2.1 3.0 2.1 2.1 3.0 4.6 2.1 3.0 4.6 2.1 4.6 2.1 3.0 4.0/4.6 2.1 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 ID (mm) 2.1 3.0 4.0/4.6 C18 97102-032130 97102-052130 97102-053030 97102-054630 97102-102130 97102-103030 97103-032130 97103-052130 97103-053030 97103-054630 97103-102130 97103-103030 97103-104630 97103-152130 97103-154630 97105-012101 97105-013001 97105-014001 97105-032130 97105-052130 97105-053030 97105-054630 97105-102130 97105-103030 97105-104630 97105-152130 97105-153030 97105-154030 97105-154630 97105-252130 97105-253030 97105-254030 97105-254630 Cat. No. 852-00 852-00 850-00 C8 – 97202-052130 – 97202-054630 97202-102130 97202-103030 – 97203-052130 – – – 97203-103030 97203-104630 – 97203-154630 97205-012101 97205-013001 97205-014001 97205-032130 97205-052130 97205-053030 97205-054630 97205-102130 97205-103030 97205-104630 97205-152130 97205-153030 97205-154030 97205-154630 – 97205-253030 97205-254030 97205-254630 aQ – 97302-052130 – 97302-054630 97302-102130 97302-103030 – 97303-052130 – – – 97303-103030 97303-104630 – 97303-154630 97305-012101 97305-013001 97305-014001 97305-032130 97305-052130 97305-053030 97305-054630 97305-102130 97305-103030 97305-104630 97305-152130 97305-153030 97305-154030 97305-154630 – 97305-253030 97305-254030 97305-254630 Phenyl – 97902-052130 – 97902-054630 97902-102130 97902-103030 – 97903-052130 – – – 97903-104630 – 97903-154630 97905-012101 97905-013001 97905-014001 97905-032130 97905-052130 97905-053030 97905-054630 97905-102130 97905-103030 97905-104630 97905-152130 97905-153030 97905-154030 97905-154630 – 97905-253030 97905-254030 97905-254630 HILIC – 97502-052130 – 97502-054630 97502-102130 97502-103030 – 97503-052130 – – – – 97503-104630 97503-152130 97503-154630 97505-012101 97505-013001 97505-014001 97505-032130 97505-052130 97505-053030 97505-054630 97505-102130 97505-103030 97505-104630 97505-152130 97505-153030 97505-154030 97505-154630 – 97505-253030 97505-254030 97505-254630 Columns for Analytical LC Silica – 97002-052130 – 97002-054630 97002-102130 97002-103030 – 97003-052130 – – – – 97003-104630 – 97003-154630 97005-012101 97005-013001 97005-014001 97005-032130 97005-052130 97005-053030 97005-054630 97005-102130 97005-103030 97005-104630 97005-152130 97005-153030 97005-154030 97005-154630 – 97005-253030 97005-254030 97005-254630 LC Columns and Accessories >> Amino – 97702-052130 – 97702-054630 97702-102130 97702-103030 – 97703-052130 – – – 97703-104630 – 97703-154630 97705-012101 97705-013001 97705-014001 97705-032130 97705-052130 97705-053030 97705-054630 97705-102130 97705-103030 97705-104630 97705-152130 97705-153030 97705-154030 97705-154630 – 97705-253030 97705-254030 97705-254630 www.thermoscientific.com/chromatography 4-083 LC Columns and Accessories Thermo Scientific Hypercarb HPLC Columns 100% porous graphitic carbon for extended separation capabilities Hydrophobicity Low High pH Range • Exceptional retention of very polar analytes 0 • Separates structurally related substances 14 Pore Size • pH stable from 0 to 14 0 250 300 Carbon Load (%) • Ideal for high temperature applications 0 100 Particle Size 3µm, 5µm, 7µm Porous Graphitic Carbon (PGC) is a unique stationary phase composed of flat sheets of hexagonally arranged carbon atoms with a satisfied valence, as in a very large polynuclear aromatic molecule. Thermo Scientific™ Hypercarb™ columns are unlike traditional silica bonded phases in both its structure and retentive properties, allowing for total pH stability and the retention and separation of highly polar species. Hypercarb columns are ideally suited to solve “problem” separations, in both reversed phase and normal phase HPLC and LC/MS applications. Schematic representation of molecular area of a planar and non-planar molecule interacting with the Hypercarb surface Thermo Scientific Chromatography Columns and Consumables 2014-2015 Retention and Resolution 4-084 The mechanism of interaction is very dependent upon both the polarity and planarity (shape) of the solute. These specific interaction mechanisms allow the successful retention and resolution of analytes that cannot be separated by typical reversed phase HPLC. Removal of complex buffering systems or ion-pair reagents, and use of increased organic modifier concentration for polar analytes allows greater compatibility with detection techniques such as MS. The overall retention on Hypercarb columns is a combination of two mechanisms: 1) Adsorption: The strength of analyte interactions with Hypercarb columns is largely dependent on the molecular area in contact with the graphite surface, and also on the type and positioning of the functional groups in relation to the graphite surface at the points of contact. The approach of a planar and a non-planar molecule to the Hypercarb surface is shown in the diagrams above. The strength of the interaction depends upon the size and orientation of the molecular area that is able to come in contact with the flat graphite surface. More planar molecules will show more retention than rigid molecules with a 3-dimensional spatial arrangement. Schematic representation of a point charge approaching the Hypercarb surface 2) Charge induced interactions of a polar analyte with the polarizable surface of graphite: The second mechanism, charge-induced dipole, is illustrated above and accounts for the strong retention exhibited by polar analytes. As the polar group with a permanent dipole approaches the surface, an induced dipole is formed, increasing the attraction between the analyte and graphite surface. These charges should not be confused with the overall ionic charge of the molecule, such as a basic compound ionized in acidic pH conditions. The charge-induced dipole mechanism is strictly due to the interaction of the electrostatic charge of the polar molecule with the graphite surface. The strong mechanisms of interaction with Hypercarb columns usually allows for shorter columns to be used during the method development process. In most cases, 100mm length columns or shorter are sufficient for a separation. PS-DVB Hypercarb Courtesy V. Coquart and M-C. Henion, J. Chrom., 1992 Retention on Hypercarb columns increases as polarity of the analyte increases, which is the opposite of typical reversed phase materials such as PS-DVB Extended pH Range One of the other key benefits of Hypercarb columns is the extreme stability of the phase to chemical or physical attack. Due to the unique characteristics of the media, it can withstand chemical attack across the entire pH range of 0 to 14, allowing applications to be run at pH levels that are incompatible with typical silica-based columns. Hypercarb columns offer more choice in buffer selection while handling both high temperature and high pressure. Hypercarb column, 5µm, 100 x 0.32mm Part Number: 35005-100365 columns Mobile Phase: Gradient: Flow Rate: Temperature: Detection: Analytes: A: H 2O + 0.1% formic acid B: ACN + 0.1% formic acid 0 to 25% B in 15 minutes 8µL/min 25°C UV at 254nm 1. Cytosine 2. Uracil 3. Guanine 4. Adenine 5. Xanthine 6. Thymine Columns for Analytical LC In typical reversed phase chromatography, the retention of an analyte is directly related to its hydrophobicity: the more hydrophobic the analyte, the longer its retention. Conversely, as the polarity of the analyte increases, analyte-solvent interactions begin to dominate and retention is reduced. This observation holds true for the majority of reversed phase systems. An exception to this rule is Hypercarb columns, for which retention may in some cases increase as the polarity of the analyte increases, illustrated to the right. This phenomenon is referred to as the “polar retention effect on graphite” (PREG). This property makes Hypercarb columns particularly useful for the separation of highly polar compounds (with logP as low as -4) that are normally difficult to retain and resolve on silica-based alkyl chain phases. The retention of very polar solutes on Hypercarb columns can be achieved without ion pair reagents or complex mobile phase conditions, as illustrated in the chromatogram below. LC Columns and Accessories >> Increased Retention of Polar Analytes Additional retention is achieved for polar compounds using a Hypercarb column compared to a polar endcapped C18. Note also the change in elution order. columns 1 1 4 2 Day 1 at pH 12 2 Day 93 at pH 12 4 Part Number: Mobile Phase: Isocratic: Flow Rate: Detection: Sample: 3 3 5 5 6 Hypercarb column, 5µm, 100 x 4.6mm 6 35005-104630 MeOH:H 2O 70:30 0.7mL/min UV at 254nm 1. Acetone 2. Phenol 3. p-Cresol 4. Anisol 5. Phenetole 6. 3,5 -Xylenol PGCpH12 0 10 20 Min 0 10 20 Min Hypercarb column stability at pH 12: retention and selectivity do not change even after 93 days of storage in 0.1M NaOH/MeOH www.thermoscientific.com/chromatography 4-085 LC Columns and Accessories Hypercarb column C18-silica Courtesy of Josef Kriz et al, 1994 Typical C18 silica Comparison of methyl and methylene group selectivity on C18 and Hypercarb columns Hypercarb Courtesy of Norman Smith, Glaxo Group Research, Greenford, 1988 1 1’ 1 Δ3 Δ2 3 5 2,2’ Free Acid 3’ 2 2’ 0 1’ Anti Isomers 3 3’ 7022 10 Min 15 20 0 5 10 Min 15 Hypercarb column, 5µm, 100 x 4.6mm Hypersil SAS, 5µm, 200 x 4.6mm Part Number: Mobile Phase: Isocratic: Flow Rate: Detector: Part Number: Mobile Phase: Isocratic: Flow Rate: Detector: 35005-104630 ACN:H 2O:MeOH:Dioxan 38:20:35:10 1mL/min UV at 254nm 20 Surface comparison between C18 bonded silica and Hypercarb porous graphitic carbon 30505-204630 MeOH:0.05M NH 4H 2PO 4 38:62 1mL/min UV at 254nm Thermo Scientific Chromatography Columns and Consumables 2014-2015 Separation of geometric isomers of Axetil: comparison of a Hypercarb and bonded silica column 4-086 Resolution of Structurally Related Compounds Ideal for Reversed Phase LC/MS of Polar Compounds By virtue of the nature of the surface and the way solute shape affects retention, Hypercarb columns can differentiate between closely related analytes such as isomers and homologous series. Where no discrimination between methylene and methyl groups is observed using a traditional C18 column, considerable resolving power is observed with Hypercarb columns, as shown above. The differentiation of analytes is based on their fit to the graphite surface, allowing for the chromatographic resolution of compounds that are very similar in structure as shown above with the resolution of diastereomers of the antibiotic Axetil. The Hypercarb column provides both a significant improvement in separation over the silica-based column originally used, as well as a change in elution order. Reversed phase-LC/MS analysis of very polar compounds is challenging because the typical hydrophobic stationary phases when combined with the most suitable mobile phases for MS detection do not provide the necessary retention to resolve and quantify these compounds. A Hypercarb column overcomes these challenges because it: • Retains and separates very polar compounds using “MS friendly” mobile phases such as 0.1% formic or acetic acid and low concentrations of volatile buffers such as ammonium acetate or ammonium formate • Can be used with high concentrations of organic modifiers in the mobile phase, which improves nebulization in atmospheric pressure ionization techniques, improving the sensitivity of the analysis. • Allows shorter column lengths and smaller diameters to be used without compromising peak capacity, often with increased sensitivity. The flow rates used with narrowbore and capillary columns are more compatible with MS techniques. • Is stable with any mobile phase and produces no phase bleed issues because the Hypercarb column’s porous graphitic surface is not modified. Particle Size (µm) 3 Format Drop-in Guard (4/pk) Length (mm) 10 HPLC Column 30 50 100 150 High Temperature HPLC Column 30 50 5 Drop-in Guard (4/pk) 10 HPLC Column 30 50 100 150 High Temperature HPLC Column 30 50 100 Javelin HTS Column Preparative HPLC Column 20 100 150 Format UNIGUARD Guard Cartridge Holder Length (mm) 10 Cat. No. 35003-011001 35003-012101 35003-013001 35003-014001 35003-031030 35003-032130 35003-033030 35003-051030 35003-052130 35003-053030 35003-054630 35003-101030 35003-102130 35003-103030 35003-104630 35003-152130 35003-153030 35003-154630 35003-032146 35003-052146 35003-054646 35003-101046 35003-102146 35003-103046 35003-104646 35005-011001 35005-012101 35005-013001 35005-014001 35005-032130 35005-033030 35005-034630 35005-051030 35005-052130 35005-053030 35005-054630 35005-101030 35005-102130 35005-103030 35005-104630 35005-151030 35005-152130 35005-153030 35005-154630 35005-032146 35005-034646 35005-052146 35005-054646 35005-102146 35005-104646 35005-022135 35005-109070 35005-109270 35005-109370 35005-159070 35005-159270 ID (mm) 1.0 2.1 3.0 4.6 Cat. No. 851-00 852-00 852-00 850-00 www.thermoscientific.com/chromatography Columns for Analytical LC 100 ID (mm) 1.0 2.1 3.0 4.6 1.0 2.1 3.0 1.0 2.1 3.0 4.6 1.0 2.1 3.0 4.6 2.1 3.0 4.6 2.1 2.1 4.6 1.0 2.1 3.0 4.6 1.0 2.1 3.0 4.6 2.1 3.0 4.6 1.0 2.1 3.0 4.6 1.0 2.1 3.0 4.6 1.0 2.1 3.0 4.6 2.1 4.6 2.1 4.6 2.1 4.6 2.1 10 21.2 30 10 21.2 LC Columns and Accessories >> Hypercarb Ordering Guide 4-087 LC Columns and Accessories Acclaim Size Exclusion Chromatography (SEC) High Performance SEC Columns for Analysis of Water Soluble Polymers • Proprietary mono-dispersed multi-pore hydrophilic resin: no inflection points in calibration curve • SEC-300 calibrated from 100 to 50,000 Daltons • SEC-1000 calibrated from 1,000 to 1,000,000 Daltons • Availability of small particle sizes packed in 4.6 x 300mm dimension allows for high-resolution analysis at reduced solvent consumption • Stable surface bonding with low column bleed and compatibility with UV, RI, MS, ELSD and Thermo Scientific™ Dionex™ Corona™ Charged Aerosol Detectors Thermo Scientific™ Acclaim™ SEC-300 and SEC-1000 are a family of resin based, high performance size exclusion chromatography columns specifically designed for the separation of water soluble polymers and oligomers. Acclaim SEC columns are available in two sizes to cover a broad molecular weight range: 5μm, 300 Å for Acclaim SEC-300 and 7μm, 1000 Å for Acclaim SEC-1000. 100,000,000 SEC-1000 Molecular Weight (Da) 10,000,000 SEC-300 1,000,000 100,000 10,000 1000 100 10 1.50 2.00 2.50 3.00 3.50 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Elution Volume (mL) 4-088 4.00 4.50 Columns: Acclaim SEC-300 Acclaim SEC-1000 Dimensions: Mobile Phases: Flow rate: Temperature: Injection Volume: Detection: Samples: 4.6 x 300mm 10mM sodium perchlorate 0.35mL/min 25°C 50μL RI (0.03% - 0.1% in mobile phase) dextran (MW 5,000,00040,000,000), PEO (MW 895,000, 580,000, 272,000, 185,000, 80,000, 43,000, and 20,000), PEG (MW 8,300, 3,000, 1,500, 600, 400 and 200), diethylene glycol (MW 106 and ethylene glycol (MW 62) 2.5 pA HPLC Conditions Column: Acclaim SEC-300 PAA Dimensions: System: Mobile Phases: 400 µg/L Isocratic: Flow rate: Injection: Temperature: Detection: 200 µg/L 100 µg/L 50 µg/L 25 µg/L -0.0 0 5 Minutes 10 Peaks: Polyacrylic acid 0.450 15 5µm, 4.6 x 300mm UltiMate 3000 RS A: Acetonitrile B: Water 10:90 0.35mL/min 35µL 30°C Corona III; evaporator 55°C, Engine 40 °C, 2 Hz, filter 5, power function 1.20 1. PAA standards in water Area [ pA *min] 0.200 0.00c0 Concentration [ µg/L] 0 125 250 Description Acclaim SEC-300 Particle Size (µm) 5 Format Guard HPLC Column Acclaim SEC-1000 7 Guard HPLC Column Length (mm) 33 150 300 33 150 300 4.6 mm ID 082740 – 079723 082739 – 079724 7.8 mm ID – 079726 079725 – 079722 079721 www.thermoscientific.com/chromatography Columns for Analytical LC Acclaim SEC Ordering Guide LC Columns and Accessories >> Polyacrylic acid using size-exclusion chromatography with charged-aerosol detection 4-089 LC Columns and Accessories Thermo Scientific Application Specific LC Columns Innovative chemistries tailored for challenging and critically important applications • Acclaim Trinity P1 and P2 for API and counterion analysis • Acclaim Organic Acid for fast organic acid analysis • Acclaim Surfactant and Surfactant Plus columns for separation of surfactants • Acclaim Explosives for separation of explosive residues • Acclaim Trinity Q1 for diquat and paraquat analysis • Acclaim Carbamate for the separation of carbamate insecticides • Acclaim Carbonyl C18 for aldehyde and ketone separation • Hypersil Green PAH for polyaromatic hydrocarbon analysis Thermo Scientific application specific columns are based on novel and unique chemistries and provide superior resolution with ease-of-use. Acclaim Trinity P1 and P2 Acclaim Trinity Q1 Application specific columns designed for the simultaneous separation of pharmaceutical drug substances and counterion analysis, as well as mixtures of acidic, basic, and neutral drugs. Unique, high-efficiency, silica-based columns designed for the separation of the herbicides diquat and paraquat. Acclaim Organic Acid Designed for separation of hydrophilic, aliphatic, and aromatic organic acids. Acclaim Surfactant and Surfactant Plus The most versatile commercially-available column specifically for the separation of all classes of surfactants. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Acclaim Explosives 4-090 Optimized column chemistry for baseline separation of all 14 explosives in EPA Method 8330, with complementary selectivity. Acclaim Carbamate A specifically column for the separation of carbamate pesticide specified in US EPA Method 531.2. Acclaim Carbonyl C18 Designed for separating DNPH derivatives of aldehydes and ketones in air, water and soil using CARB Method 1004, EPA Method 554, EPA Method 8315 and related methods. Hypersil Green PAH Specially tailored alkyl bonded silica with a high carbon loading, designed specifically for the analysis of polyaromatic hydrocarbons (PAHs), specified in EPA Method 610. Most innovative advancement in mixed mode column technology: reverse-phase, anion- and cation exchange functionality on a single support • Ideal selectivity for simultaneous separation of API and counterion • Adjustable selectivity by mobile phase ionic strength, electrolyte type, pH, and organic solvent • Low bleed; compatible with MS, CAD and ELSD. • Retention of hydrophilic ionic and ionizable analytes without ion-pairing reagents • Greater flexibility in method development: each retention mechanisms can be controlled independently The Thermo Scientific™ Acclaim™ Trinity™ P1 HPLC column is designed with unique multimode surface chemistry ideal for the simultaneous separation of drugs and their counterions. The surface chemistry concurrently provides reversed-phase, cation exchange, and anion exchange functionalities. The result is maximum flexibility in method development. Separations can be optimized easily by adjusting the chromatographic parameters (mobile phase pH, ionic strength, and organic strength). Acclaim Trinity P1 Ordering Guide Particle Size (µm) 3 Format Guard Cartridges (2/pk) HPLC Column Length (mm) 10 2.1mm ID 071391 3.0mm ID 071390 50 100 150 075565 071389 075564 071388 071387 075563 Columns for Analytical LC The Acclaim Trinity P1 stationary phase, based on this Nanopolymer Silica Hybrid (NSH) technology, consists of high-purity porous, spherical 3μm silica particles, coated with charged nanopolymer beads. The unique surface chemistry includes an inner-pore area modified with an organic layer that provides both reversed phase and anion exchange properties. The outer-pore surface, conversely, is modified with cation exchange functionality. LC Columns and Accessories >> Acclaim Trinity P1 Acclaim Guard Holder Ordering Guide Cat. No. 069580 069707 074188 Format Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Simultaneous separation of pharmaceutical counterions 300 Column: Acclaim Trinity P1, 3μm Dimensions: Mobile Phase: mV 3 1 4 2 5 8 6 7 10 9 0 0 3 6 9 Minutes 12 15 3.0 × 100mm 60/40 v/v CH 3CN/20mM (total) NH 4OAc, pH 5 Temperature: 30°C Flow Rate: 0.5mL/min Inj. Volume: 2μL Detection: Corona ultra (Gain = 100 pA; Filter = med; Neb Temp = 30°C) Peaks: (50 to 100ppm) 1. Choline 2. Tromethamine 3. Sodium 4. Potassium 5. Meglumine 6. Mesylate 7. Nitrate 8. Chloride 9. Bromide 10. Iodide www.thermoscientific.com/chromatography 4-091 LC Columns and Accessories Acclaim Trinity P2 Most innovative mixed-mode column technology; hydrophilic interaction, HILIC, anion exchange and cation exchange functionalities • Ideal for separating pharmaceutical counterions, including monovalent and divalent cations or anions • Selectivity complementary to the Trinity P1 column • Low column bleed, compatible with CAD and MS • Hydrolytically stable • High efficiency The Acclaim Trinity P2 is a unique, high-efficiency, silica-based column specifically designed for separation of pharmaceutical counterions, including monovalent and divalent cations or anions. The Acclaim Trinity P2 column is based on NSH technology, which consists of high-purity porous spherical silica particles coated with charged nanopolymer particles. The inner-pore area of the silica bead is modified with a covalently bonded organic layer that provides cation-exchange retention, while the outer surface is modified with anion-exchange nano-polymer beads. This chemistry ensures spatial separation of the anionexchange and cation-exchange regions. Acclaim Trinity P2 column is aimed to complement Acclaim Trinity P1 to provide a total solution for pharmaceutical counter ion analysis by HPLC. Pharmaceutical-related anions and cations Column: Acclaim Trinity P2, 3µm Dimensions: Mobile phase: 5.0 3.0 x 100 mm D.I. water and 100 mM NH4OFm, pH 3.65 gradient 30°C 0.60 mL/min 2μL Corona Veo Charged Aerosol Detector 0.02 – 0.10 mg/mL each in D.I. water 1. Phosphate 2. Sodium 3. Potassium 4. Chloride 5. Malate 6. Bromide 7. Nitrate 8. Citrate 9. Fumarate 10. Sulfate 11. Magnesium 12. Calcium Temperature: Flow rate: Injection volume: Detection: Samples: Peaks: 1 2 10 6 8 5 3 11 9 pA 12 Thermo Scientific Chromatography Columns and Consumables 2014-2015 4 4-092 Time (min) 7 0 0 6 18 12 24 30 Minutes H2O 0.1 M Ammonium formate, pH3.65 -10 0.760 1.474 0 80 20 2 80 20 22 0 100 30 0 100 Acclaim Trinity P2 Ordering Guide Particle Size (µm) 3 Format Guard Cartridges (2/pk) HPLC Column Length (mm) 10 2.1 mm ID 085435 3.0 mm ID 085436 50 100 085431 085432 085433 085434 Acclaim Guard Holder Ordering Guide Format Acclaim Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Cat. No. 069580 069707 074188 Optimized and application-tested for the analysis of hydrophilic organic acids • Use-tested to guarantee consistent hydrophilic organic acid separations • Compatible with 100% aqueous mobile phases • Hydrolytic stability at low-pH conditions • Ideal selectivity for separating a wide spectrum of organic acids • Excellent column efficiency and peak shapes for organic acids The Acclaim Organic Acid (OA) is a silica-based reversed-phase column designed for high-efficiency, high-throughput organic acids analysis. It offers unparalleled performance for separating hydroxyl aliphatic and aromatic organic acids. The Acclaim OA is the recommended column for determining small hydrophilic organic acids, C1 to C7 aliphatic acids, and hydrophilic aromatic acid and is also valuable for the analysis and quality assurance of food and beverage products, pharmaceutical preparations, plating baths, and manufacturing chemicals, chemical intermediates, and environmental samples. Particle Size (µm) 3 5 Format HPLC Column Guard Cartridges (2/pk) HPLC Column Length (mm) 150 10 2.1mm ID 070087 – 3.0mm ID 070086 071987 4.0mm ID – 069700 150 250 – – – – 062903 062902 Acclaim Guard Holder Ordering Guide Columns for Analytical LC Acclaim Organic Acid Ordering Guide LC Columns and Accessories >> Acclaim Organic Acid Cat. No. 069580 069707 074188 Format Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Hydrophilic organic acids Column: Acclaim OA, 5μm, 4 × 250mm 0.025 Mobile phase: AU 10 Temperature: Flow rate: Injection volume: Detection: Peaks: 2 1 3 4 567 8 12 9 11 0 0 5 10 Minutes 15 20011 100mM Na 2SO 4, pH 2.65 (adjusted with methanesulfonic acid) 30°C 0.6mL/min 5μL UV, 210nm 1. Oxalic acid 15mg/L (ppm) 2. Tartaric acid 120 3. Formic acid 180 4. Malic acid 120 5. iso-Citric acid 120 6. Lactic acid 180 7. Acetic acid 120 8. Citric acid 120 9. Succinic acid 120 10. Fumaric acid 7 11. cis-Aconitic acid * 12. trans-Aconitic acid * * 7ppm total for cis and trans isomers www.thermoscientific.com/chromatography 4-093 LC Columns and Accessories Acclaim Surfactant Excellent performance for separating a broad range of surfactants • Ideal selectivity for separation of anionic, nonionic, cationic and amphoteric surfactants • Excellent peak shapes, especially for cationic surfactants • Compatible with highly aqueous mobile phases • Improved resolution for ethoxylated surfactants • Rugged separations under a variety of conditions The Acclaim Surfactant columns are the first generation high-efficiency, silica-based columns designed specifically for separating a wide variety of surfactants, including anionic, cationic, nonionic, and amphoteric surfactants using UV, ELSD or RI detection. As a consequence of its novel chemistry, this column exhibits a unique polarity that provides significantly improved resolution for individual oligomers of ethoxylated surfactants compared with conventional C18 columns. Acclaim Surfactant columns are also resistant to dewetting under highly aqueous mobile phase conditions, and thus can be used to provide excellent resolution between strongly hydrophilic compounds, such as isomers of xylene sulfonate. Surfactants are widely used in industrial, agricultural, and pharmaceutical markets, in products as diverse as pesticides, detergents powders, petroleum products, cosmetics, and pharmaceuticals. The Acclaim Surfactant column was designed specifically for HPLC separation of these surfactants. Acclaim Surfactant Ordering Guide Particle Size (µm) 3 5 Format HPLC Column Guard Cartridges (2/pk) HPLC Column Length (mm) 150 10 2.1mm ID 070085 069693 3.0mm ID 070084 071991 4.6mm ID – 069701 150 250 068123 – – – 063201 063203 4-094 Cat. No. 069580 069707 074188 Description Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Inorganic anion, hydrotropes, cationic, nonionic, amphoteric, and anionic surfactants Column: Dimensions: Mobile Phase: 6 5 10 mV Thermo Scientific Chromatography Columns and Consumables 2014-2015 Acclaim Guard Holder Ordering Guide 3 2 4 7 1 8 13 9 12 11 0 5 10 15 20 Minutes 25 14 30 35 22793 Acclaim Surfactant, 5µm 4.6 × 150mm (A) CH 3 CN, (B) 0.1 M NH 4 OAc, pH 5.4 Gradient: 25% to 85% A in 25min, then hold 85% A for 10min Temperature: 30°C Flow Rate: 1mL/min Inj. Volume: 25µL Detection: ELS detector Peaks: 1. Chloride 2. Bromide 3. Nitrate 4. Xylene sulfonate 5. Laurylpyridinium chloride 6. Lauryldimethylbenzyl-ammonium chloride 7. Triton X-100 8. Cetyl betaine 9. Decyl sulfate 10. Dodecyl sulfate 11. C10 -LAS 12. C11-LAS 13. C12 -LAS 14. C13 -LAS Column of Choice for Surfactant Analysis using Higher Sensitivity Detection: Performance, Versatility, Throughput • Ideal selectivity for simultaneous separation of anionic, nonionic, cationic, and amphoteric surfactants • Compatible with multiple detectors including MS, CAD, ELSD and UV • Well suited for the determination of cationic surfactants • High efficiency and fast analysis • Rugged separations under a variety of conditions Acclaim Surfactant Plus is a new generation of columns offering improved performance and higher throughput for analyzing surfactants. They have similar selectivity to the Acclaim Surfactant columns, but have been designed with exceptionally low bleed for use with charged aerosol detectors (CAD) and mass spectrometers (MS). These columns can be used to separate a wide variety of surfactants including anionic, cationic, nonionic and amphoteric surfactants. Cationic surfactants 10 1 Column: Acclaim Surfactant Plus, 3μm Dimensions: System: Mobile phases: 3.0 × 150 mm ICS 3000 A: Acetonitrile B: 100mM Formic acid C: Water µS Gradient: 3 2 Time (min) -12 0 12 20 4 0 6 5 7 8 Peaks: 0 0 5 Minutes 10 %B %C 5 5 40 40 5 5 5 5 90 90 55 55 Flow Rate: Injection: Temperature: Detection: Suppressor: 5 µS %A Columns for Analytical LC Additionally, these columns can be used with evaporative light scattering detectors (ELSD), suppressed conductivity detectors (SCD), and UV-Vis detectors (UV). They are resistant to dewetting under highly aqueous mobile phase conditions, and thus can be used to provide excellent resolution between strongly hydrophilic compounds, such as isomers of xylene sulfonate. Non-metallic PEEK hardware is available for best compatibility with Dionex ion chromatography systems. LC Columns and Accessories >> Acclaim Surfactant Plus 15 0.5 mL/min 5 μL 25°C Conductivity with blank subtraction CSRS300-2mm (external water 1.0 mL/min, current = 8 mA) 1. Tetrabutylammonium 2. Tetrapentylammonium 3. Tetrahexylammonium 4. Tetraheptylammonium 5. Decyl-trimethylammonium 6. Dodecyl-trimethylammonium 7. Tetradecyl-trimethylammonium 8. Hexadecyl-trimethylammonium www.thermoscientific.com/chromatography 4-095 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-096 Acclaim Surfactant Plus Ordering Guide Particle Size (µm) 3 Format HPLC Column 5 Guard Cartridges (2/pk) HPLC Column Length (mm) 100 150 250 10 2.1mm 078955 078954 078953 078960 3.0 mm 078952 078951 – 078959 4.6 mm – 078950 – 082773 4.0 mm PEEK – – – – 250 150 – – – – 082767 082768 – 078956 Acclaim Guard Holder Ordering Guide Description Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Cat. No. 069580 069707 074188 The best solution for explosives analysis (EPA Method 8330) • Both Acclaim E1 and E2 columns provide baseline resolution of all 14 compounds targeted by EPA Method 8330 • E2 columns available in 2.2, 3 and 5μm particle size • The E1 and E2 columns have mutually complementary selectivity • Simple isocratic elution conditions • Rugged columns with good lot-to-lot reproducibility Acclaim Explosives E1 and E2 columns are specifically designed to resolve all 14 explosives listed in EPA SW-846 Method 8330: Nitroaromatics and Nitramines by HPLC. The novel and unique chemistries of these columns provide superior resolution with complementary selectivities. The Acclaim Explosives E1 is recommended for use as a direct replacement for ODS columns for the primary analysis. The Acclaim Explosives E2 may be used as either a primary or a confirmatory column. The unique selectivity and versatility of Acclaim Explosives E2 column provides a wider application range, including the analysis of explosives beyond U.S. EPA Method 8330 (ISO22478). 4 3 1 2 Acclaim Explosives E1 5 6 7 8 9 10 11 12 13 14 AU 4 3 Acclaim Explosives E2 5 0 7 2 1 Column: Eluent: Temperature: Flow Rate: Inj. Volume: Detection: Peaks: 11 6 10 9 Column: 5µm, 4.6 x 250mm 12 13 14 18 9 8 27 36 45 Minutes 22787 5µm, 4.6 x 250mm EPA 8330 mix, 50 ppm each 30 °C 1 mL/min 5µL UV, 254nm 1. HMX 2. RDX 3. 1,3,5-Trinitrobenzene 4. 1,3-Dinitrobenzene 5. Nitrobenzene 6. Tetryl 7. 2,4,6-Trinitrotoluene 8. 4-Amino-2,6-Dinitrotoluene 9. 2-Amino-4,6-Dinitrotoluene 0. 2,6-Dinitrotoluene 11. 2,4-Dinitrotoluene 12. 2-Nitrotoluene 13. 4-Nitrotoluene 14. 3-Nitrotoluene Columns for Analytical LC Complimentary baseline separation of 14 target compounds listed in EPA SW-846 method 8330 LC Columns and Accessories >> Acclaim Explosives Rapid determination of EPA 8330A explosives Column: Acclaim RSLC Explosives E2, 2.2µm 60 Dimension: HPLC System: Mobile Phases: Flow Rate: Injection Vol.: Temperature: Detection: Sample: 4 3 mAU 5 2 1 9 6 7 Peaks: 8 10 11 12 13 14 0 0 2.5 5 Minutes 7.5 2.1 × 100mm UltiMate 3000 RSLC HPG Methanol:water 48:52 (v/v) 0.34mL/min (293 bar) 1µL 31°C UV at 254nm, 10 Hz, 0.4 s resp. time Calibration mix, 25µg/mL in 50% acetontrile 1. HMX 8. 2,6-DNT 2. RDX 9. 2,4-DNT 3. 1,3,5-TNB 10. 2-NT 4. 3,5-DNB 11. 4-NT 5. NB 12. 3-NT 6. 2,4,6-TNT 13. 4-Am-2,6-DNT 7. Tetryl 14. 2-Am-4,6-DNT 10 28815 www.thermoscientific.com/chromatography 4-097 LC Columns and Accessories Acclaim Explosives Ordering Guide Acclaim Explosives E1 Ordering Guide Particle Size (µm) 5 Format Guard Cartridges (2/pk) HPLC Column Length (mm) 10 2.1mm ID – 3.0mm ID – 4.6mm ID 069702 250 – – 064305 Length (mm) 100 150 150 250 10 2.1mm ID 076225 076226 070083 – – 3.0mm ID 076227 – 070082 070081 071989 4.6mm ID – – – – 069703 250 – – 064309 Acclaim Explosives E2 Ordering Guide Particle Size (µm) 2.2 Format RSLC Column 3 HPLC Column 5 Guard Cartridges (2/pk) HPLC Column Acclaim Guard Holder Ordering Guide Description Cat. No. Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 069580 069707 074188 Acclaim Explosives Kit Ordering Guide Thermo Scientific Chromatography Columns and Consumables 2014-2015 Description E1 and E2 HPLC Columns (4.6 x 250mm) E1 and E2 Guard Cartridges (4.6 x 10mm), pkg of 2 4-098 Cat. No. 064312 For Trace Analysis of Diquat and Paraquat • Excellent resolution of diquat and paraquat • Good peak shape • Fast analysis • LC/MS compatibility • No ion-pairing reagent needed • Easy to use Acclaim Trinity Q1 columns are unique, high-efficiency, silica-based columns designed for the separation of the herbicides diquat and paraquat. These herbicides are toxic and residues are monitored in drinking water, wastewater and agricultural products. The Acclaim Trinity Q1 column is a tri-mode (WCX, WAX, RP), column based on Nano-polymer Silica Hybrid technology. It offers unmatched high-resolution and high-throughput trace analysis of the herbicides diquat and paraquat by LC-MS/MS and LC-UV methods. Diquat and paraquat 185/169 Paraquat: C-SRM 5.46E4 Mass Spectrometric Conditions System: MAX Quadrupole Mass Spectrometer Interface: Heated Electrospary Ionization with HESI II probe Spray Voltage: 1500 V Vaporizer Temp: 400 ºC Sheath Gas Pressure: 70 Aux Gas Pressure: 10 Capillary Temp: 350 ºC Quantitation Mode: Selected Reaction Monitoring (SRM) 185/170 Paraquat-IS Relative Abundance 5.47E4 193/178 Diquat: Q-SRM 9.67E5 Scan Events Paraquat Paraquat-d6 Diquat Diquat-d3 183/157 Diquat: C-SRM 3.11E5 Precursor 185 193 183 186 Quantitative SRM (CID) 169 (27) 178 (17) 157 (22) 158 (22) Columns for Analytical LC Chromatographic Conditions System: RSLC system Column: Acclaim Trinity Q1, 3µm Column Temp: Ambient Mobile Phase: 25% Ammonium Acetate (100 mM, pH 5.0); 75% Acetonitrile Flow Rate: 0.5 mL/min Injection: Paraquat: Q-SRM 1.19E5 LC Columns and Accessories >> Acclaim Trinity Q1 SRM (CID) 170 (17) 130 (31) 183/130 Diquat-IS 9.98E5 186/158 0.0 0.5 1.0 1.5 2.0 2.5 Time (min) 3.0 3.5 4.0 Acclaim Trinity Q1 Ordering Guide Particle Size 3 5 HPLC Column Guard Cartridges (2/pk) Length (mm) 50 10 3.0 mm ID 079716 079719 2.1 mm ID 079718 079720 www.thermoscientific.com/chromatography 4-099 LC Columns and Accessories Acclaim Carbamate Designed for baseline separation of carbamate pesticides specified in US EPA Method 531.2 • Baseline separation of carbamate pesticides specified in US EPA Method 531.2 • Use with either LC/postcolumn derivatization/fluorescence or LC/MS detection • Available in 2.2, 3 and 5μm particle size • Compatible with both binary (methanol/water) and ternary (acetonitrile/methanol/water) mobile phase gradients • High-efficiency, extremely low column bleed, and rugged column packing Acclaim Carbamate columns are designed for baseline separation of carbamates (N-methylcarbamate and N-methylcarbamoyloxime pesticides) specified in US EPA Method 531.2. Carbamate pesticides are widely used throughout the world. Drinking water and raw surface water is monitored for the presence of carbamate pesticides and related compounds using an established EPA Method 531.2 that uses HPLC with postcolumn derivatization. LC-MS is the method of choice for the ultimate sensitivity. Acclaim Carbamate Ordering Guide Particle Size (µm) 2.2 Format RSLC Column 3 Guard Cartridges (2/pk) HPLC Column HPLC Column 5 Length (mm) 100 150 10 2.1mm ID 075597 075596 072930 3.0mm ID – – 072929 4.6mm ID – – 072928 150 250 072927 – 072926 – 072925 072924 Acclaim Guard Holder Ordering Guide Cat. No. 069580 069707 074188 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Description Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 4-100 Carbamate standard - spiked rice samples 4000 Column: Acclaim Carbamate, 3.0 × 150mm, 3μm A 4 1 2 9 3 6 5 counts 10 7 8 11 12 1 -500 0 1.3 2.5 3.8 5 6.3 7.5 8.8 10 11.3 12.5 13.8 15 16 2000 12 B counts 1 2 3 4 2 -300 0 1 2 3 4 A: without dispersive SPE B: with dispersive SPE using PSA 5 6 7 8 Minutes 9 7 8 6 5 9 10 11 10 12 11 13 14 15 16 REF: 28175 Mobile Phase: In Gradient: Methanol–H 2 O Methanol, -4.0–0.0 min, 14%; 2.0 min, 20%; 8.0 min, 40%; 13.6–16 min, 70% Temperature: 50 ºC Flow Rate: 0.9mL/min Injection Volume: 250μL Standard Concentration: 0.4μg/L for each component Postcolumn Reagent 1: 0.2% NaOH, first reaction coil at 100 ºC Postcolumn Reagent 2: OPA regent, second reaction coil at room temperature Flow Rate of Reagents 1 and 2: 0.3mL/min Fluorescence Detection: Excitation/330nm and Emission/465nm Peaks: 1. Aldicarb sulfoxide 2. Aldicarb sulfone 3. Oxamyl 4. Methomyl 5. 3-Hydroxy carbofuran 6. Aldicarb 7. Propoxur 8. Carbofuran 9. Carbaryl 10. 1-Naphthol 11. Methiocarb 12. BDMC (I.S.) A silica-based, reversed-phase column designed specifically for separating DNPH derivatives of aldehydes and ketones • Ideal selectivity for baseline resolution of DNPH derivatives of aldehydes and ketones regulated by various official methods, including EPA 554, EPA 8315, EPA 1667, EPA TO-11, and CARB 1004 • High efficiency for UHPLC performance • Rugged columns with good lot-to-lot reproducibility • Proven robust methods Acclaim Carbonyl C18 columns are silica-based reversed phase columns designed specifically for separating DNPH derivatives of aldehydes and ketones. They exhibit superior resolution compared with other commercially available columns. Acclaim Carbonyl C18 Ordering Guide Particle Size (µm) 2.2 Format RSLC Column 3 HPLC Column 5 Guard Cartridge (2/pk) HPLC Column Length (mm) 100 150 150 250 10 2.1mm ID 077972 077973 079011 – 079012 3.0mm ID 077974 – 079010 079009 079013 4.6mm ID – – – – 079014 150 250 – – – – 079008 083214 Columns for Analytical LC Aldehydes and ketones are common pollutants in air and water. The analytical difficulties that need to be overcome include their volatility, their reactivity, and their modest UV absorption. The reaction with dinitrophenylhydrazine (DNPH) is a convenient means of trapping, stabilizing, and tagging these substances. Several standard methods have been developed to apply this chemistry to various environmental situations. Some of the better known ones include CARB 1004 for vehicle exhaust, EPA 554 for drinking water, EPA 1667 for pharmaceutical wastewater, and EPA 8315 for general wastewater. LC Columns and Accessories >> Acclaim Carbonyl C18 DNPH aldehydes and ketones 300 8 1 9 Column: Acclaim Carbonyl RSLC, 2.2μm Dimension: 2.1 × 150mm 10 11 12 7 6 2 5 3 mAU 4 EPA 554 13 14 15 16 17 19 CARB 1004 18 19 20 EPA TO-11 0 0 5 10 Minutes 15 18 Mobile Phases: A: D.I. water B: Acetonitrile Gradient (min): -4.5 0.0 8.3 15.0 18.0 %A 48 48 48 0 0 %B 52 52 52 100 100 Flow Rate: 0.400mL/min Injection: 1μL Temperature: 28°C Detection: UV at 360nm (data collection rate at 25Hz) Samples: Calibration mixes diluted in methanol Peaks: 1. Formaldehyde DNPH 2. Acetaldehyde DNPH 3. Propionaldehyde DNPH 4. Crotonaldehyde DNPH 5. Butyraldehyde DNPH 6. Cyclohexanone DNPH 7. Valeraldehyde DNPH 8. Hexanal DNPH 9. Heptanal DNPH 10. Octanal DNPH 11. Nonanal DNPH 12. Decanal DNPH 13. Acetone DNPH 14. Acrolein DNPH 15. Butanone DNPH 16. Methacrolein DNPH 17. Benzaldehyde DNPH 18. Isovaleraldehyde DNPH 19. Tolualdehyde DNPH 20. Xylylaldehyde DNPH www.thermoscientific.com/chromatography 4-101 LC Columns and Accessories Hypersil Green PAH Specially tailored alkyl bonded silica with a high carbon loading, designed specifically for the analysis of polyaromatic hydrocarbons (PAHs) • Optimized for EPA Method 610 • Rapid analysis of 16 PAHs in 4 minutes using short, fast columns • Available in 3µm and 5µm particle size and variety of column dimensions Hypersil Green PAH Ordering Guide Particle Size (µm) 3 Thermo Scientific Chromatography Columns and Consumables 2014-2015 5 4-102 Format Guard Cartridge Length (mm) 10 HPLC Column 100 HPLC Column 150 Guard Cartridge HPLC Column 10 100 150 250 ID (mm) 2.1 3.0 4.6 2.1 3.0 4.6 2.1 4.6 4.6 4.6 4.6 4.6 Cat. No. 31103-012101 31103-013001 31103-014001 31103-102130 31103-103030 31103-104630 31103-152130 31103-154630 31105-014001 31105-104630 31105-154630 31105-254630 Subscribe to our chromatography channel on YouTube www.youtube.com/chromatographyvideos DVB polymer columns for combined ion exchange and reversed-phase separations • Acid, base and solvent compatible, pH 0 to 14 • Ideal for the separation of high molecular weight organic acids • Delivers optimal separation of very hydrophobic anions • Delivers optimal separation of halogenated anions • Provides simultaneous separation of neutral and ionic species • Unique selectivity for polar and ionic organic analytes • Delivers optimal separation of organic, hydrophobic, and halogenated cations Thermo Scientific™ Dionex™ OmniPac™ PAX-100 columns are used to separate hydrophobic anionic analytes such as larger organic acids. The Dionex OmniPac PAX-500 column simultaneously separates anionic and neutral species. The Dionex OmniPac PCX-100 column separates low-molecular-weight hydrophobic cations. The Dionex OmniPac PCX-500 column simultaneously separates cationic and neutral species in a single run. Dionex OmniPac Anion Exchange Ordering Guide Description PAX-100 Porosity microporous PAX-500 macroporous Length (mm) 50 250 50 250 4.0mm ID 042151 042150 042153 042152 Length (mm) 50 250 50 250 4.0mm ID 042193 042189 042195 042191 Columns for Analytical LC The Dionex OmniPac PAX- and PCX-100 and 500 are latex-based columns. Both PAX columns have an ion exchange capacity of about 40 μeq per column, providing equivalent anion exchange separations. The PCX columns have a capacity of approximately 120 μeq per column. The PAX- and PCX-500 columns separate analytes through both ion exchange and reversed-phase mechanisms, due to their higher reversed-phase capacity relative to the PAX- and PCX-100 columns. LC Columns and Accessories >> Dionex OmniPac Dionex OmniPac Cation Exchange Ordering Guide Description PCX-100 Porosity microporous PCX-500 macroporous Gradient separation of nitrogen-containing compounds 12 9 Column: Dionex OmniPac PCX-500 (4 × 250mm) 16 13 6,7 1 2 4,5 10 11 17 8 3 0 14 15 5 10 Minutes Eluent: Acetonitrile/Sodium Chloride/Hydrochloric Acid Gradient Flow Rate: 1.0mL/min Detection: UV, 254nm Peaks: 1. Orotic Acid 2. 4-Hydroxybenzamide 3. Luminol Impurity 4. Luminol 5. Pyridine 6. PABA 7. 2,2’-Bipyridine 8. p-Phenylenediamine 9. Naphthylamine 10. Nitrobenzoic Acid 11. Tribenzylamine 15 12. p-Nitroaniline 5410 13. 2,4-Dinitroaniline 14. Dibenzylamine 15. N-Methyl-N-nitrosoaniline 16. 4-Chloro-2-nitroaniline 17. 2,6-Dichloro-4-nitroaniline www.thermoscientific.com/chromatography 4-103 LC Columns and Accessories Gradient separation of inositol mono-, di-, tri-, and tetraphosphates Column: Dionex OmniPac PAX-100 (4 x 250mm) 8 6 1 3 4 2 0 10 20 9 10 7 5 30 40 50 60 5678 Minutes Eluent: Isopropanol / Sodium Hydroxide Gradient Flow Rate: 1.0mL/min Detection: Suppressed Conductivity (Baseline Subtracted) Peaks: 1. Ins (2) P 2. Ins (1) P 3. Ins (4) P 4. Ins (1,4) P2 5. Ins (2,4) P2 6. Ins (4,5) P2 7. Ins (1,3,4) P3 8. Ins (1,4,5) P3 9. Ins (2,4,5) P3 10. Ins (1,4,5,6) P4 Dionex IonPac NS1 and NS2 Polymeric Reversed-Phase Column Ideal for the Separation of Hydrophobic, Ionizable Compounds The Thermo Scientific™ Dionex™ IonPac™ NS1-10μm and NS1-5μm columns are packed with a neutral, macroporous, high-surface-area, ethylvinylbenzene polymer crosslinked with 55% divinylbenzene. This resin makes the NS1 resistant to solvents, acids, and bases, and permits the use of eluent from pH 0 to 14. The Dionex IonPac NS1 column is the column of choice for routine ion pair chromatography. IonPac NS2 is a silica-based column for mobile-phase ion chromatography (MPIC) applications using eluents containing trifluoroacetic acid (TFA), heptafluorobutyric acid (HFBA), or tetrabutylammonium borate (TBAB). It provides high performance analysis for hydrophobic amines and hydrophobic acids using suppressed conductivity detection. • Excellent resolution • Good peak shape • Ideal for separation of large molecules that carry localized charges, such as surfactants • Compatible with acids, bases, and solvent from pH 0 to 14 • Can also be used for traditional polymeric reversed-phase applications Thermo Scientific Chromatography Columns and Consumables 2014-2015 • Utilize ion-pair chromatography for difficult separations 4-104 Dionex IonPac NS1 and NS2 Ordering Guide Description IonPac NS1 Particle Size (µm) 5 10 IonPac NS2 5 Format HPLC Column Guard Column HPLC Column Guard Column HPLC Column Length (mm) 150 35 250 35 150 250 2.0mm ID – SP4356 SP4354 – – – 4.0mm ID 039568 039567 035321 SP6907 SP6906 SP6905 A good choice for robust, general-purpose columns Hypersil BDS Ordering Guide Particle Size (µm) 2.4 Format Drop-in Guard (4/pk) Length (mm) 10 50 100 150 3 Drop-in Guard (4/pk) 10 HPLC Column 30 50 150 5 Drop-in Guard (4/pk) 10 HPLC Column 50 100 125 150 250 Format UNIGUARD Guard Cartridge Holder Length (mm) 10 C18 28102-012101 28102-014001 28102-052130 28102-054630 28102-102130 28102-104630 28102-152130 28102-154630 28103-012101 28103-013001 28103-014001 28103-032130 28103-052130 28103-053030 28103-054630 28103-102130 28103-103030 28103-104030 28103-104630 28103-152130 28103-153030 28103-154030 28103-154630 28105-012101 28105-013001 28105-014001 28105-052130 28105-053030 28105-054630 28105-102130 28105-103030 28105-104030 28105-104630 28105-123030 28105-124030 28105-124630 28105-152130 28105-153030 28105-154030 28105-154630 28105-252130 28105-253030 28105-254030 28105-254630 ID (mm) 1.0 2.1 3.0 4.0/4.6 Cat. No. 851-00 852-00 852-00 850-00 C8 28202-012101 28202-014001 28202-052130 28202-054630 28202-102130 28202-104630 28202-152130 28202-154630 28203-012101 28203-013001 28203-014001 – 28203-052130 – 28203-054630 – – – 28203-104630 – 28203-153030 – 28203-154630 28205-012101 28205-013001 28205-014001 28205-052130 28205-053030 28205-054630 28205-102130 – – 28205-104630 – – – – – 28205-154030 28205-154630 – – 28205-254030 28205-254630 Phenyl – – – – – – – – 28903-012101 – 28903-014001 – 28903-052130 – – – – – – – – – 28903-154630 – – 28905-014001 – – – – – – – – – – – – – 28905-154630 – – 28905-254030 28905-254630 CN – – – – – – – – 28803-012101 – 28803-014001 – 28803-052130 – – – – – – – – – 28803-154630 – – 28805-014001 – – – – – – – – – – – – – 28805-154630 – – – 28805-254630 www.thermoscientific.com/chromatography Columns for Analytical LC 100 ID (mm) 2.1 4.0/4.6 2.1 4.6 2.1 4.6 2.1 4.6 2.1 3.0 4.0/4.6 2.1 2.1 3.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0/4.6 2.1 3.0 4.6 2.1 3.0 4.0 4.6 3.0 4.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 LC Columns and Accessories >> Hypersil BDS 4-105 LC Columns and Accessories Hypersil Classical A global standard for many existing methods Hypersil ODS Ordering Guide Particle Size (µm) 3 Format Drop-in Guard (4/pk) Length (mm) 10 HPLC Column 50 100 125 150 250 5 Drop-in Guard (4/pk) 10 HPLC Column 50 Thermo Scientific Chromatography Columns and Consumables 2014-2015 100 4-106 125 150 200 250 ID (mm) 2.1 3.0 4.0/4.6 2.1 3.0 4.6 2.1 3.0 4.0 4.6 2.1 4.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0/4.6 2.1 3.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 2.1 3.0 4.0 4.6 2.1 4.0 4.6 2.1 3.0 4.0 4.6 Cat. No. 30103-012101 30103-013001 30103-014001 30103-052130 30103-053030 30103-054630 30103-102130 30103-103030 30103-104030 30103-104630 30103-122130 30103-124030 30103-124630 30103-152130 30103-153030 30103-154030 30103-154630 30103-252130 30103-253030 30103-254030 30103-254630 30105-012101 30105-013001 30105-014001 30105-052130 30105-053030 30105-054630 30105-102130 30105-103030 30105-104030 30105-104630 30105-122130 30105-123030 30105-124030 30105-124630 30105-152130 30105-153030 30105-154030 30105-154630 30105-202130 30105-204030 30105-204630 30105-252130 30105-253030 30105-254030 30105-254630 Particle Size (µm) 3 Format Drop-in Guard (4/pk) HPLC Column 5 Drop-in Guard (4/pk) HPLC Column Cat. No. 31603-014001 31603-054630 31603-104030 31603-104630 31603-154630 31605-014001 31605-054630 31605-104630 31605-154630 31605-254030 31605-254630 Length (mm) 10 50 100 150 250 ID (mm) 4.0/4.6 4.6 4.6 4.6 4.0 4.6 Cat. No. 30205-014001 30205-054630 30205-104630 30205-154630 30205-254030 30205-254630 Length (mm) 10 150 250 ID (mm) 4.0/4.6 4.6 4.0 4.6 Cat. No. 30305-014001 30305-154630 30305-254030 30305-254630 Length (mm) 10 150 250 ID (mm) 4.0/4.6 4.6 4.6 Cat. No. 30505-014001 30505-154630 30505-254630 150 10 50 100 150 250 Hypersil MOS (C8) Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) HPLC Column Hypersil MOS-2 (C8) Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) HPLC Column Columns for Analytical LC ID (mm) 4.0/4.6 4.6 4.0 4.6 4.6 4.0/4.6 4.6 4.6 4.6 4.0 4.6 Length (mm) 10 50 100 LC Columns and Accessories >> Hypersil ODS-2 Ordering Guide Hypersil SAS (C1) Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) HPLC Column www.thermoscientific.com/chromatography 4-107 LC Columns and Accessories Hypersil Classical continued Hypersil Phenyl Ordering Guide Particle Size (µm) 5 Length (mm) 10 50 100 150 250 ID (mm) 4.0/4.6 4.6 4.6 4.6 4.0 4.6 Cat. No. 30905-014001 30905-054630 30905-104630 30905-154630 30905-254030 30905-254630 Length (mm) 10 150 250 ID (mm) 4.0/4.6 4.6 4.6 Cat. No. 31905-014001 31905-154630 31905-254630 Length (mm) 10 150 10 150 250 ID (mm) 4.0/4.6 4.6 4.0/4.6 4.6 4.0 4.6 Cat. No. 30803-014001 30803-154630 30805-014001 30805-154630 30805-254030 30805-254630 Length (mm) 10 150 250 ID (mm) 4.0/4.6 4.6 4.6 Cat. No. 31905-014001 31905-154630 31905-254630 Format Drop-in Guard (4/pk) Length (mm) 10 HPLC Column 50 ID (mm) 2.1 4.0/4.6 2.1 4.6 2.1 2.1 4.6 3.0 4.0/4.6 3.0 4.6 4.6 4.0 4.6 Cat. No. 30703-012101 30703-014001 30703-052130 30703-054630 30703-102130 30703-152130 30703-154630 30705-013001 30705-014001 30705-103030 30705-104630 30705-154630 30705-254030 30705-254630 Format Drop-in Guard (4/pk) HPLC Column Hypersil Phenyl-2 Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) HPLC Column Hypersil CPS Ordering Guide Particle Size (µm) 3 5 Format Drop-in Guard (4/pk) HPLC Column Drop-in Guard (4/pk) HPLC Column Hypersil CPS-2 Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) HPLC Column Hypersil APS-2 Ordering Guide Thermo Scientific Chromatography Columns and Consumables 2014-2015 Particle Size (µm) 3 4-108 100 150 5 Drop-in Guard (4/pk) 10 HPLC Column 100 150 250 Particle Size (µm) 3 5 Format Drop-in Guard (4/pk) Length (mm) 10 HPLC Column 50 Drop-in Guard (4/pk) 150 10 HPLC Column 50 100 150 250 Cat. No. 30003-012101 30003-013001 30003-014001 30003-052130 30003-053030 30003-054630 30003-154630 30005-013001 30005-014001 30005-054630 30005-104630 30005-154630 30005-254030 30005-254630 ID (mm) 2.1 3.0 4.0/4.6 2.1 4.6 3.0 4.6 Cat. No. 34105-012101 34105-013001 34105-014001 34105-052130 34105-154630 34105-253030 34105-254630 Hypersil SAX Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) Length (mm) 10 HPLC Column 50 150 250 Columns for Analytical LC ID (mm) 2.1 3.0 4.0/4.6 2.1 3.0 4.6 4.6 3.0 4.0/4.6 4.6 4.6 4.6 4.0 4.6 LC Columns and Accessories >> Hypersil Silica Ordering Guide Other HPLC Columns Aquasil HPLC Columns Thermo Scientific™ Aquasil™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details. BetaBasic HPLC Columns Thermo Scientific™ BetaBasic™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details. BETASIL HPLC Columns Thermo Scientific™ BETASIL™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details. HyPurity HPLC Columns Thermo Scientific™ HyPurity™ HPLC Columns are available in limited dimensions. Please contact Customer Service for more details. www.thermoscientific.com/chromatography 4-109 LC Columns and Accessories Thermo Scientific LC Columns for Biomolecules The diversity of biological samples in terms of compound structure and properties coupled with matrix complexity demands a range of sample separation modes, column chemistries, column configurations and detection techniques for their effective characterization. Thermo Scientific addresses these needs with a range of silica and polymeric columns specifically designed to handle the unique rigors of the analysis of proteins, peptides, oligonucleotides and other biomolecules. Columns for proteins Reversed Phase Thermo Scientific™ BioBasic™ reversed-phase columns provide superior chromatography because the extra dense bonding chemistry used for these packings produce a highly stable, reproducible surface for reliable results. BioBasic reversed phase packings are available in C18, C8 and C4, chemistries. Acclaim 300 C18 features 3μm silica particles for rapid analysis of complex protein digests. Compared to 5μm column packings, the smaller particles support increased flow rates and shallower gradients on shorter columns, for faster separation analysis. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Thermo Scientific™ ProSwift™ RP monolith columns uniquely provide the advantages of high resolution at exceptionally high flow rates for fast protein separation analysis. 4-110 Ion Exchange Thermo Scientific™ ProPac™ and Thermo Scientific™ MAbPac™ ion exchange columns are based on a pellicular nonporous core particles providing exceptionally high resolution and efficiency for separations of protein variants, resolving isoforms that differ by a single charged residue. A hydrophilic layer prevents unwanted secondary interactions, and a grafted cation exchange surface provides pH-based selectivity control and fast mass transfer for high-efficiency separation and moderate capacity. ProPac WCX and MAbPac SCX columns are specifically developed for monoclonal separation and analytical characterization. Applications include protein variants in a variety of matrices, such as biopharmaceuticals and diary products. MAbPac columns are specifically designed for the analysis of monoclonal antibody variants. BioBasic AX and BioBasic SCX ion exchange columns demonstrate superior reproducibility, both column-to-column and batch-to-batch because the 5μm, 300Å silica provides high efficiency. Both phases provide superior performance for proteins, peptides and nucleic acids using protein friendly ion exchange conditions. ProSwift ion exchange monoliths provide an excellent alternative to porous or non porous ion exchange media. They offer increased loading capacity compared to pellicular phases combined with excellent resolution compared to traditional porous ion exchange media. Size Exclusion BioBasic SEC columns, based on silica with a proprietary polymeric coating, offer the mechanical stability of silica-based size exclusion columns with higher efficiencies than that of polymerbased columns. Four pore sizes (60Å, 120Å, 300Å, 1000Å) are available, making them ideal for molecular weight determination of peptides, proteins and water soluble polymers. They can also be used for sample clean-up prior to other analyses. MAbPac SEC-1 (300Å 5µm silica) is a size exclusion chromatography (SEC) column specifically designed for separation and characterization of monoclonal antibodies (MAb) and their aggregates, as well as the analysis of Fab and Fc fragments resulting from proteolysis. Hydrophobic Interaction The ProPac HIC-10 column is a high-resolution, high-capacity, 300Å, 5µm silica-based HIC column that provides excellent high resolution separations of proteins and variants for analytical and preparative applications. ProPac HIC columns provide exceptional hydrolytic stability under the highly aqueous conditions used in HIC. Affinity The MAbPac Protein A column is a unique nonporous polymeric column designed for fast, accurate determination of monoclonal antibody titer analysis from harvest cell culture. The ProPac IMAC-10 is a high-resolution analytical and semipreparative column for separation of proteins and peptides by immobilized metal affinity chromatography. It is packed with 10μm, nonporous, polymeric beads coated with a hydrophilic layer, then grafted with poly(IDA) chains. The ProSwift ConA-1S affinity monolith column is unsurpassed for fast, highly efficient enrichment and purification of Concanavalin A (Con A) binding glycans, glycopeptides, and glycoproteins containing high-mannose regions. Columns for oligonucleotides Thermo Scientific™ DNAPac™ strong anion exchange columns provide industry-leading resolution for analysis and purification of synthetic oligonucleotides. DNAPac columns can resolve full length oligonucleotides from n–1, n+1, and other failure sequences not possible with other columns. Thermo Scientific™ DNASwift™ a strong anion exchange monolith column that provides exceptionally high oligonucleotide purity. This semipreparative column incorporates the high resolution and selectivity of the DNAPac column, with increased loading capacity. Columns for carbohydrates Thermo Scientific™ GlycanPac™ AXH-1 and AXR-1 columns are HPLC columns designed for the simultaneous separation of glycans by charge, size and polarity. Separating either flourescently labeled or native glycans. Nano, capillary and micro columns Thermo Scientific™ Acclaim™ PepMap™ and PepMap RSLC columns are specially designed for high-resolution analyses of tryptic, natural, and synthetic peptides. The columns are often applied for LC-MS/MS peptide mapping for protein identification, biomarker discovery, and systems biology. Due to their high loading capacity, the columns are exceptionally suitable for the analysis of low abundant peptides in complex proteomics samples. Acclaim PepMap Trap columns are typically applied for the desalting and preconcentration of peptides before LC separation with MS detection. The columns are designed to provide the highest efficiencies for one dimensional peptide mapping experiments and 2D-LC analyses. Thermo Scientific™ PepSwift™ monolithic columns are specially designed for fast separation and identification of proteins and peptides using nano and capillary LC coupled to MS. Using highly pure chromatographic media and biocompatible, metal-free fused silica capillaries, Thermo Scientific™ EASY-Column™ capillary LC columns are produced with a focus on design simplicity and strict quality control. As a result, they deliver outstanding chromatographic performance on any nano LC system. Thermo Scientific™ EASY-Spray™ columns provide a fully integrated and temperature controlled combined column-emitter design with only a single nanoViper connection between the LC and the MS ion source. This dead volume reduction is a critical component in helping to deliver state of the art performance with ease of use. Bio Columns Selection Guide Mode of Analysis Recommended Column BioBasic SEC 4-139 MAbPac SEC-1 4-129 BioBasic AX 4-141 ProPac SCX-10, WCX-10, SAX-10, WAX-10, SCX-20 4-116 MAbPac SCX-10, MAbPac SCX-10 RS 4-127 ProSwift IEX 4-123 BioBasic 18, 8, 4 4-140 Acclaim 300 C18 4-142 Accucore 150-C18, 150-C4 4-144 ProSwift RP 4-124 ProPac HIC-10 4-121 MAbPac Protein A 4-126 ProPac IMAC-10 4-122 ProSwift ConA-1S 4-125 Acclaim PepMap 4-149 PepSwift 4-154 EASY-Column 4-155 EASY-Spray Columns 4-153 BioBasic 18, 8, 4 4-140 Acclaim 300 4-142 Preparative BioBasic Inquire Amino Acids (derivatized) Ion Exchange AminoPac PA10 Inquire Reversed phase Hypersil GOLD 4-026 Amino Acids (underivatized) Ion Exchange AminoPac PA10 Inquire Reversed phase Hypercarb 4-084 Anion Exchange Polar retention BioBasic AX 4-141 Hypercarb 4-084 BioBasic AX 4-141 DNAPac PA100, PA200, PA200 RS 4-134 DNASwift 4-137 Ligand Exchange HyperREZ XP 4-056 Ion Exchange CarboPac Inquire Mixed Mode GlycanPac AXH-1 4-032 Acclaim HILIC 4-067 Hypersil GOLD HILIC 4-037 Syncronis HILIC 4-081 Accucore 150-Amide-HILIC 4-146 Hypercarb 4-084 Size Exclusion Ion Exchange Monoclonal antibodies and Proteins Reversed Phase Hydrophobic Interaction Affinity Proteomics Peptides Analytical Nucleotides Oligonucleotides Ion Exchange Carbohydrates HILIC Polar retention View product information and application notes www.thermoscientific.com/chromatography Columns for Biomolecules Analyte LC Columns and Accessories >> Columns for proteomics 4-111 LC Columns and Accessories HPLC Phases for Biomolecules Silica-based Reversed Phase and Ion Exchange Phases Phase Particle Type Acclaim Phase 300 C18 Spherical, fully porous silica Acclaim PepMap Phases 100 C18 Spherical, fully porous silica 300 C18 Spherical, fully porous silica 100 C8 Spherical, fully porous silica 300 C4 Spherical, fully porous silica Accucore Phases 150-C18 Spherical, solid core silica 150-C4 Spherical, solid core silica 150-Amide-HILIC Spherical, solid core silica BioBasic Phases 18 Spherical, fully porous silica 8 Spherical, fully porous silica 4 Spherical, fully porous silica AX Spherical, fully porous silica SCX Spherical, fully porous silica Particle Size (µm) Pore Nominal Surface % Endcapping Carbon Size (Å) Area (m2/g) USP Code Phase Code Page 3 300 100 8 Yes L1 – 4-142 2, 3, 5 5 3, 5 5 100 300 100 300 300 100 300 300 15 9 9 3 Yes Yes Yes Yes L1 L1 L7 L26 – – – – 4-149 4-149 4-149 4-149 2.6 2.6 2.6 150 150 150 80 80 80 7 2 – Yes Yes – L1 L26 – 161 165 167 4-144 4-145 4-146 5 5 5 5 5 300 300 300 300 300 100 100 100 100 100 9 5 4 3 3 Yes Yes Yes No No L1 L7 L26 – L52 721 722 723 731 733 4-140 4-140 4-140 4-141 4-141 Columns for Protein Separations Silica-based Size Exclusion Chromatography Phases Phase SEC Type BioBasic Phases SEC 60 Aqueous SEC 120 Aqueous SEC 300 Aqueous SEC 1000 Aqueous Particle Type Particle Size (µm) Pore Exclusion Limit Size (Å) Operating Range (kDa) USP Code Phase Code Page Spherical, fully porous silica Spherical, fully porous silica Spherical, fully porous silica Spherical, fully porous silica 5 5 5 5 60 120 300 1000 – L33 L33, L59 L33 733 734 735 736 4-139 4-139 4-139 4-139 0.1 – 6 0.1 – 50 1 – 500 20 – 4000 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Silica-based Hydrophobic Interaction Chromatography Phases 4-112 Column Phase Target Applications ProPac HIC-10 Hydrophobic High resolution Interaction separations of proteins and protein variants Base Matrix Particle Pore Nominal Surface Material Size (µm) Size (Å) Area (m2/g) Spherical, porous 5 300 100 ultrapure silica with amide/ethyl surface chemistry Breakthrough Capacity 340mg lysozyme per 7.8 x 75mm column Search our database of thousands of applications www.thermoscientific.com/chromatography Solvent pH Compatibility Range Ammonium 2.5-7.5 sulfate/phosphate salts, organic solvent for cleanup Phase ProPac WCX-10 Weak Cation Exchange ProPac SCX-10 Strong Cation Exchange ProPac SCX-20 Strong Cation Exchange ProPac WAX-10 Weak Anion Exchange ProPac SAX-10 Strong Anion Exchange ProSwift Reversed RP-1S Phase ProSwift Reversed RP-2H Phase ProSwift Reversed RP-3U Phase ProSwift Reversed RP-4H Phase ProSwift Strong SAX-1S Anion Exchange ProSwift Strong SCX-1S Cation Exchange ProSwift Weak WAX-1S Anion Exchange ProSwift Weak WCX-1S Cation Exchange Target Applications High resolution separations of proteins and protein variants Base Matrix Material Ethylvinylbenzene cross linked with 55% divinylbenzene 10µm nonporous particles High resolution Ethylvinylbenzene separations of cross linked with proteins and protein 55% divinylbenzene variants 10µm nonporous particles High Resolution Divinlybenzene separations of 10μm nonporous proteins and protein particles variants High resolution Ethylvinylbenzene separations of cross linked with proteins and protein 55% divinylbenzene variants 10μm nonporous particles High resolution Ethylvinylbenzene separations of cross linked with proteins and protein 55% divinylbenzene variants 10µm nonporous particles Fast protein analysis Monolith; with high resolution polystyreneof large peptides to divinylbenzene medium proteins Fast protein analysis Monolith; with high resolution polystyreneover a wide size range divinylbenzene Fast protein analysis Monolith; with high resolution polystyrenedivinylbenzene of large proteins Fast protein analysis Monolith; with high resolution polystyrenedivinylbenzene Fast protein analysis Monolith; with high resolution polymethacrylate Functional Groups Carboxylate Breakthrough Capacity 6mg/mL lysozyme Recommended Flow Rate 0.2-2.0 mL/min Sulfonate 3mg/mL lysozyme 0.2-2.0 mL/min Sulfonic 20μg/mL Dynamic capacity 0.2-2.0 mL/min Tertiary amine 5mg/mL BSA Quaternary ammonium Solvent Compatibility 80% ACN, acetone. Incompatable with alcohols and MeOH 80% ACN, acetone, MeOH Maximum pH Backpressure Range 3000psi 2.0-12 (21 MPa) 3000psi (21 MPa) 2.0-12 50% acetonitrile 3000psi (21 MPa) 2.0-12 0.2-2.0 mL/min 80% ACN, acetone, MeOH, 3000psi (21 MPa) 2.0-12 15mg/mL BSA 0.2-2.0 mL/min 80% ACN, acetone, MeOH 3000psi (21 MPa) 2.0-12 Phenyl 5.5mg/mL Insulin 2.0-4 .0 mL/min Most common 2800psi organic (19.2 MPa) solvents 1.0-14 Phenyl 1.0mg/mL Lysozyme 1.0-10 mL/min 2800psi (19.3 MPa) 1.0-14 Phenyl 0.5mg/mL Lysozyme 1.0-16 mL/min 2800psi (19.3 MPa) 1.0-14 Phenyl 2.3mg/mL Lysozyme 0.1-0.3 mL/min 1500psi 1.0-14 Quaternary amine 18mg/mL BSA Most common organic solvents Most common organic solvents Most common organic Solvents Most common organic solvents 2.0-12 Fast protein analysis Monolith; with high resolution polymethacrylate Sulfonic acid 30mg/mL Lysozyme Fast protein analysis Monolith; with high resolution polymethacrylate Tertiary amine (DEAE) 18mg/mL BSA Fast protein analysis Monolith; with high resolution polymethacrylate Carboxylic acid 23mg/mL Lysozyme >60mg lysozyme/mL gel (4 x 250mm) 1000psi (4.6mm) 2000psi (1.0mm) Most common 1000psi 0.5-1.5 (4.6mm) organic mL/min solvents (4.6mm) Most common 1000psi 0.5-1.5 (4.6mm) mL/min organic 2000psi solvents (4.6mm) (1.0mm) 0.5-1.5mL/min Most common 1000psi (4.6mm), organic (4.6mm) 2000psi solvents 0.05-0.20 (1.0mm) 1.0mL/min EtOH, urea, 3000psi NaCl, non-ionic (21MPa) detergents, glycerol, acetic acid, guanidine HCl 0-1.0mL/min Up to 10% 2000psi methanol ProPac Immobilized High resolution separation of IMAC-10 Metal Affinity certain metalbinding proteins and peptides Polystyrene divinylbenzene 10μm nonporous particles Poly (IDA) grafts ProSwift Affinity ConA-1S Monolith; polymethacrylate Concanavalin 12-16mg of A ligands protein Concanavalin A binding (highmannose) glycans, glycopeptides and proteins 0.5-1.5 (4.6mm) Columns for Biomolecules Column LC Columns and Accessories >> Polymeric Ion Exchange, Reversed Phase and Affinity Columns 2.0-12 2.0-12 2.0-12 2.0-12 5.0-8 www.thermoscientific.com/chromatography 4-113 LC Columns and Accessories Columns for Monoclonal Antibody Separations Polymeric Ion Exchange Columns Phase IEX Type MAbPac Strong cation SCX-10 exchange (sulfonic) ProPac WCX-10 Particle Size (μm) Polymeric, Highly 3, 5, 10 crosslinked DVB Pore Size (Å) non-porous Polymeric, Non-porous DVB non-porous Particle Type Weak cation exchange (carboxylate) 10 Dynamic Capacity MAbPac SCX-10 PEEK 3µm: 60µg/mL 5µm: 40µg/mL 10µm: 20µg/mL MAbPac SCX-10 RS 5µm: 40 µg/mL 6mg/mL Recommended Flow Rate 0.2-2.0mL/min (Depending on the particle size and column pressure limits) Solvent Compatibility 50% Acetonitrile Maximum pH Page Backpressure Range 3000psi 2.0-12 4-127 (21MPa) RS columns 7000psi 0.2-2.0mL/min 80% ACN, 3000psi acetone. (21MPa) Incompatible with alcohols including methanol 2.0-12 Silica-based Size Exclusion Chromatography Columns Phase SEC Type Particle Type Particle Size (μm) Pore Size (Å) MAbPac SEC-1 Aqueous Spherical, fully porous silica 5 300 Exclusion Limit Operating Range (kDa) 1-500 USP Code Page L33, L59 4-129 Affinity Columns Phase Affinity ligand Particle Type MAbPac Protein A Protein A Polymeric Particle Size (μm) 12 Pore Size (Å) non-porous Recommended Flow Rate < 2.5 Capacity 100µg IgG/ column Maximum temperature 30 Maximum pH Backpressure Range 1000 2.5-7.5 Page 4-126 Columns for Carbohydrate Separations Thermo Scientific Chromatography Columns and Consumables 2014-2015 Polymeric Ligand Exchange Columns 4-114 Phase Particle Type HyperREZ XP Phases Spherical, polymer Carbohydrate H + Carbohydrate Pb2+ Spherical, polymer Carbohydrate Ca2+ Spherical, polymer Carbohydrate Na+ Spherical, polymer Organic Acid Spherical, polymer Sugar Alcohol Spherical, polymer Particle Size (µm) Pore Nominal Surface % Endcapping Carbon Size (Å) Area (m2/g) USP Code Phase Code Page 8 8 8 10 8 8 – – – – – – L17 L34 L19 – L17 L19 690 691 692 693 696 697 4-156 4-156 4-156 4-156 4-156 4-156 – – – – – – – – – – – – – – – – – – Silica-based HILIC and Mixed-Mode Columns Phase Particle Type GlycanPac Phases AXH-1 Spherical, fully porous silica AXH-1 Spherical, fully porous silica Accucore Phase 150-Amide-HILIC Spherical, solid core silica Particle Size (µm) Pore Nominal Surface % Endcapping Carbon Size (Å) Area (m2/g) USP Code Phase Code Page 1.9 3 175 120 220 300 - Yes Yes - - 4-132 4-132 2.6 150 80 - - - 167 4-156 Phase Particle Size (µm) Particle Type ProSwift Phases ConA-1S Concanavalin A binding (high "Monolith; mannose) glycans, glycopeptides and proteins Pore Nominal Surface % Endcapping Carbon Size (Å) Area (m2/g) USP Code Phase Code Page 175 - - 4-125 220 - Yes 5% 40µeq Hydroxide or 1.5mL/min sodium and lithium salts of Chloride or Perchlorate 0-100% 4000psi (28MPa) 2-12.5 55% 5% 40µeq Hydroxide, 1.2mL/min acetate/ or sodium and lithium salts of Chloride or Perchlorate 0-100% 4000psi (28MPa) 2-12.5 55% 5% 40µeq 0-100% 10,000 psi (69MPa) 2-12.5 70% 3% 8mg, of a 20 mer oligonucleotide Hydroxide, 1.3mL/min acetate/ hydroxide or sodium and lithium salts of Chloride or Perchlorate NaClO4 and 0.5-2.5mL NaCl Most Common Organic Solvents 1500psi, 10MPa 6.0-12.4 pH Range Maximum Backpressure Recommended Flow Rate Recommended Eluents 55% Capacity Base Matrix Material Solvent Compatibility DNASwift Latex Crosslinking DNAPac PA200 RS 13µm diameter nonporous substrate agglomerated with alkyl quaternary ammonium functionalized latex 100nm MicroBeads Improved high 8µm diameter resolution nonporous substrate separations of agglomerated with single and double alkyl quaternary stranded DNA or ammonium RNA orthogonal to functionalized latex DNAPac PA100 130nm MicroBeads UHPLC-resolution 4µm diameter separations of nonporous substrate single and double agglomerated with stranded DNA or alkyl quaternary RNA Best ammonium available functionalized latex resolution 130nm MicroBeads High resolution Monolith; separations for polymethacrylate purification of substrate agglomerated oligonucleotides with quaternary amine Highest functionalized latex latex-based capacity www.thermoscientific.com/chromatography Columns for Biomolecules DNAPac PA200 High resolution separations of single and double stranded DNA or RNA oligonucleotides Substrate Crosslinking DNAPac PA100 Target Applications Column Columns for Oligonucleotide Separations LC Columns and Accessories >> Monolithic Affinity Columns 4-115 LC Columns and Accessories ProPac WCX-10 and SCX-10 Weak and strong cation exchange columns are based on a nonporous core particle providing exceptionally high resolution and efficiency for separations of protein variants • Characterization and quality control assessment of monoclonal antibodies and other proteins • Unequalled high resolution separations • High-efficiency peaks • Unmatched column-to-column and lot-to-lot reproducibility • Useful for characterization of related protein variants including deamidation and MAb lysine truncation variants • ProPac WCX-10 is a weak cation exchanger with a carboxylate functional groups • ProPac SCX-10 is a strong cation exchange column with sulfonate functional groups ProPac WCX-10 and SCX-10 columns can resolve isoforms that differ by a single charged residue. A hydrophilic layer prevents unwanted secondary interactions, and a grafted cation exchange surface provides pH-based selectivity control and fast mass transfer for high-efficiency separation and moderate capacity. ProPac WCX-10 Ordering Guide Particle Size (µm) 10 Format Guard Column HPLC Column Length (mm) 50 50 100 150 250 2.0mm ID 063480 – – – 063472 4.0mm ID 054994 074600 SP5829 SP6703 054993 9.0mm ID – – – – 063474 22.0mm ID – – – – SP5482 Length (mm) 50 250 2.0mm ID 063462 063456 4.0mm ID 079930 054995 9.0mm ID – 063700 22.0mm ID – SP5522 ProPac SCX-10 Ordering Guide Thermo Scientific Chromatography Columns and Consumables 2014-2015 Particle Size (µm) 10 4-116 Format Guard Column HPLC Column LC Columns and Accessories >> ProPac WCX-10 and SCX-10 continued MAb lysine truncation variants Column: ProPac WCX-10 , 4 x 250mm Lysine truncation variants* Eluents: K AU KK MAb sample as received Acidic charge variants Basic charge variants After digestion with carboxypeptidase B 0 10 20 Minutes 30 40 21104 (E1) 20mM MES + 115mM NaCl + 1mM EDTA, pH 5.5 (E2) 20mM MES + 145mM NaCl + 1mM EDTA, pH 5.5 Gradient: t (min) %E1 %E2 0 100 0 2 100 0 40 0 100 60 0 100 Sample: MAb Flow Rate: 1.0mL/min λ: 280nm * Peak assignment supported by data from R.J. Harris, et.al, J.Chromatogr., A 1995,705, 129–134. and Carboxypeptidase B digest. Hemoglobin variants Eluent: 2 AU 3 Gradient: 4 Flow Rate: Inj. Vol.: Detection: Sample: 0 0 5 10 15 20 Minutes 25 14067 Columns for Biomolecules Column: ProPac SCX-10, 4 × 250mm 1 1.60 A) 20mM Sodium phosphate, 4mM Potassium cyanide, pH 6 B) 1 M Sodium chloride in water C) Water Time %A %B %C Init 50 0 50 30 min 50 50 0 1mL/min 10μL 220nm 1. Fetal hemoglobin 2. Hemoglobin 3. Sickle cell hemoglobin 4. Hemoglobin C ProPac Kits Ordering Guide Part Number SP5731 SP5732 SP5729 SP5730 SP5512 SP5513 SP5727 SP5728 Phase Description ProPac WAX-10 Lot Select Column Set ProPac WAX-10 Lot Select Column Set ProPac SAX-10 Lot Select Column Set ProPac SAX-10 Lot Select Column Set ProPac WCX-10 Lot Select Column Set ProPac WCX-10 Lot Select Column Set ProPac SCX-10 Lot Select Column Set ProPac SCX-10 Lot Select Column Set Set Contents 3 columns from 1 lot of resin 3 lots of resin, 1 column from each lot 3 columns from 1 lot of resin 3 lots of resin, 1 column from each lot 3 columns from 1 lot of resin 3 lots of resin, 1 column from each lot 3 columns from 1 lot of resin 3 lots of resin, 1 column from each lot Column Dimensions 250 x 4mm 250 x 4mm 250 x 4mm 250 x 4mm 250 x 4mm 250 x 4mm 250 x 4mm 250 x 4mm www.thermoscientific.com/chromatography 4-117 The ProPac SCX-20 column is a strong cation exchange (SCX) column designed specifically to provide highresolution separations of proteins. The stationary phase is composed of 10μm, non-porous, solvent compatible resin beads that are uniformly coated with a highly hydrophilic layer to reduce non-specific interactions between the bead surface and the biopolymer. • Grafted cation-exchange surface provides pH-based selectivity control • Fast mass transfer for high-efficiency separation Snake venoms from naja naja and Russell’s viper Column: ProPac SCX-20, 4 × 250mm 400 Eluent A: Eluent B: Gradient: Inj. Volume: Temperature: Detection: Samples: A 300 200 mAU LC Columns and Accessories ProPac SCX-20 Columns 20mM Tris pH 7.3 0.5M NaCl in Eluent A 1–100% B in 10 min 10μL 30°C UV, 214nm A. Snake Venom (Naja naja) 1mg/mL B. Snake Venom (Russell’s viper) 1mg/mL 100 -50 350 B mAU 200 100 -50 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0 4-118 5 Minutes 10 15 17 ProPac SCX-20 Ordering Guide Particle Size (µm) 10 Description Guard Column HPLC Column Length (mm) 50 250 4.0mm ID 074643 074628 Weak and strong anion exchange columns are based on a nonporous core particle providing unequalled high resolution and efficiency in the separations of protein variants • Unequalled resolution • High-efficiency peaks • Useful for characterization and quality control assessment of closely-related protein variants • Supports separation of proteins that differ by as little as one amino acid residue • Neutral hydrophilic coat that eliminates protein-resin hydrophobic interactions • Superior lot-to-lot and column-to-column reproducibility • ProPac WAX-10 column is a weak anion exchanger with a tertiary amine functional group • ProPac SAX-10 is a strong anion exchange column with quaternary amine functional group These columns can resolve protein isoforms that differ by as little as one charged residue. A hydrophilic layer prevents unwanted secondary interactions and a grafted anion exchange surface provides pH based selectivity control and fast mass transfer for high peak efficiency and resolution separations. Column: ProPac SAX-10, 4 x 250mm Eluents: mAU Gradient: Flow Rate: Inj.Volume: Detection: Sample: Ovalbumin + alk. phos. Ovalbumin 0 2 4 6 8 Minutes 10 12 14 A) Water B) 2.0 M NaCl C) 0.1 M Tris/HCl (pH 8.5) Time %A %B %C Init 80 0 20 15 min 67.5 12.5 20 1.0mL/min 30μL 214nm Ovalbumin before and after alkaline phosphatase treatment Columns for Biomolecules Resolution of phosphorylation variants of ovalbumin LC Columns and Accessories >> ProPac WAX-10 and SAX-10 16 16445 Effect of sialylation on transferrin chromatography Columns: ProPac SAX-10, 4 × 250mm Eluents: mAU HOLO HOLO + neur Flow Rate: Inj. Volume: Detection: Samples: APO Lot #1 APO Lot #1+neur APO Lot #2 APO Lot #2 + neur 0 4 8 12 16 Minutes 20 24 28 Gradient: A) Water B) 2.0 M NaCl C) 0.2 M Tris/HCl (pH 9) Time %A %B %C Init 87 3 10 30 min 83 7 10 1.0mL/min 50μL 214nm HOLO (iron rich) and APO (iron poor) human transferrin samples before and after neuraminidase treatment. Digestions were carried out overnight at 37 C in sodium acetate buffer at pH 5. 32 16446 www.thermoscientific.com/chromatography 4-119 LC Columns and Accessories ProPac WAX-10 Ordering Guide Particle Size (µm) 10 Format Guard Column HPLC Column Length (mm) 50 250 2.0mm ID 063470 063464 4.0mm ID 055150 054999 9.0mm ID – 063707 22.0mm ID – SP5598 Length (mm) 50 250 2.0mm ID 063454 063448 4.0mm ID 054998 054997 9.0mm ID – 063703 22.0mm ID – SP5594 ProPac SAX-10 Ordering Guide Particle Size (µm) 10 Format Guard Column HPLC Column ProPac PA1 For hydrophilic anionic protein separations • Good for hydrophilic proteins and peptides • Ideal for high-resolution separations of proteins with pI values from 3 to 11 • Available in semipreparative format • Pellicular packing ensures high-efficiency and fast mass transport The ProPac PA1 column supports the analysis and purification of hydrophilic anionic proteins and peptides. ProPac PA1 Ordering Guide 4-120 Format Guard Column HPLC Column Length (mm) 50 250 4.0mm ID 039657 039658 9.0mm ID – 040137 Gradient separation of protein standards 2 3,4 Column: ProPac PA1 (4 × 50mm) 5 Eluent: 0 Flow Rate: Detection: Peaks: 6 1 mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 Particle Size (µm) 10 50 Seconds 100 150 3828 10 to 350mM NaCl in 1.0mM Tris, pH 8.2 5mL/min UV, 280nm 1. Horseheart Myoglobin 33μg 2. Contaminant – 3,4. Conalbumin 66 5. Ovalbumin 66 6. Soybean Trysin Inhibitor 66 LC Columns and Accessories >> ProPac HIC-10 Hydrophobic Interaction Chromatography columns for the high resolution separation of proteins and peptides • High-resolution HPLC separation of proteins, protein variants and peptides • Proteins are separated under non-denaturing conditions • High protein loading capacity for protein purification applications • Wide range of applications • Based on 5μm ultra high purity spherical silica gel particles with 300Å pores The ProPac HIC-10 column is a high-resolution, high-capacity, silica-based HIC column that provides excellent separations of proteins and variants for analytical and preparative applications. ProPac HIC columns provide exceptional hydrolytic stability under the highly aqueous conditions used in HIC. ProPac HIC-10 Ordering Guide Format HPLC Column Length (mm) 75 100 250 2.1mm ID – 063653 – 4.6mm ID – 063655 074197 7.8mm ID 063665 – – Protein mixture Column: 7.5 × 75mm (Competitors A and B); 7.8 × 75mm (ProPac HIC-10) 600 mAU 1 Flow: WVL: Eluents: 1.0mL/min 214nm (A) 2 M (NH4)2 SO4 in 0.1 M NaH2PO4 (pH 7.0) (B) 0.1 M NaH 2PO 4 (pH 7.0) Sample: Mixture of proteins (1mg/mL each final after 1:1 dilution with eluent A) Inj. Volume: 20μL Order of elution: 1. Cytochrome c 2. Myoglobin 3. Ribonuclease A 4. Lysozyme 5. Chymotrypsinogen 100 4 5 2 3 0 Competitor A Competitor B -300 0 10 20 30 50 40 60 Minutes 70 21286 Columns for Biomolecules Particle Size (µm) 5 Monoclonal antibody 1000 Column: ProPac ® HIC-10, 4.6 × 100mm 10% CH3CN in eluent B 100 Eluents: 70 mAU Gradient: Flow: Inj. Volume: Detection: Sample: %B (A) 0.5 M (NH 4) 2 SO 4 in 0.1M NaH 2PO 4 (pH 7.0) (B) 0.1 M NaH 2PO 4 (pH 7.0) 70–100% B in 15 min 1mL/min 5μL (25μg) UV, 214nm MAb 50μL (50mg/mL) + 450μL Eluent B 0 -100 0 2.0 4.0 6.0 Minutes 8.0 10.0 12.2 21538 www.thermoscientific.com/chromatography 4-121 LC Columns and Accessories ProPac IMAC-10 Immobilized Metal Affinity Chromatography (IMAC) column for analytical and semipreparative protein and peptide applications • High-purity separations of metal-binding proteins • State-of-the-art technology for reusable columns with metal tailored specificity • Resolve target proteins using a single column in a high-resolution gradient run • Retention control by imidazole concentration or pH gradient • High loading capacity for protein purification applications • Wide range of metal-specific applications The ProPac IMAC-10 is a high-resolution analytical and semipreparative column for separation of proteins and peptides by immobilized metal affinity chromatography ProPac IMAC-10 Ordering Guide Particle Size (µm) 10 Format Loading Column Length (mm) 50 250 50 250 HPLC Column 1.0mm ID – – 063617 – 2.0mm ID – – 063272 063274 4.0mm ID 063667 – 063276 063278 9.0mm ID 063710 063718 063615 063280 Standard proteins with surface-exposed histidines 0.020 2 Column: ProPac IMAC-10 (4mm x 250mm) 3 1 mAU Proteins with surface exposed histidine 4-122 ribonuclease A 0 5 15 20 25 Minutes 10 30 myoglobin 35 40 20416 Prion-related peptides 0.070 3 1 2 Column: ProPac IMAC-10 (4 × 250mm) 4 Eluent: 7 (E1) 20mM HEPES + 0.5 M NaCl, pH = 7.5 (E2) E1 + 50mM imidazole, pH = 7.5 t (min) %A 0 95 10 0 curve 7 30 0 Inj. Volume: 15µL Sample: Prion-derived peptides Flow Rate: 0.5mL/min Wavelength: 280nm Peak # Sequence 1 PHGGGWGQ 2 PHGXGWGQ 3 KKRPKP(PHGGGWGQ) 2 4 KKRP(PHGXGWGQ) 2 5 KKRPKPWGQ(PHGGGWGQ) 4 6 WGQ(PHGGGWGQ) 4 7 WGA(PHGGGWGA) 4 X = sarcosine (N-methyglycine) Gradient: mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 0.000 -0.002 E1: 20mM MES + 0.5 M NaCl, pH = 7.5 E2: E1 + 100mM imidazole, pH = 7.5 Gradient: t (min) %E1 0 96 15 0 curve 7 40 0 Inject Vol: 15µL Sample: 1. 0.25mg/mL ribonuclease A 2. 1.00mg/mL myoglobin 3. 1.50mg/mL carbonic anhydrase Flow Rate: 0.5mL/min Wavelength: 280nm 5 5i 3i 4i 0.000 1 6 6i 4 repeats 2 repeats 1 repeat -0.007 0 5 10 15 20 Minutes 25 30 20420 2 3 4 5 6 7 22.0mm ID – – – 063282 Monolith IEX columns for high-resolution and fast protein analysis ProSwift IEX monoliths provide excellent resolving power at high mass loading combined with fast analysis capability • High resolution • High loading capacity • Fast analysis • Wide range of operational flow rates • Excellent stability over a wide pH range • Outstanding reproducibility and ruggedness ProSwift Ion Exchange Monolith Ordering Guide Functional Group WAX-1S WCX-1S SAX-1S SCX-1S Length (mm) 50 50 50 50 1.0mm ID 066642 066643 068459 071977 4.6mm ID 064294 064295 064293 066765 Columns for Biomolecules ProSwift polymer monolith (poly(Meth)acrylate) media are uniquely suited for separation of proteins. Each monolith is a single cylindrical, sponge-like polymer rod containing an uninterrupted, interconnected network of flow-through channels of a specific pore size. These large channels combined with the monolith’s nonporous surfaces result in fast mass-transfer, high-resolution, and fast protein separations. The unique globular morphology of the polymer medium provides its high capacity. LC Columns and Accessories >> ProSwift IEX Dynamic protein loading capacity of ProSwift WCX-1S 1 × 50mm Column: ProSwift WCX-1S, 1 × 50mm 2 Eluents: 3 Gradient: mAU 1 Flow Rate: Inj. Volume: Temperature: Detection: Sample: Peaks: 72µg 48µg 24µg 12µg 6µg 3µg 0 4 Minutes A. 10mM Sodium phosphate (pH 7.6) B. 1 M NaCl in eluent A 0% B for 2 min, 0–85% B in 7.5 min, 85% B for 3 min 0.2mL/min 1–24μL 30°C UV, 280nm Protein mix, 1mg/mL each 1. Ribonuclease A 2. Cytochrome C 3. Lysozyme 12 8 Protein separation Column: ProSwift WAX-1S, 1 × 50mm Eluents: Gradient: mAU Flow Rate: Temperature: Detection: Inj. Volume: Sample: 0 5 10 Minutes A. 10mM Tris, pH 7.6 B. 1 M NaCl in 10mM Tris, pH 7.6 5 to 55% of B in 13 min, hold for 2 min 0.2mL/min 30°C UV, 280nm 1.3μL 1.25mg/mL E. coli protein 15 24270 www.thermoscientific.com/chromatography 4-123 LC Columns and Accessories ProSwift RP Reversed-phase monolith columns that uniquely provide the advantages of high resolution at exceptionally high flow rates for fast protein separations and analysis Deliver the outstanding resolving power of nonporous analytical media combined with faster separations and analysis than any bead based columns available. • High resolution at high speed • Highest operational flow rates available • High throughput and improved productivity • Excellent stability over a wide pH range of 1 to 14 • Outstanding reproducibility and ruggedness • High stringent wash compatible, for example, 1 M NaOH • High loading capacity ProSwift polymer reversed-phase monolith media are (polyStyrene-co-DVB) uniquely suited for the separation of proteins. Each monolith is a single cylindrical polymer rod containing an uninterrupted, interconnected network of flow-through channels of a specific pore size; ranging from small channel (1S), medium size channels (2H & 4H) to very large channel (3U) sizes. These channels and the monolith’s nonporous surfaces result in fast mass transfer for high-resolution and fast protein separations. The channels also produce low backpressure, allowing the use of higher linear velocities with minimal loss of resolution. ProSwift RP Ordering Guide Functional Group RP-1S RP-2H RP-3U RP-4H RP-4H Length (mm) 50 50 50 50 250 1.0mm ID – – – 069477 066640 4.6mm ID 064297 064296 064298 – – Proteins Column: ProSwift RP-2H (4.6 × 50mm) Flow Rate: Eluents: 600 4-124 4mL/min 2mL/min 1 1mL/min 3 mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 8mL/min 4 2 -300 0 1 2 3 4 5 6 Minutes 7 5 8 9 10 11 22903 1, 2, 4, or 8mL/min (A) DI H2O/CH3CN (95:5; V/V) + 0.1% TFA (B) DI H 2O/CH 3CN (5:95; V/V) + 0.1% TFA Gradient: 1mL/min: 1-75% B in 12 min 2mL/min: 1-75% B in 6 min 4mL/min: 1-75% B in 3 min 8mL/min: 1-75% B in 1.5 min UV Detection: 214nm Sample: Mixture of fi ve proteins Inj. Volume: 5μL Order of Elution: 1. Ribonuclease A 1.5mg/mL 2. Cytochrome C 0.5 3. BSA 1.5 4. Carbonic anhydrase 0.9 5. Ovalbumin 1.5 Competitive comparison 450 700 0 1 2 2 mL/min 1 -100 0 4 2 Minutes 3 4 5 1. ProSwift RP-2H, 4.6 × 50mm 2. Competitor A, 4.6 × 100mm, 15μm Eluents: (A) DI H 2O/CH 3CN (95:5; V/V) + 0.1% TFA (B) DI H2O/CH 3CN (5:95; V/V) + 0.1% TFA 2mL/min: 1-75% B in 6 min 8mL/min: 1-75% B in 1. min 2 or 8mL/min 5μL 30°C UV, 214nm Mixture of fi ve proteins 1. Ribonuclease A 1.5mg/mL 2. Cytochrome C 0.5 3. BSA 1.5 4. Carbonic anhydrase 0.9 5. Ovalbumin 1.5 Gradient: 8 mL/min ProSwift RP-2H 0 Columns: ProSwift RP-2H Competitor A 8 22904 Flow Rate: Inj. Volume: Temperature: Detection: Sample: Peaks: For the highly efficient enrichment and purification of Con A binding glycans and glycoconjugates The ProSwift ConA-1S affinity monolith column is unsurpassed for fast, highly efficient enrichment and purification of Concanavalin A (Con A) binding glycans, glycopeptides, and glycoproteins. • Highly efficient enrichment and purification through high-mannose-Con A binding • Highly purified glycan and glycoconjugate products • High capacity and ligand density • High sample recovery • Low elution volumes • Fast separations • HPLC compatible • Reusable for over one hundred purifications ProSwift ConA-1S Ordering Guide Functional Group ConA-1S Length (mm) 50 5.0mm ID 074148 Columns for Biomolecules The HPLC compatible ProSwift ConA-1S affinity column maintains the specificity and selectivity for Con A binding glycans and glycoconjugates. The high capacity monolith (poly(meth)acrylate) and ligand density of the ProSwift ConA-1S column facilitates the highly efficient enrichment of samples and can purify up to 2mg of glycoproteins. The high peak efficiency monolith (poly(meth)acrylate) column produces sharp, narrow peaks resulting in low elution volumes of highly concentrated products. The column is reusable, and many enrichments and purifications are possible with minimal loss of capacity. LC Columns and Accessories >> ProSwift ConA-1S Fast, highly efficient enrichment and purification of a group of fluorescently labeled glycans Column: ProSwift ConA-1S (5 x 50mm) Flow rate: Eluent A: Eluent B: Gradient: mV Temperature: Sample: Detection: 0 1 2 3 Minutes 4 5 1.0mL/min 50mM NaOAc, 0.2M NaCl, pH 5.3, with 1mM CaCl2 and 1mM MnCl2 100mM α-methyl mannoside in eluent A 0% B for 2 min, 0-100% B in 0.5 min, 100% B for 3.5 min 25°C 2-AB labeled serum protein N-glycans Fluorescence, EM at 420nm 6 27456 Glycopeptide enrichment Column: ProSwift ConA (5 × 50mm) Eluent A: Eluent B: mAU Flow Rate: Temperature: Detection: Sample: Inj. Volume: Gradient: 0 4 8 Minutes 12 50mM NaOAc, 0.2 M NaCl, pH 5.3, with 1mM CaCl2, 1mM MgCl2 100mM α-methyl-mannopyranoside in eluent A 0.5mL/min 25°C UV 214nm Trypsin digested HRP peptides 40μL 100% A from 0–5 min, 100% B from 6–15 min, 100% A from 16–25 min 16 27452 www.thermoscientific.com/chromatography 4-125 LC Columns and Accessories MAbPac Protein A Column Fast MAb titer analysis The Thermo Scientific MAbPac Protein A column is an affinity column designed to provide fast monoclonal antibody (MAb) titer analysis of samples such as harvest cell cultures (HCC). This HPLC column offers high throughput and accurate analysis through a combination of low back pressure and high efficiency. The MAbPac Protein A column format allows rapid automation of loading, binding, elution and collection using Thermo Scientific biocompatible systems. The column is based on a novel non-porous polymeric resin consisting of a divinylbenzene core and a hydrophilic surface, optimized for affinity separation. • High efficiency column • Rugged • Excellent sample recovery • Designed for ease of use and automation Harvest cell culture titer analysis Column: MAbPac Protein A, (4 × 35mm) Unbound 5,200 Flow Rate: Eluent A: Area,mAu.min 40 4,000 Eluent B: 30 Gradient: 20 10 0 3,000 20 mAu 0 40 60 80 100 Amount rabbit IgG, μg 2,000 IgG 1,000 -200 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Minutes 4-126 MAbPac Protein A Ordering Guide Particle Size (µm) 12 Format HPLC Column Length (mm) 35 4.0 mm ID 082539 1.8 2.0 Temperature: Detection: Injection volume: Sample: 2 mL/min 50mM Sodium Phosphate, 150mM NaCl, 5% acetonitrile, pH 7.5 50mM Sodium Phosphate, 150mM NaCl,5% acetonitrile, pH 2.5 0% B for 0.2 mins, 100% B for 0.60 mins,0% B for 1.20 mins 30°C 280 nm 10μL MAb B, 5mg/mL Harvest Cell Culture Strong cation exchange column designed specifically for the high-resolution, high efficiency analysis of monoclonal antibodies and associated variants • Exceptionally high resolution for monoclonal antibody variants • High efficiency • Ideal for characterization and quality control assessment of monoclonal antibodies • Unmatched column-to-column and lot-to-lot reproducibility • Hydrophobic interactions essentially eliminated • Ideal for stability studies • Meets the regulatory requirements for biopharmaceutical characterization The unique nonporous pellicular resin provides exceptionally high resolving power, permitting the separation of monoclonal antibody variants that differ by as little as one charged residue. Hydrophobic interactions with the resin are essentially eliminated for very efficient peaks. MAbPac SCX-10 Ordering Guide Format HPLC Column HPLC Column 10 Guard Column HPLC Column Length (mm) 50 50 150 250 50 50 150 250 2.0mm ID – – – – 075749 – – 075604 4.0mm ID 077907 078656 085198 078655 074631 075603 075602 074625 9.0mm ID – – – – – – – SP6866 Columns for Biomolecules Particle Size (µm) 3 5 LC Columns and Accessories >> MAbPac SCX-10 High resolution separations of monoclonal antibody variants 6 Column: MAbPac SCX-10 (4 × 250mm) Eluents: 8 Gradient: Flow Rate: Temperature: Inj. Volume: Detection: Sample: Peaks 1–5: Peaks 6, 8, 11: Peaks 12–17: mAU 11 4 1 5 7 23 9 10 12 1314 15 0 0 10 20 30 Minutes 16 17 40 50 A. 20mM MES (pH 5.6) + 60mM NaCl B. 20mM MES (pH 5.6) + 300mM NaCl 15–36% B in 50 min 1mL/min 30°C 10μL UV at 280nm MAb B, 5mg/mL Acidic variants C-Terminal Lys variants Basic variants 58 27562 Baseline resolution of C-terminal lysine variants of a monoclonal antibody Column: MAbPac SCX-10 (4 × 250mm) 27.5 mAU 6 Eluents: 6 7 2 1 3 0 1 23 0 10 20 45 A. 20mM MES (pH 5.6) + 60mM NaCl B. 20mM MES (pH 5.6) + 300mM NaCl 15–36% B in 50 min 1mL/min 30°C 5μL UV at 280nm 1. MAb B, 900μg in 100μL (no carboxypeptidase) 2. MAb B, 900μg in 100μL + carboxypeptidase, 50μg, incubation at 37°C for 3 h Both Chromatograms: Peaks 1–5: Acidic variants Sample 1: Peaks 6-8: C-Terminal lysine truncation variants of main peak Peaks 9–11: C-Terminal lysine truncation variants of minor variant peak Sample 2: Peak 6 results from peaks 6, 7, and 8 after CBP treatment. Peak 9 results from peaks 9, 10, and 11 afer CBP treatment Gradient: Flow Rate: Temperature: Inj. Volume: Detection: Samples: 8 45 9 9 10 30 Minutes 40 2 11 1 50 58 27564 www.thermoscientific.com/chromatography 4-127 LC Columns and Accessories MAbPac SCX-10 RS Columns BioRS, strong cation exchange column designed for monoclonal antibodies and associated variants • UHPLC, high throughput analysis • Specially developed bio-inert PEEK lined stainless steel column hardware • High pressure compatibility • Suitable for operation up to 7,000 psi Higher resolution and throughput of MAb charge variant UHPLC separations can be achieved using the small particle MAbPac strong cation-exchange phase with specially developed bio-inert PEEK lined stainless steel column hardware. These columns take advantage of smaller resin size as well as longer column length to maximize the resolution of MAb variant separation, and are suitable for operation up to 7,000 psi. Higher pressure compatibility of the column hardware allows use of high flow rates for faster separation Improved MAb resolution 40.0 Peak resolution is shown for lysine truncation peaks. Equivalent linear velocity and sample load was used. Lysine variants 2.00 30.0 mAU 2.44 3.55 2.94 20.0 n.a. n.a. 10.0 ic cid ts ian var B A 0.0 0.0 4.0 8.0 Other basic variants 12.0 16.0 20.0 24.0 Minutes 28.0 A 32.0 36.0 40.0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 MAbPac SCX-10 RS Ordering Guide 4-128 Particle Size (µm) 5 Format BioRS Column Length (mm) 50 150 250 4.6 mm ID 082674 085209 082673 44.0 46.0 Column: MAbPac SCX, 5μm Dimension: Sample: Injection Volume: Eluent A: Eluent B: Flow Rate: Column backpressure: A: B: 4.6 × 250mm MAb 5mg/mL 15μL 20 mM MES pH 5.6 + 60 mM 20 mM MES pH 5.6 + 300 mM NaCl 1.5 mL/min ~8900Psi Gradient: 33-53% B in 30 min Gradient: 33-53% in 20 min A size exclusion chromatography (SEC) column specifically designed for the high resolution separation and characterization of monoclonal antibodies (MAb) and their aggregates • Anaylsis of monoclonal antibodies (MAb) and their aggregates • Analysis of MAb Fab and Fc fragments. • Hydrophilic bonded layer for minimal nondesired interactions between the biomolecules and the stationary phase • Nonmetallic and Biocompatible PEEK housing eliminates metal contamination from the column hardware • Stable surface bonding leads to low column bleed and compatibility with MS, ELSD and Corona CAD detection • Reproducible and rugged • Superior performance for the analysis of monoclonal antibodies, even using high and low-salt concentrations The stationary phase is packed in bio-inert PEEK format and is compatible with different eluent conditions containing both low and high concentrations of salt in mobile phases, and mass spectrometry friendly volatile eluents. Particle Size (µm) 5 Format Guard Column HPLC Column Length (mm) 50 150 300 Columns for Biomolecules MAbPac SEC-1 Ordering Guide 4.0mm ID 074697 075592 074696 Monoclonal antibody aggregate separation 200 LC Columns and Accessories >> MAbPac SEC-1 Column: MAbPac SEC-1, 5μm Dimension: Mobile Phase: MAb Monomer mAU Temperature: Flow Rate: Inj. Volume: Detection: Sample: 4.0 × 300mm (PEEK) 0.3 M NaCl in 50mM phosphate buffer pH 6.8 30°C 0.20mL/min 2μL UV, 280nm MAb (10mg/mL) Aggregates 0 0 5 10 15 Minutes 20 25 27614 www.thermoscientific.com/chromatography 4-129 LC Columns and Accessories Monoclonal Antibody Characterization and Analysis Kits MAb Charge Variant Analysis IEX Column Kit MAb Charge Variant Analysis IEX Column Kit includes two ion-exchange (IEX) specialty columns for MAb charge variants analysis. This kit is a convenient starter kit for researchers at the beginning of MAb analysis projects, and facilitates the screening of two columns for determination of the best column for their specific monoclonal antibody sample. Included in the Kit: • ProPac WCX-10 Analytical column, 4 × 250mm (P/N 054993), a weak cation-exchange column, the industry standard for high-resolution, high-efficiency analysis of monoclonal antibodies and associated variants • MAbPac SCX-10 Analytical column, 4 × 250mm (P/N 074625), a strong cation-exchange column designed specifically for high-resolution, high-efficiency analysis of monoclonal antibodies and associated variants MAb Charge Variants Kit Ordering Guide Description MAb Charge Variants Analysis IEX Column Kit Cat. No. 076196 MAb Analysis IEX and SEC Column Kit The MAb Analysis IEX and SEC Column Kit includes two columns: an ion-exchange (IEX) column and a size-exclusion (SEC) column. This kit is a convenient starter and column replacement kit for MAb analysis projects. Included in the kit: • MAbPac SCX-10 Analytical column, 4 × 250mm (P/N 074625), a strong cation-exchange column designed specifically for high-resolution, high-efficiency analysis of monoclonal antibodies and associated variants. • MAbPac SEC-1 Analytical column, 4 × 300mm (P/N 074696), a size-exclusion column designed for separating monoclonal antibody (MAb) monomers, aggregates, and fragments. MAb Analysis Kit Ordering Guide Thermo Scientific Chromatography Columns and Consumables 2014-2015 Description MAb Analysis IEX and SEC Column Kit 4-130 Cat. No. 076197 LC Columns and Accessories >> Thermo Scientific pH Gradient Buffers Simple method development for charge variant characterization The Thermo Scientific pH gradient platform accelerates method development and facilitates method transfer to QA/QC for a wide range of protein and MAb charge variants through a generic LC-based approach to charge variant characterization. • Patented buffer formulations enable fast, robust and reproducible pH gradients that are simple to optimize and easily automated • Ready to use with existing LC columns and systems, without the need for time consuming mobile phase adjustments • Applicable to the majority of MAbs Thermo Scientific pH buffer concentrates used in the pH gradient platform can be purchased individually or as a pair, in quantities of 125mL or 250mL. For added convenience, the 125mL buffers can also be bundled with columns in a number of specifically preconfigured kits • The CX-1pH gradient starter kit contains 125mL each of buffers A and B, plus a MAbPac SCX-10, 10μm, 4 × 250mm column Columns for Biomolecules • The CX-1 pH gradient high throughput kit contains 125mL each of buffers A and B, plus a MAbPac SCX-10, 5μm, 4 × 50mm column • The CX-1 pH gradient high resolution kit contains 125mL each of buffers A and B, plus a MAbPac SCX-10, 5μm, 4 × 250mm column For the ultimate flexibility, the preconfigured kits are also available as platforms, including the pH Designer Software.The options are listed in the table below: pH Gradient Buffers Ordering Guide Cat. No. 083273 083275 083274 085346 085348 085349 083381 083378 083272 083380 083376 083270 085022 Description CX-1 pH Gradient Buffer A (pH 5.6), 125mL CX-1 pH Gradient Buffer B (pH 10.2), 125mL CX-1 pH Gradient Buffer Kit (pH 5.6 to 10.2), 125mL CX-1 pH Gradient Buffer A (pH 5.6), 250mL CX-1 pH Gradient Buffer B (pH 10.2), 250mL CX-1 pH Gradient Buffer Kit (pH 5.6 to 10.2), 250mL CX-1 pH Gradient Starter Kit (pH 5.6 to 10.2), 125mL CX-1 pH Gradient High Throughput Kit (pH 5.6 to 10.2), 125mL CX-1 pH Gradient High Resolution Kit (pH 5.6 to 10.2), 125mL CX-1 pH Gradient Starter Platform (pH 5.6 to 10.2), 125mL CX-1 pH Gradient High Throughput Platform (pH 5.6 to 10.2), 125mL CX-1 pH Gradient High Resolution Platform (pH 5.6 to 10.2), 125mL pH Designer Software Optimiztion of MAb charge variant separation using a linear pH gradient: 25% B (pH 6.75) to 50% B (pH 7.9) 16.0 8.00 pH trace 10.0 7.50 7.25 pH Absorbance [mAU] 7.75 5.0 7.00 -2.0 0 5 10 15 20 25 30 35 40 6.60 Minutes www.thermoscientific.com/chromatography 4-131 LC Columns and Accessories GlycanPac AXH-1 Column High-Resolution Columns for Glycan Analysis • Unique glycan selectivity based on charge, size and polarity • Excellent resolution for both native and labeled glycans • Useful for both high-resolution glycan profile characterization and easy quantification of glycans based on charge • Compatible with fluorescence and MS detection methods • High chromatographic efficiency and excellent column stability Thermo Scientific™ GlycanPac™ AXH-1 is a high-performance, silica-based HPLC column for simultaneous separation of glycans by charge, size and polarity. It is designed for high-resolution and high-throughput analysis with unique selectivity for biologically important glycans, either labeled or native, by LC-fluorescence and LC-MS methods. Separation of 2AB labeled N-glycans from bovine fetuin by charge, size and polarity Column: GlycanPac AXH-1 (1.9μm) trisialylated Dimension: Mobile Phase 19 7E5 2.1 × 150mm A: Acetonitrile (100%) B: Water C: Ammonium formate (100mM, pH = 4.4) 0.4mL/min 50 Pmoles 30 °C Fluorescence at 320/420nm 2AB labeled N-glycan from bovine fetuin 20 Flow Rate: Injection Vol.: Temperature: Detection: Sample: disialylated Fluorescence Counts 14 11 monosialylated 5 neutral 1 2 3 4 7 8 15 tetrasialylated 12 13 17 9 10 18 21 22 23 16 6 24 25 26 0 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0 4-132 10 pentasialylated 20 Minutes Time (min) %A %B %C Flow (mL/min) Curve -10 78 20 2 0.4 5 0 78 20 2 0.4 5 30 70 20 10 0.4 5 35 60 20 20 0.4 5 40 50 20 30 0.4 5 40 30 GlycanPac AXH-1 Ordering Guide Particle Size (µm) 1.9 Format UHPLC Column 3 Guard Cartridges (2/pk) HPLC Column Length (mm) 100 150 250 10 2.1 mm ID 082473 082472 082521 082476 3.0 mm ID – – – 082475 4.6 mm ID – – – 082474 150 082470 082469 082468 Acclaim Guard Holder Ordering Guide Description Acclaim Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Cat. No. 069580 069707 074188 LC Columns and Accessories >> GlycanPac AXR-1 Ultra-high resolution UHPLC columns for glycan analysis • Excellent glycan selectivity based on hydrophobicity, charge, size, and isomerization • High resolution for isomeric glycans • Compatibility with fluorescence and MS detection methods • High column efficiency and stability • Ideal tool for qualitative, quantitative and structural analysis of glycans The GlycanPac AXR-1 column, based on 1.9µm, high-purity and spherical silica substrates, combines both weak anion-exchange (WAX) and reversed-phase (RP) retention mechanisms for optimal selectivity and ultra-high resolution for glycan separation. The WAX functionality separates glycans based on charge, and RP property facilitates high resolution for glycans of the same charge according to their hydrophobicity, branching and isomerization. As the result, the GlycanPac AXR-1 column provides unparalleled resolutions for complex charged glycans. No. of detected peaks: ~80 Fluorescence Counts 7E6 disialylated trisialylated Column: GlycanPac AXR-1, 1.9µm Dimension: Mobile phase: 2.1 × 150mm A: acetonitrile B: D.I. water C: ammonium formate (100 mM, pH =4.4) 0.4 mL/min 40 °C 100 pmoles fluorescence at 320/420nm 2AB labeled N-glycan from bovine fetuin Flow Rate: Temperature: Injection Vol.: Detection: Sample: tetrasialylated monosialylated Time % A %B %C Curve (min) Time %A %B %C (min) -10 0 95 5 5 0 0 0 95 5 5 5 -10 95 1 0 95 5 5 0 0 95 5 30 1 74 25 5 1 0 95 65 20 50 30 5 5 2AB neutral 0 10 20 30 Minutes 40 50 60 65 Columns for Biomolecules Separation of 2AB labeled N-glycans from bovine fetuin using GlycanPac AXR-1 Curve 5 5 5 30 1 74 25 5 65 20 50 30 5 GlycanPac AXR-1 Ordering Guide Particle Size (µm) 1.9 Format UHPLC Column Length (mm) 150 250 2.1 mm ID 088136 088135 www.thermoscientific.com/chromatography 4-133 LC Columns and Accessories LC Columns and Accessories >> DNAPac PA100 A strong anion exchange column developed to provide high-resolution analysis and purification of synthetic oligonucleotides • High-resolution oligonucleotide separations • Capable of n, n-1 resolution for oligonucleotides • Resolves oligonucleotides with secondary structures • Compatible with solvent, high pH or high temperatures • Analyzes phosphorothioate-based clinical samples • Provides easy scale-up from 2.0mm to 22mm ID column (>100x) The DNAPac PA100 is a high-resolution anion-exchange column that provides single-base resolution. It is stable under denaturing conditions, rugged, reliable, and can be readily scaled up. The DNAPac PA100 is a 13 μm pellicular, nonporous polymeric resin with bound quaternary aminefunctionalized Thermo Scientific™ Dionex™ MicroBeads™. The rapid mass-transport characteristics of this resin result in very high-resolution oligonucleotide separations. DNAPac PA100 Ordering Guide Particle Size (µm) 13 Format Guard Column HPLC Column Length (mm) 50 250 2.0mm ID SP4016 SP3686 4.0mm ID 043018 043010 Oligonucleotides Column: DNAPac PA100 0.004 Eluent: Thermo Scientific Chromatography Columns and Consumables 2014-2015 mAU Flow Rate: Detection: Sample: 4-134 -0.001 5 10 15 Minutes 20 25 10954 410-510mM NaCl in 25mM Tris-Cl, pH 8.0 1.5mL/min UV, 260nm pd(A) 40–60 9.0mm ID SP4511 043011 22.0mm ID SP4513 SP2091 Columns for Biomolecules DNAPac PA100 can resolve full length from n–1, n+1, and other failure sequences. LC Columns and Accessories >> DNAPac PA200 The DNAPac PA200 is a strong anion exchange column developed to provide bestresolution for analysis and purification of synthetic oligonucleotides • Industry leading resolution for oligonucleotide separations • Achieve n, n-1 resolution for oligonucleotides • Resolve oligonucleotides with secondary structures • Assay phosphorothioate purity • Selectivity control with eluent pH, salt, and solvent • Resolve RNA with aberrant (2’, 5’) links from normal SS-RNA • Separate individual phosphorothioate diastereoisomers • HR/AM AXLC/MS via automated desalting Columns for Biomolecules The DNAPac PA200 is packed with a pellicular anion-exchange resin composed of an 8 μm diameter nonporous polymeric substrate to which quaternary amine-functionalized Dionex MicroBeads are bound. The rapid mass transport characteristics of this resin result in high-resolution oligonucleotide separations. DNAPac PA200 can resolve full length from n−1, n+1, and other failure sequences not possible with other columns. DNAPac PA200 Ordering Guide Particle Size (µm) 8 Format Guard Column HPLC Column Length (mm) 50 250 2.0mm ID 063423 063425 4.0mm ID 062998 063000 Target, failure and trityl-on oligonucleotides 9.0mm ID 063419 063421 22.0mm ID SP6731 SP6734 Separation of oligonucleotides by length 15 Dx98 Full-length Trityl-off 30.1% Oligonucleotide length: 21 22 23 24 25 87.5% Dx96 Dx94 Dx91 Dx90 Dx88 Dx86 Dx87 D84 D80 80 mA260 mA260 55.8% Failure 1.25 M NaCl: sequences 22.8% Trityl-off Full-length Trityl-on 22.8% 0.33 M NaClO4: 22.8% 0 0 8 Minutes 16 24 21022 0 3 4 5 Minutes 6 7 21024 Column: DNAPac ® PA200 Eluent: Flow Rate: Inj. Volume: Detection: NaClO 4, pH 6.5 with 20% CH 3CN 1.2mL/min 8µL UV, 260nm www.thermoscientific.com/chromatography 4-135 LC Columns and Accessories UHPLC Solutions for Nucleic Acid Analysis DNAPac PA200 RS columns are packed with smaller, 4 μm particles, for improved resolution and better performance. The smaller particles also allow shorter columns to provide significantly higher throughput. These columns are packed in bioinert PEEK-lined stainless steel (SST) bodies, designed to protect from unwanted interactions of eluents and analytes with metals, while maintaining 10,000 psi stability. They columns offer exceptional resolution of oligonucleotides, including isomer separations; and are able to resolve full length oligonucleotides from n-1 and n+1 oligonucleotides and other failure sequences. • Provide single base resolution of oligonucleotides • Higher efficiency to improve resolution • Improved throughput • Ruggedness consistent with the DNAPac PA200 column line. • 10,000 psi stable PdA26 PdA27 PdA40 PdA41 PdA42 PdA43 PdA44 PdA45 PdA46 PdA47 PdA48 PdA49 PdA50 PdA51 PdA52 PdA53 PdA54 PdA55 PdA56 PdA57 PdA58 PdA59 PdA60 PdA28 Column: DNAPac PA200 RS, 4.6 x 50mm 20 mM Tris pH 8 A + 1.25 M NaCl 30 °C 1.30 mL/min 2.5 μL 28–43% B in 4 CV* (2.56 min) curve 3** Sample: PdA12–30, 40–60 *CV = column volumes **Curve 3 indicates continuously changing gradient, asymptotically approaching a maximum salt concentration. Programed in Chromeleon 6.8. PdA30 Eluent: A Eluent B: Temp: Flow Rate: Inj. (μL): Detection: dA12 PdA13 mA 260 PdA12 PdA29 PdA14 PdA15 PdA16 PdA17 PdA18 PdA19 PdA20 PdA21 PdA22 PdA23 PdA24 PdA25 16 4-136 dA20 dA21 dA22 dA23 dA24 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0 0 1 2 3 Minutes DNAPac PA200 RS Ordering Guide Particle Size (µm) 4 Format BioRS column Length (mm) 50 150 250 4.6 mm ID 082508 082509 082510 4 Columns for Biomolecules Partial resolution of 46 oligonucleotides LC Columns and Accessories >> DNAPac PA200 RS A strong anion exchange monolith column that provides improved capacity and industry-leading oligonucleotide yield-purity performance. DNASwift SAX-1S Ordering Guide Functional Group SAX-1S Length (mm) 150 5.0mm ID 066766 Tritylated oligonucleotide Mixed-Base 20-mer 2800 Tritylated Mixed-Base 20-mer Column: DNASwift SAX-1S, 5 × 150mm Mobile Phases: A: 15mM Tris, pH 8 B: 15mM Tris, pH 8, 1.25 M NaCl 8–64% B in 10 min 1.5mL/min 20µL UV at 260nm Prep Cell (2mm path length) Derivatized mixed-base 20mer, 20mg/mL mAU Gradient: Flow Rate: Inj. Volume: Detection: Sample: Columns for Biomolecules This semipreparative column incorporates selectivity control of the DNAPac column, providing unsurpassed purity and yields. • Micromole purifications in a 5cm column body • Substantial capacity in a small format • Tunable selectivity control, the DNAPac columns, for high resolution • Compatible with high pH mobile phases, solvents, or high temperatures • Ideal for therapeutic and diagnostic oligonucleotide research • Purify difficult oligonucleotide products • Use in combination with the DNAPac for industry leading purification and characterisation of oligonucleotide products The DNASwift column is a unique porous polymer monolith coated with functionalized latex nanobeads, optimized for oligonucleotide separations. The monolith, a polymer cylinder with interconnected flow through channels, provides fast mass transfer, low back pressure (for increased flow rates), very high capacity, and refined selectivity control. LC Columns and Accessories >> DNASwift SAX-1S 0 0 4 8 12 27019 Minutes Purification of a 21-base RNA Sample with aberrant 2' -5" linkages at the 1 and 3 positions from the 3" end Column: Eluents: 1200 A Gradient: Flow Rate: Inj. Amount: Temperature: Detection: mAU Dio-11: 5´-AUG AAC UUC AGG GUC AGC* UU*G-3´ -200 DNASwift SAX-1S, 5 × 150mm A. 40mM Tris, pH 7 B. 40mM Tris, pH 7 + 1.25 M NaCl 26–42% B in 10 column volumes 1.5mL/min 125µg 30°C UV at 260nm Fr 20 0 5 10 Minutes 15 20 www.thermoscientific.com/chromatography 4-137 4-138 Thermo Scientific Chromatography Columns and Consumables 2014-2015 LC Columns and Accessories Notes LC Columns and Accessories >> BioBasic SEC Columns Superior separation of water soluble compounds • Covers separation of analytes over a wide molecular weight range • Long column life and high column efficiencies • Simple mechanism of interaction based on molecular size and shape • Ideal for sample clean-up • Straightforward method development, simple mobile phases BioBasic SEC Ordering Guide Pore Size (Å) BioBasic SEC 60 60 Description ID (mm) Length (mm) Cat. No. Guard HPLC Column HPLC Column 7.8 7.8 7.8 30 150 300 73305-037821 7.8 7.8 7.8 30 150 300 73405-037821 7.8 7.8 7.8 30 150 300 73505-037821 7.8 7.8 7.8 30 150 300 73605-037821 73305-307846 Columns for Biomolecules BioBasic SEC 120 120 Guard HPLC Column HPLC Column BioBasic SEC 300 300 Guard HPLC Column HPLC Column BioBasic SEC 1000 1000 Guard HPLC Column HPLC Column 73305-157846 73405-157846 73405-307846 73505-157846 73505-307846 73605-157846 73605-307846 1/2 4 3 60Å 1 Columns: BioBasic SEC, 5µm, 300 x 7.8mm 2 4 3 120Å 4 2 1 300Å Eluent: Flow: Detector: Injection: Sample: 0.1M KH 2PO 4 pH 7 1.0mL/min UV at 254nm 20µL 1. Thyroglobulin (MW669,00) 2. Ovalbumin (MW 45,000) 3. Angiotensin II (MW 1,046) 4. PABA (Vm) (MW 137) 3 3/4 1 1000Å 0 2 4 6 2 8 10 MIN Molecular weight calibration curves Effect of pore size on SEC resolution www.thermoscientific.com/chromatography 4-139 LC Columns and Accessories BioBasic 18 Outstanding separation of small to medium peptides • 300Å pore size for maximum performance with biomolecules • High peak capacity stationary phase • Outstanding reproducibility, efficiency and column lifetime • Excellent for LC-MS separations Hydrophobicity Low High pH Range 0 2 to 9 14 Pore Size 0 300 Carbon Load (%) 0 25 9 Particle Size 5µm, 10µm USP L1 BioBasic 8 Optimized for the separation of a wide range of peptides 9 • 300Å pore size for improved biomolecule analysis • An excellent starting column for protein and peptide analysis • Outstanding reproducibility, efficiency and column lifetime • Excellent for LC-MS separations 5µm L7 BioBasic 4 • Based on 300Å silica for outstanding biomolecule performance • Lower carbon load for optimal retention of larger peptides and proteins Thermo Scientific Chromatography Columns and Consumables 2014-2015 • Outstanding reproducibility, efficiency and column lifetime 4-140 9 • Excellent for LC-MS separations 5µm L26 Optimized for the separation of proteins, peptides, other anionic species and polar molecules • Weak anion exchange phase for multiple charged species • 300Å pore size for enhanced protein and peptide separations • Suitable for HILIC retention and separation of highly polar molecules • Superb stability under demanding pH conditions N/A 5µm L52 BioBasic SCX For the separation of proteins, peptides, and other cationic species • Strong cation exchange phase based on sulfonic acid chemistry • Separation and retention of basic and other cationic species • 300Å pore size for enhanced protein and peptide separations • Outstanding stability under demanding pH conditions BioBasic Ordering Guide Particle Size (µm) 5 Format Drop-in Guard (4/pk) HPLC Column Length (mm) 10 50 100 150 250 Format UNIGUARD Guard Cartridge Holder ID (mm) C18 C8 C4 AX SCX 1.0 2.1 3.0 4.0/4.6 1.0 2.1 3.0 4.6 1.0 2.1 3.0 4.6 1.0 2.1 3.0 4.6 1.0 2.1 4.6 72105-011001 72105-012101 72105-013001 72105-014001 72105-051030 72105-052130 – – 72105-101030 72105-102130 72105-103030 72105-104630 72105-151030 72105-152130 – 72105-154630 72105-251030 72105-252130 72105-254630 72205-011001 72205-012101 72205-013001 72205-014001 72205-051030 72205-052130 – – 72205-101030 72205-102130 72205-103030 72205-104630 72205-151030 72205-152130 – 72205-154630 72205-251030 72205-252130 72205-254630 72305-011001 72305-012101 72305-013001 72305-014001 72305-051030 72305-052130 – – 72305-101030 72305-102130 72305-103030 72305-104630 72305-151030 72305-152130 – 72305-154630 72305-251030 72305-252130 72305-254630 73105-011001 73105-012101 73105-013001 73105-014001 73105-051030 73105-052130 73105-053030 73105-054630 73105-101030 73105-102130 73105-103030 73105-104630 73105-151030 73105-152130 73105-153030 73105-154630 73105-251030 73105-252130 73105-254630 73205-011001 73205-012101 73205-013001 73205-014001 73205-051030 73205-052130 73205-053030 73205-054630 73205-101030 73205-102130 73205-103030 73205-104630 73205-151030 73205-152130 73205-153030 73205-154630 73205-251030 73205-252130 73205-254630 Length (mm) ID (mm) 10 1.0 2.1 3.0 4.0/4.6 Columns for Biomolecules 5µm LC Columns and Accessories >> BioBasic AX Cat. No. 851-00 852-00 852-00 850-00 www.thermoscientific.com/chromatography 4-141 LC Columns and Accessories Acclaim 300 C18 Hydrophobicity High-resolution reversed-phase separation of proteins and peptides Low 0 • Designed for high-resolution peptide mapping and protein separations 14 • High-efficiency 3μm spherical silica substrate 0 • High-performance bonding chemistry on 300Å pore silica 300 Carbon Load (%) • Application tested for suitability in peptide mapping • Reproducible for dependable results 0 • LC-MS compatible Particle Size 3µm 8 • Ultrapure silica with low metal content and exhaustive bonding and endcapping Acclaim 300 C18 Ordering Guide Particle Size (µm) 3 Format HPLC Column 5 Guard Cartridges (2/pk) Length (mm) 50 150 10 2.1mm ID 060263 060264 069690 3.0mm ID – 063684 075721 Cat. No. 069580 069707 074188 Description Acclaim SST Guard Cartridge Holder V-2 Acclaim Guard Kit (Holder and coupler) V-2 Guard to Analytical Column Coupler V-2 Budesonide and related substances Column: Acclaim 300 C18 3.0µm, 4.6 x 150mm 7 Thermo Scientific Chromatography Columns and Consumables 2014-2015 2.5 to 7.5 Pore Size Acclaim Guard Holder Ordering Guide 4-142 High pH Range 0–24 mAU FS 1 2 0 3 4 5 Pump: Mobile Phase: 8 Flow: Temperature: Injection: Detector: Sample: 6 9 1 0 Peaks: 0–400 mAU FS 0 5 10 Minutes 15 20 21523 Summit P580 HPG/4 (A) Acetonitrile:ethanol 15:1 (B) 0.1% phosphoric acid Isocratic 66% B 1.0mL/min 30°C ASI-100 autosampler, 15μL UVD 340U; UV at 240nm Budesonide, 500μg/mL after three days 7, 8. Budesonide epimers, 99% Reference: Hou S, Hindle M, Byron PR; J. Pharm. Biomed. Analy. 2001 24:371-80. 4.6mm ID 060265 060266 069697 25 USP L1 Ultimate Core Performance for Biomolecule Separations The range of Accucore HPLC columns packed with 150Å pore diameter particles allow biomolecule separations to benefit from the superb resolution and high speed enabled by the Core Enhanced Technology™ that Accucore columns are based on. The 150Å pore diameter solid core particles used in the Accucore 150-C18, 150-C4 and 150-Amide-HILIC columns are designed specifically to provide the optimum combination of retention and resolution for biomolecules. The chromatograms below show the improved resolution and peak shapes achieved with Accucore 150-C18 compared to a C18 solid core phase with an 80Å pore diameter. 3.76 7.09 8.74 13.46 8.91 11.20 Columns for Biomolecules Accucore 150-C18 1.12 LC Columns and Accessories >> Accucore Columns for Biomolecules Conventional Solid Core C18 1.01 8.52 6.98 3.60 13.13 10.84 Improved resolution Improved peak shape Peak Capacity 150 140 130 120 110 100 90 Peak Capacity Comparison Sharper peaks result in a significantly higher peak capacity for 150-C18 compared to the conventional solid core C18 column. 80 70 60 Accucore 150-C18 Conventional Solid Core C18 Stationary Phase Accucore 2.6µm 150Å HPLC and nanoLC Columns Optimum Conditions and Ratings Column ID 75µm 2.1mm 3.0mm 4.6mm Optimum Flow Rate 300nL/min 400µL/min 800µL/min 1800µL/min Backpressure Rating 800 bar 1000 bar 1000 bar 1000 bar Temperature Rating 70°C 70°C 70°C 70°C www.thermoscientific.com/chromatography 4-143 LC Columns and Accessories Accucore 150-C18 Hydrophobicity • Designed for the separation of peptides Low pH Range • Outstanding resolution • 150Å pore diameter material 0 Higher peak capacities facilitate increased peptide identifications. Accucore 150-C18 provides much narrower peak widths, therefore significantly higher peak capacity than a column packed with <2μm wide pore fully porous C18. Precision of retention times is critical for reliable analysis. The Accucore 150-C18 column exhibits excellent retention time reproducibility. 0 0 Mobile phase A: Mobile phase B: Gradient: 180000 Flow: Temperature: Injection: Detection: Peaks: 160000 mAU 140000 1 120000 100000 80000 2 60000 4 3 7 5 6 40000 20000 0 -20000 -40000 0 1 2 3 4 5 6 7 8 Minutes 9 10 11 12 13 14 15 150Å 7 Particle Size 2.6µm Column: Accucore 150-C18 2.6µm, 100 x 2.1mm 240000 200000 Thermo Scientific Chromatography Columns and Consumables 2014-2015 14 300 Carbon Load (%) Peptide separations 220000 4-144 1 to 11 Pore Size 0.1% TFA in 10:90 acetonitrile:water 0.1% TFA in 70:30 acetonitrile:water 0-50% B over 15 min; hold for 2 min; drop to 0% in 0.1 min; hold at 0% B for 5 min 300µL/min 35°C 5µL UV (220nm) 1. Glycine-Tyrosine 2 . Valine-Tyrosine-Valine 3. Met-Enkephalin 4. Angiotensin III 5. Leu-Enkephalin 6. Ribonuclease A 7. Insulin 25 USP L1 Hydrophobicity • Significantly lower hydrophobic retention than C18 Low pH Range • Ideal for retention of proteins and larger peptides 0 2 to 9 14 Pore Size Accucore 150-C4 provides significantly sharper and higher peaks than a column packed with 5μm wide pore fully porous C4, thus offering better resolution and sensitivity. The Accucore 150-C4 also performs better than a column packed with <2μm wide pore fully porous C4 and generates only a fraction of the backpressure. 0 150Å 300 Carbon Load (%) 0 2 Particle Size 2.6µm 25 USP L26 Intact proteins Column: Accucore 150-C4 2.6µm, 100 x 2.1mm 130000 Mobile phase A: Mobile phase B: Gradient: 120000 110000 5 100000 3 80000 mAU 70000 60000 2 50000 6 40000 1 30000 * 20000 * 4 10000 0 -10000 0 1 2 3 4 5 6 7 8 9 Columns for Biomolecules Flow: Temperature: Injection: Detection: Peaks: 90000 0.1% TFA in 30:70 acetonitrile:water 0.1% TFA in 98:2 acetonitrile:water 0-30% B over 8 mins 30-95% B over 2 mins 95% B hold for 1 min 0% B hold for 4 mins 400µL/min 40°C 2µL 10pmol/µL solution UV (214 and 280nm) 1. Insulin 2. Cytochrome C 3. Lysozyme 4. Myoglobin 5. Carbonic anhydrase 6. Ovalbumin Carbonic anhydrase impurity LC Columns and Accessories >> Accucore 150-C4 10 Minutes www.thermoscientific.com/chromatography 4-145 Low High pH Range 2 to 8 0 14 Accucore 150-Amide-HILIC 0 Hydrophobicity 300 150 Å • Amide phase bonded onto 150 Å pore diameter solid core particles Particle Size 2.6µm Low High pH Range • High retention of a broad range of hydrophilic analytes in HILIC mode • Recommended for hydrophilic biomolecules such as glycans 2 to 8 0 14 Pore Size The amide bonded phases provide strong hydrogen bonding interaction between the stationary phase and the analytes, resulting in unique selectivity compared to other HILIC phases. Combined with larger pore size of the solid core particles, Accucore 150-Amide-HILIC is well suited for separating a variety of hydrophilic molecules, including carbohydrates and peptides. As a result the Accucore 150-Amide-HILIC is an excellent choice for glycan separations. 0 300 150 Å Particle Size 2.6µm 2-AB labelled dextran ladder 1200000 1 A 2 1100000 1000000 300000 3 11 250000 900000 12 200000 13 150000 800000 4 700000 Counts 10 14 15 16 100000 17 18 50000 600000 Column: Accucore 150-Amide-HILIC, 2.6μm, 100 x 2.1mm B 350000 Mobile phase A: Mobile phase B: Gradient: 19 20 21 0 500000 5 400000 30 300000 35 Minutes 40 45 6 200000 7 100000 8 9 10 11 0 -100000 -200000 5 10 15 20 25 30 35 40 45 Minutes acetonitrile 50 mM ammonium formate, pH 4.5 20–50 % B in 40.0 minutes 50 % B for 5.0 minutes 50–20 % B in 0.5 minutes 50 % B for 4.5 minutes 500 μL/min Flow: 1 Backpressure at 1200000 starting conditions: 110 bar 2 1100000 Run time: 50 minutes 1000000 Temperature: 60 °C Injection: 2μL to 5μL of sample.900000 Injection wash 800000 solvent: 80:20 (v/v) acetonitrile:water. 700000 Fluorescence detector 600000 acquisition parameters: 330nm excitation wavelength; 420nm 500000 emission wavelength; acquisition start after 3 min from 400000 gradient start. A 300000 11 200000 12 13 150000 4 14 15 16 100000 17 18 50000 19 20 21 0 5 30 35 Minutes 40 45 6 200000 7 100000 (B) 5 µL injection of sample, zoomed-in to the later part of the gradient rise. A further 10 glycans were detected. 10 250000 300000 (A) 2 µL injection of sample, where 11 glycans were separated. B 350000 3 Counts LC Columns and Accessories Pore Size 8 0 9 10 11 -100000 -200000 5 10 15 20 25 30 35 Minutes 4-146 mAU Thermo Scientific Chromatography Columns and Consumables 2014-2015 2-AB labelled dextran ladder 12 11 10 9 8 7 6 5 4 3 2 1 0 (A) 2 µL injection of sample, where 11 glycans were separated. Column: Accucore 150-Amide-HILIC 2.6μm, (B) 5 µL injection of sample, zoomed-in to the later part of the gradient rise. 150mm x 75μm 1 Mobile phase A: Mobile phase B: Gradient: 3 2 4 5 6 25 30 35 40 Minutes 7 8 9 10 11 12 13 14 15161718 45 50 55 Flow: Temperature: Backpressure: Sample Pick-up: Sample Loading: Detection: A further 10 glycans were detected. 98:2 (v/v) acetonitrile: water 2:98 (v/v) acetonitrile: water 0–50 %B in 50 minutes 50 % B for 8 minutes 200 nL/min 40 °C 60 bar (100% A) 0.5μL at 20 μL/min 1μL at 280 bar UV (240 and 33 nm) 35 40 45 Particle Size (µm) 2.6 Format Defender Guard (4/pk) Length (mm) 10 HPLC Column 30 50 100 150 Format UNIGUARD Guard Cartridge Holder Length (mm) 10 150-C18 16126-012105 16126-013005 16126-014005 16126-032130 16126-033030 16126-034630 16126-052130 16126-053030 16126-054630 16126-102130 16126-103030 16126-104630 16126-152130 16126-153030 16126-154630 – ID (mm) 2.1 3.0 4.6 Cat. No. 852-00 852-00 850-00 150-C4 16526-012105 16526-013005 16526-014005 16526-032130 16526-033030 16526-034630 16526-052130 16526-053030 16526-054630 16526-102130 16526-103030 16526-104630 16526-152130 16526-153030 16526-154630 – 150-Amide-HILIC 16726-012105 16726-013005 16726-014005 – – – 16726-052130 16726-053030 16726-054630 16726-102130 16726-103030 16726-104630 16726-152130 16726-153030 16726-154630 16726-252130 www.thermoscientific.com/chromatography Columns for Biomolecules 250 ID (mm) 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 4.6 2.1 LC Columns and Accessories >> Accucore Columns for Biomolecules Ordering Guide 4-147 LC Columns and Accessories Nano, Capillary and Micro LC Columns Introduction Nano, capillary and micro columns are ideal in cases where the sample amount is limited. The (theoretical) sensitivity increase of these LC columns is obtained by introducing the limited sample amounts in an environment with a low internal volume. This will result in a sensitivity gain that is dependent on the internal diameter, as shown in the table below. The best known use is the application of nano LC columns in proteomics, but certainly the concentrating effects of capillary and micro columns are beneficial in e.g. metabolomics or facilitation of LC-MS interfacing. Loadability Name Conventional Narrowbore Micro Capillary Nano Internal Diameter 4.6mm 2.1mm 1.0mm 300µm 75µm Theoretical Sensitivity 1 5 21 235 3761 Predominant application area Small molecules, pharma DMPK, Metabolomics, LC-MS Protein prefractionation Peptide prefractionation Proteomics Sensitivity Thermo Scientific Chromatography Columns and Consumables 2014-2015 Formats 4-148 Our columns come in a variety of formats. We offer most chemistries in nano (75µm), capillary (300µm) and micro (1000µm) internal diameter. In addition, different lengths are available to allow tuning for throughput with short micro columns or high resolution and sensitivity with 50cm long nano columns. However most importantly, Thermo Scientific™ Dionex™ nanoViper™ fingertight fitting system has now become the standard column format for our nano, capillary, and micro LC columns. nanoViper fingertight connections eliminate the assembly of PEEK sleeve connections, allowing tool-free nano-LC connections. They are capable of withstanding pressures up to a 1000 bar, offering the same levels of performance with improved ease of use. Our trap columns come in two formats as well, the nano trap design, features a single capillary with the trap column at the end. These are ideal for high resolution separation of relatively clean samples or samples with hydrophilic peptides. The cartridge based trap columns are use at higher flow rates and have a wider ID. These are ideal for large sample amounts and when sample quality demands more robustness. For more details on trap columns, please check the RSLCnano standard applications manual. Thermo Scientific EASY-Spray columns minimize the impact of dead volumes through the use of specifically designed columns in which the separation column, column heater, high-voltage electrode, and emitter are integrated in a ready-made assembly which incorporates a nanoViper connector. Eliminating imperfect connections makes it far easier to achieve state-of-the-art nanospray performance and data. Chemistries The most common “sample limited” sources are from biological origin and involve the analysis of peptides and proteins. It is for that reason that our nano, capillary and micro column portfolio has a strong focus on column chemistries for peptides and proteins. The Acclaim PepMap range (C18, C8, C4) has been the benchmark for nano LC peptide analysis for many years. The Acclaim PepMap RSLC (C18, 2µm particle) range builds on this heritage improving peptide separation capabilities. Acclaim PepMap columns are available in 100 and 300Å pore size to accommodate a wide range of peptides and proteins. In addition to these hallmark columns our range also includes monolithic columns that are capable of separating both proteins and peptides with very high efficiency. Our portfolio is completed by a selection of columns that are applicable as the first dimension in a multidimensional approach. The standard for peptide separations in proteomics • High resolution protein identification for biomarker discovery, verification or any other analysis • Highest sensitivity in LC-MS • Quality control by factory testing all columns Acclaim PepMap and Acclaim PepMap RSLC columns are specially designed for high-resolution analyses of tryptic, natural, and synthetic peptides. The columns are often applied for LC-MS/MS peptide mapping for protein identification, biomarker discovery, and systems biology. Due to their high loading capacity, the columns are exceptionally suitable for the analysis of low abundant peptides in complex proteomics samples. Protein mixture digest Column: Acclaim PepMap C18 3µm (75um ID x 50cm) 10.0 Flow Rate: Mobile Phase: 8.0 mAU 4.0 2.0 0.0 -1.5 15 30 40 50 60 70 80 Minutes 90 100 110 120 130 Columns for Biomolecules Gradient: Detection: Sample: Temperature: 6.0 300 nL/min A: water + 0.1% formic acid B: water/acetonitrile 20/80 v/v% + 0.08% formic acid 4–55% B in 120min, 5 min wash UV, 214nm Protein mixture digest 35°C LC Columns and Accessories >> Acclaim PepMap C18 100Å Acclaim PepMap C8, 100Å An excellent alternative to Acclaim PepMap 100Å C18, when separating very hydrophobic peptides (e.g., non-tryptic peptides) Acclaim PepMap C4, 300Å Ideal for the separation of larger hydrophobic peptides and proteins, providing higher recoveries Acclaim PepMap C18, 300Å Wide pore version allows larger peptides and proteins Accucore 150-C18 Designed to give narrow peaks and high peak capacity for peptide separations Accucore 150-C4 Ideal for high resolution protein separations Accucore 150-Amide-HILIC Well suited for separating a variety of hydrophilic molecules, including carbohydrates and peptides www.thermoscientific.com/chromatography 4-149 LC Columns and Accessories Separation Columns Acclaim PepMap C18 100Å Ordering Guide Particle size (µm) 2 3 5 Length (mm) 50 150 250 500 50 150 250 500 50 150 250 50µm ID 164561 164562 164709 164710 164712 164713 164714 164715 – – – nanoViper Column 75µm ID 300µm ID 164563 164560 164534 164537 164536 – 164540 – 164567 164716 164568 164571 164569 – 164570 – – 164901 – 164902 – – Classic Column 75µm ID – – – – 1000µm ID 164454 164711 – – 164717 164572 – – 164899 164900 – 160321 164261 – – 160323 160326 1000µm ID 164723 Classic Column 75µm ID 161185 1000µm ID 164721 Classic Column 75µm ID 163579 1000µm ID 164719 Classic Column 75µm ID 163574 Acclaim PepMap C8 100Å Ordering Guide Particle size (µm) 3 Length (mm) 150 50µm ID – nanoViper Column 75µm ID 300µm ID 164706 164722 Acclaim PepMap C4 300Å Ordering Guide Particle size (µm) 5 Length (mm) 150 50µm ID – nanoViper Column 75µm ID 300µm ID 164707 164720 Acclaim PepMap C18 300 Å Ordering Guide Particle size (µm) 5 Length (mm) 150 50µm ID – nanoViper Column 75µm ID 300µm ID 164708 164718 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Accucore 150-C18 Ordering Guide 4-150 Particle size (µm) 2.6 Length (mm) 150 500 nanoViper Column 75µm ID 16126-157569 16126-507569 Accucore 150-C4 Ordering Guide Particle size (µm) 2.6 Length (mm) 150 500 nanoViper Column 75µm ID 16526-157569 16526-507569 Accucore 150-Amide-HILIC Ordering Guide Particle size (µm) 2.6 Length (mm) 150 nanoViper Column 75µm ID 16726-157569 Acclaim PepMap C18 100Å Ordering Guide Type nanoViper Column Classic Column Part number 164535 164705 164564 164197 164199 164213 Particle Size 3 3 5 5 5 5 ID 75 75 100 100 100 200 Bed Length (mm) 20 20 20 10 20 20 Total Length 150 70 150 150 150 150 Quantity 2 2 2 2 2 2 Note: 164705 is a shorter total length used for vented column set up for example with EASY nLC 1000 Trap Columns – Cartridge (Set of 5) Acclaim PepMap C18 100Å Ordering Guide 300µm ID 160454 – 1000µm ID 160434 160438 Columns for Biomolecules Length (mm) 5 15 Acclaim PepMap C8 100Å Ordering Guide Length (mm) 5 300µm ID 161194 Acclaim PepMap C4 300Å Ordering Guide Length (mm) 5 LC Columns and Accessories >> Trap columns – nano trap design 300µm ID 163591 Acclaim PepMap C18 300Å Ordering Guide Length (mm) 5 300µm ID 163589 Accucore 150-C18 Ordering Guide Length (mm) 5 300µm ID 16126-900379 Accucore 150-C4 Ordering Guide Length (mm) 5 300µm ID 16526-900379 Accucore 150-Amide-HILIC Ordering Guide Length (mm) 5 300µm ID 16726-900379 μ-Precolumn™ holder, 5mm, with 30μm ID connecting tubing, nanoViper fi ttings Length (mm) 5 300µm ID 164649 www.thermoscientific.com/chromatography 4-151 LC Columns and Accessories 1st dimension columns for 2D separations Improving separation efficiency can be achieved by adding a column to the setup for multidimensional analysis. There are dedicated application kits which utilize either SCX or RP as the first dimension. A TiO2 trapping column can be added to the application for phosphopeptide analysis. Check the RSLCnano standard applications manual on the Thermo Scientific website for more information on 2D separations. 1st dimension columns Ordering Guide Part Number 164565 Chemistry SCX Description 300μm ID x 10cm, packed with Poros 10 S, nanoViper column 1.0mm ID x 15cm, packed with Polysulfoethyl ASP, 5μm, 300Å, nanoViper column 300μm ID x 15cm, packed with Acclaim PA2, nanoViper column 100µm ID x 1cm packed with TiO2 200µm ID x 1cm packed with TiO2 164566 RP 164592 TiO2 164205 164215 Application 2D Salt plugs (6720.0325) Automated off-line 2D LC of Peptides, micro SCX x nano RP (6720.0330) Automated off-line 2D LC, Cap RP (basic) x Nan RP (acidic) (6720.0335) Phosphopeptide analysis Phosphopeptide analysis Isolation of two synthethic phosphopeptides from a BSA tryptic digest x10 8 4 Breakthrough A 3 2 1 0 x10 8 4 Bound fraction B SVENLPEAGlpTHEQR 1758.8 Da 3 2 ENIMRpSENSESQLTSK 1931.8 Da 1 Thermo Scientific Chromatography Columns and Consumables 2014-2015 0 4-152 5 10 15 20 25 Minutes 30 35 40 45 23731 Trap Column: 200μm ID x 1cm, packed with TiO2 , 5μm 100μm ID x 1cm, packed with Acclaim PepMap C18, 5μm Separation Column: Acclaim PepMap C18, 3μm Dimensions: 75μm x 15cm Loading Solvent: 0.05% HFBA in DI H 2O Wash Solvent: 0.01% HFBA in DI H 2O Mobile Phases: (A) 0.05% TFA in DI H 2O (B) 0.04% TFA in acetronitrile/ DI H 2O (80:20 v/v) Gradient: 3–40% acetronitrile in 30 min TiO 2 Trap Eluent: 250mM NH 4HCO 3 in DI H 2O, pH 9.0 Flow Rate: 300nL/min Loading Flow: 8μL/min Inj. Volume: 5μL Detection: MS Nanoflow LC-MS utilizes very narrow fused silica columns and as a result, even the slightest inconsistency in column connection can result in leaks and dead volumes and are a frequent source of poor data, causing unstable spray and a significant loss in performance. Trouble-shooting connection issues can be difficult and time consuming, even for experts. Thermo Scientific™ EASY-Spray™ columns address the aforementioned issues by incorporating high-performance nanobore columns and integrating them within a radical new design. Packed with either C18 for peptide separations or with selected wide pore phases and monoliths for the separation of intact proteins, e.g. top-down proteomics or the structural identification of biotherapeutics, the “plug and spray” approach delivers state of the art performance in a simple to use format, making research routine. Acclaim PepMap C18 100Å Ordering Guide Particle size (µm) 2 50µm ID ES801 – – – 75µm ID – ES802 ES803 ES800 – 75µm ID ES811 Columns for Biomolecules 3 Length (mm) 150 250 500 150 Accucore C4 150Å Ordering Guide Particle size (µm) 2.6 Length (mm) 150 Acclaim PepMap C18 300Å Ordering Guide Particle size (µm) 5 Length (mm) 150 – LC Columns and Accessories >> EASY-Spray Columns 75µm ID ES812 PepSwift Ordering Guide Length (mm) 250 – 200µm ID ES810 EASY-Spray Emitters EASY-Spray emitters are integrated devices consisting of a sprayer coupled with a transfer line. The EASY-Spray benefits of pre-made connections, as well as the easy installation in the MS ion source can now be exploited in cases where no chromatography is required, for example when tuning the mass spectrometer. EASY-Spray Emitters Ordering Guide Description EASY-Spray emitter, nanoflow (Emitter ID 7µm, Transfer line ID 20µm, Transfer line length 500mm) EASY-Spray emitter, microflow (Emitter ID 20µm, Transfer line ID 75µm, Transfer line length 500mm) Cat. No. ES791 ES792 www.thermoscientific.com/chromatography 4-153 LC Columns and Accessories PepSwift and ProSwift (PS-DVB) Capillary and Micro HPLC Columns • High-resolution for protein identification, biomarker discovery, and systems biology • High-speed peptide and protein separations (<15 min) • Highest sensitivity for LC/MS • Highest column-to-column reproducibility • Wide range of column IDs and lengths available • Superior lifetime • nanoViper fittings for easy column installation PepSwift and ProSwift monolithic columns are specially designed for fast and high-resolution LC/MS analysis in protein identification, biomarker discovery, and systems biology. Based on a polystyrene divinylbenzene copolymer, the monolithic structure offers a high-quality alternative to traditional microparticulate sorbents, providing important advantages to the chromatographic separation. High-sensitivity proteomics and biotech applications are easily performed using these columns. PepSwift Precolumns can be used for preconcentration and desalting of samples consisting of peptides and proteins without negative impact on the chromatographic performance or recovery of the compounds. Various ion-pairing agents can be used in the loading solvent and/or mobile phases to change the selectivity of the separation or improve the trapping efficiency. PepSwift and ProSwift, nanoViper Ordering Guide Thermo Scientific Chromatography Columns and Consumables 2014-2015 Length (mm) 5 50 250 4-154 100µm ID – 164584 164543 PepSwift 200µm ID 164558 164557 164542 500µm ID – 164585 – ProSwift 1000µm ID – 164586 – Excellence in nanoscale separations • Compatible with any nanoscale HPLC system • Optimized for online LC-MS • Quality control on every column • Simple, flexible design Using highly pure chromatographic media and biocompatible, metal-free fused silica capillaries, Thermo Scientific™ EASY-Column™ capillary LC columns are produced with a focus on design simplicity and strict quality control. As a result, EASY-Column capillary LC columns deliver outstanding chromatographic performance on any nano LC system. EASY-Columns Ordering Guide Quantity 3 1 3 1 Cat. No. SC001 SC200 SC2003 SC600 1 SC601 1 SC602 30 SC603 www.thermoscientific.com/chromatography Columns for Biomolecules Description EASY-Column, 2cm, ID 100µm, 5µm, C18-A1 (Trap column) EASY-Column, 10cm, ID 75µm, 3µm, C18-A2 (Analytical column) EASY-Column, 10cm, ID 75µm, 3µm, C18-A2 (Analytical column) HPLC-to-Column Connector kit Zero-dead-volume union (1/32in OD tubing), 10x SC603 A/B mixing tee & Venting Tee for two-column setup Nanoliter-dead-volume tee (1/32in OD tubing), 10 sleeves for 360µm OD fused silica (10xSC603). Connector Kit for two-column setup Zero-dead-volume union (1/32in OD tubing), Nanoliter-dead-volume tee (1/32 inch OD tubing), 10x SC603 Sleeves (2cm, 1/32in OD) for 360µm OD fused silica LC Columns and Accessories >> EASY-Columns 4-155 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-156 HyperREZ XP Polymer-based columns for carbohydrate analysis • Designed for the determination of carbohydrates, saccharides, 8µm, 10µm organic acids, and alcohols • Efficient and reproducible monodisperse particles • Stable for long column lifetimes even at low pH and high temperatures Thermo Scientific™ HyperREZ™ XP Carbohydrate columns are based on a monodisperse resin with a 4 or 8% divinylbenzene content, and provide an ideal medium for the analysis of carbohydrates and organic acids. Unlike silica based columns they are stable at low pH, allowing the use of dilute acid as a mobile phase. The columns can also be run at elevated temperatures, for faster analysis and improved resolution of some closely eluting analytes. The columns can easily be regenerated for increased column lifetime. Control of the degree of cross-linking of the gel provides a size exclusion mode of operation in addition to the ligand exchange interactions with the metal ion associated with the sulfonated resin. Selectivity differences arise from the interactions of the different counter-ion forms with the hydroxyl groups on the analyte molecules. HyperREZ XP columns are available in H+, Ca2+, Pb2+, and Na+ forms, enabling you to choose the appropriate counter-ion to meet your application requirements. Refer to the tables below to help choose the best column based on application area or retention times. HyperREZ XP columns are also available in dedicated organic acid and sugar alcohol forms. Phase HyperREZ XP Carbohydrate H+ Counter-ion HyperREZ XP Carbohydrate Pb2+ Counter-ion HyperREZ XP Carbohydrate Ca2+ Counter-ion HyperREZ XP Carbohydrate Na+ Counter-ion HyperREZ XP Organic Acids HyperREZ XP Sugar Alcohols Column Type HyperREZ XP Ca2+ HyperREZ XP Pb2+ HyperREZ XP H+ HyperREZ XP Na+ 12 + HyperREZ XP Organic Acids, 8µm, 100 x 7.7mm HyperREZ XP Carbohydrate Na , 10µm, 300 x 7.7mm Part Number: Eluent: Flow Rate: Temperature: Detector: Sample: Part Number: Eluent: Flow Rate: Temperature: Detector: Sample: 3 69608-107780 0.0001mM H 2SO 4 0.7mL/min 57°C RI 1. Maltotriose 2. Maltose 3. Glucose 4. Succinic Acid 5. Lactic Acid 6. Glycerol 7. Acetic Acid 8. Ethanol 69310-307780 H 2O 0.3mL/min 80°C RI 1. Dp8 2. Dp7 3. Dp6 4. Dp5 5. Dp4 6. Dp3 7. Dp2 8. Dpl (Glucose) 9. Fucose 1 8 8 4 5 6 7 7 23 4 6 9 5 REZ-9023 REZ-9025 0 4 MIN Products of fermentation, including organic acids, sugars and alcohols, can be separated using a HyperREZ XP Organic Acids column Particle Size (µm) 8 8 8 10 8 8 Porosity 8% cross linkage 8% cross linkage 8% cross linkage 4% cross linkage 8% cross linkage 8% cross linkage Application Areas Adulteration of food & beverages, confectionary, disaccharides, food additives Alcohols, dairy products, fermentation, wine Anomer separation Fruit juice, monosaccharides Alcohols, dairy products, fermentation, wine Oligosaccharides, glycoprotein constituents, organic acids, fermentation products Corn syrup 0 10 20 30 MIN Analysis of sports drink using a HyperREZ XP Carbohydrate Na+ column HyperREZ XP Carbohydrate H+ Particle Size (µm) 8 HyperREZ XP Carbohydrate Ca2+ 8 HyperREZ XP Carbohydrate Pb2+ 8 HyperREZ XP Carbohydrate Na+ 10 HyperREZ XP Organic Acids 8 HyperREZ XP Sugar Alcohols 8 Format HyperRez Guard Cartridge Holder Length (mm) 5 50 300 5 50 300 5 50 300 5 50 300 5 – 100 5 – 300 ID (mm) 3.0 7.7 7.7 3.0 7.7 7.7 3.0 7.7 7.7 3.0 7.7 7.7 3.0 – 7.7 3.0 – 7.7 Cat. No. 69008-903027 69008-057726 69008-307780 69208-903027 69208-057726 69208-307780 69108-903027 69108-057726 69108-307780 69310-903027 69310-057726 69310-307780 69008-903027 Inquire 69608-107780 69208-903027 Inquire 69708-254030 Length (mm) 5 ID (mm) 3 Cat. No. 60002-354 Retention Times of Common Saccharides (min) Saccharide Adonitol Arabinose Erythritol Fructose Fucose Galactose Glucose Glycerol Lactose Maltose Maltotriose Mannitol Mannose Raffinose Sorbitol Sucrose Xylose H+ 11.5 11.4 12.7 10.6 12.2 1.07 9.9 14.1 8.6 8.4 7.7 11.0 1.5 8.2 11.1 9.8 10.6 Ca2+ 14.9 13.6 15.6 13.5 13.7 12.2 11.1 16.1 9.7 9.5 8.7 17.3 12.5 8.6 20.7 9.4 12.0 Pb2+ 20.4 19.4 20.3 19.3 17.1 15.6 13.9 19.5 12.8 12.5 11.9 28.9 16.7 11.4 N/A 11.9 15.0 Conditions: Column: 300 x 7.7mm Mobile Phase: H2O Flow Rate: 0.6mL/min Detection: RI Temperature: 75°C (H +) 85°C (Ca 2+) 80°C (Pb2+) Note: partial hydrolysis may occur with some saccharides using H +. www.thermoscientific.com/chromatography Columns for Biomolecules Format Guard Cartridge (2/pk) Guard Column HPLC Column Guard Cartridge (2/pk) Guard Column HPLC Column Guard Cartridge (2/pk) Guard Column HPLC Column Guard Cartridge (2/pk) Guard Column HPLC Column Guard Cartridge (2/pk) Guard Column HPLC Column Guard Cartridge (2/pk) Guard Column HPLC Column LC Columns and Accessories >> HyperREZ XP Ordering Guide 4-157 LC Columns and Accessories Thermo Scientific LC Accessories Viper Fingertight Fittings Provides ease of use and dead-volume free plumbing of every conventional HPLC and UHPLC system • Provides virtually zero-dead volume fingertight connections • Supports operating pressures up to 1250 bar (18,130 psi) • Available in different lengths: 65mm and from 150 to 950mm in 100mm steps • Available in different inner diameters: 0.1mm (0.004in), 0.13mm (0.005in) or 0.18mm (0.007in) • Easy to use due to stainless steel or biocompatible MP35N™ capillaries (1/32in OD) and fingertight design • Works with virtually any valve and column from any manufacturer • Fits narrow connections such as 10-port valves and enables mixed use with different designs The Thermo Scientific™ Dionex™ Viper™ fingertight fitting system provides ease of use and virtually dead-volume free plumbing of every conventional HPLC and modern UHPLC system. Together with flexible stainless steel (SST) or biocompatible MP35N® capillaries, it opens a new dimension in liquid chromatography. The Viper system improves chromatographic results, independent of various different connection geometries and system backpressures. Connecting LC modules, valves, and columns quickly and easily without tools is simple with the Viper system. Extra column volumes in HPLC have the most detrimental effects on the separation efficiency of an LC system and must be minimized. Conventional fittings tightened by hand or using tools have considerable drawbacks which can compromise efficiency. The Viper fitting system overcomes these drawbacks by design, working without ferrules to reduce the dead volume of any fluidic connection to virtually zero. The Viper system unifies robust performance, ease of use, acceptable lifetime, and universal compatibility with virtually all different valves and columns for HPLC system users. All Thermo Scientific™ Dionex™ UltiMate™ 3000 XRS, RS, BioRS and SD systems are equipped with Viper fingertight fitting system as a standard. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Viper Fingertight Fitting Systems – SST and Biocompatible MP35N 4-158 Length (mm) 65 150 250 350 450 550 650 750 850 950 0.1mm ID, SST 6040.2207 6040.2215 6040.2225 6040.2235 6040.2245 6040.2255 6040.2265 – – – 0.1mm ID, MP35N 6042.2306 6042.2320 6042.2330 6042.2340 6042.2350 6042.2360 – – – – 0.13mm ID, SST 6040.2307 6040.2315 6040.2325 6040.2335 6040.2345 6040.2305 6040.2310 6040.2320 6040.2330 6040.2350 Viper Accessories Description Plug, SST Union, SST Cat. No. 6040.2303 6040.2304 Quantity 1 Each 1 Each 0.18mm ID, SST 6040.2357 6040.2360 6040.2385 6040.2375 6040.2365 6040.2355 6040.2395 6040.2370 6040.2380 6040.2390 0.18mm ID, MP35N – – 6042.2315 6042.2327 6042.2337 6042.2365 – 6042.2355 – 6042.2375 LC Columns and Accessories >> LC Accessories Viper Fingertight Fittings Kits for UltiMate 3000 Systems Viper Fingertight Fitting Kits for UltiMate 3000 systems Description Viper Capillary Kit for ISO, LPG or DGP pumps Viper Capillary Kit for HPG pumps Viper Capillary Kit for biocompatible RSLC systems On-line SPE Solution Kit for x2 Dual Standard or RSLC systems Tandem Operation Solution Kit for x2 Dual Standard or RSLC systems Application Switching Solution Kit for x2 Dual Standard or RSLC systems Parallel Setup Solution Kit for x2 Dual Standard or RSLC systems Inverse Gradient Kit for x2 Dual Standard or RSLC systems Automated Method Scouting Solution Kit for Standard or RSLC systems MS Connection Kit for MS-frontends with WPS autosampler, excluding UV detection MS Connection Kit for MS-frontends with WPS autosampler, including UV detection MS Connection Kit for MS-frontends with HPG-RS pump and OAS autosampler excluding UV detection MS Connection Kit for MS-frontends with HPG-RS pump and OAS autosampler including UV detection SD Systems 6040.2302 6040.2309 – 6040.2802 6040.2804 6040.2806 6040.2810 6040.2819 6040.2808 6720.0355 6720.0365 – – Description Viper Capillary Kit for standalone system with with WPS autosampler Viper Capillary Kit for standalone system with with OAS autosampler MS Connection Kit for MS-frontends with WPS autosampler excluding UV detection MS Connection Kit for MS-frontends with WPS autosampler including UV detection MS Connection Kit for MS-frontends with LPG-XRS pump and OAS autosampler excluding UV detection MS Connection Kit for MS-frontends with LPG-XRS pump and OAS autosampler including UV detection XRS Systems 6043.2301 6845.2301A 6720.0380 6720.0385 6720.0372 6720.0377 RS Systems 6040.2301 6040.2308 6841.2301 6040.2801 6040.2803 6040.2805 6040.2809 6040.2820 6040.2807 6720.0370 6720.0375 6720.0372 6720.0377 An overview of all UltiMate 3000 systems can be found in the Instruments spare parts section on 4-209. www.thermoscientific.com/chromatography 4-159 LC Columns and Accessories nanoViper Fingertight Fittings The Thermo Scientific™ Dionex™ nanoViper™ fingertight UHPLC fitting system is virtually dead volume-free by design. It offers nano LC connections that never fail and brings a peace of mind appreciated by novices, as well as the most experienced users. • Provides virtually zero-dead-volume connections • Compatible with backpressures up to 1000 bar (14,500 psi) • Suitable for temperatures up to 80°C • Easy to use 1/32" PEEK sheathed fused silica and fingertight design • Works with virtually any valve, and any column, from any manufacturer • Paves the way for the easy, tool-free nano, capillary and micro LC setup of any application • nanoViper fingertight fittings allow you to configure your nano, capillary, and micro LC application completely tool free. Classical connections in nano LC involve either PEEKsil™ or fused silica in PEEK™ sleeves. These connections consist of a nut, ferrule, sleeve, and fused silica, which all need to be assembled correctly. The sealing is at the ferrule, which theoretically leaves a large potential dead volume. Apart from the risk of dead volume (an extreme case indicated above), there is also a risk of damaging the fused silica tubing or column at the point where the ferrule is compressed. The ferrule has to hold the tubing in the sleeve and seal of the connection. Thermo Scientific Chromatography Columns and Consumables 2014-2015 The nanoViper connections come preassembled and high pressure tested. The fittings do not “grab” the tubing at a single point. It seals at the tip and not with a ferrule. Nor does it “grab” the tubing at a single point. It is this feature that gives nanoViper fittings their UHPLC, virtually dead-volume-free capabilities. The theoretical performance effect between the two example chromatograms below is about 20% increase in peak capacity. 4-160 Length (mm) 70 150 250 350 450 550 650 750 950 Orange 20µm ID 6041.5120 6041.5121 – 6041.5240 – 6041.5260 6041.5275 6041.5280 6041.5122 Brown 50µm ID 6041.5123 6041.5124 – 6041.5540 – 6041.5560 6041.5575 6041.5580 6041.5125 Black 75µm ID 6041.5126 6041.5127 6041.5730 6041.5735 – 6041.5760 6041.5775 6041.5780 6041.5128 Red 100µm ID 6041.5820 6041.5811 6041.5812 6041.5813 6041.5814 6041.5815 – 6041.5816 – Purple 150µm ID 6041.5817 6041.5818 6041.5819 6041.5820 6041.5821 6041.5822 – 6041.5823 – Fitting ID (µm) 30 75 Cat. No. 164649 164650 Trap Column Cartridges Holders with nanoViper Fittings Description µ-Precolumn holder for trap cartridges with 2 x 100mm length nanoViper fittings Cartridge Length (mm) 5 15 Sample Loops with nanoViper Fittings Cat. No. 6826.2401 6826.2405 6826.2410 6826.2420 6826.2450 6826.2412 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each LC Accessories Volume (µL) 1 5 10 20 50 125 LC Columns and Accessories >> nanoViper Fingertight Fittings nanoViper Application Kits nanoViper fittings are also included in tubing and application kits designed for use with the UltiMate 3000 RSLCnano systems. nanoViper Fitting Kits for UltiMate 3000 RSLCnano Systems Description Tubing Samples Trap Column Separation Column Direct Injection nano LC Kit Direct Injection capillary LC Kit Preconcentration nano LC kit Preconcentration capillary LC kit Preconcentration monolithic LC kit 2D salt plugs kit Automated off line SCX-RP peptides kit Automated off line RP-RP peptides kit Tandem nano LC kit FLM nanoViper tubing kit (All tubing for nano LC preconcentration) MS connection kit EASY-Spray connection kit (Supports direct injection as well as preconcetration) Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y – – – Y Y Y Y Y Y Y – Y Y Y Y Y Y Y Y Y – RSLCnano Systems 6720.0300 6720.0305 6720.0310 6720.0315 6720.0320 6720.0325 6720.0330 6720.0340 6720.0335 6041.5100 Y Y – Y – Y – – 6720.0345 6720.0395 www.thermoscientific.com/chromatography 4-161 LC Columns and Accessories SLIPFREE Connectors Universal self-adjusting connections • Void-free and leak-free by pushing tubing and ferrule SLIPFREE Ferrules into the end-fitting • Compatible with all column end-fittings • Stainless-steel threads • Fingertight connections to 10,000psi Material PEEK Vespel Kel-F Cat. No. 36023 36024 36025 Quantity 1 Each 1 Each 1 Each SLIPFREE Connectors, Single Length (cm) 0.005in ID 0.010in ID 0.020in ID 6 10 20 30 50 30106 30110 30120 30130 30150 31106 31110 31120 31130 – 32106 32110 32120 32130 – 10.5 15 28 40 50 60 30111-FLEX 30115-FLEX 30128-FLEX 30140-FLEX 30150-FLEX 30160-FLEX 39111-FLEX 39115-FLEX 39128-FLEX 39140-FLEX – – – – – – – – 6 10 20 30306 30310 30320 31306 31310 31320 32306 32310 32320 10 20 30510 30520 31510 31520 – 32520 Length (cm) 0.005in ID 0.010in ID 0.020in ID 6 10 20 30 40 30206 30210 30220 30230 30240 31206 31210 31220 31230 – 32206 32210 32220 32230 – 10.5 15 28 40 30211-FLEX 30215-FLEX 30228-FLEX 30240-FLEX 39211-FLEX 39215-FLEX 39228-FLEX 39240-FLEX – – – – 6 10 20 30406 30410 30420 31406 31410 31420 32406 32410 32420 10 20 31710 31720 32710 32720 – – Single Single Flexible Single PEEK Collared Single Long-neck SLIPFREE Connectors, Double Thermo Scientific Chromatography Columns and Consumables 2014-2015 Double 4-162 Double Flexible Double PEEK Collared Double Long-neck LC Columns and Accessories >> PTFE One-Piece Column Connector Excellent for high-throughput screening and quick connection • Fingertight, leak-free connection of analytical and guard columns with 10-32 threads • Minimizes dead volume • Inert and biocompatible material PEEK One-Piece Column Connector Description One Piece Coupler Cat. No. 60170-370 Quantity 1 Each Solvent Inlet Filters Feature a large surface area for a long lifetime LC Accessories • 10μm inlet filters for longer lifetime • No tools required for replacement Solvent Inlet Filters for UltiMate 3000 Systems Description Filter frit, porosity 10µm, V4A stainless steel Filter frit, porosity 10µm, titanium Filter holder Solvent supply line, 1.5 x 3.0 x 1000mm Model UltiMate 3000 pumps Cat. No. 6268.0110 Quantity 10 Each UltiMate 3000 pumps UltiMate 3000 pumps UltiMate 3000 pumps 6268.0111 6268.0115 6030.2548 10 Each 6 Each 1 Each Bottom-of-the-Bottle solvent filters • Efficient draw • 100% PTFE polymer, including 2µm filters • Built-in helium sparge port and frit Solvent Filters for HPLC Systems Type For Use with Cat. No. Quantity Stainless Steel Fit 1/16in OD tube to 1/8in OD plastic tubing A-302 1 Each Stainless Steel Fit to 1/8in OD plastic tubing using 1/8in PP nut A-302A 1 Each Bottom-of-the-Bottle 3/16in OD plastic tubing A-436 1 Each Bottom-of-the-Bottle 1/8in OD tubing A-437 1 Each www.thermoscientific.com/chromatography 4-163 LC Columns and Accessories High Pressure Stainless Steel Nuts and Ferrules Accommodate a wide range of configurations • Designed for 10-32 port configurations • Burr and contaminant free Thermo Scientific High Pressure Stainless Steel Nuts and Ferrules Type Cat. No. Quantity 10-32 thread nut with ferrule F-190 1 Each Replacement PEEK Ferrules F-192x 10 Pack Male hex nut U-400x 10 Pack Universal ferrules, 0.625in U-401x 10 Pack Valco male hex nut, 10-32 thread U-320x 10 Pack Valco ferrules, 0.625in U-321x 10 Pack Male hex nut, Waters compatible U-410x 10 Pack Reducing Union for Preparative Columns Connects 30 to 50mm ID preparative columns to 1/16in tubing • Stainless steel construction • 1.0mm bore • Without frit Thermo Scientific Chromatography Columns and Consumables 2014-2015 Reducing Union for Preparative Column 4-164 Description 1/8in to 1/16in Reducing Union for Preparative Column Cat. No. 60182-357 Quantity 1 Each LC Columns and Accessories >> PEEK Fingertight Fittings Machined for reliability and ease of use • Resist cracking, breaking, thread stripping and leaking in both low and high pressure applications • Biocompatible for a broad range of applications PEEK Fingertight Fittings Type Cat. No. Quantity One-piece Fingertight Fitting, 1/16in, 0.37in head F-120x 10 Pack One-Piece Long Fingertight Fitting, 1/16in, 0.37in head F-130x 10 Pack One-Piece PEEK Fingertight Fitting, 1/32in, 0.25in head M-645x 10 Pack Two-Piece Fingertight Wing Nut with Ferrule, 1/16in F-300x 10 Pack Replacement PEEK Ferrules F-142x 10 Pack Column End Plugs, 1/16in, 10-32 coned, Delrin, Black Column End Plugs, 1/16in, 10-32 coned, Delrin, Red U-467BLKx U-467Rx 10 Pack 10 Pack LC Accessories Viper Unions Suitable only for direct connection of two Viper capillaries Description Viper union, stainless steel Cat. No. 6040.2304 Quantity 1 Each Stainless Steel Unions, Tees and Crosses Well-suited to high pressure applications • Absolute zero or low dead volume formats • Includes two stainless steel nuts and ferrules Stainless Steel Unions, Tees and Crosses Description Through Hole (in) Swept Volume (µL) Cat. No. Quantity Union, stainless steel, Upchurch Scientific/Parker fittings compatible, includes 2 stainless steel nuts and ferrules 0.010 0.025 U-435 1 Each Union, stainless steel, Upchurch Scientific/Parker fittings compatible, includes 2 stainless steel nuts and ferrules 0.020 0.134 U-402 1 Each Union, stainless steel, Upchurch Scientific/Parker fittings compatible, includes 2 stainless steel nuts and ferrules 0.050 0.836 U-437 1 Each Union, stainless steel, Upchurch Scientific/Parker fittings compatible, includes 2 stainless steel nuts and ferrules 0.062 ~0.0 U-438 1 Each Union, stainless steel, Waters fittings compatible, includes 2 stainless steel nuts and ferrules 0.020 0.129 U-412 1 Each Union, stainless steel, Valco fittings compatible, includes 2 stainless steel nuts and ferrules 0.020 0.103 U-322 1 Each Tee, stainless steel, 10-32 fittings for use with 1/16in OD tubing 0.020 0.57 U-428 1 Each Cross, stainless steel, 10-32 fittings for use with 1/16in OD tubing 0.020 0.72 U-430 1 Each www.thermoscientific.com/chromatography 4-165 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-166 PEEK Unions, Tees and Crosses Well-suited to high pressure applications • Absolute zero or low dead volume formats • Biocompatible PEEK and PEEK Lined Unions, Tees and Crosses Description Through Hole (in) Swept Volume (µL) Cat. No. Quantity Union, PEEK polymer, includes two PEEK 2-piece fittings 0.010 0.070 P-742 1 Each Union, PEEK polymer, includes two PEEK 2-piece fittings 0.020 0.28 P-704 1 Each Tee, PEEK, 10-32 fittings for use with 1/16in OD tubing, includes three 10-32 PEEK double-winged nuts 0.020 0.57 P-727 1 Each PEEK, 10-32 fittings for use with 1/16in OD tubing, includes four 10-32 PEEK double-winged nuts 0.020 0.72 P-729 1 Each Excellent chemical compatibility and very low carryover • Precision-bore fused silica tubing coated with 1/16in OD PEEK covering • Usable in most standard chromatography systems • Withstands high pressures • Smooth internal surface for excellent flow characteristics • Tubing is stiff: not recommended for uses requiring tubing bends • Precut lengths only: cutting in the lab may damage tubing PEEKsil Capillary Tubing ID (in) 0.002 0.007 Cat. No. 60182-500 60182-501 60182-502 60182-503 60182-504 60182-505 60182-506 60182-507 60182-508 Quantity 5 Pack 5 Pack 2 Pack 5 Pack 5 Pack 2 Pack 5 Pack 5 Pack 2 Pack Applications: • HPLC • LC-MS LC Accessories 0.004 Length (cm) 10 20 50 10 20 50 10 20 50 LC Columns and Accessories >> PEEKsil Capillary Tubing PEEK Sleeves for Fused Silica Capillary Tubing Withstands high pressures 1/16in OD PEEK Sleeves for Fused Silica Capillary Tubing ID (in) 0.008 0.010 0.012 0.015 0.021 0.030 Color Yellow Blue Natural Orange Natural Natural Cat. No. F-227 F-228 F-229 F-230 F-231 F-232 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each www.thermoscientific.com/chromatography 4-167 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-168 316 Stainless Steel Capillary Tubing Cleaned, polished, passivated and ready-to-use • Suitable for ultra high pressure applications • Wide chemical compatibilty • Prefinished, square, burr-free ends and interiors to minimize dead volume connections • Not recommended for biological samples • Rough internal surface may lead to sample carryover 316 Stainless Steel Capillary Tubing ID (in) Length (cm) Color 1/16in OD Precut Tubing 0.005 5 Red 10 Red 20 Red 30 Red 50 Red 100 Red 0.007 5 Black 10 Black 20 Black 30 Black 50 Black 100 Black 0.010 5 Blue 10 Blue 20 Blue 30 Blue 50 Blue 100 Blue 1/32in OD Precut Tubing with 1/16in Sleeves 0.005 10.5 Red 15 Red 28 Red 40 Red 0.007 10.5 Yellow 15 Yellow 28 Yellow 40 Yellow Cat. No. Quantity U-152 U-153 U-154 U-155 U-156 U-157 U-126 U-127 U-128 U-129 U-130 U-131 U-111 U-112 U-113 U-114 U-132 U-133 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 30011-FLEX 30015-FLEX 30028-FLEX 30040-FLEX 39011-FLEX 39015-FLEX 39028-FLEX 39040-FLEX 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. U-158 U-108 U-106 U-105 U-107 U-144 U-151 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1/16in 316 Stainless Steel Tubing, 5-Foot Coil ID (in) 0.005 0.007 0.010 0.020 0.030 0.040 0.046 Pre-cut and color-coded for easy identification and use • Broad chemical compatibility • Biocompatible • Easily cut to desired length • Appropriate for many HPLC applications • Resistant to most organic solvents, but nitric acid, sulfuric acid, dichloromethane, THF and DMSO are not recommended 1/16in OD Precut PEEK Tubing ID (in) 0.003 0.007 0.010 0.020 Color Natural Natural Natural Natural Natural Natural Red Red Red Red Red Red Yellow Yellow Yellow Yellow Yellow Yellow Blue Blue Blue Blue Blue Blue Orange Orange Orange Orange Orange Orange Cat. No. 37003-5 37003-10 37003-20 37003-30 37003-50 37003-100 37005-5 37005-10 37005-20 37005-30 37005-50 37005-100 37007-5 37007-10 37007-20 37007-30 37007-50 37007-100 37010-5 37010-10 37010-20 37010-30 37010-50 37010-100 37020-5 37020-10 37020-20 37020-30 37020-50 37020-100 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. 37003 37005 37007 37010 37020 37030 37040 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each LC Accessories 0.005 Length (cm) 5 10 20 30 50 100 5 10 20 30 50 100 5 10 20 30 50 100 5 10 20 30 50 100 5 10 20 30 50 100 LC Columns and Accessories >> PEEK Capillary Tubing 1/16in OD PEEK Tubing, 5-Foot Coil ID (in) 0.003 0.005 0.007 0.010 0.020 0.030 0.040 www.thermoscientific.com/chromatography 4-169 LC Columns and Accessories Polymer Tubing Cutter Produces a flat, 90°, burr-free end • Compatible with rigid polymeric tubing • Guide holes for 1/16in and 1/8in tubing Polymer Tubing Cutter Description Polymeric Tubing Cutter Replacement blades Cat. No. A-327 A-328 Quantity 1 Each 5 Pack Terry Tool Tubing Cutters Produce clean, 90° cuts of stainless steel tubing Terry Tool Tubing Cutters Thermo Scientific Chromatography Columns and Consumables 2014-2015 Description 1/16in stainless steel tubing 1/8in stainless steel tubing 4-170 Cat. No. 60182-509 60182-510 Quantity 1 Each 1 Each Cat. No. 6720.0016 Quantity 1 Each Fused Silica Cutter Fused Silica Cutter Description Cutter for fused silica capillaries Allow continuous flow between the load and inject positions to protect against pressure shock • Stainless steel construction • Make-Before-Break (MBB) design • Can use partial filling for zero sample waste or complete filling for better reproducibility • Inject 1µL to 5mL with high accuracy and precision • 7725i features a position sensing switch for a reproducible start signal Rheodyne 7725 and 7725i Sample Injectors Model 7725 7725i Mode Dual Dual Features Continuous flow Continuous flow, position sensing switch Cat. No. 7725 7725i Quantity 1 Each 1 Each Allow continuous flow between the load and inject positions to protect against pressure shock • Biocompatible PEEK construction LC Accessories Rheodyne 9725 and 9725i Sample Injectors LC Columns and Accessories >> Rheodyne 7725 and 7725i Sample Injectors • Make-Before-Break (MBB) design • Can use partial filling for zero sample waste or complete filling for better reproducibility • Inject 1µL to 5mL with high accuracy and precision • 9725i features a position sensing switch for a reproducible start signal Rheodyne 9725 and 9725i Sample Injectors Model 9725 9725i Mode Dual Dual Features Continuous flow Continuous flow, position sensing switch Cat. No. 9725 9725i Quantity 1 Each 1 Each Rheodyne 8125 Low-dispersion Microscale Injector Designed for use with 1 and 2mm ID HPLC columns • Can use partial filling for zero sample waste or complete filling for better reproducibility • Position sensing switch provides reproducible start signal • Suitable for use with 5 to 50µL sample loops Rheodyne 8125 Low-dispersion Microscale Injector Model 8125 Mode Dual Features Continuous flow Cat. No. 8125 Quantity 1 Each www.thermoscientific.com/chromatography 4-171 LC Columns and Accessories Rheodyne 7010 Sample Injector Single-mode sample injector designed for the complete filling method Rheodyne 7010 Sample Injector Model 7010 Mode Single Features Complete filling method Cat. No. 7010 Quantity 1 Each Rheodyne 9010 Sample Injector Single-mode sample injector designed for the complete filling method • Compatible with sample loop sizes 5µL to 10mL • PEEK stator • Position sensing switch provides a reproducible start signal Rheodyne 9010 Sample Injector Model 9010 Mode Single Features Continuous flow, position sensing switch Cat. No. 9010 Quantity 1 Each Rheodyne Ports for Injectors Thermo Scientific Chromatography Columns and Consumables 2014-2015 Suitable for popular Rheodyne injector models 4-172 Rheodyne Ports for Rheodyne Injectors Models 7010 and 9010 For Use with Rheodyne Model 7010 Filler Port, Stainless Steel 9010 Filler Port, PEEK 9010 Needle Port, PEEK Cat. No. 7012 9012 9013 Quantity 1 Each 1 Each 1 Each Compatible with: sample loop sizes 5µL to 20mL For Rheodyne sample injectors in stainless steel or biocompatible PEEK PEEK and Stainless Steel Sample Loops Description Sample loops for 7010 and 7125 injectors, Stainless Steel Sample loops for 7725 and 7725i injectors Stainless Steel Sample loops for 9010 and 9725 injectors, PEEK Sample loops for 9725 and 9725i injectors, PEEK ID (mm / in) 0.18 / 0.007 0.30 / 0.012 0.30 / 0.012 0.51 / 0.020 0.51 / 0.020 0.76 / 0.030 0.76 / 0.030 0.76 / 0.030 1.0 / 0.040 0.18 / 0.007 0.30 / 0.012 0.30 / 0.012 0.51 / 0.020 0.20 / 0.008 0.20 / 0.008 0.25 / 0.010 0.30 / 0.012 Internal 0.18 / 0.007 0.25 / 0.010 0.25 / 0.010 0.51 / 0.020 0.51 / 0.020 0.51 / 0.020 0.76 / 0.030 0.76 / 0.030 0.76 / 0.030 0.18 / 0.007 0.25 / 0.010 0.25 / 0.010 0.51 / 0.020 Cat. No. 7020 7021 7022 7023 7024 7025 7026 7027 7029 7755-020 7755-021 7755-022 7755-023 8020 8021 8022 8023 7755-015 9055-020 9055-021 9055-022 9055-023 9055-024 9055-025 9055-026 9055-027 9055-029 9055-020 9055-021 9055-022 9055-023 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. 3725-999 7010-999 7125-999 7410-999 7520-999 7725-999 8125-999 9125-999 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each LC Accessories Sample loops for 8125 injectors, Stainless Steel Volume 5μL 10μL 20μL 50μL 100μL 200μL 500μL 1mL 5mL 5μL 10μL 20μL 50μL 5μL 10μL 20μL 50μL 2µL 5µL 10µL 20µL 50µL 100µL 200µL 500µL 1mL 5mL 5μL 10μL 20μL 50μL LC Columns and Accessories >> Rheodyne Sample Loops RheBuild Kits Maintain Rheodyne valves and injectors RheBuild Kits For Use with Rheodyne Models 3725/3725i/3725-038/3725i-038 7010/7000 7125/7126 7410 7520/7526 7725/7725i/7726 8125/8126 9125/9126 www.thermoscientific.com/chromatography 4-173 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-174 Rheodyne Suction Needle Adapter For use with Rheodyne sample injectors Rheodyne Suction Needle Adapter For Use with Rheodyne Injector Models 9725 and 9725i Cat. No. 9125-076 Quantity 1 Each Rheodyne Replacement Rotor Seals for Injectors Suitable for popular Rheodyne injector models Rheodyne Replacement Rotor Seals for Injectors For Use with Rheodyne Models Vespel Seals 7000/7010/7040/7067 7030 7060/7066 7125/7126 7410 7413 8125/8126 Tefzel Seals 7000/7010/7040 7030 7060/7066/9060 7410 7125/7126 8125 9010 9125 PEEK Seals 3725/3725i/3725-038/3725i-038 Cat. No. Quantity 7010-039 7030-003 7060-070 7125-047 7410-038 7413-013 8125-038 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 7010-071 7030-015 7060-074 7410-075 7125-079 8125-097 9010-051 9125-082 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 3725-018 1 Each Suitable for popular Rheodyne injector models Rheodyne Stators For Use with Rheodyne Models 7000/7010/7030/7040/7125 7010-087/7125-081 7060/7066 7410/7413 9010/9030/9125 9060 7725 8125/8126 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. 3725-039 7125-067 8125-074 8125-094 9060-015 7725-026 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. 7125-054 Quantity 1 Each Cat. No. 7160-010 7160 Quantity 1 Each 1 Each Rheodyne Stator Face Assemblies For Use with Rheodyne Models 3725/3725i/3725-038/3725i-038 7125 8125 9125/9010/9030 9060 9725 LC Accessories Cat. No. 7010-040 7010-066 7060-039 7410-041 9125-043 9060-016 7725-010 8125-098 LC Columns and Accessories >> Rheodyne Stators Rheodyne Injection Port Needle Cleaner For use with Rheodyne sample injectors Rheodyne Injection Port Needle Cleaner For Use with Rheodyne injectors Rheodyne Valve Mounting Brackets For use with Rheodyne sample injectors Rheodyne Injection Port Needle Cleaner For Use with Angle bracket Mouting panel www.thermoscientific.com/chromatography 4-175 LC Columns and Accessories RheFlex High Pressure Fittings Precision machined from 316 stainless steel RheFlex High Pressure Fittings Type Short Fittings Set Short Fittings Set Long Fittings Set Long Fittings Set Extra Long Fittings Set Extra Long Fittings Set 1/16in Ferrule 1/16in Ferrule 0.5mm Ferrule for Model 8125 Cat. No. 6000-109 6000-209 6000-111 6000-211 6000-162 6000-262 6000-110 6000-210 8125-084 Quantity 5 Pack 10 Pack 5 Pack 10 Pack 5 Pack 10 Pack 5 Pack 10 Pack 1 Each RheFlex Two-Piece PEEK Fittings Provide inert, metal-free connections • Slotted back-side of the ferrule is squeezed down onto the tube by the mating conical surface of the nut • May be used on 1/16in metal or plastic tubing reliably up to 5000 psi • Reusable ferrule and nut Thermo Scientific Chromatography Columns and Consumables 2014-2015 RheFlex Two-Piece PEEK Fittings 4-176 Type Fitting set, standard length Fitting set, short Fitting set, X-long Replacement ferrules Cat. No. 6000-054 6000-055 6000-066 6000-051 Quantity 5 Pack 5 Pack 1 Each 5 Pack Can be used as an injector or switching valve • 1/16in fittings • 0.010in ports • Available in 6-port or 10-port configurations • Available with manual or microelectric actuator Cheminert Model C2 Microbore Injector Description Model C2 injector, N60 stainless stator, 5μLloop, manual Model C2 injector, N60 stainless stator, 5μL loop, microelectric actuator Sample injector loops, stainless steel Sample injector loops, PAEK Cat. No. C2-1006 C2-1000 C2-1006EH C2-1000EP CSL2 CSL5 CSL10 CSL20 CSL50 CSL100 C2-1346EH C2-1340EP CZSL5PK CZSL10PK CZSL50PK CZSL100PK Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Volume 20μL loop 20μL loop – 2μL 5μL 10μL 20μL 50μL 100μL Cat. No. C6W EPC6W SSAC6W SL2CW SL5CW SL10CW SL20CW SL50CW SL100CW Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. VISF-1 VISF-2 ZN1-10 LZN1-10 ZF1-10 Quantity 1 Each 1 Each 10 Pack 10 Pack 10 Pack LC Accessories Model C4 injector, PAEK stator, 5μL loop, microelectric actuator Sample Volume 6 ports 10 ports 6 ports 10 ports 2μL 5μL 10μL 20μL 50μL 100μL 6 ports 10 ports 5μL 10μL 50μL 100μL LC Columns and Accessories >> Cheminert Model C2 Microbore Injector Valco Injector Model C6W Description Model C6W injector, six 0.016in ports, manual Model EPC6W injector, six 0.016in ports, microelectric actuator Replacement rotor Sample injector loop, stainless steel Sample injector loop, stainless steel Sample injector loop, stainless steel Sample injector loop, stainless steel Sample injector loop, stainless steel Sample injector loop, stainless steel Valco Accessories Description Valco syringe ports Valco syringe ports Valco Nuts and Ferrules Valco Nuts and Ferrules Valco Nuts and Ferrules Volume 22 ga. needles; 1/16in fittings 22 ga. 2in needles 1/16in standard nut 1/16in long nut 1/16in SS ferrule www.thermoscientific.com/chromatography 4-177 LC Columns and Accessories HPLC Syringes Syringes and needles for Thermo Scientific LC instruments Syringes for UltiMate 3000 Instruments Volume (µL) 25 100 250 500 1000 Instrument All WPS-3000 variants All WPS-3000 variants All WPS-3000 and ACC-3000 variants, except the WPS-3000TXRS All WPS-3000 variants, except the WPS-3000TXRS All WPS-3000 and ACC-3000 variants, except the WPS-3000TXRS Volume (µL) 250 500 250 500 2500 Needle Type Removable Removable – – – Instrument LCQ, XP, DECA, Advantage LCQ MS LCQ XSQ Thermo Scientific Chromatography Columns and Consumables 2014-2015 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each Cat. No. 365ILT21 365JLT41 365ILT91 365JLT61 365LLT81 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each Syringes for Thermo Scientific Instruments Length (mm) 50 50 – – – Gauge 22 22 – – – Instrument LCQ LCQ AS1000, AS3000 AS1000, AS3000 AS3000, AS3500 Mass Spectrometry Replacement ESI Probe Needles 4-178 Cat. No. 6822.0001 6822.0002 6822.0003 6822.0004 6822.0005 Cat. No. 365RNLT1 365RNLT2 365RNLT3 Quantity 1 Each 1 Each 1 Each Easy, accurate and reproducible manual injection • Square tip to prevent damage to the injector • Wide range of volumes • Precision made from borosilicate glass and stainless steel • Robust design and easy-to-read markings Gas Tight Syringes for Rheodyne / Valco Injectors Volume (µL) 10 25 50 100 250 500 Needle Type Removable Removable Removable Removable Removable Removable Length (mm) 50 50 50 50 50 50 Gauge 22 22 22 22 22 22 Tip Style 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End Cat. No. 365RNL15 365RNL25 365RNL25 365RNL25 365RNL25 365RNL25 Quantity 5 Pack 5 Pack 5 Pack 5 Pack 5 Pack 5 Pack Cat. No. 365DLG21 365FLG31 365GLG41 365HLG51 365ILG61 365JLG71 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Replacement Needle for Syringe 365DLG21 365FLG31 365GLG41 365HLG51 365ILG61 365JLG71 LC Accessories Replacement Needles LC Columns and Accessories >> LC Syringes for Manual Injection valves Gas Tight Syringes for Rheodyne / Valco Injectors Volume (µL) 10 25 50 100 250 500 Needle Type Fixed Fixed Fixed Fixed Fixed Fixed Length (mm) 50 50 50 50 50 50 Gauge 22 22 22 22 22 22 Tip Style 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End Cat. No. 365DL263 365F6315 365G6316 365H6317 365ILG61 365J6319 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Gauge 22 22 22 22 22 22 22 Tip Style 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End 90° Blunt End Cat. No. 365CL221 365DL231 365FL241 365GL251 365HL261 365IL271 365JL281 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Syringes for Rheodyne / Valco Injectors Volume (µL) 5 10 25 50 100 250 500 Needle Type Fixed Fixed Fixed Fixed Fixed Fixed Fixed Length (mm) 50 50 50 50 50 50 50 www.thermoscientific.com/chromatography 4-179 LC Columns and Accessories Other LC Syringes • Instrument specific syringes • Luer-Lok syringes ideal for priming • Macro volume sampling syringes suitable for liquid or gas samples Syringes for CTC Instruments Volume (µL) 10 25 50 100 Needle Type Fixed Fixed Fixed Fixed Length (mm) 50 50 50 50 Cat. No. 365DL991 365FL715 365GL810 365HL331 Quantity 1 Each 1 Each 1 Each 1 Each Tip Style Bevel 90° Blunt End Cat. No. 365BL443 365BL447 Quantity 1 Each 1 Each Gauge – – – – – Tip Style – – – – – Cat. No. 365KL531 365LL541 365ML551 365NL561 365PL571 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each Gauge 22 Tip Style 90° Blunt End Cat. No. 365RNL22 Quantity 2 Pack Gauge 22s 22s 22s 22s Tip Style Bevel Bevel Bevel Bevel Cat. No. 365K3051 365LL375 365M5212 365N5214 Quantity 1 Each 1 Each 1 Each 1 Each Gauge 22 22 22 22 Tip Style – – – – Cat. No. 365RP532 365RP922 Quantity 1 Each 1 Each Length (mm) 70 70 Gauge 26s 26s Length (mm) – – – – – Replacement Plungers Replacement Plunger for Syringe 365DL991 365FL715 Syringes for Waters Instruments Volume (µL) 1 1 Needle Type Fixed Fixed PTFE Luer-Lok Syringes Volume (mL) 1 2.5 5 10 25 Needle Type – – – – – Needles for Luer-Lok Priming Syringes Thermo Scientific Chromatography Columns and Consumables 2014-2015 Length (mm) 50 4-180 Syringes for Macro Volume Sampling Volume (mL) 1 2.5 5 10 Needle Type Fxed Fxed Fxed Fxed Length (mm) 50 50 50 50 Detector Lamps for UltiMate 3000 Instruments Description Deuterium Lamp Tungsten Lamp Quantity 1 Each 6074.2000 1 Each Model SP8400/SP8430/SP8440/SP8450/SP8480/SP8490 Cat. No. DSP-901 Quantity 1 Each SP8480XR/SP8773XR Linear UV100/UV200/UV1000/UV2000/UV3000/ Focus/Spectrochrom DSP-907 DSP-908 1 Each 1 Each Cat. No. DHP-901 Quantity 1 Each DHP-902 DHP-903 DHP-909 DHP-910 DHP-911 DHP-912 DHP-913 DHP-906 DHP-910LL DHP-911LL 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Detector Lamps for Thermo Scientific Instruments Description Deuterium Lamp Detector Lamps for Agilent Instruments Description Deuterium Lamp Xenon Lamp LL Deuterium Lamp Model Agilent HP1040/HP1050 (G1306A) DAD/HP 1050 DA (1050 MWD)/ HP MW (79854A) / HP 1090 (75880A) DAD HP 1080/HP 1081/HP1081B/HP1082B/HP1084/HP1084B HP 1050 VW (79853C) HP 8450/8450A HP 1100 (G1314) VW HP 1100 (G1315A) DAD HP 8453 HP 8452 A DAD/HP 8452A Opt 002 HP 1046/HP1046A Agilent 1100 VWD long life Agilent 1100 DAD long life LC Accessories Cat. No. 6074.1110 Model MWD-3000(RS), DAD-3000(RS), VWD-3100 and VWD-3400RS MWD-3000(RS), DAD-3000(RS), VWD-3100 and VWD-3400RS LC Columns and Accessories >> Detector Lamps Detector Lamps for Merck-Hitachi Instruments Description Deuterium Lamp Model 101/102/111 Cat. No. DHI-901 Quantity 1 Each DHI-902 1 Each DHI-903 1 Each Xenon Lamp 100-10/100-40/100-50/100-60 150-20/200/220/300/330/340/2000/3000/4000/ L2500/L3000/L4000/L-4500 L4200/L4250/L4500 LaChrom L4720/L4520/L7400/L450 Hitachi fluorescence detectors F1000/2000/4000 Series DHI-908 DHI-910 DHI-911 1 Each 1 Each 1 Each Our applications team is regularly presenting at key events, download one of our recent posters. www.thermoscientific.com/chromatography www.thermoscientific.com/chromatography 4-181 LC Columns and Accessories Detector Lamps for PerkinElmer Instruments Description Deuterium Lamp Model Lambda 3/7/9 Cat. No. DPE-903 Quantity 1 Each Tungsten Lamp 360/460/560 Integral 2000/Integral 4000/LC55/LC65/LC85/LC95 LC-90/LC-290 Lambda 2/2S/10/11 and others Series 200 DAD Lambda 2/2S/10/11 and others DPE-906 DPE-911 DPE-913 DPE-914 DPE-915 DPE-908 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Model UV120/UV160/UV160A/UV240/UV260/UV265 Cat. No. DSH-901 Quantity 1 Each SPD-2A/SPD-3/SPD-4 D300L/UV200S SPD 6A/SPD-6AV SPD 10A/SPD 10AS/SPD-10AV/SPD-10AVP SPD-M10AVP PDA Shimadzu RF530/RF510 Shimadzu RF540/RF535/RF551/RF500 Shimadzu RF1501.5301/5000 RF10A RF10AX DSH-902 DSH-903 DSH-916 DSH-917 DSH-918 DSH-912 DSH-913 DSH-914 DSH-915 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Shimadzu SPD-10 Series long life DSH-918LL 1 Each Model UV 2050 Cat. No. DVA-901 Quantity 1 Each UV 50/Varichrom UV100/UV200 UV5/2550 LC5000/LC5500 Star 9050 ProStar 340/345 UV/Vis DVA-903 DVA-904 DVA-905 DVA-906 DVA-907 DVA-909 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Detector Lamps for Shimadzu Instruments Description Deuterium Lamp Xenon Lamp LL Deuterium Lamp Detector Lamps for Varian Instruments Thermo Scientific Chromatography Columns and Consumables 2014-2015 Description Deuterium Lamp 4-182 Detector Lamps for Waters Instruments Description Mercury Lamp Model 440/441/490 Cat. No. DWA-901 Quantity 1 Each Deuterium Lamp 480/481/480LC/481LC/Lambda Max/LC1 484 486 2486 996 PDA/2996 990/991/994 PDA 2487 Dual Wavelength/2488 RI/R401/R403/R404 440/441/490 440/441/490 470/475/2475 Lamp only 474 Waters 996 Waters Alliance 2487/2488 DWA-910 DWA-915 DWA-918 DWA-918LC DWA-921 DWA-926 DWA-930 DWA-911 DWA-912 DWA-913 DWA-923 DWA-929 DWA-921LL DWA-930LL 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Tungsten Lamp Cadmium Lamp Zinc Lamp Xenon Lamp LL Deuterium Lamp Pump Spares for UltiMate 3000 Instruments Model Cat. No. Quantity ISO-3100SD LPG-3400SD DGP-3600SD HPG-3x00SD LPG-3400RS DGP-3600RS HPG-3x00RS LPG-3400XRS 6040.1950 6040.1951 6040.1952 6040.1953 6040.1954A 6040.1955A 6040.1956A 6043.1954 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each SD, RS, BM pumps LPG-3400XRS pump 6040.0042 6043.0169 2 Each 1 Each SD, RS pumps SD, RS pumps LPG-3400XRS pump 6040.0304 6040.0306 6043.0295 2 Each 2 Each 1 Each SD, RS, BX, BM pumps SD, RS, BX, BM pumps LPG-3400XRS pump 6041.2300 6041.2301 6043.0145 1 Each 1 Each 1 Each Model Cat. No. Quantity Accela pump Accela pump 00201-11328 00950-01-00126 2 Each 2 Each Accela pump Surveyor LC Surveyor LC Surveyor LC Surveyor MS Surveyor MS Surveyor MS 00950-01-00128 SFS-220 SFS-220G SFS-230 SFS-320 SFS-320U SFS-330 2 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Accela pump Accela pump Surveyor LC Surveyor LC Surveyor MS 00950-01-00131 00950-01-00130 SFS-3001 SFS-3002 SFS-6001C 1 Each 1 Each 1 Each 1 Each 1 Each LC Accessories Description Maintenance Kits Maintenance Kit for ISO-3100SD Maintenance Kit for LPG-3400SD Maintenance Kit for DGP-3600SD Maintenance Kit for HPG-3x00SD Maintenance Kit for LPG-3400RS Maintenance Kit for DGP-3600RS Maintenance Kit for HPG-3x00RS Maintenance Kit for LPG-3400XRS Pistons Pistons, sapphire Pistons, sapphire Piston Seals Piston Seals, RP Piston Seals, NP Piston Seal Check Valves Check valve, sapphire, cardridge type Check valve, ZrO ceramics, cardridge type Check valve, sapphire, cardridge type LC Columns and Accessories >> Pump Spares Pump Spares for Thermo Scientific Instruments Description Pistons Piston Piston, sapphire Piston Seals Secondary piston seal (wash seal) Piston Seal Black Piston Seal Yellow Wash Seal White Piston Seal Black Piston Seal Clear Wash Seal clear Check Valves Outlet Check Valve Inlet Check Valve Inlet Check Valve Assembly – Cartridge Type Outlet Check Valve Assembly – Cartridge Type Inlet/Outlet Check Valve Cartridge www.thermoscientific.com/chromatography 4-183 LC Columns and Accessories Pump Spares for Agilent Instruments Description Pistons Piston Assembly Sapphire Piston Assembly Sapphire Piston Seals Piston Seal Yellow Piston Seal Black Check Valves and Spares Replacement Inlet/Outlet Check Valve Cartridge Inlet/Outlet Check Valve Assembly Model Cat. No. Quantity 1090 1050 and 1100 SHP-200 SHP-400 1 Each 1 Each 1050, 1090 and 1100 1050 and 1100 SHP-220G SHP-420K 1 Each 1 Each 1090 1090 SHP-5002 SHP-5001 1 Each 1 Each Model Cat. No. Quantity Series 200, 400, 410, 620, Model 250, Integral 4000 Series 200, 400, 410, 620, Model 250, Integral 4000 SOT-PE600 SOT-PE500 1 Each 1 Each Series 200, 400, 410, 620, Model 250, Integral 4000 Series 200, 400, 410, 620, Model 250, Integral 4000 Series 200, 400, 410, 620, Model 250, Integral 4000 Series 200, 400, 410, 620, Model 250, Integral 4000 SOT-PE220 SOT-PE220G SOT-PE320 SOT-PE320G 1 Each 1 Each 1 Each 1 Each Series 200, 400, 410, 620, Model 250, Integral 4000 Series 200, 400, 410, 620, Model 250, Integral 4000 SOT-PE3001 SOT-PE3002 1 Each 1 Each Model Cat. No. Quantity 5000, 5500, 5600 2010, 2210, 2510 SOT-VA200 SOT-VA400 1 Each 1 Each 5000, 5500, 5600 2010, 2210, 2510 2010, 2210, 2510 SOT-VA220 SOT-VA320 SOT-VA320G 1 Each 1 Each 1 Each 2010, 2210, 2510 2010, 2210, 2510 SVA-3001 SVA-3002 1 Each 1 Each Pump Spares for PerkinElmer Instruments Description Pistons HP Piston Assembly Sapphire HP Piston Assembly Sapphire Piston Seals HP Piston Seal Grey HP Piston Seal Yellow LP Piston Seal Black LP Piston Seal Yellow Check Valves and Spares Inlet/Intermediate Check Valve Assembly Outlet Check Valve Assembly Thermo Scientific Chromatography Columns and Consumables 2014-2015 Pump Spares for Varian Instruments 4-184 Description Pistons Piston Assembly Sapphire Piston Assembly Sapphire Piston Seals Piston Seal Black Piston Seal Black Piston Seal Yellow Check Valves and Spares Inlet Check Valve Assembly Outlet Check Valve Assembly Pump Spares for Shimadzu Instruments Cat. No. Quantity LC-10 AS, LC-6, LC-6A LC-9, LC-10AD, LC-600 SOT-SH200 SOT-SH202 1 Each 1 Each LC-10 AT LC-10 AT LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS LC-9, LC-10AD, LC-600 LC-10 ATvp SOT-SH-100-01 SOT-SH-100-02 SOT-SH220 SOT-SH220G SOT-SH520G SOT-SH420 SOT-SH520 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS LC-3, LC-4, LC-5, LC-6, LC-6A, LC-10 AS LC-9, LC-10AD, LC-600 LC-9, LC-10AD, LC-600 SOT-SSH3001 SOT-SSH3002 SSH-6001 SSH-6002 1 Each 1 Each 1 Each 1 Each Model Cat. No. Quantity M510, M590, M600, M610 M6000 M510, M590, M600, M610 M6000 M45, M501 M515 Alliance 2690 SWA-WA200 SWA-WA200R SWA-WA205 SWA-WA800 SWA-WA900 1 Each 1 Each 1 Each 1 Each 1 Each M45, M501, M510, M590, M600, M610 M6000 M45, M501, M510, M590, M600, M610 M6000 M510EF, M590EF, M600EF, M610EF, M6000EF M515 M515 Alliance 2690 Alliance 2690 SWA-WA220 SWA-WA220G SWA-WA600S SWA-WA820 SWA-WA820G SWA-WA920 SWA-WA920G 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each M45, M501, M510, M590, M600, M610 M6000 M45, M501, M510, M590, M600, M610 M6000 M45, M501, M510, M590, M600, M610 M6000 M510, M590, M600, M610 M6000 M45, M501, M510, M590, M600, M610 M6000 M45, M501, M510, M590, M600, M610 M6000 M510EF, M590EF, M600EF, M610EF, M6000EF M510EF, M590EF, M600EF, M610EF, M6000EF M515 M515 Alliance 2690 SWA-3201 SWA-3202 SWA-3202B SWA-3212 SWA-3402 SWA-3402B SWA-4107 SWA-4123 SWA-8001 SWA-8002 SWA-9001 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each 1 Each Pump Spares for Waters Instruments Description Pistons Piston Assembly Sapphire Piston Assembly Ruby Piston Assembly Sapphire Piston Assembly Sapphire Piston Assembly Sapphire Piston Seals Piston Seal Black Piston Seal Yellow Piston Seal Grey Piston Seal Black Piston Seal Yellow Piston Seal Black Piston Seal Yellow Check Valves and Spares Inlet Check Valve Assembly Outlet Check Valve Assembly Actuator Type Outlet Check Valve Inlet Check Valve Repair Kit Outlet Check Valve Assembly Actuator Type Outlet Check Valve Assembly Ball & Seat Type Inlet Check Valve Assembly Inlet Check Valve Repair Kit Inlet Check Valve Assembly Outlet Check Valve Assembly Check Valve Cartridge www.thermoscientific.com/chromatography LC Accessories Model Description Pistons Piston Assembly Sapphire Piston Assembly Sapphire Piston Seals Piston Seal Yellow Wash Seal White Piston Seal Grey Wash Seal White Piston Seal Yellow Piston Seal Grey Piston Seal Black Check Valves and Spares Inlet Check Valve Assembly Outlet Check Valve Assembly Inlet Check Valve Assembly – Cartridge Type Outlet Check Valve Assembly – Cartridge Type LC Columns and Accessories >> Pump Spares continued 4-185 LC Columns and Accessories Dionex ICS-900 Ion Chromatography Systems Routinely Analyze Multiple Anions or Cations in 10-15 Minutes • Sensitive, stable, heated conductivity detection for precise results • Compatibility with a broad range of polymeric separation columns for unparalleled application flexibility and reliability • Thermo Scientific™ Dionex™ Chromeleon™ SE allows control of a single Thermo Scientific Dionex ICS-900 with an autosampler The Dionex ICS-900 is an integrated, single-channel ion chromatography system designed to run specific isocratic anion and cation applications. The system uses Thermo Scientific™ Dionex™ MMS™ 300 membrane suppression with Displacement Chemical Regeneration (DCR) technology for low noise and stable baselines. Each ICS-900 system has an all-polymeric flow path with a reliable dual piston pump, high-pressure pulse damper, electrically actuated PEEK™ valve, and a temperature-controlled conductivity cell. Dionex ICS-900 Key Features: Thermo Scientific Chromatography Columns and Consumables 2014-2015 • 4-186 Dionex MMS 300 membrane suppression with DCR technology for drift-free baseline and ease-of-use • Wide pump flow rate range to support 2, 3, and 4mm isocratic anion and cation columns • Dual-piston, serial-pumping system with PEEK™ flow path for low maintenance costs and maximum up-time • All-polymeric flow pathway to eliminate contamination and corrosion • Chromeleon Chromatography Data System software for full control, quality integration, and versatile reports to exceed all your data processing needs • USB connectivity for fast, trouble-free instrument connection and configuration Dionex ICS-900 Physical Specifications Dimensions (h × w × d): 33 × 24 × 40cm (13 × 9.5 × 15.75in) Weight 10kg (22lbs) Power Requirements: 100 - 240 VAC, 50–60Hz, autoranging The AS-DV autosampler is a low-cost, metalfree, rugged, automated sample loading device designed especially for ion chromatography applications. The random access and sample preparation capabilities provide easily automated sample introduction to the chromatograph. Its new software control provides high flexibility to select the optimum injection parameters for filling injection loops or loading concentrator columns. AS-DV Key Features: • Full loop and concentrator loading • Optional 6-port or 10-port valve for automated sample preparation or sample injection • Random access • Chromeleon control provides high flexibility to select the optimum injection parameters • 0.5mL and 5.0mL polymeric vials with optional filter caps • Automatic switching power supply for universal input voltage • Sample Overlap injections for increased productivity AS-DV Autosampler Specifications Dimensions (h × w × d): 23 × 45 × 56cm (9 × 17.5 × 22in) Weight: 16 kg (35 lbs) Power Requirements: 100 - 240 VAC, 50–60Hz, autoranging Description Dionex ICS-900 IC System for Anions with AS-DV Autosampler and Chromeleon Software Dionex ICS-900 IC System for Cations with AS-DV Autosampler and Chromeleon Software Cat. No. 078028 078029 Contact Customer Service for details of replacement columns and reagents for the Dionex ICS-900 The Anion Bundle includes: • • • • The Cation Bundle includes: • • • • Dionex ICS-900 Ion Chromatography System AS-DV Autosampler Chromeleon 7 SE Software CS12A 4mm Consumables Bundle: 1- Dionex IonPac CS12A 4mm, 046073 1- Dionex IonPac CG12A 4mm, 046074 1- Thermo Scientific™ Dionex™ CMMS™ 300 Cation MicroMembrane Suppressor 4mm, 064560 1- Dionex CMMS III Regenerant Concentrate, 4 pack, 057556 1- Dionex ICS-900 DCR Cation Regenerant 2L Bottle, 057713 1- CS12A Eluent Concentrate, 057562 www.thermoscientific.com/chromatography LC Accessories Dionex ICS-900 Ion Chromatography System AS-DV Autosampler Chromeleon 7 SE Software AS22 4mm Consumables Bundle: 1- Dionex IonPac AS22 4mm, 064141 1- Dionex IonPac AG22 4mm, 064139 1- Thermo Scientific™ Dionex™ AMMS™ 300 Anion MicroMembrane™ Suppressor 4mm, 064558 1- Dionex AMMS III Regenerant Concentrate, 4 pack, 057555 1- Dionex ICS-900 DCR Anion Regenerant 2L Bottle, 057712 1- AS22 Eluent Concentrate, 0670 LC Columns and Accessories >> Dionex ICS-900 Ion Chromatography System Bundles 4-187 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-188 HOT POCKET and COOL POCKET Column Temperature Controllers Wrap-around column temperature control systems • Easy to install and use with a variety of column lengths • Dual display of both actual and set point temperature • HOT POCKET range from just above ambient to 85°C • COOL POCKET range from 5°C to 55°C • Explore sample selectivity and stability on both sides of ambient Column Heating and Cooling in an Efficient, Compact Design The Thermo Scientific™ HOT POCKET™ and Thermo Scientific™ COOL POCKET™ Column Temperature Controllers have a unique, space saving design for the efficient control of HPLC column temperature using a novel, soft, wraparound sealing mantle. The mantle is wrapped directly onto the column, in situ, in horizontal, vertical, or slant position. The standard size accepts column lengths up to 300mm, and columns up to 150mm can be used with the short HOT POCKET model. The inserts also allow the use of guard columns or the optional eluent pre-heater. The inserts are modular, allowing them to be easily removed or rearranged for your specific column configuration. Special inserts are available for larger or smaller diameter columns. The temperature is set on the Temperature Controller Unit, which is permanently attached to the heater/cooler. Both the actual temperature and the user selected set point are simultaneously displayed on the LED controller display. HOT POCKET Column Heater The HOT POCKET Column Heater has a temperature range of just above ambient to 85°C with excellent control, allowing validation of HPLC methods at accurate temperatures. HPLC method ruggedness can be investigated by exploring the sensitivity of a separation to temperature changes. The HOT POCKET is available in a standard size to accommodate column combinations up to 300mm in length, and a short version for columns up to 150mm. It is easy to install a column into the HOT POCKET or COOL POCKET. Depending upon column length and auxiliary fixtures such as a guard column or eluent pre-heater, some of the inserts may have to be rearranged or removed through the special slot at one end. Rotate the inserts so that the groove in each is positioned in the open part of the channel. Columns are simply placed into the inserts, which are then rotated to lock the column into the channel. The insulated mantle is wrapped around the column with a Velcro™ closure. COOL POCKET Temperature Controller The COOL POCKET Temperature Controller provides efficient control of the temperature of HPLC columns both above and below ambient, with an operational temperature range of 5°C to 55°C. The COOL POCKET Temperature Controller is ideal for chiral applications where a lower temperature may give better separation of enantiomers or other closely related compounds. It also allows you to validate HPLC methods at accurate temperatures near ambient and check HPLC method ruggedness by exploring the sensitivity of your separation to temperature changes on both sides of ambient. Column heating or cooling in a compact, efficient design • The Eluent Pre-Heater provides a dramatic improvement in chromatography as demonstrated by the USP method for enalapril maleate at 80°C • For preheating or precooling the mobile phase before it enters column • Use in temperatures above 50°C or below 15°C • 0.005in ID Product Specifications Controller Dimensions Mantle Dimensions Power Cord Weight Power With Eluent Pre-heater Without Eluent Pre-heater 80°C 80°C oven4 0 1 2 LC Equipment HOT POCKET COOL POCKET 5°C above ambient to 85°C 5°C to 55°C Dual LED displays of actual and set point temperatures in °C ± 2°C over entire range ± 2°C over entire range ± 1°C ± 1°C ± 0.1°C ± 0.1°C 85°C in less than 30 minutes 55°C in 25 minutes, 5°C in 20 minutes Standard: up to 3/8in OD and up to 300mm in length and end-fittings up to 19mm OD (250mm length column with guard or eluent pre-heater in addition to column) Short (HOT POCKET only): up to 150mm total length (100mm column plus guard or pre-heater) 2.8 x 4.0 x 6.5in 2.8 x 4.0 x 6.5in Standard: 1.5 x 1.5 x 17in Standard: 1.5 x 4.0 x 17in Short: 1.5 x 1.5 x 12in 3 foot retractable coil cord 3 foot retractable coil cord 1lb enclosure (3lb total with power supply) 2lb enclosure (4lb total with power supply) 24 VAC, 25 Watts maximum 15 VDC, 20 Watts maximum Operating Range Display Temperature Accuracy Temperature Repeatability Temperature Stability Time to Stabilization (from ambient) Column Capacity LC Columns and Accessories >> HOT POCKET Column Heaters, Eluent Preheater/Precooler and COOL POCKET Chiller 3 4 5 MIN oven3 0 1 2 3 4 5 MIN Effect of eluent pre-heater on efficiency Data courtesy of Dr. Richard F. Myer, Quantitative Technologies, Inc., Whitehouse, NJ HOT POCKET Column Heaters, Eluent Preheater/Precooler and COOL POCKET Chiller Description HOT POCKET Column Heater HOT POCKET Column Heater – short version COOL POCKET Column Chiller Eluent Preheater/Precooler Cat. No. 92016 92016-150 92017 92018 Quantity 1 Each 1 Each 1 Each 1 Each www.thermoscientific.com/chromatography 4-189 LC Columns and Accessories ColumnOven Exact temperature control for LC columns • Temperature control from ambient to 90°C • Fits up to four columns in one compartment • Compatible with CTC autosamplers and Thermo Scientific Mass Spectrometer ion sources Product Description Options The Thermo Scientific ColumnOven delivers efficient temperature control of HPLC columns. With exact temperature control to +/- 0.1°C, ColumnOven provides the highest levels of reproducibility and stability. Temperatures up to 90°C can significantly reduce backpressure when using columns packed with sub 2µm particle sizes, allowing the use of higher flow rates to accelerate the chromatography. ColumnOven 200 can accommodate up to 4 analytical HPLC columns with lengths up to 150mm while ColumnOven 300 can accommodate up to 4 analytical columns with lengths up to 250mm. Optional mounts allow ColumnOven to be secured to a CTC PAL® autosampler and many Thermo Scientific Mass Spectrometer ion sources. The optional software package includes a standalone version (Windows™ compatible) and drivers for Thermo Scientific™ XCalibur™ and other instrument control software. An optional heat exchanger can be used as a connection / transfer line between injection port and column. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Product Specifications 4-190 Description Column oven dimensions: Column oven dimensions (with mounted heat shield): Thermostated column compartment: Remote control Power supply Temperature control: Temperature stability: ColumnOven 200 212mm x 40mm x 46mm 212mm x 62mm x 52mm 200mm x 21mm x 21mm LAN or serial port 110 - 240 VAC Ambient to 90°C in 0.1°C increments +/- 0.1°C Thermo Scientific ColumnOven Description ColumnOven 200 CTC Mount Thermo MS Mount ABI MS Mount Heat Exchanger Heat Exchanger (low volume) Software drivers ColumnOven 300 Cat. No. 66001-020 66001-021 66001-022 66001-023 66001-024 66001-025 66001-026 66001-030 ColumnOven 300 312mm x 40mm x 46mm 312mm x 62mm x 52mm 300mm x 21mm x 21mm LAN or serial port 110 - 240 VAC Ambient to 90°C in 0.1°C increments +/- 0.1°C Reduce mobile phase consumption by up to 90% • Continuously monitors the output signal of the chromatographic detector, recycling the mobile phase to the solvent reservoir when the baseline is below a certain preset threshold • Easy-to-use software is provided to configure system parameters, perform on-line monitoring and audit trail facilities • No power adapter is required as the solvent saver is powered directly from the chromatography data system PC through a USB connection • Recycles the mobile phase only if switched on: in case of power failure the valve remains in the waste position and the mobile phase in the reservoir remains uncontaminated • Analog input allows unipolar or bipolar operation of the device within a range of ±1V with an analog-to-digital converter • TTL/contact closure for the device can be configured as start, auto-zero or valve position control input (a) - At start of 1st waste run (b) - After delay Operational Principle LC Equipment • If the input signal level exceeds this threshold value, the SRS Pro redirects the eluent flow LC Columns and Accessories >> SRS Pro Solvent Recycling System to waste (a), taking account of the transport time from the detector to the switching valve • When the signal returns below the threshold (b), the SRS Pro again waits for the transport delay and then switches the mobile phase back to the reservoir • Autosampler injection marker connected to the SRS Pro zeroes signal input at the moment of injection Product Specifications Data Rate Wetted Material Power Source Max. Pressure Requires Compatible with: 1Hz PEEK USB port of PC 30psi/0.2MPa 1 free USB port, MS-Windows XP/2000/Vista Any HPLC detector SRS Pro Solvent Recycling System Description SRS Pro solvent recycling system Cat. No. 66001-001 Quantity 1 Each www.thermoscientific.com/chromatography 4-191 LC Columns and Accessories SDG Pro Solvent Degasser For gas-free HPLC solvents • High efficiency in-line system • Reliable, continuous operation • Quick equilibration and short startup times • Removes dissolved gases from solvents • Used to degas the mobile phase for HPLC and can be employed in other applications where gases may interfere with the use of the system (such as an autotitrator) Product Specifications General Channels Mode of Degassing Maximum Flow Rate Degassing Capacity Dead Volume Materials Contacting Solvents Power Power Requirement if using supplied AC Adapter Power Requirement if not using supplied AC Adapter Wall Sockets Thermo Scientific Chromatography Columns and Consumables 2014-2015 Installation Over-Voltage Category Validation Output Signal 4-192 Accuracy Operating Conditions Ambient Temperature Ambient Relative Humidity (RH) Altitude Indoor vs. Outdoor Use Pollution Degree Storage Conditions Ambient Temperature Ambient Relative Humidity Altitude Physical Dimensions Weight 4 independent Gas permeation through a fluoropolymer tube 10mL/min ~2ppm at 1mL/min ~480µL per channel for standard channel PEEK, Glass-filled PTFE, Teflon AF 100 – 240 VAC (±10%), 1A, 50 to 60Hz (±3Hz) 15 – 24 VDC at 0.85 A maximum (0.5 A typical) 4 supplied with AC adapter, interchangeable: North America/Japan, U.K., Continental Europe, Australia II 5mVDC / 1mm Hg absolute from 20 to 800mm Hg (0.100 VDC at 20mm Hg; 4.000 VDC at 800mm Hg) ±1.0% of reading ±0.010 VDC from 20 to 800mm Hg 10 to 35°C 20 to 80 % RH (without condensation) 0 to 2000 Meters Indoor 2 -20 to +60°C 20 to 80% RH (without condensation) 0 to 12000M Height: 127mm (5.0in) Width: 73mm (2.8in) Depth: 250mm (9.8in) 2.7kg (6lb). SDG Pro Description SDG Pro degasser Cat. No. 66001-010 Quantity 1 Each Introduction to HPLC Ion Pair Reagents High-purity reagents with the selectivity needed for good separation. In the past, reverse-phase HPLC analysis of highly charged acidic and basic compounds was frustrating and resulted in poor resolution. Important biomolecules such as amino acids, peptides, organic acids, polyamines and catecholamines had to be separated by ion exchange or by suppression techniques. By using the correct ion pair reagents, you achieve: Two principal theories have been proposed to explain reverse-phase ion pair chromatography. In the first theory, small polar ion pair reagents react with the ionized solute, forming neutral ion pairs. The second theorizes that an active ion exchange surface is produced in which long chain, nonpolar anions and cations are absorbed by the hydrophobic stationary phase. To optimize chromatographic separations in ion pair elution systems, high-purity reagents of exceptional optical transparency are needed. Ion Pair Reagents are specially purified for ion pair chromatography and provide the selectivity needed for good separations. LC Reagents Thermo Scientific Ion Pair Reagents enable you to quickly and efficiently analyze charged compounds using reverse-phase techniques. Our ion pair reagents are simply dissolved in the HPLC solvent system, resulting in the formation of stable chromatographic complexes that can be separated using reverse-phase columns. Reverse-phase ion pair chromatography theories LC Columns and Accessories >> Thermo Scientific Liquid Chromatography Reagents • Increased or decreased retention, permitting controlled selectivity • Resolution of complex ionic mixtures without using ion exchange columns • Improved peak symmetry www.thermoscientific.com/chromatography 4-193 LC Columns and Accessories Derivatization and Visualization Reagents for HPLC Designed to provide selectivity and improve sensitivity. The lack of a universal HPLC detector that provides high sensitivity (as well as some degree of selectivity) established the need for suitable derivatization procedures. Derivatization is the chemical modification of an existing compound, producing a new compound that has properties more suitable for a specific analytical procedure. It is an analytical tool that can be used to provide both selectivity and improved sensitivity. There are several requirements for derivatization protocol: 1. At least one acidic, polar functional group must be available for reaction on the parent compound. 2. A single derivative should be formed per parent compound. 3. The reaction should be reproducible under the given time and reaction conditions. 4. The reaction should proceed quickly and easily under mild conditions. Thermo Scientific Chromatography Columns and Consumables 2014-2015 5. The reaction byproducts (if any) should not interfere with the chromatography, or with detection of the sample. 4-194 Pre- and post-chromatographic techniques are both used in HPLC derivatization. In addition, off-line and on-line reactions have been employed with both techniques. Prechromatographic (or pre-column techniques) offer more than greater selectivity and sensitivity in detection. Pre-column techniques can be used to enhance stability, improve resolution, improve peak symmetry and increase or decrease retention of solutes. FDAA (Marfey’s Reagent) allows separation and quantification of optical isomers of amino acids (Figure 2). Post-chromatographic (or post-column) techniques are used primarily to provide selectivity and improve sensitivity. We offer a variety of HPLC detection reagents for pre- and post-chromatographic techniques. All compounds and formulations are purified for chromatography, minimizing artifact formation. Separation of D- and L-DOPA on 100mm x 4.6mm C18 Conditions: A – 0.05 M triethylamine phosphate, pH 3.0; B – acetonitrile. Linear gradient: 10 to 40% B in 45 minutes, 2.0mL /minute, 25˚C, 340nm Functional Group Carboxylic Acid Page 4-201 Comments Formulation of 1.0mmol/ml p-bromophenacyl bromide and 0.005mmol/ml crown ether in acetonitrile; pre-column; nanomole detection levels: λmax = 260nm1-7 Vis 4-204 4-N, N-dimethylaminoazobenzene-4’-sulfonyl chloride (dabsyl chloride); pre-column; nanomole detection levels: λmax = 436nm8-14 UV 4-200 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide (FDAA); pre-column; nanomole detection levels: λmax = 340nm. For chiral separations of amino acids.15, 28-29 Vis 4-203 UV 4-204 Phenylisothiocyanate (PITC); pre-column; picomole detection levels: λmax = 254nm23-24 EC, UV 4-201 2,4,6-Trinitrobenzene-sulfonic acid (TNBSA); pre- or post-column; nanomole detection levels with EC and UV, GC - 0.85V; λmax = 250nm25-26 Ninhydrin (see structure above) Vis 4-203 PITC (see structure above) UV 4-204 O O R C OH Primary Amine R Br C CH2 Br Dabsyl Chloride p-Bromophenacylate MW 277.94 N H N H O N S N O Cl Dabsyl Chloride MW 323.80 FDAA, Marfey’s Reagent O N – O N + O– O H N NH 2 + O F Ninhydrin FDAA O (Marfey's Reagent) MW 272.19 OH OH Ninhydrin MW 178.14 Post-column; nanomole detection levels: λmax = 570nm22 O PITC N TNBSA O – O O S O Secondary Amine R NH R' N+ PITC Edman‘s Reagent MW 135.19 O N O S OH N+ – C + O– TNBSA MW 293.17 LC Reagents Detection* UV Description p-Bromophenacylate LC Columns and Accessories >> Derivatization Reagents for HPLC O Post-column; nanomole detection levels: λmax = 440nm22 Phenylisothiocyanate (PITC); pre-column; picomole detection levels: λmax = 254nm23-24 *EC = electrochemical; F = fluorescence; UV = ultraviolet; Vis = visible. References 1. Durst, H.D., et al. (1975). Anal. Chem. 47, 1797. 2. Borch, R.F., et al. (1975). Anal. Chem. 47, 2437. 3. Grushka, E. et al. (1975). J. Chromatogr. 112, 673. 4. Fitzpatrick, F.A. (1976). Anal. Chem. 48, 499. 5. Nagels, L., et al. (1980). J. Chromatogr. 190, 411. 6. Ahmed, M.S., et al. (1980). J. Chromatogr. 192, 387. 7. Pierce Technical Bulletin: Preparation of Phenacyl and p-Bromophenacyl Derivatives for HPLC. 8. Stocchi, V., et al. (1985). J. Chromatogr. 349, 77-82. 9. Chang, J.Y., et al. (1981). Biochem. J. 199, 547-555. 10. Chang, J.Y., et al. (1982). Biochem. J. 203, 803-806. 11. Vendrell, J., et al. (1986). J. Chromatogr. 358, 401-413. 12. Chang, J.Y., et al. (1984). J. Chromatogr. 295, 193-200. 13. Lin J.K., et al. (1980). Anal. Chem. 52, 630-635. 14. Chang, J.Y., et al. (1981). FEBS Letters 132, 117-120. 15. Marfey, P. (1984). Carlsberg Res. Comm. 49, 591-596. 16. Jones, B.N. and Gilligan, J.P. (1983). Amer. Biotech. Lab. 12, 46-51. 17. Fiedler, H.P., et al. (1986). J. Chromatogr. 353, 201-206. 18. Fried, V.A., et al. (1985). Anal. Biochem. 146, 271-276. 19. Jones, B.N., et al. (1983). J. Chromatogr. 266, 471-482. 20. Seaver, S.S., et al. (Sept./Oct. 1984). Biotechniques 254-260. 21. Böhlen, P., et al. (1979). Anal. Biochem. 94, 313-321. 22. Stein, W.H. and Moore, S. (1949). Cold Spring Harbor Symp. Quant. Bio. 14, 179. 23. Heinrickson, R.L. and Meredith, S.C. (1984). Anal. Biochem. 137, 65-74. 24. Scholze, H. (1985). J. Chromatogr. 350, 453-460. 25. Caudill, W.L., et al. (1982). J. Chromatogr. 227, 331. 26. Caudill, W.L., et al. (1982). Bioanalytical System’s Current Separations 4(4), 59. 27. Andrews, J.L., et al. (1982). Arch. Biochem. Biophys. 214, 386-396. 28. Aberhart, D.J., et al. (1985). Anal. Biochem. 151, 88-91. 29. Szokan, G., et al. (1988). J. Chromatogr. 444, 115-122. www.thermoscientific.com/chromatography 4-195 LC Columns and Accessories Developments in Amino Acid Analysis Improvements in amino acid analysis by ion exchange chromatography have involved the analytical system, as well as the instrumentation. Systems have been developed (by varying buffer pH or ionic strength) that work to displace the amino acids into discrete bands. The buffer systems are compatible with single- or two-column analysis of amino acids found in protein hydrolyzates or physiological fluids. Buffer systems are determined by the counter ion used (sodium or lithium) and by the method of buffer changes introduced to the resin (step changes or gradient elution).9-15 The most commonly used buffering component, citrate, is suitable for solutions below pH7.16 Buffers are prepared either with citric acid or an alkali salt. Unfortunately, for high-sensitivity work, citric acid is a significant contributor to amino acid contamination. Therefore, to achieve consistent analyses, it is essential to use high-purity reagents for buffer preparation. Alternatives to ion exchange are available for the separation of amino acids. Because amino acid analysis is one of the basic protein chemistry tools available, more rapid and sensitive methods for separation and quantitation are desirable.17 Several precolumn derivatization methods using reverse-phase HPLC have been developed. Two of the most widely used of these methods involve the formation of dansyl18-19 or o-phthalaldehyde (OPA)20-24 derivatives of amino acids prior to HPLC analysis. Both methods offer greater sensitivity and shorter analysis time than post-column derivatization techniques. Other methods include the quantitative derivatization of amino acids with phenylisothiocyanate (PITC) and the separation and quantitation of the resulting phenylthiocarbonyl derivatives via HPLC. These derivatives are stable enough to eliminate in-line derivatization. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Note: Please refer to page 3-122 for details of references 4-196 The extraction and purification of proteins play an important role in determining amino acid content. These methods are based on one or more of their physical characteristics (e.g., solubility, molecular size, charge, polarity and specific covalent or noncovalent interactions). The techniques commonly used to separate proteins and peptides include: • Reverse-phase HPLC • Polyacrylamide gel electrophoresis • Gel filtration • Ion exchange chromatography • Affinity chromatography • The table below provides a more detailed list of methods for fractionating peptide mixtures.25 Hydrolysis Most protein samples require some form of chemical treatment before their component amino acids are suitable for analysis. Protein and peptide samples must be hydrolyzed to free amino acids from peptide linkages. Acids (usually HCI) are the most widely used agents for hydrolyzing proteins. A simplified hydrolysis procedure involves refluxing the protein with excess HCI, then removing the excess acid in vacuum.26 The lyophilized protein then is suspended in constant boiling 6 N HCI and introduced into the hydrolysis tube. The sample is frozen by immersing the tube in dry ice and acetone. Before sealing, the tube is evacuated to avoid formation of cysteic acid, methionine sulfoxide and chlorotyrosine.27 This procedure minimizes decomposition of reduced S-carboxymethylcysteine and analyzes S-carboxymethylated proteins. Hydrolysis is conducted at 110°C (with the temperature accurately controlled) for 20-70 hours by Moore and Stein’s method.28 After hydrolysis, residual HCI is removed in a rotary evaporator. The residue is dissolved in water and brought to the appropriate pH for addition to the analyzer column.28 Technique Centrifugation Size exclusion chromatography Ion exchange chromatography Paper electrophoresis Paper chromatography Thin layer electrophoresis Thin layer chromatography Polyacrylamide gel electrophoresis High-performance liquid chromatography (HPLC) Gas chromatography Counter-current extraction Affinity chromatography Covalent chromatography or irreversible binding References 1. Braconnot, H. (1820). Ann. Chem. Phys. 13, 113. 2. Mulder, G.J. (1848). J. Prakt. Chem. 38, 294. 3. Kjeldahl, J.G.C.T. (1883). Z. Anal. Chem. 22, 366. 4. Rattenbury, J.M. (1981). Amino Acid Analysis, Ellow Horwood, publisher, Chicester, England, Chapter 1. 5. Martin, A.J.P. and Synge, R.L.M. (1941). Biochem. J. 35, 1358. 6. Elsden, S.R. and Synge, R.L.M. (1944). Proc. Biochem. Soc. IX. 7. Synge, R.L.M. (1944). Biochem. J. 38, 285. 8. Stein, W.H. and Moore, S. (1949). Cold Spring Harbor Symposia Quant. Biol. 14, 179. 9. Liao, T.H., et al. (1973). Anal. Chem. 45, 2286. 10. Hare, P.E. (1972). Space Life Sciences 3, 354. 11. Dus, K., et al. (1966). Anal. Biochem. 14, 41. Properties of Peptide Molecules Exploited Solubility Size Charge, with some influence of polarity Charge and size Polarity Charge and size Polarity Charge and size Polarity Volatility of derivatives Polarity; sometimes specific interactions Specific interactions Disulfide bond formation; reactivity of homoserine lactone 12. Thomson, A.R. and Miles, B.J. (1964). Nature 203, 483. 13. Robinson, G.W. (1975). J. Chromatogr. 3, 416. 14. Benson, J.V. (1972). Anal. Biochem. 50, 477. 15. Benson, J.V. and Patterson, J.A. (1965). Anal. Chem. 37, 1108. 16. Bates, R.G. and Pinching, G.D. (1949). J. Amer. Chem. Soc. 71, 1274. 17. Jones, B.N. and Gilligan, J.P. (1983). American Biotech. Lab. December, 46-51. 18. Weidmeier, V.T., et al. (1982). J. Chromatogr. 231, 410-417 19. DeJong, C., et al. (1982). J. Chromatogr. 241, 345-359. 20. Lindroth, P. and Mopper, K. (1979). Anal. Chem. 51, 1667-1674. 21. Jones, B.N., et al. (1981). J. Liq. Chromatogr. 4, 565-586. LC Reagents Methods for the fractionation of peptide mixtures. LC Columns and Accessories >> Sample Preparation and Hydrolysis 22. Turnell, D.C. and Cooper, J.D.H. (1982). CIin. Chem. 28, 527-531. 23. Umagat, H., et al. (1982). J. Chromatogr. 241, 324-334. 24. Jones, B.N. and Gilligan, J.P. (1983). J. Chromatogr. 266, 471-482. 25. AlIen, G. (1981). Laboratory Techniques in Biochemistry and Molecular Biology, T.S. Work and R.H. Burdon, Eds., Elsevier/North-Holland, Biomedical Press. 26. Moore, S. and Stein, W.H. (1963). Meth. Enzymol. 6, 819. 27. Eveleigh, J.W. and Winter, G.D. (1970). Protein Sequence Determination, S.B. Needleman, Ed., Springer-Verlag, pp. 92-95. 28. Moore and Stein, op. cit. www.thermoscientific.com/chromatography 4-197 LC Columns and Accessories HPLC Ion Pair Reagents Heptafluorobutyric Acid Ion-pair reagent for the reverse-phase HPLC separation of proteins and peptides • Typical purity is 99.7% by GC; <0.1% water • Sequencing reagent for classical and automated Edman degradation of peptides and proteins • Density: 1.645 • B.P. 120°C • Packaged under nitrogen in amber glass ampules or bottles • Clear, colorless liquid Heptafluorobutyric Acid Description Heptafluorobutyric Acid, Sequencing Grade Heptafluorobutyric Acid, HPLC Grade Quantity 100mL 10 x 1mL ampules X Cat. No. TS-25003 TS-53104 Quantity 1 Each 1 Pack 1. Hearn, M.T.W. and Hancock, W.S. (1979). Trends Biochem. Sci. 4, N58-N62. 2. Bennett, H.P.J., et al. (1980). J. Liquid Chromatogr. 3, 1353-1366. 3. Bennett, H.P., et al. (1981). Biochemistry. 20, 4530-4538. X in the ordering table indicates that hazardous shipping charges apply. Triethylamine (TEA) 2(a);0.000M TEA 2(b);0.010M TEA 2(c);0.020M TEA 4-198 0 Min 3 2(c);0.020M TEA 2,4-dimethylaniline 2,4-dimethylaniline 2,4-dimethylaniline Triethylamine is an ion-pairing reagent that alters selectivity in reverse-phase HPLC separations. By pairing with peptides, it effectively sharpens peaks, resulting in improved peak resolution. • 99.5% triethylamine purity, allowing sensitive peptide detection at low UV wavelengths in reverse-phase HPLC peptide separation systems • Packaged in amber glass bottles with protective PTFE-lined fluorocarbon caps for reagent integrity • Has a low UV absorbance to provide the most sensitive detection across 2(a);0.000M TEA 2(b);0.010M TEA all wavelengths 0 Min 3 2(d);0.030M TEA H N • Molecular weight 101.19 • Density 0.726g/mL 2,4-dimethylaniline • Molecular formula C 6 15 Triethylamine (TEA) Description Triethylamine, HPLC Grade Triethylamine, Sequencing Grade 0 Min 0 Min 3 Quantity 25g 100g 2,4-dimethylaniline • Alternate names TEA, Diethylethanamine X X in the ordering table indicates that hazardous shipping charges apply. 2,4-dimethylaniline Properties of Triethylamine 2,4-dimethylaniline Thermo Scientific Chromatography Columns and Consumables 2014-2015 Ideal for HPLC separation and analysis of peptides 3 Cat. No. TS-53101 TS-25108 0 Min Quantity 1 Each 1 Each 3 0 Min 3 0 Min 3 Well-suited for HPLC and mass spectrometry applications Formic acid is a component found in reverse-phase mobile phases to provide protons for LC/MS analysis. The presence of a low concentration of formic acid in the mobile phase is also know to improve the peak shapes of the resulting separation. Unlike trifluoroacetic acid (TFA), formic acid is not an ion-pairing reagent, and it does not suppress MS ionization of polypeptides when used as a mobile phase component. • Prescored, nitrogen-flushed, amber glass to protect formic acid from light and moisture • 99% purity for consistent LC baselines and no interference introduced into LC and mass spectrometry applications • Convenient format simplifies preparation of gradient and isocratic mobile phases containing 0.1% (v/v) formic acid in water or acetonitrile • Contents of a single vial in a final volume of 1L solvent yields a mobile phase of the most common formic acid concentration Description Formic Acid 99+% Quantity 10 x 1mL ampules Cat. No. TS-28905 Quantity 1 Each For complex peptide separations, the key to success can be to vary selectivity. Varying mobile phase composition on the same column can change selectivity enough to resolve peptide that would otherwise overlap. The TFA concentration is usually specified as 0.1% for reverse-phase HPLC of peptides. For reproducible separations from run-to-run or from lab-to-lab, it is essential to make concentrations the same. www.thermoscientific.com/chromatography LC Reagents Formic Acid Ampules LC Columns and Accessories >> Formic Acid Ampules 4-199 LC Columns and Accessories Derivation and Visualization Reagents for HPLC Trifluoracetic Acid (TFA) Routinely used ion-pairing agent in reversed-phase peptide separations • Purity: >99.5% TFA and exceptional clarity allows sensitive, nondestructive peptide detection at low UV wavelengths • High-performance packaging: Packaged under nitrogen in amber glass with protective TFE-lined fluorocarbon caps to ensure TFA integrity • Choice of formats for convenience: 1mL ampules can prepare 1L of 0.1% v/v TFA solution for the mobile phase in reverse-phase chromatography in moments Trifluoracetic Acid (TFA) Description Trifluoracetic Acid, Sequencing Grade Trifluoracetic Acid, Sequencing Grade Trifluoracetic Acid, Sequencing Grade Trifluoracetic Acid, Sequencing Grade Quantity 500mL 100g 10 × 1mL 1g X X X X Cat. No. TS-28901 TS-28903 TS-28904 TS-28902 Quantity 1 Each 1 Each 1 Pack 1 Each X in the ordering table indicates that hazardous shipping charges apply. FDAA, Marfey’s Reagent Makes separation and quantitation of optical isomers of amino acids by reverse-phase chromatography quick and easy • Optical isomers of amino acids derivatization complete in just 90 minutes • Derivatives have an absorption coefficient of ~3 x 104 • Derivatives can be detected by UV at 340nm with picomole sensitivity 4-200 10 20 30 D-Phenyalanine D-Methionine L-Phenylalanine Quantity 50mg D-Alanine L-Methionine L-Aspartic Acid L-Glutamic Acid D-Aspartic Acid D-Glutamic Acid L-Alanine Excess Reagent Description FDAA, Marfey’s Reagent Absorbance @ 340 nm Thermo Scientific Chromatography Columns and Consumables 2014-2015 FDAA, Marfey’s Reagent 40 Cat. No. TS-48895 Quantity 1 Each Gives quantitative yields with few or no side reactions • Premixing of phenacylbromide and crown ether is not necessary • Derivatization is both rapid and quantitative, with yields of >95% in 15 to 20 minutes at 80°C • Excess reactants do not interfere • Large excess of alkylating reagent is not necessary • Small amounts of water or alcohol do not interfere • If isolation is desired, products are usually crystalline p-Bromophenacylate Reagent Description p-Bromophenacylate Reagent Cat. No. TS-48891 Quantity 10mL Quantity 1 Each 1. Durst, H.D., et al. (1975). Anal. Chem. 47, 1797. 2. Borch, R.F., et al. 1975). Anal. Chem. 47, 2437. 3. Grushka, E., et al. (1975). J. Chromatogr. 112, 673. 4. Fitzpatrick, F.A., et al. (1976). Anal. Chem. 48, 499. LC Columns and Accessories >> p-Bromophenacylate Reagent LC Reagents TNBSA (Trinitrobenzene Sulfonic Acid) An excellent choice for spectrophotometric detection • Couples with primary amines, sulfhydryls and hydrazides in aqueous solution at pH 8, without undesirable side reactions • Excellent for solution or solid phase analysis • Suitable for qualitative and quantitative estimation of biomolecules; including amino acids, eptides or proteins • Chromogenic, O = 335nm max • Reacts readily with primary amino groups of amino acids in aqueous format at pH 8 to form yellow adducts • No colored derivatives are formed with secondary amino acids proline and hydroxyproline • Colored derivatives are monitored at 345nm and have extinction coefficients in range of 1-1.5 x 104 TNBSA Description TNBSA Quantity 100mL X Cat. No. TS-28997 Quantity 1 Each 1. Goodwin, J.F. and Choi, S-Y. (1970). Clinical Chemistry. 16, 24-31. 2. Snyder, S.L. and Sobocinski, P.Z. (1975). Anal. Biochem. 64, 284-288. 3. Drozdovskaya, N.R., et. al. (1982). FEBS Lett. 150, 385. 4. Takahashi, S., et al. (1984). Chem. Lett. (Jpn). 1, 127 X in the ordering table indicates that hazardous shipping charges apply. www.thermoscientific.com/chromatography 4-201 LC Columns and Accessories Hydrolysis Reagents Constant Boiling (6N) Hydrochloric Acid Sequencing-grade reagent for total protein hydrolysis • Hydrolyzes peptides in 6 hours at 150°C • Specially purified to give ninhydrin-negative blank on hydrolysis • Packaged in prescored ampules to eliminate contamination and ensure product integrity Constant Boiling (6N) Hydrochloric Acid Description Hydrochloric Acid 6N Quantity 10 × 1mL Cat. No. TS-24308 Quantity 1 Pack 1. Eveleigh, J.W. and Winter, G.D. (1970). Protein Sequence Determination, Ed Needleham, S.B., Springer-Verlag, pp. 92-95. 2. Blankenship, et al. (1989). High-sensitivity amino acid analysis by derivatization with o-Phthaldialdehyde and 9-Fluorescence detection: applications in protein structure determination. Anal. Biochem. 178, 227-232. 3. Hurley, J.B., et al. (1984). Isolation and characterization of a cDNA clone for the subunit of bovine retinal transducin. Proc. Natl. Acad. Sci. USA. 81, 6948-6952. 4. Lee, K. et al. (1979). Derivatization of cysteine and cystine for fluorescence amino acid analysis with the o-Phthaldialdehyde/2mercaptoethanol reagent. J. Biol. Chem. July 25, 6248-6251. Amino Acid Standard H High-purity calibration standard for protein hydrolysates • Uses L-form configuration to permit standardization of microbial and other assays • Molar concentration verified by conventional amino acid analysis methods • With the exception of cystine, each amino acid is supplied at a concentration of 2.5µmoles/mL in 0.1N HCl The following amino acids are included in Amino Acid Standard H: Thermo Scientific Chromatography Columns and Consumables 2014-2015 L-alanine, Ammonia [(NH4)2SO4], L-Arginine, L-Aspartic Acid, L-Cystine, L-Glutamic Acid, Glycine, L-Histidine, L-Isoleucine, L-Leucine, L-Lysine•HCl, L-Methionine, L-Phenylalanine, L-Proline, L-Serine, L-Threonine, L-Tyrosine, L-Valine. 4-202 Amino Acid Standard H Description Amino Acid Standard H Quantity 10 × 1mL Cat. No. TS-20088 Quantity 1 Pack When kept frozen, an unopened vial has an indefinite storage life. Once the seal is broken, the reagent has a maximum storage life of six months. Store frozen between uses. Ninhydrin The reagent of choice for detection of amino acids • Used in amino acid chromatography • Offers superb color response and low blank • Indefinitely stable and requires no refrigeration Ninhydrin Description Ninhydrin Quantity 500g Cat. No. TS-21003 Quantity 1 Each 1. Stein, W.H. and Moore, S. (1949). Cold Spring Harbor Symp. Quant. Biol. 14, 179. 2. Moore, S. (1968). Amino acid analysis: aqueous dimethyl sulfoxide as solvent for the ninhydrin reaction. J. Biol. Chem. 243(23), 6281-6283. 3. James, L.B. (1978). Amino acid analysis: ninhydrin reaction with titanous chloride. J. Chromatogr. 152, 298-300. Indefinitely stable. No refrigeration required. Keep bottle tightly sealed. Avoid exposure to direct sunlight and ammonia. LC Columns and Accessories >> Amino Acid Detection Reagents LC Reagents www.thermoscientific.com/chromatography 4-203 LC Columns and Accessories High-Purity Pre-Column Derivatization Reagents Dabsyl Chloride Recrystallized twice • For the precolumn derivatization and detection of amino acids in visible light down to sub-picomolar levels • Analysis of 10-30ng of protein hydrolysates • Analysis of peptides and determination of C-terminal sequence of polypeptides • Analysis of phosphoamino and amino acid amides • Analysis of amino acid neurotransmitters in mouse brain Dabsyl Chloride Description Dabsyl Chloride (4-N,N Dimethylaminoazobenzene-4sulfonyl chloride) Quantity 500g X Cat. No. Quantity TS-21720 1 Each 1. Chang, J.Y., et al. (1981). Biochem. J. 199, 547-555. 2. Chang, J.Y., et al. (1982). Biochem. J. 199, 803-806. 3. Chang, J.Y. (1984). J. Chromatogr. 295, 193-200. 4. Chang, J.Y., et al. (1981). FEBS Lett. 132, 117-120. 5. Vendrell, J. et al. (1986). J. Chromatogr. 358, 401-413. 6. Lin, J.K., et al. (1980). Clin. Chem. 26, 579-583. 7. Chang, J.Y., et al. (1983). Methods. Enzymol. 92, 41-48. 8. Stocchi, V., et al. (1985). J. Chromatogr. 349, 77-82. X in the ordering table indicates that hazardous shipping charges apply. PITC (Phenylisothiocyanate) High-purity reagent for pre-column quantitative derivatization of amino acids by reverse-phase HPLC • Also known as Edman’s Reagent • Reacts readily with amino acids in 5 to 10 minutes at room temperature • Resulting phenylthiocarbamyl derivatives can be separated and quantified in 30 minutes Thermo Scientific Chromatography Columns and Consumables 2014-2015 using reverse-phase HPLC to produce stable products with all amino acids including proline 4-204 PITC (Phenylisothiocyanate) Description PITC (Edman’s Reagent) Quantity 10 × 1mL 1. Heinrikson, R.L. and Meridith, S.C. (1984). Anal. Biochem. 136, 65-74. 2. Scholze, H. (1985). J. Chromatogr. 350, 453-460. 3. Janssen, et al. (1986). Chromatogr. 22(7-12). Evert, R.F. (1986). Anal. Biochem. 154, 431-435. Cat. No. TS-26922 Quantity 1 Pack Ultrapure solvents are carefully packed for thorough protection • Distilled in glass, filtered through 0.2µm TFE membranes and packed in solvent-rinsed, amber glass bottles • TFE-lined screw caps seal bottles Dimethylsulfoxide (DMSO), Sequencing Grade, Physical Properties • UV Cutoff: 190nm • C2H6OS • Optical Absorbance: <0.02 at 220nm • Purity (GC): >99.5% • Refractive Index at 25°C: 1.342 • MW: 78.13 Water, HPLC Grade, Physical Properties • Density: 1.101 • UV Cutoff: 190nm • Water: ≤0.2% • Optical Absorbance: <0.005 at 220nm Pyridine • Refractive Index at 25°C: 1.332 • C5H5N Dimethylformamide (DMF), Sequencing Grade, Physical Properties • Purity (GC): ≥99% • HCON(CH3)2 • Density: 0.978 • Purity (GC): ≥99% • B.P. 115°C LC Reagents Acetonitrile, HPLC Grade, Physical Properties LC Columns and Accessories >> HPLC and Spectrophotometric Grade Solvents • MW: 79.10 • MW: 73.09 • Density: 0.944 • B.P. 153°C • Water: 0.1% HPLC and Spectrophotometric Grade Solvents Description Acetonitrile Water Dimethylformamide (DMF) Dimethyl sulfoxide (DMSO) Pyridine Quantity 1L 1L 50mL 950mL 100g X X X X Cat. No. TS-51101 TS-51140 TS-20673 TS-20688 TS-25104 Quantity 1 Each 1 Each 1 Each 1 Each 1 Each X in the ordering table indicates that hazardous shipping charges apply. www.thermoscientific.com/chromatography 4-205 LC Columns and Accessories Peptide Standards Peptide Retention Standard Allows accurate prediction of elution time for peptides of known amino acid composition up to 20 residues in length Save time in peptide purification • Simplify identification of specific peptides in a complex mixture • Increase the efficiency of predicting peptide elution profiles • Determine the relative order of peptide elution of a complex mixture • Predict the HPLC retention time for peptides of known amino acid composition on reverse phase HPLC columns • Monitor column performance: efficiency, selectivity and resolution during column ageing • Compare reverse-phase columns from different manufacturers • Evaluate reverse-phase supports of varying n-alkyl chain lengths and ligand densities • Retention times are predicted by totaling the values that represent the contribution in minutes of each amino acid residue and the peptide terminal groups. Retention time is dependent upon the molecular weight of the peptide. The effect on retention is relatively unimportant with a small peptide, but it increases with the size of the molecule. The accuracy of predicting peptide retention time significantly decreases beyond 20 residues. To ensure accuracy, a peptide standard is used to correct for instrument variation, column aging, n-alkyl chain length variation and ligand density. Peptide Retention Standard Description Peptide Retention Standard, S1-S5 Quantity 1 vial Cat. No. TS-31700 Quantity 1 Each 1. Guo, D., et al. (1985). Proceedings of the Ninth American Peptide Symposium, Published by Thermo Fisher Scientific, Rockford, Illinois, page 23. 2. Guo, D., et al. (1986). J. Chromatogr. 359, 499-517. 3. Guo, D., et al. (1986). J. Chromatogr. 359, 519-532. 4. Mant, C.T. and Hodges, R.S. (1986). L.C. Magazine Liq. Chrom. and HPLC 4(3), 250. 5. Guo, D., et al. (1987). J. Chromatogr. 386, 205-222. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Peptide Retention Time Calibration Mixture 4-206 Heavy peptide mixture for column assessment and prediction of peptide retention times • Assessment of chromatography and MS instrument performance • Prediction of peptide retention across multiple instrument platforms • Prediction of peptide retention time from sequence using calculated hydrophobicity factor • Optimization of scheduled MS acquisition windows for improving quantification and increased multiplexing • Internal standard to normalise for variation in retention times and peak intensities between runs Thermo Scientific Peptide Retention Time Calibration Mixture Description Peptide Retention Time Calibration Mixture, 0.5pmol/µL Peptide Retention Time Calibration Mixture, 5pmol/µL Quantity Cat. No. Quantity 50µL TS-88320 1 Each 200µL TS-88321 1 Each • The Dionex IonPac ready-to-use ion standards are designed for routine anion or cation determinations • All standards are traceable to NIST Standard Reference Materials • Shipped with a Certificate of Analysis verifying the concentration Description Combined Five Anion Standard Combined Seven Anion Standard I Combined Seven Anion Standard II Fluoride Standard (1000mg/L) Chloride Standard (1000mg/L) Sulfate Standard (1000mg/L) Combined Six Cation Standard-I Combined Six Cation Standard-II Volume (mL) 100 50 100 100 100 100 50 50 Cat. No. 037157 056933 057590 037158 037159 037160 040187 046070 • MS standards for validation and calibration of the MSQ System Description Kit of 2 standards for performance validation in ESI and APCI modes Standard for Mass Calibration (All MSQ Models) Cat. No. 061496 062917 LC Reagents MS Standard Concentrates LC Columns and Accessories >> Dionex Ion Standard Concentrates Perchlorate Internal Standard • For the quantification of perchlorate at low parts-per-trillion levels using mass spectrometric detection. Contains stable-labeled perchlorate (1mg/L). Description Perchlorate-ISTD. 18O Internal Standard for Perchlorate Cat. No. 062923 www.thermoscientific.com/chromatography 4-207 LC Columns and Accessories Haloacetic Acid Internal Standards • Ready-to-use internal standards for haloacetic acid analysis using electrospray-mass spectrometric detection • Stable-labeled internal standards prepared in MTBE (methyl-t-butyl ether), 1mL ampule plus empty vial Description Monochloroacetic Acid -2-13C Internal Standard, 1000mg/L in MtBE Monobromoacetic Acid -1-13C Internal Standard, 1000mg/L in MtBE Dichloroacetic Acid -2-13C Internal Standard, 1000mg/L in MtBE Trichloroacetic Acid -2-13C Internal Standard, 1000mg/L in MtBE Thermo Scientific Chromatography Columns and Consumables 2014-2015 Cat. No. 069406 069407 069408 069409 Ion Pairing Reagents • Highly purified ion-pairing reagents are used in mobile phase ion chromatography (MPIC), combining reversed-phase ion-pair chromatography with chemical suppression Description Tetrabutylammonium hydroxide (TBAOH) Tetrapropylammonium hydroxide (TPAOH) Hexanesulfonic acid (HSA) Octanesulfonic acid (OSA) 4-208 Volume (mL) 1 1 1 1 Volume (mL) 500 500 500 500 Cat. No. 035360 035363 035361 035362 Our liquid chromatographs and mass spectrometers advance scientific knowledge, enable drug discovery, improve manufacturing processes, and protect people and the environment. When your instrument is maintained with Thermo Scientific parts, you can expect the best results and highest productivity, keeping your research or processes moving smoothly. LC Columns and Accessories >> LC and LC-MS Instrument Accessories, Consumables and Spare Parts LC and LC-MS Instrument Parts www.thermoscientific.com/chromatography 4-209 LC Columns and Accessories Thermo Scientific Dionex UltiMate 3000 Systems Portfolio Basic Automated System The Thermo Scientific™ Dionex™ UltiMate™ 3000 Basic Automated System is optimized for reliability and ease-of-use with routine LC applications, and also offers full UHPLC compatibility. The ACC-3000 Autosampler Column Compartment is at the heart of the system. Its unique instrument design combines a rugged sample injection principle with a powerful column oven. Standard (SD) LC Systems Thermo Scientific Chromatography Columns and Consumables 2014-2015 UltiMate 3000 Standard (SD) Systems are designed to meet current and future challenges. System components provide low extra-column and gradient delay volume for high separation efficiency and low gradient response times, as well as superior mixing performance. They offer full support of all HPLC applications and provide UHPLC compatibility – allowing you to move to UHPLC whenever you are ready. 4-210 Rapid Separation (RS) LC Systems UltiMate 3000 Rapid Separation (RS) Systems provides unrivaled performance and flexibility in UHPLC. Precision-engineered instrumentation, advanced data processing, and highly optimized chemistries meet all chromatographic performance challenges. With an outstanding product portfolio, UltiMate 3000 RS Systems offers industry leading versatility covering the maximum range of UHPLC applications, while keeping fully compatible to conventional LC. UltiMate 3000 Biocompatible Rapid Separation (BioRS) Systems are powered by UltiMate 3000 Rapid Separation technology to support the high pressures required for the separation of bioanalytes on high resolution bio UHPLC columns. This state-of-the-art technology combined with a biocompatible, low-dispersion flow path, provides the highest peak capacity and sensitivity for complex samples, whether proteins, peptides, or biotherapeutics. Bio UHPLC enables faster gradients and shorter run times compared to conventional Bio LC analysis. This results in increased sample throughput in typical modes of biochromatography. x2 Dual Rapid Separation LC Systems Advanced chromatographic techniques such as parallel, tandem, and two-dimensional LC become a realistic and practical option, even at ultrahigh speed. Operate conventional or ultrahigh speed applications, increase sample throughput, achieve higher chromatographic resolutions, or automate tedious sample preparation steps such as analyte enrichment or matrix elimination. LC and LC-MS Instrument Parts UltiMate 3000 x2 Dual LC Systems are the ideal choice to maximize sample throughput of conventional HPLC and UHPLC separations by combining the flexibility of ternary solvent selection with the productivity and performance of x2 Dual technology. LC Columns and Accessories >> Biocompatible Rapid Separation (BioRS) LC Systems XRS LC Systems UltiMate 3000 XRS Systems are highly LC/MS optimized quaternary UHPLC systems and the best choice for ultrahigh resolution and high speed UHPLC MS-frontend solutions. This system is the ideal solution for high sample throughput and ultra high resolution chromatography. www.thermoscientific.com/chromatography 4-211 LC Columns and Accessories Accessories, Consumables and Spare Parts for UltiMate 3000 Instruments Accessories for the Use with UltiMate 3000 Instruments Description Drainage kit for UltiMate 3000 systems Menu pen Installation tool for Viper capillaries with torque toothing HPLC Troubleshooting Guide poster Quantity 1 1 1 1 Cat. No. 6040.0005 6300.0100 6040.2314 6040.0050 UltiMate 3000 Solvent Racks Consumables and Spare Parts for UltiMate 3000 Solvent Racks Description Solvent supply line (analytical) Solvent supply line from analytical degasser to pump head Bottle cap for solvent reservoirs Caps and retaining guides for solvent bottles (5 caps and 10 retaining guides) Degasser Channel Upgrade Kit for SRD External power supply unit Quantity 1 1 4 Cat. No. 6030.2548 6030.2546 6270.0013 1 6030.9101 1 1 6035.0089 6510.0004 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Also use our HPLC Troubleshooting Guide as a Mobile App for iPhone, Android and Windows phones. 4-212 Accessories for the Use with UltiMate 3000 Pumps Description Quantity Cat. No. Filter holder for solvent filter Filter holder for solvent filter Filter frit (SST, porosity: 10µm) Filter frit (Ti, biocompatible, porosity: 10µm) Filter frit (PEEK, porosity: 10µm) Solvent lines kit with filter frits (SST, porosity: 20µm) 6 2 10 10 10 1 6268.0115 6268.0116 6268.0110 6828.0111 6828.0117 6043.8109 SD pumps X RS pumps X X X X For use with BM pumps X XRS pumps BX pumps X X X X X Spare Parts for UltiMate 3000 Pumps Description RS pumps For use with BM pumps X X XRS pumps BX pumps Quantity Cat. No. 1 1 6041.2301 6043.0145 X 2 2 2 6040.0042 6040.0842 6043.0169 X 2 1 6040.0306 6040.9010 X 2 2 1 6040.0304 6266.0305 6025.2012 X 1 6040.9010 X 1 6040.9011 X 1 6043.0295 2 1 6040.0012 6025.2012 1 6040.9010 X 1 6040.9011 X X X X X X X X X X X X LC and LC-MS Instrument Parts Check Valves Check valve, cartridge (ceramics) for the inlet and outlet valve Check valve cartridge for the inlet and outlet valve Pistons Pistons (sapphire) Pistons (ceramics) Pistons (sapphire) Piston Seals Piston seal (in plate of seal wash system) Normal Phase Piston seal kit (in plate of seal wash system) Reversed Phase (2 seals and 1 support ring) Piston seal (main piston seal) Reversed Phase Piston seal (main piston seal) Reversed Phase Piston seal kit (main piston seal) Reversed Phase (2 seals and 1 support ring) Piston seal kit (main piston seal) Reversed Phase (2 seals and 1 support ring) Piston seal kit (main piston seal) Normal Phase (2 seals and 1 support ring) Piston seal kit Reversed phase (2 primary piston seals and 2 secondary piston seals) Support Rings Support ring for pistons seals Support ring/pistons seal kit Reversed Phase (2 piston seals and 1 support ring) Support ring/pistons seal kit Reversed Phase (2 seals and 1 support ring) Support ring/pistons seal kit Normal phase (2 seals and 1 support ring) SD pumps LC Columns and Accessories >> UltiMate 3000 Pumps X X X X www.thermoscientific.com/chromatography 4-213 LC Columns and Accessories Manual Sample Injection Valves for the Use with UltiMate 3000 Pumps Description Manual injection valve kit, pressures > 50 MPa Manual injection valve kit, biocompatible, pressures > 34 MPa Motorized manual injection valve kit Mounting kit for manual injection valve Syringe for use with the manual injection valve 100µL syringe 5mL syringe 1mL syringe Quantity 1 1 1 1 Cat. No. 6040.0610 6042.0600 6040.0110 6040.0611 1 1 1 6035.0665 6035.0670 6040.0620 Mixing Systems for the Use with UltiMate 3000 Pumps Description Mixer for 35µL mixing volume Mixer for 100µL mixing volume Mixer for 200µL mixing volume Mixer for 400µL mixing volume Mixer for 800µL mixing volume Mixer for 1550µL mixing volume 1 except ISO-3100 Quantity 1 1 1 1 1 1 SD pumps1 6040.5000 6040.5100 Cat. No. RS pumps 6042.5000 6042.5100 6040.5110 6040.5310 6040.5750 6040.5450 Normal Phase Upgrade Kits for UltiMate 3000 Pumps Description Normal Phase Kit for HPG-3200BX Normal Phase Kit for SD pumps Quantity 1 1 Cat. No. 6040.1975 6040.1972 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Maintenance Kits for UltiMate 3000 Pumps 4-214 Description Maintenance kit for ISO-3100 Quantity 1 SD pumps 6040.1950 RS pumps – BM pumps 6042.1950 XRS pumps – BX pumps – Maintenance kit for HPG-3x00 Maintenance kit for LPG-3400 1 1 6040.1953 6040.1951 6040.1956A 6040.1954A – 6042.1951 – 6043.1954 Maintenance kit for DGP-3600 1 6040.1952 6040.1955A 6042.1952 – – 6042.1954(NP) 6042.1953(RP) – Sample Racks for the Use with UltiMate 3000 WPS Autosamplers and UltiMate 3000 ACC Autosampler Column Compartments For use with WPS ACC Quantity Cat. No. 1 1 1 1 1 1 1 1 1 6820.4086 6820.4084 6820.4070 6820.4087 6820.4096 6820.4094 6820.4090 6820.4091 6820.4097 X X X X X X X X X 1 1 1 6820.4079 6820.4083 6820.4089 X X X X X X X X X X X X Find the Thermo Fisher Scientific Vials and Closures portfolio on page 2-001. www.thermoscientific.com/chromatography LC and LC-MS Instrument Parts Description Sample Racks Sample Rack (10 position and/or normal well plates) for 10mL vials, 22.5mm×46mm Sample Rack (22 position) for 4mL vials, 15mm×45mm, cylindrical Sample Rack (40 position) for 2mL vials, 12mm×32mm, cylindrical Sample Rack (40 position and/or normal well plates) for 1.1mL vials, 12mm×32mm, conical Sample Rack (40 position) for 0.5mL Eppendorf tubes Sample Rack (40 position) for 1.5mL Eppendorf tubes Sample Rack (72 position and/or normal well plates) for 1.2mL vials, 8mm×40mm, cylindrical Sample Rack (72 position and/or normal well plates) for 0.3mL vials, 8mm OD, cylindrical Sample Rack (72 position) for 0.3mL micro dialysis vials Support Racks for Well Plates Support rack for deep well plates (34 – 46mm high) Support rack for deep well plates (30 – 36mm high, 96 or 384 wells) Support rack for deep well plates (20 – 32mm high, 96 or 384 wells) LC Columns and Accessories >> UltiMate 3000 Autosamplers 4-215 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-216 Optional Accessories for the Use with UltiMate 3000 WPS-3000 Split Loop Autosamplers For use with Description Needles Needle (SST) Viper compatible Needle (ceramic) Viper compatible Needle (SST) Needle (SST) Needle Seat Capillaries Needle seat capillary, 3.1µL, Viper (SST) Needle seat capillary, 3.1µL, Viper (PEEK) Needle seat capillary, 3.1µL, Viper (MP35N ™) Needle seat capillary, 1µL, Viper (MP35N) Needle seat capillary, 1µL, Viper (SST) Needle seat capillary, 24µL, Viper (SST) Sample Loops - SST 25µL Sample loop, Viper 100µL Sample loop, Viper 250µL Sample loop, Viper 500µL Sample loop, Viper 1000µL Sample loop Sample Loops – MP35N 25µL Sample loop, Viper 100µL Sample loop, Viper 250µL Sample loop, Viper 500µL Sample loop, Viper Sample Loops – PEEK 100µL Sample loop Buffer Loops Buffer loop, 100µL, Viper (SST) Buffer loop, 100µL, Viper (MP35N) Buffer loop, > 250µL, Viper (SST) Buffer loop, > 250µL, Viper (MP35N) Syringes 25µL Syringe 100µL Syringe 250µL Syringe 500µL Syringe 1000µL Syringe 2500µL Syringe WPS3000TXRS/ WPS-3000(T) RS Quantity Cat. No. 1 1 1 1 6820.2432 6841.2420 6820.2403 6820.2419 X/X 1 1 1 6820.2464 6827.2408 6841.2464 X/X 1 1 1 6841.2472 6820.2472 6820.2409 1 1 1 1 1 6820.2452 6820.2451 6820.2453 6820.2454 6820.2429 1 1 1 1 6841.2452 6841.2451 6841.2453 6841.2454 1 6820.2431 1 1 1 1 6820.2466 6841.2466 6820.2468 6841.2468 X/X 1 1 1 1 1 1 6822.0001 6822.0002 6822.0003 6822.0004 6822.0005 6822.0006 X/X X/X X/X X/X X/X X/X WPS3000TBRS WPS-3000(T) SL Analytical WPS3000TBSL WPS-3000(T) SL Semiprep X X X X X X X X X/– X X/X X/X –/X –/X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X Injection Volume Kits for the Use with UltiMate 3000 WPS-3000 Split Loop Autosamplers Description Quantity Cat. No. Injection volume kit (250µL), Viper Injection volume kit (250µL), Viper Injection volume kit (500µL), Viper Injection volume kit (500µL), Viper Injection volume kit (1000µL) 1 1 1 1 1 6822.2442 6841.2442 6822.2443 6841.2443 6822.2436 WPS-3000(T)RS X X For use with WPS-3000(T)SL WPS-3000TBRS Analytical X X X X WPS-3000(T)SL Semiprep X Description Quantity Cat. No. 1 1 1 1 6824.0030 6825.0030 6821.0045 6820.1427 X X 1 1 1 1 6820.3010 6820.3023 6820.3025 6820.3115 X X X2 X2 1 1 1 1 1 1 6826.2401 6826.2405 6826.2410 6826.2420 6826.2450 6826.2412 X X X X X X 1 1 6824.0019 6824.0020 1 1 1 1 1 1 6823.0016 6823.0017 6823.0018 6823.0019 6821.0032 6823.0020 X X X X2 X2 1 1 1 1 1 1 1 1 6820.0019 6821.0019 6820.0020 6821.0020 6820.0056 6821.0022 6820.0021 6821.0021 X X3 X X3 X X3 1 1 1 1 1 1 1 6822.0001 6822.0002 6822.0003 6822.0004 6822.0005 6820.0031 6821.0031 X X X4 X4 X4 X X3 1 1 1 1 6824.0015 6825.0015 6824.0016 6825.0016 WPS-3000TBFC X X X X X X X X X1 X1 X X X1 X1 X X X X X1 X1 X1 X1 X X X1 X1 X1 X1 X X X X X X X X X X X X X X X X LC and LC-MS Instrument Parts Modification Kits Nano/Cap modification kit Nano/Cap modification kit Upgrade kit PAEK Front cover, transparent Needles 2.4µL Needle, nanoViper 3.6µL Needle, Viper 15µL Needle, Viper 15µL Needle, nanoViper Sample Loops – nanoViper 1µL Sample loop, nanoViper 5µL Sample loop, nanoViper 10µL Sample loop, nanoViper 20µL Sample loop, nanoViper 50µL Sample loop, nanoViper 2 125µL Sample loop, nanoViper 2 Sample Loops – SST 50µL Sample loop 250µL Sample loop Sample Loops – PEEK 5µL Sample loop 10µL Sample loop 20µL Sample loop 50µL Sample loop 125µL Sample loop 250µL Sample loop Buffer Tubings 50 µL Buffer tubing (SST) 50 µL Buffer tubing (PEEK/PTFE) 500 µL Buffer tubing (SST) 500 µL Buffer tubing (PEEK/PTFE) 1000 µL Buffer tubing (SST) 1000 µL Buffer tubing (PEEK/PTFE) 2000 µL Buffer tubing (SST) 2000 µL Buffer tubing (PEEK/PTFE) Syringes 25µl Syringe 100µl Syringe 250µl Syringe 500µl Syringe 1000µl Syringe Syringe upgrade kit (250µL) 4 Syringe upgrade kit (250µL, biocompatible version) 4 Bridges 1µL Bridge (PEEKsil) 1µL Bridge (PEEKsil) 6.2µL Bridge (PEEK) 6.2µL Bridge (PEEK) For use with WPS-3000TBPL WPS-3000TFC Analytical WPS-3000(T)PL RS Nano/Cap LC Columns and Accessories >> Optional Accessories for the Use with UltiMate 3000 WPS-3000 Pulled Loop Autosamplers X1 X1 X X X X X X X X X1 X X X1 X X X1 X1 X X Find recommended combinations for syringes, buffer tubings and sample loops on page 4-218. 1. Can be used for Nano/Cap/Mic applications only if Nano/Cap modification kit is previously installed 2. For analytical applications only, please use the smaller sample loops for Nano/Cap 3. Can be mounted only when PAEK Upgrade Kit is previously installed 4. For analytical applications only, please use the smaller syringes for Nano/Cap www.thermoscientific.com/chromatography 4-217 LC Columns and Accessories Recommended Syringes, Buffer Tubings and Sample Loops Combinations for the Use with UltiMate 3000 WPS-3000(T)PL RS Nano/Cap, WPS-3000(T)BPL Analytical, WPS-3000TFC and WPS-3000TBFC Autosamplers Recommended Combinations Syringe Buffer Tubing Sample Loop WPS-3000(T)PL RS Nano/Cap 25µL 50µL 1µL, 5µL 100µL 500µL 10µL, 20µL 250µL 500µL 50µL, 125µL WPS-3000TBPL Analytical 100µL 500μL 5µL, 10µL, 20µL, 50µL 250µL 1000µL 125µL, 250µL WPS-3000TFC and WPS-3000TBFC Analytical (with Nano/Cap Modification Kit) 25µL 50µL 1µL, 5µL 100μL 500μL 5µL, 10µL, 20µL, 50µL 250µL 500µL 50µL, 125µL 250µL 1000µL 250µL Accessories and Spare Parts for the Use with UltiMate 3000 OAS-3x00 Open Autosamplers Description Holding loop (FEP) with needle adapter mounted Holding loop kit (FEP) Holding loop (SST) 2µL Sample loop for Cheminert injection valve 20µL Sample loop for Cheminert injection valve 100µL Sample loop for Cheminert injection valve 100µL Syringe Sample tray (VT54) for 2mL vials (54 positions per tray) Quantity 1 1 1 1 1 1 1 6 Cat. No. 6845.0034 6845.0035 6845.0029 755.CSL2 755.CSL20 755.CSL100 6845.0062 6845.0055 Quantity 1 1 1 1 1 1 1 1 1 1 Cat. No. 6820.4092 6820.2402 6820.2403 6820.2468 6830.2414 6830.2416 6830.2418 6822.0003 6822.0005 6820.1427 Thermo Scientific Chromatography Columns and Consumables 2014-2015 Accessories and Spare Parts for the Use with ACC-3000(T) Autosampler Column Compartments 4-218 Description Adapter for 5-position holder to be used for 2mL vials Adapter for vial pusher for uncapped 10mL vials Needle Buffer tubing, >250µL, Viper 20µL Sample loop, Viper 50µL Sample loop, Viper 200µL Sample loop, Viper 250µL Syringe 1000µL Syringe Front cover, transparent Accessories for the Use with UltiMate 3000 TCC-3000(RS) Thermostatted Column Compartments Description 1µL Pre-column heater, Viper (SST) 2µL Pre-column heater, Viper (SST) 7µL Pre-column heater, Viper (SST) 11µL Pre-column heater, Viper (SST) Post-column cooler insert, Viper (SST) Column clip for columns with an outer diameter < 8mm Column clip for columns with an outer diameter of 8-12mm Quantity 1 1 1 1 1 6 6 Cat. No. 6722.0510 6722.0530 6722.0540 6722.0550 6730.0008 6722.0290 6722.0280 Optional Column Switching Valves for the Use with UltiMate 3000 TCC-3000RS/SD Thermostatted Column Compartments Valve Drives Valve Drive (Actuator) for right side installation Valve Drive (Actuator) for left side installation Pods Pod, 2-position, 6-port Quantity TitanHT ™ < 103 MPa (15000 psi) TitanHP ™ < 41 MPa (6000 psi) Non biocompatible TitanHT ™ < 103 MPa <125 MPa (15000 psi) (18130 psi) 1 6730.0003 6730.0001 6730.0003 6730.0001 1 6730.0004 6730.0002 6730.0004 6730.0002 6730.0031 (Ti) 6730.0032 (Ti) 6730.0030 (Ti) 6722.9013 (SST) 6722.9023 (SST) – 1 Pod, 2-position, 10-port 1 Pod, 6-position, 7-port 1 6723.9013 (PEEK) 6723.9023 (PEEK) – – – 6722.9035 (Ti) 6730.0006 (SST) 6730.0026 (SST) 6730.0016 (SST) – – LC and LC-MS Instrument Parts Description Biocompatible TitanHP ™ < 34 MPa < 41 MPa (5000 psi) (6000 psi) LC Columns and Accessories >> UltiMate 3000 Thermostatted Column Compartments 6730.0050 (SST) www.thermoscientific.com/chromatography 4-219 LC Columns and Accessories UltiMate 3000 Detectors Flow Cells for UltiMate 3000 DAD-3000(RS), MWD-3000(RS), VWD-3x00(RS) and FLD-3x00(RS) Detectors Description Quantity Cat. No. Analytical flow cell (13µL, SST, 12 MPa) Semi-analytical flow cell (5µL, SST, 12 MPa) Semi-micro flow cell (2.5µL, SST, 12 MPa) Analytical flow cell (13µL, PEEK, 5 MPa) Semi-micro flow cell (2.5µL, PEEK, 5 MPa) Semipreparative flow cell (0.7µL, PEEK, 10 MPa) Flow cell, analytical (11µL, SST, 12 MPa) Flow cell, semi-micro (2.5µL, SST, 12 MPa) Flow cell, analytical (11µL, PEEK, 5 MPa) Flow cell, semi-micro (2.5µL, PEEK, 5 MPa) Flow cell, semi-preparative (0.7µL, PEEK, 10 MPa) Flow cell, Mic (180nL, fused silica, 20 MPa) Flow cell, capillary (45nL, fused silica, 20 MPa) Flow cell, nano (3nL, fused silica, 20 MPa) UV monitor, ultra-low dispersion (45nL, fused silica, 30 MPa) Flow cell, dummy Flow cell, analytical (8µL, SST, 2 MPa) Flow cell, micro (2µL, SST, 4 MPa) 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 6082.0100 6082.0200 6082.0300 6082.0400 6082.0500 6082.0600 6074.0250 6074.0360 6074.0200 6074.0300 6074.0320 6074.0290 6074.0280 6074.0270 6074.0285 6074.0190 6078.4230 6078.4330 DAD-3000(RS) MWD-3000(RS) X X X X X X VWD-3x00(RS) FLD-3x00(RS) X X X X X X X X X X X X Thermo Scientific Chromatography Columns and Consumables 2014-2015 Accessories and Spare Parts for UltiMate 3000 Optical Detectors 4-220 Description Quantity Cat. No. Deuterium lamp Tungsten lamp DAC board DAC cable (analog out) 1 1 1 1 6074.1110 6074.2000 6082.0305 6074.0002 DAD-3000(RS) MWD-3000(RS) X X X X VWD-3x00(RS) X X X X FLD-3x00(RS) X LC Columns and Accessories >> Accessories and Spare Parts for the Use with the UltiMate 3000 PCM-3000 pH and Conductivity Monitor Quantity 1 1 1 1 Cat. No. 6082.2030 6082.2020 6082.2042 6082.2060 Description In-Line Filter Kit with Graphite Filter Elements In-Line Filter Kit with PEEK Filter Elements Filter Element (Graphite) Filter Element (PEEK) Amperometric Cell 6041RS ultra (25nL or 50nL, glassy carbon or boron-doped diamond, 1.4 MPa) Quantity 1 1 5 5 1 Cat. No. 70-0893 70-4093 70-0898 70-3824 6070.3000 Gasket for Amperometric Cell 6041RS (25nL, boPET) Gasket for Amperometric Cell 6041RS (50nL, boPET) Working Electrode for Amperometric Cell 6041RS (boron-doped diamond, BDD) Working Electrode for Amperometric Cell 6041RS (glassy carbon, GC) Coulometric Cell 6011RS ultra (7.06µL, micro-porous graphite carbon, 4 MPa) 5 5 1 6070.2528 6070.2529 6070.3100 1 6070.3200 1 6070.2400 Potentiostat Module, dual channel DC Cell Simulator – SimulatorRS Cell Simulator – QualifierRS 1 1 1 6070.1400 6070.4100 6070.4200 Description pH flow cell, 28µL (PEEK) pH electrode for PCM-3000 Dummy electrode for PCM-3000 Conductivity flow cell, 21µL (Ti) Accessories and Spare Parts for the Use with the UltiMate 3000 ECD-3000RS Detectors LC and LC-MS Instrument Parts Accessories and Spare Parts for the Use with Charged Aerosol Detectors Description Gas Filter Assembly (HEPA/Carbon) Corona Waste Bottle Cap Assembly Bottle, 5.0 L Waste (Compatible with 70-7754 only) Filter-HPLC Inline Drain/Vent Tubing Assembly Gas Inlet Tubing (¼") Gas Exhaust Hose Assembly Maintenance Kit for Nitrogen Generator Corona ultra RS Waste Level Detector Assembly Quantity 1 1 1 1 1 1 1 1 1 Cat. No. 70-6224 70-7754 70-7751 70-4538 70-7115 70-6260 70-6261 70-6230 70-9363R Corona CAD X X X X X X X X Corona ultra RS X X X X X X X X X www.thermoscientific.com/chromatography 4-221 LC Columns and Accessories UltiMate 3000 Automated Fraction Collector Sample Racks and Accessories for the Use with the UltiMate 3000 AFC-3000 Description Sample Racks Sample Rack (10 position) for 10mL vials, 22mm × 45mm Sample Rack (22 position) for 4mL vials, 15mm × 45mm Sample Rack (40 position) vfr 2mL vials, 12mm × 32mm Sample Rack (21 position) for 50mL tubes, 30mm × 100mm Sample Rack (24 position) for 30mL tubes, 24mm × 100mm Sample Rack (40 position) for 20mL tubes, 20mm × 100mm Sample Rack (60 position) for 14mL tubes, 16mm × 100mm Sample Rack (90 position) for 8mL tubes, 13mm × 100mm Sample Rack (36 position) for Foxy vials, scintillation vials, 28mm OD Sample Rack (60 position) for Foxy vials, 1.5mL Sample Rack (72 position) for Foxy mini tubes, 18mm OD Positioning pins for Foxy racks Funnel rack Adapter for 4 WPS racks Adapter for 4 well plates Adapter for 2 well plates (Foxy Jr./R1) Accessories Diverter valve, 0.4mm ID drop former, and tubing (“Kit for low flow rates”) Drop former, 1mm ID SST, including grounding cable (“Kit for normal-phase LC”) Thermo Scientific Chromatography Columns and Consumables 2014-2015 Find the Vials and Closures portfolio on page 2-001. 4-222 Quantity Cat. No. 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 6820.4086 6820.4084 6820.4070 6702.0021 6702.0024 6702.0040 6702.0060 6702.0090 5701.2025 5701.2023 5701.2024 6702.9006 6702.1021 6702.0100 6702.0200 5701.2021 1 6702.0300 1 6702.0400 The UltiMate 3000 RSLCnano System was developed with throughput in mind. The robust, continuous direct-flow delivery is designed for interruption-free analysis. The wide flowpressure footprint enables the application of UHPLC to the nano scale, allowing analysts to tune for the highest resolution or the fastest analysis time, even for tryptic peptide samples of utmost complexity. Configurable for speed, separation power, or sensitivity, the UltiMate 3000 RSLCnano is the only system to deliver all. Consumables and Spare Parts for UltiMate 3000 RSLC Nano Systems Quantity 1 1 1 1 1 1 1 1 1 1 1 2 1 2 2 1 1 1 1 Cat. No. 6042.1951 6041.2301 6041.0004 6041.0001 6041.7901A 6041.7902A 6041.7903A 6041.0002 6041.0003 6041.0014 6041.7130 6040.0042 6040.0306 6266.0305 6040.0012 6041.1901A 6041.1902 6040.2208 6041.0011 2 6040.3023 LC and LC-MS Instrument Parts Description Maintenance kit for NCS-3500RS loading pump Check valve cartridge, ceramics (for both the inlet and outlet valve) for the NC pump and loading pump Column switching valve; 2-position, 6-port valve Column switching valve; 2-position, 10-port valve Flow meter, biocompatible with flow selector for nano LC (50 – 1000nL/min) Flow meter, biocompatible with flow selector for capillary LC (0.5 – 10µL/min) Flow meter, biocompatible with flow selector for micro LC (5 – 50µL/min) Flow selector for nano LC (50 – 1000nL/min) Flow selector for capillary LC (0.5 – 10µL/min) Flow selector for micro LC (5 – 50µL/min) Mixer kit (8µL) for capillary LC Piston (sapphire) Piston seal (in plate of seal wash system), NP, for NC pump and loading pump Piston seal (main piston seal), RP, for NC pump and loading pump Support ring for main piston seal for NC pump and loading pump Pump head, entire assembly, for NC pump Pump head, entire assembly, for loading pump Rear seal wash system, seals Column installation/capillary clips kit (6 column clips, 2 clips to attach capillaries routed from top to bottom through the column chamber) Tubing from proportioning valve to pump head, loading pump (with fittings) LC Columns and Accessories >> UltiMate 3000 RSLC Nano System EASY-nLC 1000 Liquid Chromatograph Thermo Scientific™ EASY-nLC 1000 Liquid Chromatograph provides effortless split-free, nano-flow UHPLC performance up to 1000 bar with minimal investment. Dual in-line flow sensors before mixing gives uncompromising gradient precision. Identify more peptides and increase productivity with higher pressure without compromising reliability and robustness. Common replacement parts for EASY-nLC 1000 Systems Description Pump Piston Seal Replacement Kit (contains four spring-energized piston seals and a piston seal tool) Valve rotor seal (contains one rotor seal) Autosampler needle, ASC model Column Out solvent line Waste In solvent line Flow sensor filters (contains four flow sensor filters) Cat. No. LC510 LC228 LC302 LC560 LC562 LC542 www.thermoscientific.com/chromatography 4-223 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-224 Accessories, Consumables and Spare Parts for Thermo Scientific Accela High Speed LC Accela Accessory Kits Description Accela Pump accessory kit LC/MS system solvent interconnect kit Kit, maintenance, Accela Pump Kit, Seal, Accela Pump Accela Autosampler Accessory Kit Accela PDA Detector accessory kit Quantity 1 1 1 1 1 1 Cat. No. 60157-62001 F5050-010 60157-62002 60157-62003 60357-62001 60257-62001 Quantity 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Cat. No. 60357-60017 60053-60102 60053-60041 A3692-010 60053-60014 60053-20002 A4258-010 00301-01910 60357-62002 60053-62043 60053-62009 00101-08-00001 00101-08-00002 1611-0030 00101-07-00001 60053-20031 F1100-010 F1100-020 F1100-030 F1123-010 F1123-020 F1123-030 00301-07527 60053-10035 60053-62002 60053-60006 00109-99-00007 00109-99-00008 00109-99-00009 00109-99-00010 00109-99-00011 00109-99-00012 00109-99-00013 00109-99-00014 F1034-010 00110-03-00013 00109-99-00021 00110-03-00019 60053-60005 Accela Autosampler Consumables Description Assembly, needle, inert Assembly, needle tubing Assembly, wash bottle kit Assembly, syringe valve Assembly, transfer tube, 0.012in ID Cooling Adapter, 96 well Filter, flush solvent Grease, Silicon/Teflon (for lead screw) Kit, maintenance Accela Autosampler Kit, needle tubing Kit, wash bottle Ferrule Front, Swagelok Ferrule Back, Swagelok Lubricant, Triflow Nut, compression, long, 10-32, 1/16in OD tube Port, needle Syringe, concentric, 100µL Syringe, concentric, 250µL Syringe, concentric, 500µL Plunger, replacement, concentric, 100µL Plunger, replacement, concentric, 250µL Plunger, replacement, concentric, 500µL Reservoir vials, 16mL Retainer, needle port (injection port) Syringe, standard, 2500µL (with needle tubing ext) Syringe, 2.5mL Sample Loop, 5µL Sample Loop, 10µL Sample Loop, 20µL Sample Loop, 25µL Sample Loop, 50µL Sample Loop, 100µL Sample Loop, 500µL Sample Loop, 1000µL Stripper Valve, injector Seal, Rotor Rotor, Valco injection valve Tubing assembly, 2.5mL syringe Cat. No. 00950-01-00311 00950-01-00317 00950-01-00322 00950-01-00323 00950-01-00324 00950-01-00325 00950-01-00326 00950-01-00327 00950-01-00328 00950-01-00330 00950-01-00334 00950-01-00335 00950-01-00336 00110-03-00019 00950-01-00337 Kit Description Kit, asp tube, Accela Open A/S, RoHS PAL DilAspKit Loop, hldg, dlw, Accela Open A/S, RoHS PAL DLWLoop Screw, syringe, Accela Open A/S, RoHS PAL DLW Screw Tool, needle, Accela Open A/S, RoHS PAL DLW NdTool Holder, needle, dlw, Accela Open A/S, RoHS PAL DLWNHA Syringe, dlw, Accela Open A/S, RoHS SyrC DLW 100-R Plunger, syringe, dlw, Accela Open A/S, RoHS PLG DLW 100 Holder, needle, dlw, Accela Open A/S, RoHS PAL DLWPlg Needle kit, dlw, 3PK, Accela Open A/S, RoHS PAL DLWNdl Kit, dlw replament, Accela Open A/S, RoHS PAL DLW Option Kit, insert, dlw, Accela Open A/S, RoHS PAL DLW Insert Tube, waste, dlw, Accela Open A/S, RoHS PAL TubeWaste Valve, inject, 18K, Accela Open A/S, RoH N/A Rotor, Inj Valve, VALCO 18KPSI, RoHS C72-16R6 Stator Inj Valve, VALCO 18KPSI, RoHS C72V-6C96 Accela PDA Accessories Quantity 1 Cat. No. 803264 1 1 1 1 1 1 803265-01 803237-01 802259 2522-0285 803260 703950 Quantity 4 2 1 1 1 2 2 Cat.No. 00950-01-00122 00950-01-00129 00950-01-00131 00950-01-00130 00950-01-00120 00950-01-00126 00950-01-00128 Accela Pump Liquid Displacement Assembly Parts Description Seal, film, PEEK, Accela pump Seal, high pressure, GFP, Accela pump Valve, check, outlet, Accela pump Valve, check, inlet, Accela pump Cartridge, blind, inlet, Accela pump Piston, sapphire Secondary piston seal (wash seal) Material PEEK PTFE/GFP/Ti Ti/ruby/sapphire Ti/ruby/sapphire PEEK Sapphire PE/SS LC and LC-MS Instrument Parts Description Filter Wheel for linearity calibration, (5 position; 1 cuvette with perchloric acid blank and 4 cuvettes with different concentrations of potassium dichromate in perchloric acid solution, NIST traceable) 10mm flowcell assembly, with inlet/outlet tubing and fittings (1cm LightPipe) Flowcell Assembly, with inlet/outlet tubing and fittings (5cm Lightpipe) Backpressure regulator FingerTight PEEK Ferrule Nuts Inlet tubing, with insulation, PEEK 1/16 x 0.005in ID (red) Outlet tubing, PEEK 1/16 x 0.01in ID (blue) LC Columns and Accessories >> Accela Open Autosampler Consumables Accela 1250 Pump Liquid Displacement Assembly Description UHP PEEK sealing ring Check valve, Titanium and ruby Piston unit, sapphire Secondary piston seal (GFP) Primary piston seal (GFP) Quantity 5 2 2 2 2 Cat. No. 00950-01-00284 00110-05110 00950-01-00126 00950-01-00129 00950-01-00129 Quantity 1 1 1 2 2 2 5 2 Cat. No. 00950-01-00286 60157-62008 00950-01-00292 00107-18110 00201-11324 00110-05110 00950-01-10013 00107-18114 Accela 600 Pump Spare Parts Description O-ring, 2.69mm x 1.14mm, perfluor Kit, inlet tubing, Accela 600 pump Mixer, static, 65µL, Accela 600 pump, RoHS Seal, H/P Piston, 1/8", TZP Valve, check Seal, face, PEEK, check valve Seal, piston, secondary www.thermoscientific.com/chromatography 4-225 LC Columns and Accessories Accessories, Consumables and Spare Parts for Thermo Scientific LC-MS Systems Ion Traps and Orbitrap Series Thermo Scientific™ Orbitrap™ LC-MS systems are the recognized standards for high-resolution, accurate-mass measurements. Orbitrap LC-MS technology routinely delivers the superior mass resolution and mass accuracy necessary to reduce analysis times and increase confidence in results. This makes it the platform of choice for the most confident protein and metabolite identification, characterization and quantitation. Combined with superior dynamic range and unsurpassed sensitivity, Orbitrap systems are the only mass spectrometers capable of providing all four benefits at the same time, without compromise. With market and technology leadership for over 25 years, we offer ion trap LC-MS systems for every application and budget. Ion trap LC-MS systems offer unique capabilities such as MSn and datadependent analysis along with excellent full-scan sensitivity to provide routine detection and rapid identification of low-level analytes. Ion trap systems support a variety of applications from compound identification and routine HPLC detection through the most demanding analyses of low-level components in complex biological matrices. TSQ Series Triple Quadrupole LC-MS Robust triple-stage quadrupole LC-MS systems enable ultra-sensitive quantitation of target compounds in complex matrices. Thermo Scientific Chromatography Columns and Consumables 2014-2015 TSQ Series triple-stage quadrupole LC-MS systems offer a wide range of capabilities. From the value-conscious Thermo Scientific™ TSQ Endura™ LC-MS to the precise Thermo Scientific™ TSQ Quantiva™ LC-MS, our mass spectrometers couple perfectly with our HPLC, UHPLC, and nano-LCs to easily accommodate complex matrices encountered in a variety of applications. 4-226 MSQ Plus Single Quadrupole LC-MS Our single quadrupole LC-MS system offers superior ease-of-use and modest price and space requirements. The system can be used with both HPLC and IC systems in a wide range of applications and methodologies. API Source Components Description USI ESI probe USI APCI probe Capillary heater cage assembly, for LTQ & LTQ XL Capillary heater cage assembly, for the LXQ & LCQ Fleet Fittings, Ferrules, Sample Loops and Tubing Cat. No. OPTON-20011 OPTON-20012 97055-60040 97055-60181 Accessory Kits Description TSQ Quantum forepump kit (used with LTQ series) Hose and accessories kit, LTQ & LTQ XL Hose, single mechanical pump accessory, LXQ & LCQ Fleet LTQ series ship kit Accessory Kit, LTQ/LTQ XL Accessory Kit, LXQ & LCQ Fleet Cat. No. 70111-62014 97055-62007 97055-60135 70111-62033 97055-62044 97055-62045 Description LCQ chemicals kit, for LTQ & LCQ series Met-Arg-Phe-Ala, 20mg Ultramark 1621 Caffeine, 1mg/mL Reserpine, 1g Cat. No. 97000-62042 00301-07709 00301-12200 00301-12310 00301-12901 Pressure Gauges Description Ion gauge Ion gauge o-ring, 0.737id 3/32 THK Viton Cat. No. 00105-01525 00107-10056 Metal Needle Kits Description 32-Gauge metal needle kit for high flow LC flow rates between 5µL/min to 400µL/min Uses 32-gauge needle, (P/N 00950-00954) 34-Gauge metal needle kit for low flow LC flow rates between 500nL/min to 10µL/min Uses 34-gauge needle, (P/N 97144-20040) Cat. No. OPTON-53003 OPTON-30004 Cat. No. 00101-18080 00101-18081 00101-18195 00101-18198 00101-18200 00101-18075 2522-0066 2522-3830 00101-18114 00101-18116 00101-18120 00101-18196 00101-18199 00101-18182 00101-18202 00101-18204 00110-22026 00110-22012 00110-22028 207180 00110-22018 00110-22020 00110-22022 00106-10498 00106-10499 00106-10502 LC and LC-MS Instrument Parts Chemicals Description Fitting, HPLC, adaptor 10-32 x 1/4 PEEK Fitting, nut, fingertight, HPLC, 10-32, PEEK Fitting, fingertight, 2, Upchurch Fitting, flangeless, 1/8in, Delrin, green Fitting, flangeless, 1/8in, Delrin, blue Fitting, plug, 1/4-28, Tefzel Nut, LC, 1/16in, ss, Rheodyne Ferrule, LC, 1/16in, ss, Rheodyne Ferrule, 0.008in ID, Kel-F HPLC Ferrule, 0.012in ID, Kel-F HPLC Ferrule, 0.016in ID, PEEK, HPLC Ferrule, Fingertight 2, Upchurch Ferrule, 1/8in, Tefzel Fitting, grounding union, 1/16, ss Fitting, LC union, 0.010in orifice, PEEK Fitting, LC TEE union, 0.020in orifice, PEEK 5µL sample loop, ss 10µL sample loop, ss 20µL sample loop, ss 50µL sample loop, ss 100µL sample loop, ss 500µL sample loop, ss 1mL sample loop, ss Tubing, 0.15mm ID x 0.39mm OD fused-silica capillary for APCI sample tube Tubing, 0.10mm ID x 0.19mm OD fused-silica capillary for ESI sample tube Tubing, 0.05mm ID x 0.19mm OD fused-silica capillary for ESI sample tube for low flow up to 200µL/min Tubing, 0.1mm ID x 0.363mm OD fused-silica capillary for infusion line Tubing, 0.075mm ID x 0.3193mm OD fused-silica capillary for low flow applications instead of metal needle Teflon tube, .03in ID x 1/16in OD for syringe adapter assembly Tubing, 1in ID x 3/16in, Tygon Hose, 1.5in ID, PVC reinforced LC Columns and Accessories >> Ion Traps and Orbitrap Series 00106-10504 00106-10511 00301-22915 00301-22922 00301-24163 www.thermoscientific.com/chromatography 4-227 LC Columns and Accessories TSQ Series Triple Quadrupole LC-MS Chemicals Description Cesium iodide TSQ Quantum AM calibration kit Reserpine Polytyrosine-1,3,6 calibration solution (liquid) Polytyrosine-1,3,6 calibration standard (solid) FC43 calibration liquid Syringe Adapter Kit for infusion of calibrant Quantity 1 x 1g vial 1 1 x 1g vial 1 x 20mL 1 1 vial 1 Cat. No. HAZMAT-01-00004 70111-62029S 00301-12901 00301-22924 00301-22925 50010-30059 70005-62011 Quantity 1 1 1 1 1 1 1 1 1 Cat. No. 80100-20641 70005-20606 70111-20972 97055-20199 70111-20396 70111-20100 70005-20423 70005-20922 97055-20442 Ion Transfer Tubes, Seals and O-rings Description TSQ Quantiva TSQ Endura TSQ Quantum Access TSQ Quantum Ultra, Ultra AM & Ultra EMR TSQ Quantum Discovery Max TSQ Quantum, Quantum AM & Quantum Discovery TSQ Vantage, Vantage AM & Vantage EMR Seal for ion transfer tube (TSQ Quantiva) Graphite vespel o-ring (not TSQ Quantiva) Thermo Scientific Chromatography Columns and Consumables 2014-2015 Metal Needle Kits 4-228 Description Metal Needle Insert Standard Flow (TSQ Quantiva and TSQ Endura) Metal Needle Insert Low Flow (TSQ Quantiva and TSQ Endura) 32-Gauge Metal Needle Insert LC Flow rates between 5μL/min to 2000μL/min Contains Metal Needle PN 70005-20434 34-Gauge Metal Needle Insert LC Flow rates between 1μL/min to 10μL/min Contains Metal Needle PN 70005-20483 High flow metal needle kit Low flow metal needle kit High flow metal needle kit Low flow metal needle kit High flow metal needle kit Low flow metal needle kit H-ESI metal needle kit Quantity 1 Cat. No. 80000-60317 1 1 80000-60152 OPTON-53010 1 OPTON-53011 1 1 1 1 1 1 1 OPTON-20004 OPTON-20005 OPTON-20016 OPTON-20017 OPTON-53003 OPTON-30004 OPTON-20034 Description MS maintenance kit Ferrule, 0.008 ID, Kel-F (for fused silica sample tube) Tubing, fused-silica, 0.10 ID x 0.19 OD (fused silica sample tube) O-ring, Viton, 0.125 ID x 1/16 (for source mount assembly gas connections) Fitting, HPLC, 10-32, short one-piece, 10/pk, RoHS Oil, vacuum pump, 1 liter Syringe, 500µL, Gastight Tubing, PEEK, 0.005in ID x 1/16in OD, red Tube, Teflon, 0.03in ID x 1/16in OD (for syringe adapter) Polytyrosine-1,3,6 calibration standard (liquid) Screw, 6-32 x 3/8 (used to secure front panels to chassis) Screw, 6-32 x 5/16 (used to secure side panels and EMI shield to chassis) Screw, 4 x 6mm (used to secure PS2 and PS3 to chassis) Screw, 8-32 x 3/8 (used to secure top cover to chassis) O-ring, graphite Vespel (for ion transfer tube) Quantity 1 2 6ft 2 3 3 1 3ft 0.1ft 1 2 2 2 2 1 Cat. No. 70111-62032 00101-18114 00106-10499 00107-02550 00109-99-00016 00301-15101 00301-19016 00301-22912 00301-22915 00301-22924 00405-63266 00407-63205 00407-90000 00415-83206 97055-20442 Description ESI needle ESI needle seal H-ESI needle H-ESI needle seal Quantity 1 1 1 1 LC and LC-MS Instrument Parts ESI Needles and Needle Seal Cat. No. 00950-00990 00950-00952 97055-20273 97055-20271 Viper Fittings (for grounded union to sprayer/probe) Description Viper Capillary, 65 µm x 150 mm PEEK Viper Capillary, 130 µm x 150 mm PEEK Viper Kit, 65 µm x 150mm PEEK (5 capillaries) Viper Kit, 130 µm x 150mm PEEK (5 capillaries) LC Columns and Accessories >> MS Maintenance Kit Cat. No. 6041.5615 6041.5616 6041.5601 6041.5602 Pump Oil Description Pump oil for Sogevac sv65 mechanical pump Cat. No. HAZMAT-01-00063 www.thermoscientific.com/chromatography 4-229 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-230 MSQ Plus Single Quadrupole LC-MS MSQ Plus Kits Description Annual Maintenance Kit* Probe Heater Repair Kit – insulating teflon parts Surveyor MSQ Test Kit Quantity 1 1 1 Cat. No. 60111-62014 60111-62010 FM104284 * The Consumables Kit contains the non-liquid parts required for the upkeep of your MSQ Plus MS Detector including source O-rings, entrance cones, esi/apci capillaries, cone wash assembly, esi probe repair supplies, turbo oil wick Consumables and General Spares Description Probe repair supplies: APCI probe capillary ESI probe capillary PEEK tube insert For API probe Graphite 1/16in Vespel ferrule ESI ceramic sleeve Calibration solution MSQ version 1.4 and higher Calibration solution MSQ classic version 1.0 Titanium entrance cone complete with O-ring Cone wash nozzle assembly Cone wash nozzle O-ring Entrance cone spare O-ring Exit (Extraction) cone Swage connector for 6mm nitrogen tubing 6mm x 1/4in NPT nitrogen fitting 6mm Teflon tubing for nitrogen supply Pfeiffer turbo replacement oil wick Edwards vacuum pump oil ESI probe complete APCI probe complete Probe heater assembly Vacuum exhaust hose, blue, 1in ID Hexapole screws – 3 required USB cable 2m APCI corona pin needle Quantity 1 1 1 pk (12) 1 pk (10) 1 1 1 1 1 1pk (10) 1pk (10) 1 1 1 Per foot 1 1 liter 1 1 1 Per foot 1 1 1 Cat. No. FM102594 FM102598 FM102591 6070119 FM103394 60111-01001 FM104285 60111-60049 FM102521 FM101464 5711000 FM102263 00101-02-00006 00103-02-00001 00109-99-00004 00950-01116 00301-15102 FM102595 FM102587 60111-60023 00301-08301 60111-20055 00302-99-00008 70005-98033 Software Description Upgrade to MSQ 2.0 with Xcalibur 2.0 includes LC Devices 2.02 Validation MSQ Plus on Xcalibur 2.0 – MS only Cat. No. OPTON-20432 OPTON-09015 Method Transfer to Accucore 2.6 μm Columns Containing solid core particles, which are engineered to a diameter of 2.6μm and a very narrow particle size distribution; Accucore HPLC columns allows high speed, high resolution separation, with back pressures significantly lower than those associated with sub-2μm fully porous particles. An understanding of some practical calculations can help to achieve the correct scaling and maintain a consistent assay profile between the original and transferred method. An identical approach can be used when transferring a conventional HPLC analysis to a UHPLC method using columns packed with sub-2μm fully porous particles such as Hypersil GOLD 1.9μm and Syncronis 1.7μm. There are three main considerations when transferring a method to a shorter column using different particles: Scaling the flow rate, adjusting the injection volume and adjusting the gradient profile. These are discussed in more detail below.* 3. Adjust the Gradient Profile To maintain an equivalent separation when transferring a method it is important to keep the linear velocity constant between the original and new method. The linear velocity is related to the flow rate, internal diameter of the column and particle size. A simple equation can be derived to calculate the flow rate (F2) required for the new method. This is shown below, normalized for particle size. Geometrical transfer of the gradient requires calculation of the number of column volumes of mobile phase in each segment (time interval) of the gradient in the original method to ensure that the new calculated gradient takes place over the same number of column volumes, for the new column. F2 = F1 x (dc22 / dc12) x (dp1 / dp2) F1 – original flow rate (mL/min) dc1 – original column internal diameter (mm) dp1 – original column particle size (µm) dc2 – new column internal diameter (mm) dp2 – new column particle size (µm) 2. Adjust the Injection Volume Because sub-3μm solid core based methods are most often transferred to smaller volume columns, the same injection volume will take up a larger proportion of the new column, possibly leading to band broadening or potentially overloading the column. It is therefore important to scale down the injection volume to match the change in column volume. Once again, a simple equation can be used to calculate the injection volume (Vi2) required for the new method. Vi2 = Vi1 x (dc22 x L2 / dc12 x L1) Vi1 – original injection volume (µL) dc1 – original column internal diameter (mm) L1 – original column length (mm) Vi2 – new injection volume (µL) dc2 – new column internal diameter (mm) L2 – new column length (mm) The following calculation should be performed for each time segment of the gradient, including column re-equilibration. It takes into consideration the void volume of each column (Vc, calculation described below), the flow rate in the original method and the flow rate in the new method (calculated in step 1 above) and the time segment in the original method. tg2 = tg1 x (Vc2 /Vc1) x (F1/F2) tg1 – Time segment in original gradient (min) tg2 – Time segment in new gradient (min) Vc1 – Original column void volume (mL) Vc2 – New column void volume (mL) F1 – Original flow rate (mL/min) F2 – New flow rate (mL/min) Technical Information When transferring methods from conventional HPLC to a solid core column several approaches can be taken, depending on the analytical needs. If column dimensions are maintained and the particle characteristics, design and diameter, are changed then an improvement in efficiency and, therefore, sensitivity, resolution and peak capacity is obtained. A second approach is to change not only particle characteristics but also column dimensions, which has the benefit of further reducing analysis time and increasing sensitivity. 1. Scale the Flow Rate LC Columns and Accessories >> LC Technical Information The void volume of the column is the volume that is not taken up by the stationary phase (approximately 68% of the column volume): Vc = 0.68 x π x r2 x L Vc – column volume (mL) L – column length (cm) r – column radius (cm) An example of a method transferred following steps 1 to 3 above is illustrated in the following table: www.thermoscientific.com/chromatography 4-231 LC Columns and Accessories Column: Fully porous 5 µm, 150 x 4.6mm Flow rate (mL/min) Inj. volume (µL) Gradient Time (min) 0.0 10.0 11.0 17.0 Backpressure Resolution Run Time (minutes) Solvent Used (mL) 1.00 1 %B 35.0 60.0 35.0 35.0 59 2.6 17.0 17 To obtain the best data using fast chromatography it is critical that the LC instrument system is optimized to operate under these conditions. All system components for the assay should be considered. System volume (connecting tubing ID and length, injection volume, flow cell volume in UV) must be minimized, detector time constant and sampling rate need to be carefully selected, and when running fast gradients pump dwell volume needs to be minimal. Minimizing System Volume Thermo Scientific Chromatography Columns and Consumables 2014-2015 0.4 1.4 %B 35.0 60.0 35.0 35.0 218 2.5 6.0 2.4 Method transfer conditions from HPLC (150 x 4.6mm, 5μm columns) to Accucore (100 x 2.1mm, 2.6μm and 50 x 2.1mm, 2.6μm columns). *We offer a convenient method transfer calculator at the Chromatography Resource Center (www.thermoscientific.com/crc) System Considerations 4-232 Column: Accucore RP-MS 2.6 µm, 100 x 2.1mm Flow rate (mL/min) Inj. volume (µL) Gradient Time (min) 0.0 3.5 3.8 6.0 Backpressure Resolution Run Time (minutes) Solvent Used (mL) Excess system volume gives rise to band broadening, which has a detrimental effect on the chromatographic performance. This can arise from the column, the autosampler, the tubing connecting the column to injector and detector and in the detector flow cell. The extra column effects become more significant for scaled down separations because of the smaller column volumes and for less retained peaks which have a lower peak volume making it even more critical to minimize extra column dispersion. Detector Sampling Rate With 1.9µm particles, operating parameters can be optimized to give fast analysis. This results in narrow chromatographic peaks which may be of the order of 1-2 seconds or less in width. It is important to scan the detector (whether it is UV or MS) fast enough to achieve optimum peak definition, otherwise resolution, efficiency and analytical accuracy will be compromised. Dwell Volume The HPLC pump dwell volume is particularly important when running high speed applications using fast gradients, typical of high throughput separations on small particle packed columns. This is because the pump dwell volume affects the time it takes for the gradient to reach the head of the column. If we consider a method using a flow rate of 0.4mL/min and a fast gradient of 1 minute, the theoretical gradient reaches the column immediately. A pump with a 65µL dwell volume will get the gradient onto the column in 9.75 seconds. A traditional quaternary pump with a dwell volume of 800µL will take 2 minutes to get the gradient to the column. When running rapid gradients this is too slow and it may become necessary to introduce an isocratic hold at the end of the gradient to allow elution of the analytes. Column: Accucore RP-MS 2.6µm, 50 x 2.1mm Flow rate (mL/min) Inj. volume (µL) Gradient Time (min) 0.0 1.7 1.9 3.0 Backpressure Resolution Run Time (minutes) Solvent Used (mL) 0.4 0.7 %B 35.0 60.0 35.0 35.0 120 1.5 3.0 1.2 Reasons to Scale Down a HPLC or LC/MS Method Typical Flow Rates for Analytical, Narrowbore, Capillary and Nanobore Columns (5µm Particles) There are applications where it is desirable to scale down a method without transferring the method to U-HPLC. These reasons may be to: Column ID (mm) 4.6 3.0 2.1 1.0 0.5 0.32 0.18 0.1 0.075 • Maximize sensitivity when small amounts of sample are available • Make flow rate compatible with ionization technique in MS detection • Reduce costs by reducing solvent consumption Transfer Method to a Narrower Column If all other method parameters (column length and particle size, column chemistry, mobile phase composition, gradient range and time, separation temperature) are kept unchanged, the change to a narrower column only requires adjustment of the flow rate. F2 = F1 x (dc2/dc1)2 F1 – original flow rate (to be reduced) F2 – new flow rate dc1 – original column internal diameter dc2 – new column internal diameter This is applicable to both isocratic and gradient methods. The new method should produce a chromatogram with identical resolution and identical run time. If small changes in retention times and resolution are observed this is generally caused by system dwell volume (discussed below). Optimum Flow Rate1 (µL/min) 1250 500 250 50 12 5 2 0.5 0.3 Recommended Injection Volume2 (µL) 30 10 5 1 0.35 0.15 0.05 0.015 0.01 API Source APCI or High flow ESI APCI or High flow ESI APCI or Micro-ESI Micro-ESI Micro-ESI Micro-ESI Micro-ESI Nanospray Nanospray 1. Recommended for good efficiency and moderate pressure. Higher flow rates may lead to column voids. Lower flow rates are recommended for washing column bed or changing solvents. 2. Estimates based on negligible loss of efficiency and isocratic elution with sample solvent identical to mobile phase. Larger volumes can be introduced under gradient conditions or using weaker sample solvent. Transfer Method to a Shorter Column Dwell Volume In gradient elution, the simplest way to reduce the method cycle time is to reduce the column length. If all other method parameters (column ID and particle size, column chemistry, mobile phase composition, gradient range, flow rate, separation temperature) are kept unchanged the only requirement is to change the gradient time using the equation below, where gradient time is reduced by the same factor as the reduction in column volume. Dwell volume is just as important when scaling down a method as for method transfer to U-HPLC. The effect of dwell volume on the separation is more significant when narrow columns are used at low flow rates. For instance, if the system has a dwell volume of 2.0mL and a 4.6mm ID column is run at 1mL/ min, it takes 2 minutes for the gradient to reach the head of the column; however, if a 2.1mm ID column is used with a 0.4mL/min flow rate it will take 5 minutes for the gradient to reach the column. In high throughput gradient separations using small volume columns, dwell volume causes an increase in run times and longer reequilibration time between runs. tg1/Vc1= tg2/Vc2 tg1 – gradient time in original method (min) tg2 – gradient time in new method (min) Vc1 – original column volume (mL) Vc2 – new column volume (mL) Column volume Vc (mL) can be estimated using: Vc = 0.68 x π x r2 x L Vc – column volume (mL) L – column length (cm) r – column radius (cm) Technical Information Reducing the scale of a separation by reducing the column internal diameter may be necessary when transferring a method from UV to MS detection, or when only very small amounts of sample are available, such as in drug discovery or proteomics. In the first case ionization technique or source design determines the best flow rate range (see table above) and in the latter case, method sensitivity is maximized because solutes elute in more concentrated chromatographic bands. Flow Rate Range (µL/min) 1000 – 1500 400 – 600 200 – 300 40 – 60 10 – 25 4 – 10 1–3 0.4 – 1 0.2 – 0.5 LC Columns and Accessories >> Scaling Down a Method Several approaches can be taken to minimize these effects: • Select a pump with a small gradient delay volume (e.g., Thermo Scientific Accela high speed LC system has a delay volume of only 65µL); • Delay sample injection until gradient has reached the head of the column; • Set the pump at a higher flow rate and split the flow before the column. www.thermoscientific.com/chromatography 4-233 LC Columns and Accessories Scaling Up a Method Reasons to Scale Up an HPLC Method Column Backpressure • Increase method capacity Column operating backpressure is affected by column length, internal diameter, media particle size, temperature, solvent properties and solvent flow rate. It can also be affected by the use of gradients, where the pressure may vary with solvent composition. Typical operating backpressure for columns or cartridges can be calculated using the following equation: Pressure (atm) = 2.1 x Φ x L x η d p2 x d 2 • Isolation and purification of target compounds • Increase sample throughput Analytical methods may require scale up to preparative sizes to isolate and purify compounds from mixtures. In choosing the best column and packing material for your preparative application, consider the selectivity and loadability of the media as well as column dimensions, to give the results you need most quickly or economically. We have established a strong reputation for the manufacture and supply of high quality preparative silicas and bonded phases, designed to give the same levels of performance and reproducibility as our popular analytical silica ranges such as the Thermo Scientific Hypersil phases. Thermo Scientific Chromatography Columns and Consumables 2014-2015 Scale up is easiest when starting from an analytical column packed with smaller particle size media offering the same selectivity as the larger particle size preparative media. The leading families of Thermo Scientific phases are offered in various sizes to complement lab scale operations and facilitate the scale up to preparative chromatography. Scout columns, typically 250 x 4.6mm packed with the media of interest can also be used to develop the separation. Once the method is finalized on the smaller column, a scaling factor can be applied. 4-234 Mobile phase viscosity changes with composition Scaling Up to a Preparative Column Flow rate and column load scaling are only required when changing the internal diameter of the column. The scaling of flow rates allows peak retention times to remain relatively constant between columns with different internal diameters. The typical solvent flow rate through a column is dependent on its internal diameter and the particle size of the column packing material. This scaling factor can also be used to estimate the loading capacity of a given column. Assuming column length is a constant, the scale factor can be calculated using the following formula: Φ = column impedance (1000 for 4.6mm ID columns) L = column length (mm) dp = particle diameter (µm) d = column diameter (mm) η = mobile phase viscosity (centipoises) The mobile phase viscosity varies with composition. As an example, the figure above shows how water viscosity varies with the addition of methanol or acetonitrile. This variability is a critical component in maximizing throughput with respect to the chromatography instrumentation being used. Scale Factor = dc22/dc12 dc1 – original column internal diameter (mm) dc2 – new column internal diameter (mm) The column loading capacity and flow rate required for the new larger ID columns can be calculated using this factor. Discover eVol XR for precise and accurate dispensing PAGE 1-062 Peak Shape – Hypersil GOLD Media Media selection for your preparative separation is important. Choose media that has a narrow particle size distribution which will provide high efficiency columns with a low back pressure, since there are no ‘fines’ to block frits or impede flow. The uniformly spherical particles, with narrow pore size distribution, apparent in Thermo Scientific preparative columns, provide reproducible performance and a longer column life. Media that is available in a range of particle sizes offers choice for scale up applications with controlled selectivity. We offers a range of choices for preparative media in several particle sizes to tailor the media to your application. In analytical HPLC, the use of packings based on highly pure silica has been shown to improve peak shape. Our highly developed and reproducible manufacturing processes ensure that our leading analytical brand of Hypersil GOLD media is also available in a range of particle sizes suitable for preparative LC without compromise on performance. High Load and High Retention – HyperPrep HS Often when dealing with very polar compounds, achieving sufficient retention can be a challenge. We are able to offer a variety of choices to overcome this common problem: The polar endcapping on Hypersil GOLD aQ provides a controlled interaction mechanism by which moderately polar compounds can be retained. Hypersil GOLD AX can be used in HILIC mode to provide retention of polar compounds. Hypercarb columns offer truly orthogonal selectivity to C18 in reversed phase LC and can be used to retain highly polar compounds. Hypercarb columns can also be used to differentiate between very closely related compounds including geometric and positional isomers. When it comes to the analysis of peptides, the correct selection of packing material becomes ever more important. When deciding on which pore size of packing material to use in the analysis of a polypeptide mix, molecular weight and hydrophobicity of the peptides must be taken into account. Our breadth of silica offerings allow the chromatographer to obtain the best resolution using materials with pore diameters in the 100 to 300Å range. A general rule is that hydrophilic peptides with a molecular weight of less than 2000 daltons can be analyzed using a lower pore volume media, such as Hypersil GOLD media. Above this molecular weight, access to small pores is restricted, and separations tend to be inefficient. For hydrophobic peptides with a molecular weight greater than 2000, a 300Å media such as Thermo Scientific BioBasic is recommended. For the separation of small or hydrophilic peptides, a 100Å material such as HyperPrep HS may give better resolution. www.thermoscientific.com/chromatography Technical Information Materials with higher surface area can offer increased loadability. This drive to maximize surface area must be undertaken in a considered manner particularly with regard to particle pore diameter and pore volume. Too high a pore volume will compromise stability and robustness of the bed and too small a pore diameter will influence mass transfer at the expense of efficiency. The high surface area provides enhanced retention of polar compounds. A high carbon loading gives a robust, stable phase. Please contact Technical Support for more information on Thermo Scientific HyperPrep columns and media. Polar Compounds and Isomers – Hypercarb Columns, Hypersil GOLD aQ Peptides and Proteins – BioBasic and Hypersil GOLD Media LC Columns and Accessories >> Selecting the Media 4-235 LC Columns and Accessories Thermo Scientific Chromatography Columns and Consumables 2014-2015 4-236 HPLC Troubleshooting Before you start any troubleshooting, it is essential to observe safe laboratory practices. Know the chemical and physical properties of any solvents used and have the appropriate Material Safety Data Sheets (MSDSs) readily available. All electrically powered instruments should be shut down and unplugged before starting. Eye protection should also be worn. The following table lists common HPLC problems encountered, the possible causes and solutions for your quick reference. Symptom Cause Pressure Related Problems Low Low viscosity mobile phase. Pressure Piston seals leaking. Leak in system. Air in solvent lines or pump. High High viscosity mobile phase. Pressure Pump flow-rate malfunction. Tubing blocked. Guard blocked. Sample precipitation. Detector blockage. Baseline Related Problems Fluctuating System not equilibrated. Baseline Bubbles in flow cell. Contaminated guard. Contaminated column. Detector contamination. Contaminated solvents. Old detector lamp. Sloping Contaminated solvents. Baseline Gradient mobile phase. System not equilibrated. Leak in system. Temperature fluctuations. Contaminated column. Pump not mixing solvents properly. Blocked solvent reservoir frits. Old detector lamp. Peak Shape Problems Broad System not equilibrated. Peaks Injection solvent too strong. Injection volume too high. Injected mass too high. Extra column volume too high. Temperature fluctuations. Old guard cartridge. Old column. Double Peaks Negative Peaks Contaminated column. Voided column. Old guard cartridge. Contaminated column. Voided column. Contaminated solvents. Wrongly wired detector. Unbalanced RI detector optics. Ion pair method. Action Confirm expected pressure using the Kozeny-Carmen or similar equation. Check for evidence of leaking or wear and replace where necessary. Check for and replace any leaking tubing or fittings. Ensure that the reservoirs and solvent lines are fully primed and the purge valve is fully closed. Confirm expected pressure using the Kozeny-Carmen or similar equation. Contact manufacturer. Working backwards from detector outlet, check source of blockage and replace item as necessary. Replace guard cartridge. Consider sample clarification steps such as filtration or SPE. Clean the flow cell according to the manufacturer’s instructions. Equilibrate the column with 10 volumes of mobile phase. Degas the mobile phase and pass degassed solvent through the flow-cell. Do not exceed the cell’s pressure limit. Replace the guard cartridge. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Clean the flow cell according to the manufacturer’s instructions. Use freshly prepared solvents of HPLC grade. Replace the lamp, particularly when this has been in use for > 2000 hours. Use freshly prepared solvents of HPLC grade. Consider purer solvents or higher wavelengths. Otherwise, this is normal. Equilibrate the column with 10 volumes of mobile phase. Check for and replace any leaking tubing or fittings. Use a thermostatically controlled column oven. Wash the column using an appropriate solvent. Ensure that a gradient method has a wash period at the end. Where being used, ensure that the proportioning valve is mixing the solvents correctly. If the method is isocratic, blend the solvents manually. Ultrasonicate the reservoir frits in water and then methanol. Replace the lamp, particularly when this has been in use for > 2000 hours. Equilibrate the column with 10 volumes of mobile phase. Ensure that the injection solvent is the same or weaker strength than the mobile phase. Reduce the injection volume to avoid overload. Typically injection volumes of < 40% of the expected peak width should be used. Reduce the sample concentration to avoid mass overload. Reduce diameter and length of connecting tubing. Reduce the volume of the flow cell where possible. Use a thermostatically controlled column oven. Higher temperatures will produce sharper peaks. Replace the guard cartridge. Do not use columns that have been used with ion-pair reagents for reverse-phase methods. Old columns give much lower efficiencies than new columns. Replace the column if necessary. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Replace the column. Do not use outside the recommended pH range. Replace the guard cartridge. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Replace the column. Do not use outside the recommended pH range. Use freshly prepared solvents of HPLC grade. Check the signal polarity from the detector to the recording device. Refer to manufacturer’s instructions. Inject the sample in the mobile phase. Injected mass too high. Old column. Contaminated column. Voided column. Old or damaged column. Fronting Peaks Peak Size and Retention Problems Small Degraded sample. Peaks Low analyte concentration. Detector set-up. No wash solvent. Missing Peaks Extra Peaks Old detector lamp. Degraded sample. Immiscible mobile phase. Fluctuations in pH. Degraded sample. Contaminated solvents. Immiscible mobile phase. Fluctuations in pH. Contaminated guard cartridge. Contaminated column. Varying Retention System not equilibrated. Leak in system. Temperature fluctuations. Contaminated column. Blocked solvent reservoir frits. Pump not mixing solvents properly. Contaminated solvents. Different dwell volume. Piston seals leaking. Air in solvent lines or pump. Reduce the sample concentration. Check the detector attenuation and re-zero. Replace the guard cartridge. Ensure that the injection solvent is the same or weaker strength than the mobile phase. Reduce the injection volume to avoid overload. Typically injection volumes of < 40% of the expected peak width should be used. Reduce the sample concentration to avoid mass overload. Do not use columns that have been used with ion-pair reagents for reversed phase methods. Old columns give much lower efficiencies than new columns. Replace the column if necessary. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Replace the column. Do not use outside the recommended pH range. Replace the column. Inject a fresh sample. Increase the analyte concentration. Check the detector attenuation and re-zero. Check that the solvent wash reservoir is filled with a miscible solvent and that the injector is set to wash between injections. Replace the syringe. Check injector loop size and that no more than 50% of this volume is used for partial loop injections. Reduce syringe draw-time. Replace the lamp, particularly when this has been in use for > 2000 hours. Fill sample vial. Check for and replace any leaking tubing or fittings. Where being used, ensure that the proportioning valve is mixing the solvents correctly. If the method is isocratic, blend the solvents manually. Replace the syringe. For gradient methods, check that the dwell volume of any new system is not very different from any previous system. Apply a final hold time if necessary. Replace the lamp, particularly when this has been in use for > 2000 hours. Inject a fresh sample. Check that any solvent already in the column is miscible with the mobile phase. Flush with propan-2-ol or ethanol where necessary. Buffer the mobile phase so that retention of ionizable compounds is controlled. Inject a fresh sample. Use freshly prepared solvents of HPLC grade. Gradient methods often show ‘ghost-peaks’. Check that any solvent already in the column is miscible with the mobile phase. Flush with propan-2-ol or ethanol where necessary. Buffer the mobile phase so that retention of ionizable compounds is controlled. Replace the guard cartridge. Wash the column using an appropriate solvent. If this does not resolve the problem , replace the column. Equilibrate the column with 10 volumes of mobile phase. Check for and replace any leaking tubing or fittings. Use a thermostatically controlled column oven. Wash the column using an appropriate solvent. If this does not resolve the problem, replace the column. Ultrasonicate the reservoir frits in water and then methanol. Where being used, ensure that the proportioning valve is mixing the solvents correctly. If the method is isocratic, blend the solvents manually. Use freshly prepared solvents of HPLC grade. For gradient methods, check that the dwell volume of any new system is not very different from any previous system. Apply a final hold time if necessary. Check for evidence of leaking or wear and replace where necessary. Ensure that the reservoirs and solvent lines are fully primed and that the purge valve is fully closed. Technical Information No Peaks Damaged or blocked syringe. Incorrect amount injected. Viscous injection solvent. Old detector lamp. Sample vial empty. Leak in system. Pump not mixing solvents properly. Damaged or blocked syringe. Different dwell volume. Action LC Columns and Accessories >> Symptom Cause Peak Shape Problems Flat topped Detector overload. Peaks Detector set-up. Tailing Old guard cartridge. Peaks Injection solvent too strong. Injection volume too high. For more information, please request Successful HPLC Operation – A Troubleshooting Guide, TG20094. www.thermoscientific.com/chromatography 4-237 LC Columns and Accessories HPLC Definitions and Equations Backpressure The pressure required to pump the mobile phase through the column. It is related to mobile phase viscosity (η), flow rate (F), column length (L) and diameter (dc), and particle size (dp) by the following equation: Pressure Drop (psi) = L η F dp dc = = = = = 250 L ηF dp2 dc2 column length (cm) viscosity flow rate (mL/min) particle diameter (µm) column internal diameter (cm) Capacity Factor (k) Expression that measures the degree of retention of an analyte relative to an unretained peak, where tR is the retention time for the sample peak and t0 is the retention time for an unretained peak. A measurement of capacity will help determine whether retention shifts are due to the column (capacity factor is changing with retention time changes) or the system (capacity factor remains constant with retention time changes). k= t R-t 0 t0 Efficiency (N) Thermo Scientific Chromatography Columns and Consumables 2014-2015 Also number of theoretical plates. A measure of peak band spreading determined by various methods, some of which are sensitive to peak asymmetry. The most common are shown here, with the ones most sensitive to peak shape shown first: 4-238 5-Sigma N = 25(tR /W)2 W = peak width at 4.4% peak height 4-Sigma or Tangential N = 16(tR /W)2 W = tangential peak width or 13.4% peak height Half-Height N = 5.54(tR /W)2 W = peak width at 50% peak height Elution Volume (VR ) Refers to the volume of mobile phase required to elute a solute from the column at maximum concentration (apex). V R = F • tR where F is flow rate in volume/time and tR is the retention time for the peak of interest. HETP Height equivalent to a theoretical plate. A carryover from distillation theory: a measure of a column’s efficiency. For a typical wellpacked HPLC column with 5µm particles, HETP (or H ) values are usually between 0.01 and 0.03mm. HETP = L/N where L is column length in millimeters and N is the number of theoretical plates. Linear Velocity The flow rate normalized by the column cross section. This effects column performance and is directly related to column pressure. Linear velocity is given by the following equation where L is column length and t 0 is the breakthrough time of an unretained peak: µ= L t0 Resolution (Rs ) The ability of a column to separate chromatographic peaks. Resolution can be improved by increasing column length, decreasing particle size, changing temperature, changing the eluent or stationary phase. Rs = It can also be expressed in terms of the separation of the apex of two peaks divided by the tangential width average of the peaks: Rs = (t2 – t1) 0.5(W1 + W2) Selectivity (α) A thermodynamic factor that is a measure of relative retention of two substances, fixed by a certain stationary phase and mobile phase composition. Where k1 and k2 are the respective capacity factors. α= k2 k1 Tailing Factor (T) A measure of the symmetry of a peak, given by the following equation where W0.05 is the peak width at 5% height and f is the distance from peak front to apex point at 5% height. Ideally, peaks should be Gaussian in shape or totally symmetrical. T = W0.05 /2f van Deemter Equation An equation used to explain band broadening in chromatography. The equation represents the height equivalent of a theoretical plate (H) and has three terms. The A term is used to describe eddy diffusion, which allows for the different paths a solute may follow when passing over particles of different sizes. The B term is for the contribution caused by molecular diffusion or longitudinal diffusion of the solute while passing through the column. The C term is the contribution of mass transfer and allows for the finite rate of transfer of the solute between the stationary phase and mobile phase. u is the linear velocity of the mobile phase as it passes through the column. B H = A + u + Cu A partial list of common buffers and their corresponding pH values is shown in the Common Buffer Systems table. Perhaps the most common HPLC buffer is some form of phosphoric acid. Remember that a true buffer should have the ability to resist pH change when a sample is introduced at a different pH, and that buffer capacity is only 100% at the pK value of the acid or base. At pH 4, phosphate is a poor buffer and would change rapidly toward one of its pKa values if a more acidic or basic sample were introduced. Common Buffer Systems Buffer TFA Phosphate Citrate Formate Acetate Tris Base (Trizma, THAM) Ammonia Borate Diethylamine Carbonate pK1 pK 2 Triethanolamine When control at a lower pH (2-3) is desired, phosphate, or stronger organic acids such as TFA or acetic acid, are commonly used. If control at pH 4-5 is desired, an organic acid buffer such as acetate or citrate should be considered in place of phosphate. 2 3 Useful pH Range MS-Compatible 0.30 2.1 7.2 12.3 3.1 4.7 5.4 3.8 4.8 8.3 9.2 9.2 10.5 6.4 10.3 7.80 1.1 – 3.1 6.2 – 8.2 11.3 – 13.3 2.1 – 4.1 3.7 – 5.7 4.4 – 6.4 2.8 – 4.8 3.8 – 5.8 7.3 – 9.3 8.2 – 10.2 8.2 – 10.2 9.5 – 11.5 5.4 – 7.4 9.3 – 11.3 Yes No No No No No No Yes Yes Yes Yes No Yes Yes Yes Yes A: pH 2.1 1 B: pH 2.5 2 3 7 Part Number: Eluent: 7 1 BETASIL C18, 5µm, 50 x 4.6mm Flow Rate: Detector: Sample: The figure to the right shows the importance of choosing the correct pH for a separation. Even slight changes in pH, either from measuring errors, mixing complications with the pump, or atmospheric water adsorption into the mobile phase, can alter any method if not properly buffered. 4 4 5 5 6 6 701-085 0 70105-054630 50% ACN / 50% 25mM KH 3PO 4 at pH indicated 0.8mL/min UV @ 220nm 1. Uracil 2. Tolmetin 3. Naproxin 4. Fenoprofen 5. Diflunisal 6. Indometacin 7. Ibruprofen Technical Information As a rule, one should work within ±1pH unit of the buffer pKa value for good pH control of the mobile phase. Adequate buffer concentrations for HPLC tend to be in the 10-100 millimolar level depending on the size and nature of the sample, as well as the column packing material. Phases based on highly pure silica with robust bondings such as the Hypersil GOLD range, are often more compatible with dilute buffers than traditional packings. Care should be taken when choosing a buffer and organic modifier mixture to ensure that a solution of the two does not produce a solid salt which could cause blockages and system contamination. pK1 pK 2 pK 3 pK1 pK 2 pK 3 pKa LC Columns and Accessories >> Selecting the Right Buffer 6 MIN 701-086 0 6 MIN Effect of small changes in pH on the separation of mildly ionizable compounds Buffers should always be flushed from the analytical column and instrument after use to avoid salts being deposited on delicate frits etc. Buffer Selection for LC/MS Buffer choice will be very dependent on the analyte and the instrumentation used. Ideally, LC/MS applications should use a volatile buffer as this will not form a contaminating deposit on the source. Inorganic acids, involatile buffers and ion-pair reagents should all be avoided. Typical LC/MS buffers include: • Ammonium acetate/formate/hydrogen carbonate (< 50mM) • Formic/acetic acid (0.01 – 1% v/v) • Trifluoroacetic acid (< 0.1% v/v) • Trialkylamine (< 0.1% v/v) and aqueous ammonia type bases • TRIS • BIS-TRIS propane Note: There are LC/MS instruments available, for example the Thermo Scientific Surveyor MSQ LC/MS, which incorporate a self-cleaning mechanism to reduce the build up of inorganic buffers on the source during routine use. Care should still be taken not to purposefully over-contaminate the instrument source as this will lead to operating difficulties. www.thermoscientific.com/chromatography 4-239 LC Columns and Accessories Preparation of Mobile Phases Correct solvent preparation is very important. It can save vast amounts of time spent troubleshooting spurious peaks, baseline noise etc. Quality Buffers All reagents and solvents should be of the highest quality. HPLC grade reagents may cost slightly more than lower grade reagents, but the difference in purity is marked. HPLC grade reagents contain no impurities to produce spurious peaks in a chromatogram baseline whereas lower grade reagents do contain trace levels of impurities, which may produce spurious baseline peaks. All buffers should be prepared freshly on the day required. This practice ensures that the buffer pH is unaffected by prolonged storage and that there is no microbial growth present. Changes in pH and microbial growth will affect chromatography. Ensure that any water used in buffer preparation is of the highest purity. Deionized water often contains trace levels of organic compounds and therefore is not recommended for HPLC use. Ultra pure HPLC water (18mΩ resistivity) is generated by passing deionized water through an ion exchange bed. Modern water purification instruments use this mechanism to produce water of suitable quality in high volumes. Preferably, HPLC grade water can be purchased from solvent suppliers. Important: Do not store HPLC grade water in plastic containers. Additives in the plastic may leach into the water and contaminate it. Always store HPLC grade water in glass containers. 4-240 Degassing Before the freshly prepared mobile phase is pumped around the HPLC system, it should be thoroughly degassed to remove all dissolved gasses. Dissolved gas can be removed from solution by: If buffer solutions are stored, be aware that they have a finite lifetime. Refer to pharmacopoeia monographs or similar for further guidance on buffer shelf life. Buffer reagents can contain a stabilizing agent, for example, sodium metabisulphite. These stabilizing agents often affect the optical and chromatographic behavior of buffer solutions, so it is often worth buying reagents that contain no stabilizer. Containers of solid reagent are easily contaminated by repeated use. For this reason, we recommend that reagents be purchased in low container weights. • Bubbling with helium • Sonication • Vacuum filtration If the mobile phase is not degassed, air bubbles can form in the low pressure of the detector cell resulting in problems with system instability, spurious baseline peaks etc. The most efficient form of degassing is bubbling with helium or another low solubility gas. If this method is available, we recommend that the mobile phase is continually degassed at very low levels throughout the analysis. This will inhibit the re-adsorption of gases over the analysis time. Filtration Ideally, all HPLC solvents should be filtered through a 0.45µm filter before use This removes any particulate matter that may cause blockages. After filtration, the solvents should be stored in a covered reservoir to prevent re-contamination with dust etc. Mobile Phase – Not Degassed 20 0 -20 Note: Ensure that the solvent reservoir has a vent to the atmosphere to prevent the build up of pressure inside the reservoir. Mobile Phase – Degassed 40 Response (Millivolts) Response (Millivolts) Thermo Scientific Chromatography Columns and Consumables 2014-2015 40 Filtering HPLC solvents will benefit both your chromatography and the wear and tear of the HPLC system. Pump plungers, seals and check valves will perform better and lifetimes will be maximized. 20 0 -20 0 2 4 6 Minutes Baseline noise from gas in mobile phase 8 0 2 4 6 Minutes 8 LC Columns and Accessories >> Solvent Properties (vs Silica Gel) and Miscibility Technical Information www.thermoscientific.com/chromatography 4-241 LC Columns and Accessories Chromophore Detection Wavelengths Chromophores are light absorbing groups. Their behavior is used to allow the detection of analytes. They have one or more detection wavelengths, each of which has a molar adsorbtivity associated with it. The information contained in the following table is intended as a guide to common chromophores. It is not an exhaustive list. Chromophore 175 – 180 210 280 – 300 195 190 285 – 400 184 202 255 6,000 Strong 11 – 18 2,800 5,000 3 – 25 46,700 6,900 170 -C==C-CHO Amine Azidin Azo Benzene -NH2 > C=N-N=N- Carboxyl Ester Ether Ethylene Ketone -COOH -COOR -O-C=C> C=O 200 – 210 205 185 190 195 270 – 285 220 275 312 50 – 70 50 1,000 8,000 1,000 18 – 30 112,000 175 5,600 -ONO2 -(C=C)-2 acyclic -(C=C) 3 C=C-C=C C=C-C=N C=C-C=O C=C-NO2 -C==N -ONO 270 210 – 230 260 219 220 210 – 250 300 – 350 160 220 – 230 300 – 400 210 302 190 174 195 251 12 21,000 35,000 6,500 23,000 10,000 – 20,000 Weak – 1,000 – 2,000 10 Strong 100 5,000 80,000 6,000 1,700 180 210 194 1,600 195 Very strong 1,500 4,600 Nitrate Nitrile Thermo Scientific Chromatography Columns and Consumables 2014-2015 εmax (L/m/cm) Acetylide Aldehyde Napthalene 4-242 λmax (nm) Nitro Nitroso Oxime Pyridine -NO2 -N=O -NOH N Sulfone Sulfoxide Thioether Thiol -SO2> S-O -S215 -SH 1,400 Testing of column performance can be undertaken using the experimental conditions in the test certificate provided with the column. The column efficiency, capacity factor, etc. should be measured at the start and end of the clean-up procedure to ensure that it has been performed successfully and has improved the column performance. In all instances, the volume of solvent used is 40 – 60 column volumes unless otherwise stated. Ensure that no buffers or samples are present on the column and that the solvent used prior to the clean up is miscible with the first wash solvent. After the clean up, ensure that the test mobile phase is miscible with the last solvent in the column. Normal Phase Media Protein Size Exclusion Media There are two wash/regeneration procedures associated with the removal of contaminants from protein size exclusion media. 2. Flush with 50mL methanol Strongly Retained Proteins 1. Flush for 60 minutes using a 100% water to 100% acetonitrile gradient 4. Flush with 50mL 0.1M hydrochloric acid Porous Graphitic Carbon 7. Flush with 50mL dichloromethane There are four wash or regeneration procedures associated with porous graphitic carbon. The one(s) used will depend on the analytes and solvents that have been used with the column 3. Flush with tetrahydrofuran 1. Invert the column 4. Flush with methylene chloride 2. Flush with 50mL tetrahydrofuran:water (1:1) containing 0.1% trifluoroacetic acid Reversed Phase Media 1. Flush with HPLC grade water; inject 4 aliquots of 200µL DMSO during this flush 2. Flush with methanol 3. Flush with chloroform 4. Flush with methanol 3. Flush with 50mL tetrahydrofuran:water (1:1) containing 0.1% triethylamine or sodium hydroxide 4. Flush with 50mL tetrahydrofuran:water (1:1) containing 0.1% trifluoroacetic acid 5. Flush column with 70 column volumes of THF Anion Exchange Media 6. Flush with methanol/water (95:5) to re-equilibrate 1. Flush with HPLC grade water 7. Re-invert the column 2. Flush with gradient of 50mM to 1M appropriate buffer solution Author: R. Plumb – Glaxo, UK 4. Flush with methanol Strong Organic Regeneration Suitable for applications involving polar and/ or ionized species analyzed in aqueous mobile phases. 5. Flush with chloroform 1. Flush with 50mL acetone 3. Flush with HPLC grade water Cation Exchange Media 1. Flush with HPLC grade water; inject 4 aliquots of 200µL DMSO during this flush 2. Flush with tetrahydrofuran 3. Flush with 50mL water 5. Flush with 50mL water 6. Flush with 50mL methanol 8. Flush with mobile phase until equilibrated Author: A. Karlsson – Uppsala, Sweden Removal of TFA and DEA TFA and DEA have the potential to adsorb to the surface of porous graphitic carbon; after using these additives in the mobile phase, regeneration of the column should be undertaken to ensure the original Hypercarb selectivity and optimum performance will always be achieved. The regeneration is as follows: 1. Removal of TFA: Flush column with 70 column volumes of THF. 2. Removal of DEA: Set column oven to 75°C and flush column with 120 column volumes of ACN. Technical Information 2. Flush with methanol 5. Flush with benzene-free n-hexane 1. Flush with 50mL dichloromethane Weakly Retained Proteins 1. Flush with 30mL 0.1M pH 3.0 phosphate buffer Acid/Base Regeneration Suitable for ionized species analyzed in strongly aqueous mobile phases. 1. Flush with tetrahydrofuran Normal Phase Regeneration Suitable for applications running predominantly in normal phase mobile phases. LC Columns and Accessories >> Column Cleaning and Regeneration Polymeric Media with Metallic Counter Ions There are three types of regeneration available for polymeric columns with metal counter ion. Details of each procedure are listed in the following table. 2. Flush with 120mL dibutylether 3. Flush with 50mL acetone 4. Flush with aqueous mobile phase until equilibrated Column Type Metal Contamination Organic Contamination Column Cleaning Hydrogen Counter Ion Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min with 20:80 ACN: H 2 O @ 25°C for 4 hr with 20:80 ACN: 0.01M H 2SO 4 @ 65°C with 0.1M H2SO 4 @ 25°C for 4 to 16 hr for 4 hr Calcium Counter Ion Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min with 20:80 ACN:H2 O @ 25°C for 4 hr with 20:80 ACN:H2 O @ 25°C for 4 hr with 0.1M Ca(NO 3) 2 @ pH 6.3 and 85°C for 4 to 16 hr Sodium Counter Ion Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min with 20:80 ACN:H2 O @ 25°C for 4 hr with 0.1M NaNO 3 @ 85°C for 4 to 16 hr with 20:80 ACN:H2 O @ 25°C for 4 hr Lead Counter Ion Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min Pump in reverse flow mode at 0.1mL/min with 20:80 ACN: H 2 O @ 25°C for 4 hr with 20:80 ACN: H2 O @ 25°C for 4 hr with 0.1M Pb(NO 3) 2 @ pH 5.3 and 85°C for 4 to 16 hr www.thermoscientific.com/chromatography 4-243 Resources for Chromatographers Chromatography Resource Center Our web-based resource center provides technical support, applications, technical tips and literature to help move your separations forward. 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