Compaq | Netelligent 1108 | KTApurifier chromatography systems

datafile
Liquid chromatography
ÄKTApurifier chromatography systems
High-performance purification
and characterization of proteins,
peptides and nucleic acids
Fast and easy start to purification
•
method wizard for easy programming
Speed and reliability for every run
•
fast optimization and peak identification, accurate result
comparisons, improved peak purity, minimal procedure times
with column knowledge built-in
Increased reproducibility every day
•
automatic buffer preparation, evaluation and report
procedures
Flexibility to change with your needs
•
•
easy system modification
increased productivity and throughput
ÄKTAdesign support
Introduction
ÄKTA™purifier (Fig 1) is a liquid chromatography system
designed for fast and reliable separations of proteins,
peptides and nucleic acids.
Fig 1. ÄKTApurifier, from the ÄKTApurifier chromatography
system series.
ÄKTApurifier runs all chromatography techniques, from µg
to mg scale at flow rates 0.001 ml/min – 10 ml/min and
pressures 0–25 MPa or alternatively at flow rates 0.01–100
ml/min and pressures 0–10 MPa. With operational simplicity
and the method wizard ÄKTApurifier incorporates many
features to minimize preparation and run times and
maximize reliability and reproducibility of results.
ÄKTApurifier is essential to guarantee fast, reproducible
runs everyday, as shown in Figure 2 and Figure 3.
ÄKTApurifier systems are part of ÄKTAdesign, a range of
liquid chromatography systems for academic and industrial
research laboratories. ÄKTAdesign systems are controlled
by UNICORN™ , a control, evaluation and reporting
platform which also controls production systems at
kilogram scale. Designed for Windows NT™/Windows
2000, UNICORN ensures quick, simple communication
between systems and users and meets the stringent control
and data handling procedures of modern laboratories.
Optimized methods are transferred easily to production
scale.
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Liquid chromatography
▲
Run
Fast and easy start
to purification
▲
Simple, direct start for any run
Select method
BufferPrep mode
Protein mixture
mAU
RESOURCE Q
10 ml/min
100
pH 5.0
pH 6.5
pH 8.0
5
Speed and reliability
for every run
50
%cond.
Automatic pH scouting using
BufferPrep
0
0
2
0
2
0
2
Time (min)
mAU
Sephasil™ Peptide C18 5 µm ST 4.6/250
,
Tryptic digest RVP ovalbumin
Flow rate 1 ml/min
CH3CN/TFA
100
Increased reproducibility
every day
50
Reproducible, high resolution results
0
0
20
40
60
80
100
120 ml
Fig 2. Working with ÄKTApurifier–for the shortest route to a pure product
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Liquid chromatography
mAU
mAU
mS/cm
mS/cm
KIPDPPQPDFPQLNSDWVPS-amide
YADKITEDLK
400
1000
1.2
300
2.5
800
1.0
600
200
0.8
2.0
400
100
0.6
200
1.5
0
0
0.0
10.0
20.0
0.0
30.0 ml
mAU
mS/cm
mAU
20.0
40.0
60.0
ml
KIPDPPQPDFPQLNSDWVPS-amide
%B
YADKITEDLK
100
3.0
30.0
80
100
20.0
2.5
10.0
2.0
60
40
50
20
0.0
1.5
0
0
0.0
2.0
mAU
4.0
6.0
0
0.0
ml
4.0
mAU
mS/cm
YRSRKYSSWYVALKR
2.0
6.0
mS/cm
ASVSVTAEDEGT
1.20
150
600
ml
2.0
1.00
100
400
0.80
1.5
200
50
0.60
0
1.0
20.0
mAU
0
40.0
60.0
10.0
ml
%B
100
YRSRKYSSWYVALKR
80
mAU
20.0
30.0
ml
%B
ASVSVTAEDEGT
100
20.0
80
40.0
60
15.0
60
40
10.0
40
20
5.0
20
0
0.0
20.0
0
0.0
2.0
4.0
6.0
ml
0
0.0
2.0
4.0
6.0
ml
Fig 3. Purification of 4 peptides (small chromatograms) and purity analysis (large chromatograms) on one system using the
experimental conditions below.
Examples shown are from a series of
synthetic peptide purifications and
analyses. Routine purity checks were
speeded up using Autosampler A-900.
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18-1119-48 AC, 2001-08 • p3
Experimental conditions
Purifications
Crude material
Purity analyses
Collected fractions
Eluents
Sephasil Peptide C18 5 µm ST 4.6/100 or 4.6/250
1 ml/min, 214 nm
µRPC C2/C18 3 µm SC2.1/100
0.15 ml/min, 214 nm
pH 2.5 CH3CN/TFA: aii, bi, ci, cii, dii
pH 7.0 CH3CN/phosphate buffer: ai, bii, di
Liquid chromatography
Fast and easy start to purification
Users are able to create complete purification schemes for a
range of biomolecules by using the method wizard. This
makes ÄKTApurifier extremely easy to use and speeds up
routine purifications. It reduces the need for expertise in
programming, in chromatography and in time-consuming
literature searches, as all this knowledge is built into
ÄKTApurifier.
Each method created contains certain parameters as
variables, e.g. flow rate, sample volume, column
equilibration, for simple changes before each run if required
(Fig 5).
The method wizard in ÄKTApurifier provide a unique
combination of flow configuration and methods support for
all chromatographic techniques:
•
•
•
•
•
•
Gel filtration (size exclusion) chromatography
Anion exchange chromatography
Cation exchange chromatography
Hydrophobic interaction chromatography
Affinity chromatography
Reversed phase chromotography
The method wizard is based on extensive and welldocumented work in our laboratories and can be used
directly. Using input parameters as for example, components
in use, flow rate, sample volume or gradient, the wizard builds
up the method to fit your needs.
Select technique
Fig 5. Simple input of chromatographic parameters.
The method wizard eliminate time consuming and repetitive
method programming. The method wizard uses column
volume as a programming base for easy use with any size of
column. Figure 6 shows the simplicity of working with
ÄKTApurifier systems once an established method is created
and saved.
Select column
▲
▲
▲
Run
▲
Fig 4. A new method is created in less than 3 min.
Select method
Fig 6. Start established method immediately.
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Liquid chromatography
Speed and reliability for every run
Routine runs quickly optimized
To quickly establish fast routine runs, the method wizard
supports programming of method scouting (Fig 7). These
scouting methods make it easy to systematically vary run
parameters in repeated, automated procedures, reducing
optimization times.
mAU
%B
100
RESOURCE Q
1 ml column
1 ml/min
CH3CN/TEAA
A648
A260
1000
500
Fig 7. Automatic scouting during purification development
and optimization. Example shows scouting for flow rate,
sample volume, and gradient length.
0
0.0
10.0
20.0
30.0
ml
Fig 8. Purification and on-line detection of Cy™5-labelled
(CyDye™) 20-mer oligonucleotide
Fast peak identification
User selected multi-wavelength monitoring enables on-line
detection of specific labelled products (Fig 8) and can be
used to detect contaminants at higher sensitivity wavelengths
(e.g. 280/214/254 nm)
Accurate result comparisons
All data, including running conditions, are stored
permanently for every run within the run log e.g. flow,
pressure, UV, conductivity, temperature. This makes it
extremely easy to compare runs and determine a cause for
any deviations that may be seen.
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Liquid chromatography
Improved peak purity
Accurate peak fractionation can help to improve the purity
of a selected peak. Figure 9 shows the accuracy of peak
fractionation in ÄKTApurifier, by re-injection of daughter
fractions from the original digest. Peak fractionation can be
based on peak detection using slope or level sensing.
Fraction marks and fraction numbers make it easy to
identify fractions and peaks (Fig 8).
Minimal procedure times
User selected and defined WATCH functions (Fig 10) are
entered into a method to instruct specific events to occur
when preset conditions are reached. For example, the
WATCH function will wait for a return to normal baseline
level before instructing the next run to begin, minimizing the
time spent on column re-equilibration between runs.
mAU
Fig 10. User defined WATCH functions for improved automatic
event control.
Sephasil Peptide C18 5 µm ST 4.6/250
,
Tryptic digest RVP ovalbumin
Flow rate 1 ml/min
CH3CN/TFA
60
ml
Fig 9. Accuracy of automatic peak fractionation in ÄKTApurifier.
Column knowledge – built-in
ÄKTApurifier ensures every column is run optimally and is
protected by ÄKTApurifier from damaging events such as
over-pressure. The user selects a pre-packed column from
the extensive column library (Fig 11) and ÄKTApurifier
automatically sets the optimal running parameters for that
column (e.g. column volume, flow rate and pressure limit)
into the method. The user adjusts running conditions (if
required) by simple method editing. New columns can be
added to the library and will be run in the same secure way.
All column specifications
▲
Create your own library
▲
▲
Choose any column
Methods are automatically
adapted to your new column
Fig 11. Comprehensive column library ensures optimum running conditions.
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Liquid chromatography
A
mAU
Mono Q HR 5/5
1 ml column
A: 30% CH3CN/Phosphate pH 7.0
B: as A + 0.75 NaCl
C
% cond.
80
mAU
40.0
1000
60
% cond.
RESOURCE RPC
3 ml column
A: NH4HCO3 pH 8.9
B: 60% CH3CN
80
60
20.0
40
500
40
20
0.0
0
0.0
20
0
10.0
20.0
30.0
40.0
ml
0.0
50
100
150
ml
D
B
mAU
Mini Q PE 4.6/50
0.8 ml column
A: 30% CH3CN/Phosphate pH 7.0
B: as A + 0.75 NaCl
% cond.
500
40.0
RESOURCE RPC
3 ml column
A: NH4HCO3 pH 8.9
B: 60% CH3CN
80
80
60
20.0
400
60
300
40
200
20
0
0.0
% cond.
mAU
40
0.0
20
0
10.0
20.0
ml
0.0
50
100
150
ml
Fig 12. Two-step purification of a synthetic peptide using ion-exchange, with automatic peak fractionation. RPC is used for the
final purification of the collected fractions.
The selection of the correct column for each purification step
is essential for optimum results. Figure 12 shows the
improved resolution of Mini Q™ (Fig 12B) compared to
Mono Q™ (Fig 12A) yielding a higher purity product after
only a single step, shown by the corresponding purity
checks on RESOURCE™ RPC (Fig 12C and 12D).
Optimum elution and accurate peak fractionation in
ÄKTApurifier ensure a minimum number of purification
steps.
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Liquid chromatography
Increased reliability and reproducibility
from day to day
Accurate, automatic buffer preparation
The unique function, BufferPrep, eliminates the timeconsuming manual buffer preparation and titration needed
for every pH change in chromatography.
The user selects the buffers to be used from the buffer
library and defines the pH required (Fig 13). BufferPrep
prepares correct buffer pH on-line from four stock solutions
(e.g. water, salt, buffer, acid/base) and controls the pH
throughout the run, automatically compensating for changes
in temperature and salt concentration. New buffer
substances can be added to the buffer library and used by
BufferPrep to create the required elution pH conditions
cytochrome c
pH 5.5
pH 4.8
pH 4.65
pH 4.5
▲
pH 3.9
Select from
buffer library
Fig 14. Small changes in pH significantly change
chromatographic results. Above, a change of 0.15 pH units
alters an elution profile.
Fig 13. Simple programming for automatic buffer preparation.
The accuracy of pH is crucial in many separations, as shown
in Figure 14. The accuracy of BufferPrep (Fig 15) greatly
increases the reproducibility and reliability of results from
day to day by eliminating and correcting errors which occur
when producing buffers manually.
BufferPrep can be used for pH scouting in rapid method
optimization. The method wizard makes programming of
automatic eluent pH scouting for ion exchange
chromatography easy and safe. Figure 16 shows BufferPrep
used on ÄKTAexplorer 100 at 60 ml/min to test seven pH
conditions to find the optimal pH for elution of a pure Fab
fragment.
Automatic evaluation and reporting
ÄKTApurifier includes pre-programmed evaluation and
report procedures which can be included within run
methods. User- defined evaluation procedures and
customized report layouts are quickly created (Fig 17).
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18-1119-48 AC, 2001-08 • p8
Fig 15. Constant pH, accurately maintained by BufferPrep,
as salt concentration increases. Results shown from tests
covering pH 5.3–9.7.
Liquid chromatography
pH
7
Run 3
Run 2
6.5
Run 4
5.5
4.5
6
Run 5
5
mAU 280 nm
Run 6
➊ Pump P-903
➋ Monitor pH/Cond-900
➌ Monitor UV-900
➍ Autosampler A-900
➍
Run 7
4.5
4
mS/cm
RESOURCE S
6 ml column
Fab fragment
80.0
250
➌
➊
▲
pH 4.0
150
➋
▲
60.0
▲
200
▲
Run 1
START CONDITIONS
pH 4.5
40.0
pH 5.0
100
pH 5.5
pH 6.0
50
20.0
pH 6.5
pH 7.0
0
2.0
4.0
6.0
Fig 18. ÄKTApurifier 10XT for increased throughput.
8.0 Time (min)
Fig 16. BufferPrep for automatic, rapid selection of pH.
To maximize productivity ÄKTApurifier 10XT is supplied
with Autosampler A-900 (Fig 18). This allows automatic
injection of up to 96 samples from standard vials or 160
samples from micro-vials.
An extended evaluation package, UNICORN analysis
module is included, providing:
•
•
•
•
Quantification by internal or external standards
Determination of molecular size by gel filtration
Quantification of sample amount and sample concentration
Recovery calculations
The autosampler is fully controlled through UNICORN
control system and the method wizard includes the use of
the autosampler in ÄKTApurifier 10XT, ensuring simple and
easy start-up before leaving the system to run, evaluate, and
report the samples automatically.
Fig 17. Automatic user defined evaluation and report
procedures.
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Liquid chromatography
ÄKTApurifier 10XT configuration with
optional functions as displayed on screen
Air sensors
Column selection valves
▲
▲
▲
▲
▲
▲
Sample pump
Flow direction valve
Optional valves
Buffer selection valve
Fig 19. Additional equipment is easily added to an ÄKTApurifier chromatography system.
Easy system modification
New functions to enhance system capabilities or create new
system configurations are easily added (Fig 19). Functions,
consisting of hardware components and software
instructions, are selected, installed and implemented by the
user. UNICORN controls and monitors the new functions in
the same way as all other ÄKTAdesign components. Further
details of these optional functions are described on page 14
and include:
•
Room 1
ÄKTAexplorer 100 A
ÄKTAFPLC™
ÄKTApurifier 10 XT
Local PC
Remote PC
Room 2
Automatic sample application (large volumes or high
Buffer preparation unit
throughput)
•
•
•
•
ÄKTAexplorer 10 XT
Ultrafiltration unit
Automatic column selection
Automatic buffer selection
Increased security for programmed events - using air sensors
Server
BioProcess System 1
PC Network
Share data
Local PC
BioProcess System 2
Flexible control of external equipment
ÄKTAdesign support
Fig 20. Users control and monitor chromatography systems
from network-connected computers.
ÄKTAdesign networks
•
•
•
•
Faster communication
High security
Complete overview of operations
Maximum compliance with regulatory needs
UNICORN control system has full networking capability,
with high security and maximum compliance with
regulatory needs. Network support allows control of any
ÄKTAdesign system from any connected UNICORN
workstation (Fig 20).
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System control and data access via a PC network is
extremely easy to use and gives complete overview of
operations.
With defined access rights, operators connect to and control
systems from any remote control or local PC. The user
interface is exactly the same. Results can be automatically
saved on a server and evaluation and generation of reports
can be made locally or at remote PCs.
Liquid chromatography
ÄKTAdesign service
•
•
•
•
•
Controlled costs
System description
General description
Traceable results
Regulatory requirements support
Maximum productivity
Continuous operation at optimal performance
Regular, planned maintenance increases productivity by
keeping a system in top working condition and minimising
downtime. A service agreement lets users budget for all
service and maintenance costs, giving full cost control and
eliminating unexpected expenses.
Service agreements will greatly simplify compliance with
regulatory and quality demands.
Scheduled preventive maintenance carried out by qualified
engineers using traceable test equipment and the in-built
service diagnostics of ÄKTAdesign, provide users with
support to meet the requirements of Good Laboratory
Practice (GLP).
ÄKTAdesign systems are supported with a comprehensive
range of after-sales service agreements that can be
customized to individual needs. All ÄKTAdesign instruments
are supported in the laboratory by Labcrew™ equipment
specialists (Fig 21). Labcrew is dedicated to minimizing
Fig 22. The swivel platform
gives easy access to any side
of the system.
operational disruption and ensuring functionality of ÄKTA
equipment. Your local Amersham Biosciences
representative will be pleased to discuss the options with
you.
Figure 22 shows an ÄKTApurifier chromatography system.
All systems are supplied assembled and tested. Requiring
minimal bench space, each system is mounted on a swivel
platform for easy access to every side. Columns are snapped
in place on the outside of the system, either individually or as
groups held in column holders.
Fig 21. Labcrew specialists offer service support for all
ÄKTAdesign instruments.
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18-1119-48 AC, 2001-08 • p11
Liquid chromatography
Figure 23 shows the configuration of ÄKTApurifier
chromatography systems providing:
•
easy pump wash for quick solvent exchange without disturbing
the column
•
•
automatic on-line buffer preparation from stock solutions
automatic sample application
– fixed volume loops from 100 µl to 1 ml, volumes smaller than
100 µl are applied by partial loop filling
– Superloop™ 10 ml, Superloop 50 ml, and Superloop 150 ml
for the range 1–10 ml, 1–50 ml and 1–150 ml respectively.
– sample throughput is increased by the autosampler in
ÄKTApurifier 10XT
•
•
•
•
on-line UV-detection of up to three wavelengths simultaneously
on-line monitoring of conductivity
UNICORN control system is used from laboratory to full
scale production and allows real time control of
ÄKTAdesign systems. As a single platform UNICORN
ensures quick and simple communication between
ÄKTAdesign systems and users. The same interface and
methods used to run routine applications can be used to
optimize a purification in research and development and
applied at production scale.
UNICORN software is installed on the PC prior to delivery
and controls all ÄKTAdesign instrument settings and
functions.
For further information on the latest version of UNICORN
system control, please contact your local Amersham
Biosciences representative.
optional on-line monitoring of pH
System components
fraction collection options ranging from microtitre plates to
ÄKTApurifier has three main instrument components, a
number of ancillary items, and the option of a fraction
collector and autosampler. All instrument settings and
functions are under the direct control of UNICORN.
large vessels
•
System control – UNICORN
routine, automated system wash
Flow configuration
At every start-up, each instrument automatically performs a
self-diagnostic test and calibrates appropriate settings. This
ensures top performance throughout day-to-day operation.
Autosampler A-900
BufferPrep
C
A
B
All wetted materials are inert, fully biocompatible and
resistant to all eluents commonly used in liquid
chromatography, ensuring high recovery and no loss of
biological activity.
ÄKTApurifier 10
0.001–10 ml/min,
25 MPa with
Pump P-903
D
ÄKTApurifier 100
0.01–100 ml/min,
10 MPa with
Pump P-901
Injection Valve
pH,Cond
Fractionation
Fig 23. The flow configuration of ÄKTApurifier and
ÄKTApurifier 10XT.
UV-Vis
Pumps P-901 and P-903
The P-900 series pumps have been designed to meet the
needs of all chromatographers working with biomolecules.
A unique new pump design has been developed to overcome
many of the problems associated with traditional high
performance pumping systems, and to produce accurate,
reproducible flow and gradient formation over a wide flow
rate range. The pressure and flow rate ranges covered by the
two pumps are shown in Figure 24.
The wide flow rate range of Pump P-901 makes it suitable
for both laboratory and pilot scale applications, running at
optimal flow rates without changing pump modules or
pump heads.
The lower flow rates and higher pressure specification of
Pump P-903 are ideal for running laboratory scale
separations and high-resolution analyses.
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18-1119-48 AC, 2001-08 • p12
Liquid chromatography
ÄKTApurifer 100 systems include a UV-900 monitor with a
2 mm pathlength flow cell (internal volume 2 µl). The flow
cell in ÄKTApurifer 10 systems has a pathlength 10 mm
(internal volume 9 µl).
Pressure range (MPa)
25
20
P-903
Further information: Please ask for Monitor UV-900
Data File No. 18-1111-17.
10
Ancillary equipment
Standard
P-901
0.001
0.01
0.1
1
10
Flow rate range (ml/min)
100
Fig. 24. Flow rates and pressure ranges of P-900 pumps.
Both pumps can be used for loading samples larger than
50 ml, but for easy, routine handling of large sample
volumes the Sample Pump P-950 is recommended.
Further information: Please ask for Pump P-900 Series
Data File No. 18-1119-49.
Monitor pH/Cond-900
Monitor pH/C-900 is a combined monitor for accurate,
on-line monitoring of pH, conductivity and temperature
recording real time information about chromatographic
conditions.
Valves
ÄKTApurifier systems have one seven port injection valve,
INV-907, for sample application and one eight port
fractionation valve, IV-908, for diversion to fraction
collector or waste. Two switch valves work together with a
P-900 pump in BufferPrep mode.
Mixer M-925
Mixer M-925 is a dynamic, single chamber mixer powered
and controlled from a P-900 pump. Each system is delivered
with interchangeable mixing chambers to allow optimal
mixing over the entire flow rate range (0.6 and 2 ml for
ÄKTApurifier 10 and 2, 5, and 12 ml for ÄKTApurifier 100).
An accurate response, coupled with high precision over a
wide measuring range, make this monitor ideal for use in all
chromatography techniques, from reversed phase with very
low conductivity eluents to hydrophobic interaction
chromatography in high salt solutions.
The pH/C-900 consists of a control unit, a flow cell for
conductivity and temperature, a flow cell with a holder for
the pH electrode and the pH electrode.
Further information: Please ask for Monitor pH/C-900
Data File No. 18-1111-19.
Monitor UV-900
Monitor UV-900 is a high sensitivity multi-wavelength
UV-Vis monitor, recording up to three wavelengths
simultaneously for optimal detection in the range
190–700 nm.
The use of advanced fibre optics together with the unique
flow cell design ensures a high signal-to-noise ratio for
maximum performance. Maintenance costs are minimized by
use of a unique Xenon flash lamp, giving a siginificant
increase in lamp lifetime.
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18-1119-48 AC, 2001-08 • p13
Fig 25. Frac-950
Optional
Fraction collector Frac-950
Frac-950 (Fig 25) offers two independent ways to prevent
sample loss across the entire flow rate range of ÄKTAdesign
systems, from 0.001 ml/min to 100 ml/min, and the highest
flexibility in the choice of collection mode. Fractions are
collected in microplates (over 384 fractions, up to 2.5 ml per
fraction), a variety of test tube sizes, or in 20 large vessels
(up to 250 ml per fraction) or even funnels for unlimited
fraction size. Collection is in volume or time mode and
different fraction sizes can be collected during different
stages of a separation.
Liquid chromatography
Automatic peak fractionation, based on peak detection using
slope or level sensing, minimizes peak dilution and crosscontamination. Event marks correlate the fractions with the
chromatogram. A recycle function enables collection from
repetitive runs.
Further information: Please ask for Frac-950
Data File No. 18-1153-57.
Fraction collector Frac-901
Frac-901 fraction collector can collect up to 175 fractions in 12
mm diameter tubes, up to 95 fractions in 10–18 mm diameter
tubes and up to 40 fractions in 30 mm diameter tubes.
Fixed volume fractionation or automatic peak fractionation
is possible. Peak fractionation is based on peak detection
using slope sensing. Fraction marks and fraction numbers
make it easy to identify fractions and peaks.
Automatic multiple sample injection (large volumes or high
throughput)
Sample Pump P-950, together with a motorized valve
PV-908, is used for automatic multiple sample injection.
The flow rate range (up to 50 ml/min), low pulsation and
pressure specification (up to 1 MPa) allow rapid sample
application directly onto a column or via a Superloop.
Large sample volumes can be applied smoothly and safely.
The pump is controlled by UNICORN and requires no
calibration before use.
Autosampler A-900 is available with or without Peltier
cooling option and increases system throughput by allowing
automatic injection of up to 96 samples from standard vials
or 160 samples from microvials. Control of the autosampler
is through UNICORN control system.
For further information: Please ask for
Data File No. 18-1152-89
Automatic column or buffer selection
Motorized 8-port valves can be used to select a specified
column or buffer allowing automatic column, media or buffer
scouting.
Increased security in programmed events
A maximum of three air sensors can be positioned within the
system and controlled through UNICORN to increase
security in programmed events. For example, UNICORN is
programmed to ensure that a chromatographic elution
begins only when the completion of sample application has
been detected by an air sensor. Variable sample volumes are
applied with no risk of air entering the system or column.
Flow control
Motorized valves INV-907, IV-908, or PV-908 can be added
to allow additional programmable functions such as
automated reverse flow through a column or flow diversion.
Flexibility in connecting external equipment
AD-900 is an analog/digital converter for connecting an
external instrument to the UNICORN control system in an
ÄKTAdesign system.
The module has one high-resolution analog input for
monitoring e.g. pressure, UV monitor signals, or other
signals available as voltage outputs.
System specifications
ÄKTApurifier 10
Flow rate range setting 0.001–10 ml/min
Pressure range
0–25 MPa
ÄKTApurifier 100
Flow rate range setting 0.01–100 ml/min
Pressure range
0–10 MPa
Conductivity range
1 µS/cm–999.9 mS/cm
(RPC – IEX – HIC gradients)
pH range
2–12
Temperature range
4 °C–40 °C
Solvent compatibility
all commonly used chromatographic
solvents
Size
500 mm × 460 mm × 620 mm
(W × D × H)
Weight
75 kg
Operating data
P-903
Flow rate range
Increment
Pressure range
Pressure limits
Internal volume
Viscosity
P-901
Flow rate range
Increment
Pressure range
Pressure limits
Internal volume
Viscosity
18-1119-48 AC, 2001-08 • p14
0.01–100 ml/min
0.01 ml/min
0–10 MPa (100 bar, 1450 psi)
programmable upper and lower limit
< 1800 µl/pump module
maximum 5cP
Monitor UV-900
Wavelength range
Bandwidth
Wavelength accuracy
Wavelength
reproducibility
Linearity
Noise* (at 230 nm)
Drift (at 254 nm)
Max. pressure
df
0.001–10 ml/min
0.001 ml/min
0–25 MPa (250 bar, 3625 psi)
programmable upper and lower limit
< 600 µl/pump module
maximum 5cP
190–700 nm in step of 1 nm,
3 wavelengths simultaneously
4 nm
± 2 nm
± 0.01 nm
< 2% deviation up to 2 AU at
260 nm with Uracil at pH 2
< 6 × 10–5 AU
< 2 × 10–4 AU/h
2 MPa (20 bar, 290 psi)
Liquid chromatography
Monitor pH/C-900
Conductivity unit
Conductivity range
Noise
pH unit
pH range
Accuracy
Stability
A/D Converter AD-900
1 µS/cm–999.9 mS/cm
±0.5% of full-scale calibrated range
0–14 (specifications valid
between 2–12)
±0.1 pH unit, temperature compensated
max 0.1 pH units deviation/10 h
Fraction collection
With Frac-950
Flow rate range
Fraction capacity
Optional racks
Rack A
Rack B
Rack C
Rack D
Rack E
Rack F
Rack G
Prep Mode
With Frac-901
Tube capacity
0.001 ml/min–100 ml/min
120 × 18 mm tubes
8 × 30 mm tubes
240 × 12 mm tubes
4 microplates (4 × 96 wells per plate)
8 × 30 mm tubes
45 × 30 mm tubes
80 × 30 mm tubes
20 × 250 ml vessels
30 funnels
Conversion Kit
95 in Tube Rack 18 mm
40 in Tube Rack 30 mm (optional)
175 in Tube Rack 12 mm (optional)
Voltage range
Resolution
Input impedance
Accuracy
Max. input voltage
Power requirement
Dimensions incl. attachment
(W x D x H)
Weight
± 2V DC
1 µV, 20 bit
> 1 M⍀
> ± 0.5 % or ± 0.05 mV
±5V
18–36 DC through the UniNet
2 connection
45 x 175 x 85 mm
0.4 kg
Computer requirements
Pentium II/333 MHz or later (minimum Pentium/90 MHz)
One system: 64 Mb RAM (minimum 32 Mb)
Two or more systems: 92 Mb RAM (minimum 64 Mb)
1 Gb available hard disk(150 Mb to run UNICORN),
NTFS file system
Colour monitor, 1024 × 768 with 64 k colours
1 ISA slot/connected system
Mouse
CD-ROM drive
1.44 Mb (3.5²) diskette drive
Software requirements for UNICORN 4.0
* Measured with water at 1 ml/min, time constant 1 s, 10 mm flow cell.
Windows NT™ Workstation 4.0 + Service Pack 4 or later or
Windows 2000.
Sample Pump P-950
Network requirements
Flow rate range
Operating pressure
Viscosity
0.1–50 ml/min
0–1.0 MPa
5 cP
10 cP at reduced flow rate
Autosampler
Sample capacity
Injection volume
Injection valve
switch time
Precision
Full loop injections
Partial loopfill injections
µl pick-up
Sample viscosity
Programming
Dimensions
96 standard vials (1.5 ml),
160 micro vials (0.5 ml)
5–1000 µl, flushed loop
1–500 µl, partial loop fill
1–470 µl, µl pick up
<100msec
rsd ≤ 0.3%
rsd ≤ 0.5%
for injection volumes > 5 µl
rsd ≤ 1.0% for volumes > 5 µl
0–10 cP
through UNICORN control only
280 mm × 540 mm × 440 mm
(W × D × H)
A-900 with cooling only
Cooling capacity
>11.5 °C below ambient
temperature (T) for 16 °C <T <40 °C
Typical cooling times
34 min from 23 °C to 4 °C
(at 45% relative humidity)
60 min from 32 °C to 4 °C
(at 35% relative humidity)
Programming through UNICORN control only
df
18-1119-48 AC, 2001-08 • p15
Supported network cards:
3COM Etherlink III
Compaq™ Netelligent 10/100 TX Embedded UTP Controller
Compaq Integrated NetFlex-3 Controller
AMD PCNET PCI Ethernet Adapter (Integrated)
Novell™ NetWare™ version 4.50.819 or later, alternatively or
Windows NT Server 4.0
A valid network connection
Liquid chromatography
Ordering information
Code No.
18-1400-00
ÄKTApurifier 100 (fraction collector not included) 18-1404-00
Computer*
* Your local Amersham Biosciences representative will provide details of the fully
installed computer to be supplied with the system.
Frac-950 fraction collector
complete with Rack A
OR
Frac-901 fraction collector
complete with 18 mm tube rack
Increased automation: automatic column,
media or buffer scouting
Buffer selection
Valve IV-908, including one UniNet cable
(one valve for pump A)
Column selection
Valve PV-908, including one UniNet cable
(2 valves required)
18-6083-00
18-1118-97
Sample application
Superloop 10 ml (ÄKTAdesign), load 1–10 ml
18-1113-81
Superloop 50 ml (ÄKTAdesign), load 1–50 ml
18-1113-82
Superloop 150 ml, load 1–150 ml
18-1023-85
(requires union 1/16″ female – M6 male fitting, code no. 18-1112-57)
Additional items
UNICORN analysis module
AD-900, including a UniNet cable 0.7 m,
a Mini-DIN cable 1.5 m
18-1134-74
18-1148-62
Related product literature
18-1108-42
18-1108-41
Autosampler A-900
18-1116-61
OR
Autosampler A-900, cooled version
18-1144-61
(both include 200 x 1.5 ml sample vials and caps)
ÄKTAdesign XT kit
18-6083-19
(includes tubing and UNICORN analysis module)
Sample Pump P-950
Valve PV-908
18-1108-40
18-1108-42
18-1108-41
18-6083-01
18-1108-41
Increased security in programmed events – air sensors
Air-900 control box, includes one UniNet
2 cable, (controls up to 3 air sensors)
18-1121-22
Purge valve (required for buffer inlet
position only)
18-1126-33
Air sensor Air-925 (2.5 mm i.d.)
18-1121-24
Air sensor Air-912 (1.2 mm i.d.)
18-1121-23
Product
Data Files
ÄKTAFPLC™
ÄKTAbasic Systems
ÄKTAexplorer Systems
Monitor UV-900
Pump P-900 series
Monitor pH/C-900
ÄKTAprime
UNICORN Control System
Frac-950
Autosampler A-900
Application Notes
Purification of peptides from natural sources
Efficient isolation of protein fragments
for structural analyses
Purification of synthetic peptides
Code No.
18-1128-41
18-1128-19
18-1124-09
18-1111-17
18-1119-49
18-1111-19
18-1136-91
18-1111-20
18-1153-57
18-1152-89
18-1119-52
18-1119-53
18-1119-51
to order:
Asia Pacific Tel: +852 2811 8693 Fax: +852 2811 5251 Australasia Tel: +61 2 9899 0999 Fax: +61 2 9899 7511 Austria Tel: 01 576 0616 20 Fax: 01 576 0616 27 Belgium Tel: 0800 73 888 Fax: 03 272 1637
Canada Tel: 1 800 463 5800 Fax: 1 800 567 1008 Central, East, South East Europe Tel: +43 1 982 3826 Fax: +43 1 985 8327 Denmark Tel: 45 16 2400 Fax: 45 16 2424 Finland & Baltics Tel: +358 (0)9 512 3940
Fax: +358 (0)9 512 1710 France Tel: 0169 35 67 00 Fax: 0169 41 9677 Germany Tel: 0761 4903 401 Fax: 0761 4903 405 Italy Tel: 02 27322 1 Fax: 02 27302 212 Japan Tel: 81 3 5331 9336 Fax: 81 3 5331 9370 Latin
America Tel: +55 11 3667 5700 Fax: +55 11 3667 87 99 Middle East and Africa Tel: +30 (1) 96 00 687 Fax: +30 (1) 96 00 693 Netherlands Tel: 0165 580 410 Fax: 0165 580 401 Norway Tel: 2318 5800 Fax: 2318 6800
Portugal Tel: 21 417 7035 Fax: 21 417 3184 Russian & other C.I.S. & N.I.S. Tel: +7 (095) 232 0250,956 1137 Fax: +7 (095) 230 6377 South East Asia Tel: 60 3 8024 2080 Fax: 60 3 8024 2090 Spain Tel: 93 594 49 50
Fax: 93 594 49 55 Sweden Tel: 018 612 19 00 Fax: 018 612 19 10 Switzerland Tel: 01 802 81 50 Fax: 01 802 81 51 UK Tel: 0800 616 928 Fax: 0800 616 927 USA Tel: +1 800 526 3593 Fax: +1 877 295 8102
ÄKTA, UNICORN, FPLC, RESOURCE, Sephasil, Cy, CyDye, Mono Q, Mini Q, Superloop, and Labcrew are trademarks of Amersham Biosciences Limited.
Compaq is a trademark of Compaq Computer Corporation. Windows NT is a trademark of Microsoft Corporation. Novell and Netware are trademarks of Novell Inc.
Patents pending for Pump P-900, Sample Pump P-950, and Frac-950. Amersham Biosciences AB Björkgatan 30, SE-751 84 Uppsala, Sweden. Amersham Biosciences UK Limited Amersham Place, Little
Chalfont, Buckinghamshire HP7 9NA, England. Amersham Biosciences Inc 800 Centennial Avenue, PO Box 1327, Piscataway, NJ 08855 USA. Amersham Biosciences Europe GmbH Munzinger Strasse
9, D-79111 Freiburg, Germany. Amersham Biosciences K.K. Sanken Building, 3-25-1, Hyakunincho, Shinjuku-ku, Tokyo 169-0073, Japan. All goods and services are sold subject to the terms and conditions
of sale of the company within the Amersham Biosciences group that supplies them. A copy of these terms and conditions is available on request. © Amersham Biosciences AB 2001 – All rights
reserved.
df
18-1119-48 AC, 2001-08 • p16
Produced by Wikströms, Sweden 1010980, 08.2001
Printed matter. Licence 341 051
Product
ÄKTApurifier 10 (fraction collector not included)
Computer*
Flow control e.g. for reverse flow, flow diversions
(up to three valves of any type)
INV-907, including one UniNet cable
IV-908, including one UniNet cable
PV-908, including one UniNet cable
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