Transcript ISET 2 - Agilent

Page 1
July 17, 2015
ISET Training
Agilent Restricted
Agilent 1290 Infinity with
Intelligent System Emulation Technology
(ISET)
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July 17, 2015
ISET Training
Agilent Restricted
Introduction
Intelligent System Emulation Technology (ISET)
Trainings Material
Course: AN-CE-LC-3-015-A (ISET)
Course: AN-CE-LC-3-015-B (ISET 2)
LC - Intelligent System Emulation Technology (ISET)
(link to video about ISET features)
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Agenda
Understand
Basic
Concepts of
ISET
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July 17, 2015
Activate and
Configure
ISET
Work with
ISET
Support
ISET
ISET 2
Update from
ISET
ISET Training
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Goal
Running traditional HPLC methods using
a 1290 Infinity LC with ISET system
obtaining very similar results!
Method transfer
Check specifications about ISET product
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Needs
Recently a number of UHPLC instruments have been introduced to the market.
These systems are typically designed for the operation of small bore columns (2.1 mm 3 mm internal diameter) packed with small particles to achieve high separation
efficiencies.
With regard to solvent delivery these systems usually differ significantly from standard
HPLC systems, in particular the dwell volumes of such systems are minimized to allow
fast gradient separations.
When the attempt is made to transfer methods developed on standard HPLC systems to
modern UHPLC systems, changes in retention times and often also in selectivity are
observed.
These changes are due to the differences in system (dwell) volumes, mixing
characteristics and also properties of the pumping system (composition accuracy).
ISET is to enable the transfer of methods from standard HPLC type instruments to
UHPLC systems without changing the original method.
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Important Parameters
Design differences between LC instrumentation – such as power range, delay volume, mixing
behavior, temperature control, extra column volume and detector cell design – all affect the
ability to transfer a method from one system to another.
Instrument to instrument method transfer is often problematic, especially in highly regulated
industries, because any modifications of the instrument and original method should be avoided.
Extra column volume
Delay volume
Now, the 1290 Infinity LC with ISET extends capability by enabling method transfer between
(U)HPLC systems with different instrument designs.
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Pump
Sampler
Column
Thermostat
Detector
July 17, 2015
Delay volume
Gradient mixing behavior
Pressure x flow rate
Retention Time
Resolution
Delay volume
Extra column volume
Injection volume
Retention Time
Resolution
Sensitivity
Temperature profile
Extra column volume
Data rate
Extra column volume
Path-length
Retention Time
Resolution
Resolution
Retention Time
Sensitivity
ISET Training
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Gradient Analyses with Different LC Systems
- Impact of delay volume and mixing behavior
DAD1 B, Sig=275,4 Ref=400,40 (B:\RIC\AGI...RIC DATA\1 PHARMA METOCLOPRAMIDE\WAD PHARMA\XBRIDGE-2000004.D)
mAU
200
8.930
1200 Series LC Quaternary Pump
9.975
12.005
12.215
12.393
10
12
9.545
50
3.049
2.747
100
8.747
8.121
8.323
150
2.744
9.937
6.872
7.480
100
7.923
150
6.328
2.884
1290 Infinity LC
6.673
mAU
200
6.088
2
4
6
8
DAD1 C, DAD1B, DAD: Signal B, 275 nm/Bw:4 nm Ref 400 nm/Bw:60 nm (B:\RIC\AGI...A\GV 1PHARMA\HPLC000001.D)
10.249
0
50
0
10.379
9.951
9.159
8.744
3.320
2.887
50
0
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2
12
The result:
• Difference in RT and Resolution
• One peak is missing!
150
100
9.325
8.538
mAU
200
10
12.395
12.630
12.763
2
4
6
8
DAD1 C, DAD1B, DAD: Signal B, 275 nm/Bw:4 nm Ref 400 nm/Bw:60 nm (B:\RIC\AGI...A\GV 1PHARMA\HPLC000003.D)
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July
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Emulation!
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Delay volumes and Mixing
What happens with a programmed gradient?
response
90%
Programmed gradient step
1200 Series Binary Pump SL
1290 Infinity Binary Pump
Gradient slope/
Mixing behavior
10%
Delay
volume
Total delay volume of the system
(sum of capillaries, mixer, cells,
valves..)
1200 Series
1290 Infinity
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time
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How it works
• Simple and flexible
(select the emulated LC system)
• No determination of the dwell volume etc. required
(perfectly covers pumps with damper and variable dwell volume)
• Requires no modification of the methods
(no conflicts in regulated environment)
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How it works 1290 to 1200
mAU
400
350
300
Injection
Same gradient conditions by moving
the gradient and by emulating the
gradient mixing behavior.
250
200
150
Programmed gradient
100
1290 gradient
50
1200 gradient
0
0
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0.5
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1
1.5
2
2.5
3
3.5
min
ISET Training
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Why 1290?
Two components
A. Best in class performance of the
1290 Infinity LC
Peak 3
Overlay of 10 runs
RSD = 0.005 %
Broad power range, ultra-low delay volume,
superior sensitivity, unmatched flow and
composition accuracy
B. The revolutionary ISET emulation
algorithm
Full characterization of target LC systems
Creation of emulation function
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Emulation – What is?
Method Transfer
- Emulate other HPLC and instruments
- Run existing HPLC methods without modifying your method or system
- Achieve very similar retention times and peak resolution for infinitely better method transfer
Check specifications about ISET product
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Emulation Models
ISET 2 updates
Generic Emulation
Waters
Alliance
2690/95
Waters
Alliance HT
2790/95/96
1220 Infinity LC
1290 Infinity LC with ISET
1100 Series LC
Added the Jet weaver V380 Characterization
1260 Infinity LC
1200 Series LC
Pump Valve Cluster Support
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Emulation - Agilent
Pumps
G1311A
G1311B
G1311C
G1312A
G1312B
G1312C
G5611A
1100 Series Quaternary Pump
1200 Series Quaternary Pump
1260 Infinity Quaternary Pump
1260 Infinity Quaternary Pump VL
1100 Series Binary Pump
1200 Series Binary Pump SL
1260 Infinity Binary Pump
1260 Infinity Binary Pump VL
1260 Infinity Bio-inert Quaternary Pump
Autosamplers
G1313A
G1329A
G5667A
1100 Series Standard Autosampler
1100 Series Thermostatted Autosampler
1200 Series Standard Autosampler
1200 Series Standard Autosampler SL
1260 Infinity Standard Autosampler
1100 Series Well Plate Sampler
1100 Well Plate Sampler
1200 Series High Performance Autosampler
1200 Series High Performance Autosampler SL
1200 Series High Performance Autosampler SL+
1260 Infinity High Performance Autosampler
1100 Micro Well Plate Sampler
1200 Series Micro Well Plate Autosampler
1260 Infinity High Performance Micro Autosampler
1260 Infinity High Performance Bio-inert Autosampler
G4288A, G4289AA, G4290AA
G4288C, G4290C
G4288B, G4290B, G4294B
1120 Compact LC
1220 Infinity LC (VL)
1220 Infinity LC
G1329B
G1367A (100 μL syringe)
G1367B (100 μL syringe)
G1367C (100 μL syringe)
G1367D (100 μL / 40 μL syringe)
G1367E (100 μL / 40 μL syringe)
G1377A
LC Systems
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Emulation - Waters
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Emulation - Generic
The generic emulation using dwell volume can be used for LC instruments not listed as ISET supported modules/systems.
In these cases the dwell volume has to be entered into the field Dwell Volume
Note:
The generic emulation using dwell volume does not compensate the following aspects:
• Different mixing behaviors
• Composition errors
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Emulation - Generic
Dwell Volume
Definition
The system dwell volume includes the volume of the system flow path from the point of mixing of the mobile
phase components to the head of the column.
It has an important effect on the gradient that the pump delivers because every gradient has an initial isocratic
segment as the mobile phase must travel through the dwell volume before any change made at the pump arrives
at the head of the column.
Any differences in dwell volume cause variations in retention times and often also in selectivity.
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ISET Training
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Emulation - Generic
Determination of Dwell Volume
1 - Replace column by a zero dead volume connection or low volume capillary.
2 - Run a linear gradient from 0 – 10 min from 5 – 95 % B at a flow rate (F) of 1 mL/min with water
(channel A) and water with 0.2 % acetone (channel B) at 263 nm detection wavelength.
3 - Determine the difference in time between the programmed and actual elution time of the gradient at
50 % of the composition.
4 - Measure tdwell (usually measured at 50 % of composition delta)
Vdwell can be calculated from:
Vdwell = tdwell × F
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Compatibility
ISET will be included in a new RC.net driver for the 1290 Infinity pumps,
thus any software supporting RC.net drivers can make use of this feature!
ISET
•
•
•
•
•
•
Rc.net Drivers A.02.05
OpenLab CDS ChemStation Edition C.01.03
OpenLab CDS EZ Edition A.04.03
ChemStation B.04.03 SP1
ICF A.01.04
Masshunter B.05.00
ISET 2
•
•
•
•
•
•
•
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Drivers Update only!
Rc.net Drivers A.02.06
OpenLab CDS ChemStation Edition C.01.03 (+ planned SP1)
OpenLab CDS ChemStation Edition C.01.04 + Rc.net A.02.06
OpenLab CDS EZ Edition A.04.03 (+ planned SP1)
OpenLab CDS EZ Edition A.04.04 + Rc.net A.02.06
ICF A.01.05 (Sep 2012)
Masshunter (planned B.05.01 Q/TOF – B.06.00 QQQ)
July 17, 2015
ISET Training
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Product structure
Product
Description
G2197AA
G2197AA ISET Kit
Price $2000
Delete option will be available to remove ISET
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Activation
Firmware >= B.06.42
Activation or re-activation will be done plugging the USB stick into Pump
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Activation Steps
1- Power off the Module
2- Plug the USB Dongle into the Infinity Pump USB Socket
3- Turn power on the G4220A/B
4- Once restarted you can choose to Remove the USB Dongle or not
Store the USB Dongle in a safe place
Must be used to re-activate ISET after a Mother Board replacement
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Configuration
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Configuration
ISET properly detected
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Configuration
New Jet Weaver v380 Mixer support
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Configuration
Importing future ISET Descriptions
(emulated hplc's)
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Graphical User Interface
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Method Parameters
2
1
3
4
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Solvent Types
1
Do not deselect!
Solvents selections
-V.0x drop down menu)
-Aqueous/Organic/Generic
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Enabling ISET
Enabling ISET to emulate target HPLC
2
ISET V1.0 still available
To keep the compatibility
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Enabling ISET
Enabling ISET to emulate target HPLC
2
ISET 2 will ask for Manufacturer:
-Agilent
-Waters
-Generic LC Instrument
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Select Target Pump
Select the target HPLC pump - Agilent
3
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Select Target Pump
Select the target HPLC pump - Waters
3
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Select Target Pump
Select the target HPLC pump – Generic
3
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Select Target Autosampler - Agilent
4
Select the target HPLC Sampler
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Select Target Autosampler - Waters
4
Select the target HPLC Sampler
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Advanced Parameters
Fine tuning about compressibility
Fine tuning to adjust chromatographic results
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Setting the Time Table - Agilent
Original Time Table
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View “Emulated” Gradient - Agilent
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ISET Time Table - Agilent
Initial conditions are updated
Not possible to modify during RUN
Note: the “Emulated” Timetable will NOT be printed
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Setting the Time Table - Waters
With Waters you HAVE to select first the “Manufacturer”
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View “Emulated” Gradient - Waters
Curve 6 means “Linear”
Enter here the original Method parameters (Waters)
…and populate here with the “Original” Waters Method
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ISET Time Table - Waters
Initial conditions are updated
Not possible to modify during RUN
Note: the “Emulated” Timetable will NOT be printed
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Method printout – ISET
Emulation Mode
Enabled:
Model:
Yes
ISET V1.0
ISET Parameter
Delay Volume Offset:
Manually Select ISET Solvent Model:
Enable Tuning:
Manual Set Sampler Parameter:
Manual Set Pump Parameter:
Emulated Pump:
Solvent Model:
Emulated Sampler:
0 µL
No
No
No
No
G1311A V1.0
Methanol (Channel A) - Water (Channel B)
G1367A - 100 µL Syringe V1.0
Pump Parameter
Typical Operating Pressure: 200.00 bar
Compressibility A:
100 10e-6/bar
Sampler Parameter
Sampler Seat Capillary:
2.30 µL
Method printout (only the ISET part is reported here)
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Method printout – ISET 2
ISET Parameter
Emulation Mode
Emulation Enabled:
Emulation Model:
Emulation Manufacturer:
Generic
Yes
ISET 2 V1.0
Generic LC Instrument
ISET Parameter
Generic Emulation Dwell Volume:
ISET Parameter
Emulation Mode
Emulation Enabled:
Emulation Model:
Emulation Manufacturer:
350 µL
Waters
Yes
ISET 2 V1.0
Waters
ISET Parameter
Emulated Pump:
Alliance 2690, 2695 V1.0
Solvent Model: Water (Channel A) - Acetonitrile (Channel B)
Emulated Sampler: Alliance 2690, 2695 (100 µL Loop) V1.0
Manually Select ISET Solvent Model:
No
Dwell Volume Offset:
0 µL
Enable Tuning:
No
Manual Set Sampler Parameter:
No
Manual Set Pump Parameter:
No
ISET Parameter
Emulated Pump:
G1311A V1.0
Solvent Model: Water (Channel A) - Acetonitrile (Channel B)
Emulated Sampler:
G1367A - 100 µL Syringe V1.0
Manually Select ISET Solvent Model:
No
Dwell Volume Offset:
0 µL
Enable Tuning:
No
Manual Set Sampler Parameter:
No
Manual Set Pump Parameter:
No
Agilent
Sampler Parameter
Sampler Seat Capillary:
Pump Parameter
Typical Operating Pressure:
Compressibility A:
2.3 µL
200.00 bar
100 10e-6/bar
Sampler Parameter
Pump Parameter
Typical Operating Pressure:
200.00 bar
Method printout (only the ISET part is reported here)
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Method changed history
The Method Resolution will appear because the
ISET parameter was not present
Method changed flag will be shown in the history
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Solvents
Best results with “Characterized Solvent” V.0x drop down
menu
Solvent lists are characterized for gradient compositions
“Aqueous”
“Organic”
when buffers or solvent mixtures are used
Aqueous (Channel A) – Organic (Channel B)
or
Organic (Channel A) – Aqueous (Channel B)
for pure solvents as appropriate
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Demo Function
The ISET will be available with each new 1290 pump
This function is temporarily available for demo purposes
Demo features will be available if a delete option is selected when ordering a New 1290 Pump
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Repair
The ISET will NOT be removed updating the Firmware
After a Mother board replacement the USB stick
(originally used for ISET)
must be used to re-activate the function
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Specifications
Retention Time (RT) Deviation
at 40°C for non Agilent instruments
RT
±5%
±0.3 min
(for RT > 6 min)
(for RT ≤ 6 min)
∆RT
±5%
±0.1 min
(for ∆RT > 2 min)
(for ∆RT ≤ 2 min)
Excluded:
• Generic Emulation
• Generic Solvent
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Labs
- Check out sample
- Modify delay volume parameters
- Understanding the emulations available
(100 lines of timetable)
- Selecting the “Emulated HPLC”
- Offline ISET
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Advanced
- ISET used to develop method for old hardware using the 1290 LC system
1100 Series LC
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1220 Infinity LC
1260 Infinity LC
1200 Series LC
ISET Training
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Advanced – Offline ISET
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Advanced – Offline ISET
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Advanced – Offline ISET
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What’s new
ISET 2 – What’s new
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What’s new
Generic Emulation
ISET 2 updates
Waters
Alliance
2690/95
Waters
Alliance HT
2790/95/96
1220 Infinity LC
1290 Infinity LC with ISET
Added the Jet weaver V380 Characterization
Pump Valve Cluster Support
No temperature compensation
1100 Series LC
Method Transfer
1260 Infinity LC
1200 Series LC
- Emulate other HPLC and instruments
- Run existing HPLC methods without modifying your method or system
- Achieve very similar* retention times and peak resolution for infinitely better method transfer
Page 59
July 17, 2015
ISET Training
Agilent Restricted
Emulation - Agilent
Pumps
G1311A
G1311B
G1311C
G1312A
G1312B
G1312C
G5611A
1100 Series Quaternary Pump
1200 Series Quaternary Pump
1260 Infinity Quaternary Pump
1260 Infinity Quaternary Pump VL
1100 Series Binary Pump
1200 Series Binary Pump SL
1260 Infinity Binary Pump
1260 Infinity Binary Pump VL
1260 Infinity Bio-inert Quaternary Pump
Autosamplers
G1313A
G1329A
G5667A
1100 Series Standard Autosampler
1100 Series Thermostatted Autosampler
1200 Series Standard Autosampler
1200 Series Standard Autosampler SL
1260 Infinity Standard Autosampler
1100 Series Well Plate Sampler
1100 Well Plate Sampler
1200 Series High Performance Autosampler
1200 Series High Performance Autosampler SL
1200 Series High Performance Autosampler SL+
1260 Infinity High Performance Autosampler
1100 Micro Well Plate Sampler
1200 Series Micro Well Plate Autosampler
1260 Infinity High Performance Micro Autosampler
1260 Infinity High Performance Bio-inert Autosampler
G4288A, G4289AA, G4290AA
G4288C, G4290C
G4288B, G4290B, G4294B
1120 Compact LC
1220 Infinity LC (VL)
1220 Infinity LC
G1329B
G1367A (100 μL syringe)
G1367B (100 μL syringe)
G1367C (100 μL syringe)
G1367D (100 μL / 40 μL syringe)
G1367E (100 μL / 40 μL syringe)
G1377A
LC Systems
Page 60
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Emulation - Waters
Page 61
July 17, 2015
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Compatibility
ISET will be included in a new RC.net driver for the 1290 Infinity pumps,
thus any software supporting RC.net drivers can make use of this feature!
ISET
 Rc.net Drivers A.02.05
 OpenLab CDS ChemStation Edition C.01.03
 OpenLab CDS EZ Edition A.04.03
 ChemStation B.04.03 SP1
 ICF A.01.04
Drivers Update only!
 Masshunter B.05.00
ISET 2
 Rc.net Drivers A.02.06
 OpenLab CDS ChemStation Edition C.01.03 (+ planned SP1)
 OpenLab CDS ChemStation Edition C.01.04 + Rc.net A.02.06
 OpenLab CDS EZ Edition A.04.03 (+ planned SP1)
 OpenLab CDS EZ Edition A.04.04 + Rc.net A.02.06
 ICF A.01.05 (Sep 2012)
 Masshunter (planned B.05.01 Q/TOF – B.06.00 QQQ)
Page 62
July 17, 2015
ISET Training
Agilent Restricted
Emulation - Generic
The generic emulation using dwell volume can be used for LC instruments not listed as ISET supported modules/systems.
In these cases the dwell volume has to be entered into the field Dwell Volume
Note:
The generic emulation using dwell volume does not compensate the following aspects:
• Different mixing behaviors
• Composition errors
Page 63
July 17, 2015
ISET Training
Agilent Restricted
Emulation - Generic
Dwell Volume
Definition
The system dwell volume includes the volume of the system flow path from the point of mixing of the mobile
phase components to the head of the column.
It has an important effect on the gradient that the pump delivers because every gradient has an initial isocratic
segment as the mobile phase must travel through the dwell volume before any change made at the pump arrives
at the head of the column.
Any differences in dwell volume cause variations in retention times and often also in selectivity.
Page 64
July 17, 2015
ISET Training
Agilent Restricted
Emulation - Generic
Determination of Dwell Volume
1 - Replace column by a zero dead volume connection or low volume capillary.
2 - Run a linear gradient from 0 – 10 min from 5 – 95 % B at a flow rate (F) of 1 mL/min with water
(channel A) and water with 0.2 % acetone (channel B) at 263 nm detection wavelength.
3 - Determine the difference in time between the programmed and actual elution time of the gradient at
50 % of the composition.
4 - Measure tdwell (usually measured at 50 % of composition delta)
Vdwell can be calculated from:
Vdwell = tdwell × F
Page 65
July 17, 2015
ISET Training
Agilent Restricted
Enabling ISET
Enabling ISET to emulate target HPLC
2
ISET 2 will ask for Manufacturer:
-Agilent
-Waters
-Generic LC Instrument
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Select Target Pump
Select the target HPLC pump - Agilent
3
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Select Target Pump
Select the target HPLC pump - Waters
3
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Select Target Pump
Select the target HPLC pump – Generic
3
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Select Target Autosampler
4
Select the target HPLC Sampler
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Select Target Autosampler
4
Select the target HPLC Sampler
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