User manual | Accelerate your Quantitative LC/MS Workflows with a Fully Integrated UHPLC

Accelerate your Quantitative LC/MS
Workflows with a Fully Integrated
UHPLC-QQQ Platform
Remco Swart
Pittcon
March 8-12, 2015
The world leader in serving science
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Outline
• Thermo Scientific™ Vanquish™ UHPLC System
• Thermo Scientific™ TSQ Endura™ Triple Quadrupole Mass
Spectrometer (MS) and Thermo Scientific™ Quantiva™ Triple
Quadrupole Mass Spectrometer
• Thermo Scientific™ Dionex™ Chromeleon™ Chromatography
Data System (CDS) Software 7.2 for LCMS
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Why a New UHPLC?
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•
More resolution
•
More sensitivity
•
Easier and less method development
•
More productivity
•
Easier interaction and operation
•
More robust and more reliable
•
More sample capacity
•
More access to experts
•
More and fast service
•
New, fresh & exciting instruments
•
Better results in shorter time
Mass Spectrometry
Perspective
Chromatography
Power Perspective
Software Perspective
The New Thermo Scientific Vanquish UHPLC System
We heard you! And we:
•
•
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Started with a complete
new design in line with
MS portfolio
Put in 30 years of
experience and
know-how
•
Developed a new and
unique technology
•
Closed gaps and
pushed beyond
previous limitations
•
Boosted robustness
•
Developed a new column
technology
• Built the best UPHLC
around the column/user
•
Further leveraged our gold
standard CDS software
•
Boosted serviceability
beyond established
concepts
Vanquish System…Provides More Solutions!
• Vanquish UHPLC is not only a system – it is a problem-solving solution philosophy
Better integration with
the world’s best mass
spectrometers
Maximum speed and
resolution with new
Thermo Scientific™
Accucore™ Vanquish™
columns
Revolutionary ease-of- More throughput capacity
use experience by new
with the new Thermo
Chromeleon CDS
Scientific Vanquish
features and workflows
Charger module
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Vanquish System is…
Confidence
Controlling the separation
with more confidence
Driving the separation with
more power
• 2 thermostatting modes
• 5 C to 120 C temperature range
• Active preheating
• 1500 bar (22,000 psi) of
pump pressure
• Flow rate up to 5 mL/min
• 2 × 3 solvent channels
Sensitivity
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Power
Accuracy
Detecting the analyte with
more sensitivity
Handling the sample with
more accuracy
• Linear up to 3000 mAU
• Noise levels down to ±3 µAU
• Lowest dispersion with Thermo
Scientific™ LightPipe™ technology
• Up to 23 well plates for up to
8832 samples
• Unsurpassed sample dosage
• Automation of workflows with
barcode reading
Vanquish Binary Pump
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Feature
Benefit
1500 bar / 22 000 psi
Up to 5000 µL/min
Flexibility for all
working conditions
New parallel dual
piston principle
very small pressure
ripple => column
and sensitivity
Adaptive Thermal
Effect Compensation
(ATEC™)
best flow and
gradient
performance => RT
stable
2 × 3 solvent
channels
9 different solvent
combinations
Fingertight check
valve design
Easy to service
Vanquish Split Sampler
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Feature
Benefit
New switching valve
no maintenance
Adjustable GDV
easy method transfer
Automatic sample
pre-compression
• reduced physical
stress on column =
longer lifetime
• better retention
time reproducibility
New air-to-air cooling
concept
no condensation
Increased sample
capacity
4 racks or well plates
Automated barcode
reader
Rack tracking (esp.
with Charger)
Vanquish Charger Module
• Fully integrated robotic unit with less
than 1 min cycle time
• Hosts up to 9 additional racks/ deep
well plates or 20 shallow well plates
• Up to 9000 samples – temperature
controlled
• Precise temperature control using
new designed air stream cooling
concept
 4 – 40 ºC
• Integrated barcode reader
23×96
wells
100s
cycle
time
2208
samples
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The Vanquish Column Compartment
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Feature
Benefit
Active Pre-Heater
Perfect temperature
control (QC)
Two thermostatting
modes
Easy method transfer
Three independent
column
compartments
Method scouting,
different temperature
zones
Vanquish Column Compartment for MS integration
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Feature
Benefit
Column compartment
is vertical
The column outlet is
closer to the MS ion
source
Position of the
column compartment
can be to the left or to
the right
Better integration with
all Thermo Scientific
Mass Spectrometers
Vanquish Diode Array Detector
• LightPipe technology for an
unmatched detection experience
 10 mm standard flow cell
 60 mm high sensitivity flow cell
• Best signal-to-noise performance
• Lowest baseline noise
• Ultra-wide dynamic range
 Up to 3000 mAU
 For simultaneous detection of
highly concentrated compounds
and trace impurities
• Variable slit width for optimum
resolution
1 – 8 nm
• Low peak dispersion
• Reduced RI and thermal effects
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Vanquish System Features & Benefits for LC-MS Users
• Improved separation performance
FEATURE
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BENEFIT
• Better resolution
• Minimal peak
dispersion
• Less matrix effects
• More consistent quantification
• More consistent sensitivity
• Faster analysis
• Improved LCMS productivity
• Rock solid retention
time stability
• More reliable identification
• More reliable quantification
• New core enhanced
1.5 µm columns
• Less method development
• Less peaks of interest in the void volume
Challenges in Quantitative Mass Spectrometry
Large Sample Numbers
Low Concentration
Triple Quads
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TSQ Endura MS and TSQ Quantiva MS
TSQ Endura MS
Extreme Quantitative Value
• Best-in-class sensitivity
• Unprecedented usability
• Ultimate robustness
TSQ Quantiva MS
Extreme Quantitative Performance
• Attogram sensitivity
• Unprecedented usability
• Exceptional robustness
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Sensitivity, Speed & Robustness for High-Throughput
Laboratories
TSQ Quantiva MS
Extreme quantitative performance
TSQ Quantiva MS
10-1850
10-3400
Max SRM Number
30,000 SRMs
30,000 SRMs
SRM/Sec
500 SRMs/sec
500 SRMs/sec
Quadrupole Design
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•
•
•
Reserpine
Specification
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Active Ion
Management (AIM)
Ion Max NG source
Electrodynamic ion
funnel
ion beam guide with
neutral blocker
6 mm HyperQuad
quadrupoles with
asymmetric RF drive
100,000 :1 S/N for 1 pg
Reserpine
Designed for the most challenging
assays.
•
For scientists needing to stay at the
forefront of analytical technology
TSQ Endura MS
Mass Range
Ion Optics
•
S- LENS with Beam
Blocker Technology
4mm Quadrupoles
with Asymmetric
RF
10,000 :1 S/N for 1
pg Reserpine
TSQ Endura MS
Extreme quantitative value
•
Designed for non-stop operation.
•
For scientist who need to run routine
samples day in and day out.
Low Sample Concentration
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Challenge
TSQ Quantiva MS
Demanding assays which require
the absolute lowest limit of
detection
Powered by AIM (Active Ion
Management) technology,
the TSQ Quantiva MS is the
world’s most sensitive triple
quadrupole MS, detecting
compounds at the ppt level.
Large Sample Numbers
Challenge
Need to analyze multi-residue
analytes in very short run times,
using UHPLC with robust
performance 24/7
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Vanquish System with Charger
and TSQ Quantiva MS
The AIM technology includes an
ultra fast collision cell designed for
fast SRMS with no loss in
sensitivity. Neutral clusters
eliminated for robustness.
Routine Quantitation
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Challenge
TSQ Endura MS
Robust and reliable quantitation
LODs and LOQs unrivaled in its
class. It delivers this best-in-class
quantitation run-after-run and dayafter-day regardless of sample
type or matrix.
TSQ Quantiva MS: Ease-of-Use
Challenge
Analysts don’t have time to waste
learning new software or
optimizing methods.
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Vanquish System and TSQ MS
with Chromeleon CDS Control
The entire software workflow
has been redesigned to remove
un-needed parameters.
Introduction
• How can you include MS in your laboratory CDS?
• Currently it’s on a workstation and there is no way to work remotely
• Chromeleon™ 7.2 Chromatography Data System is the first CDS
to support MS instrument control and data processing with all main
front-end separation techniques (GC, LC, IC) in an enterprise
environment
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Quantiva & Endura - Fully Integrated within
Chromeleon CDS
• Chromeleon 7.2 CDS supported workflows
• Compliance, scalability, suite of quantification features
• MS-specific data organization at component level
• Context menu to open data in MS applications (e.g.
Thermo Scientific™ Xcalibur™ Software)
• Key benefits
• Only need to install, validate and learn one software
package
• Can use all compliance and processing features of
Chromeleon CDS
• Enhanced data security
• Easily start analyses with eWorkflows™
• Additional MS-specific detection algorithms (Genesis,
ICIS)
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MS Data Visualization
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Component and Channel Centric Data Viewing
All channels and
components listed
and multi-selectable
Filter box to
quickly narrow list
of components or
channels
Components are
tagged with tool tips,
indicating analysis
results
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MS Data Views – Total Ion Chromatogram (TIC)
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MS Data Views – MS Quantitation Channel
Overlay of all
Quantitation XICs
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Results for
selected
component
MS Data Views – Mass Spectrum Plot
Peak apex spectrum
Click on component
name or peak and
select mass spectra
Also show leading/
tailing spectra & library
hits – stack or overlay
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MS Data Views – Component Traces
Quantitation and
confirmation ions
displayed...
Click on component
name or peak
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Data Visualization – Multiple Components / Injections
• Using SmartLink and Component Traces…
Select SmartLink...
Injections in
rows...
View
...or stacked...
overlaid
...and pin
injections /
components
...components in
columns
Quantitation and
confirmation ions
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MS Data Processing
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MS Data Processing
• With multiple traces and possibly hundreds of components,
how can you quickly and easily process your MS data?
• How can you rapidly and accurately setup your component
table?
• Existing data processing tools in Chromeleon CDS are now
complimented by a suite of MS-specific tools for fast and
efficient MS data processing, including:
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•
•
•
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Easy extracted ion chromatogram (XIC) creation
Additional MS-specific detection algorithms
Full processing control on a component-by-component basis
MS Library searching
MS Data Processing
• Apply all existing calibration modes and options to MS
• Easy extracted ion chromatogram creation
• Additional MS-specific detection algorithms
• Genesis, ICIS
• Full processing control on a component-by-component basis
• Additional MS-specific parameters in component table – quantitation and
confirming ions, CAS numbers, molecular formulas, etc.
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MS Data Processing – Extracted Ion Chromatograms
• Manual extraction of XIC
Right-click on
Channels
Channel
or
MS Plot
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MS Data Processing – Extracted Ion Chromatograms
• Graphical extraction
Click below the
axis...
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MS Data Processing – Extracted Ion Chromatograms
• Graphical extraction
... and drag
out a range
New channel
added to list and
displayed
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Integrated NIST Library Search
• Use NIST libraries for spectral library screening and ad-hoc
library search
Library screening
results
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Import from Compound Databases and NIST Libraries
• Quickly populate
processing method
component tables
• Use library searches
• NIST
• User libraries
• Use predefined
compound data
bases
• Thermo Scientific™
TraceFinder™ software
• Use Xcalibur
software raw files
with component info
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MS Data Reporting
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MS Data Reporting
• Can be a very time-consuming activity!
• Multiple components and traces make generic reporting
tricky, if not impossible
• How do you effectively report such large quantities of data?
• Several new additions to Chromeleon CDS reporting tools
• New MS based reporting objects
• Mass Spectral Plot
• Component Traces Plot
• MS related report variables and tables
• New tools for reporting of large amounts of multi-channel data
• Consolidated Report Tables
• Autorepeat Area Enhancements
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MS Data Reporting
MS peaks often only appear in certain detection channels
(filters) – how do you create a report for your peaks of interest?
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MS Data Reporting – Consolidated Report Tables
Chromeleon CDS Consolidated Report Tables
• Automatically report for the Channel with the highest response
• Useful for reporting large quantities of MS data in a generic, compact
table
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Antibiotics 1 ppb
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36 Antibiotics Targeted Screening
Accucore Vanquish C18 column, 1.5µm x 2.1mm x 100mm
Mobile phase A:
Overlaid selected reaction monitoring chromatograms showing detection
of 36 antibiotics within a 5 minute detection window, binary Vanquish
system and TSQ Vantage MS
0.1% formic
acid / Water
0.1% formic
acid / MeOH
400 µL/min
40 °C, active
preheating
2 µL
Mobile phase B:
Flow rate:
Column temperature:
Injection volume
Table 1. LC gradient conditions
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Time (min)
%B
0.000
10
4.375
90
5.000
90
5.125
10
8.750
10
47 Drugs in 4 Minutes
Overlaid selected reaction monitoring chromatograms showing detection of
47 drugs within a 4 minute detection window, binary Vanquish system and TSQ
Vantage MS
Accucore
Vanquish C18
column, 1.5µm x
2.1mm x 100mm
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Mobile phase A:
acetate in water
Mobile phase B:
methanol
Flow rate:
Column temperature:
preheating
Injection volume:
10 mM ammonium
0.1% formic acid in
500 µL/min
50 °C, with active
2 µL
Table 1. LC gradient conditions
Time (min)
%B
0.00
10
0.16
10
2.88
90
3.20
90
3.28
10
5.60
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Summary – Bringing It All Together
Chromeleon CDS has all the tools to help you process
and report your MS data in the shortest time!
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Thank You!
Any Questions?
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