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Agilent 1290 Infinity with ISET

1. Sap gt de ES 0008000588086088 O mt L O 1290 Infinity ISET 1290 Infinity ISET Installing and Configuring ISET 2 3 Switch on the G4220A B pump to start the activation procedure Do not remove the ISET USB dongle at this stage The pump configuration screen is displayed the ISET installed check box in the Options section is marked to indicate that the installation is complete You can now remove the ISET USB dongle Options Pressure Unit bar X Installed mixer Jet Weaver V35 Mixer gt V ISET installed ISET Configuration 17 2 Installing and Configuring ISET ISET Removal and Reinstallation 18 ISET functionality will not be affected by a firmware update If ISET was present before the update it will remain active after the procedure Replacement of the main board of 1290 Infinity pump will completely remove the ISET functionality Reinstalling ISET In the case of a replacement of the pump s main board you will need to reinstall ISET To reinstall ISET follow the installation procedure see above You must use the same ISET USB dongle for reinstallation as was originally used for installation The ISET USB dong
2. 1 i 333 333 IP TT l es Agilent Technologies Notices Agilent Technologies Inc 2011 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Manual Part Number 64220 90310 Edition 09 11 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn This product may be used as a com ponent of an in vitro diagnostic sys tem if the system is registered with the appropriate authorities and com plies with the relevant regulations Otherwise it is intended only for gen eral laboratory use Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connection with the furnishing use or perfor mance of this document or of any information contained herein Should Agilent and the
3. user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 or as a commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 June 1987 or any equivalent agency regulation or contract clause Use duplication or dis closure of Software is subject to Agilent Technologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 June 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 June 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in any technical data Safety Notices CAUTION A CAUTION notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhere
4. 0 This chapter explains in detail how ISET works Agilent Technologies 29 4 Understanding ISET Functionality Understanding ISET Functionality 30 The transfer of a method from one liquid chromatography system to another usually involves a direct transfer of all instrument parameters for example flow rate mobile phase composition and gradient timetable injection volume column temperature and UV detection wavelength However there can also be subtle differences such as system delay or dwell volume the volume of the system flow path from the point of mixing of the mobile phase components to the top of the column the design of the autosampler and column compartment flow paths and temperature detector cell design and the extra column volume of the system detector data collection rates and response times and the sensitivity of the detector A closer investigation into the comparative performance of different designs of LC gradient pumps reveals that other hidden factors have an effect on how the pump delivers the gradient Principally this is characterized by the mixing behavior of the pump which in turn is determined by the basic concept and design of the flow path the volume and design of the mixer the efficiency of the solvent mixing and the behaviour of the piston movement under different loads and compressibilities of solvents This behaviour can be visualized by using tracer experiments 0 2 acetone in channel B
5. 220A B pumps with firmware revision B 06 42 and later Unless the demonstration trial version is upgraded to a registered version the ISET function will be deactivated when the trial period has elapsed 1290 Infinity ISET 13 1 Whatis ISET The solution for instrument to instrument method transfer 1290 Infinity LC with ISET 14 1290 Infinity ISET 1290 Infinity ISET Oo e Oo 3 m o o e0 90 20 dia and Configuring ISET Mal O we e il Installing and Configuring ISET 16 e ISET Removal and Reinstallation 18 This chapter gives detailed step by step instructions for installing a new system and upgrading an existing system Agilent Technologies 2 Installing and Configuring ISET Installing and Configuring ISET 16 e ISET USB dongle G4220A B pump with firmware rev B 06 42 or later e RC Net drivers rev A 02 04 or later e Either OpenLAB CDS ChemStation Edition c 01 03 or OpenLAB CDS EZChrom Edition A 04 04 The installation of ISET in the trial version is already complete if the prerequisits are met and the components have been properly installed To upgrade to the registered version you need to follow these steps 1 Switch off the G4220A B pump 2 Plug the ISET USB dongle into the USB port on the back of the G4220A B pump USB Port Y GO 35 SEO Y O 26 T FLASH CAN DC QUT GAH CAN i O ies _ pe ass Zm Z gt DCL C a a C
6. ISET ISET V1 0 v View Emulation Set Model Parameter Emulated Pump G1311A V1 0 gt manually select ISET solvent model Compressibility 108 bar Emulated Sampler G1367A 100 Syringe V1 0 x manually set Enable manual fine tuning Figure 2 Model Parameter section 23 3 Setting Up ISET Parameters The Model Parameter section enables you to configure the ISET emulation by selecting the original hardware module that you used during the development of your original method Currently the ISET library holds parameters only for the pump and the sampler since these are the major components that influence the mobile phase composition and its transient parameters as seen on the head of the column 6 Click the Emulated Pump down arrow and select the pump used for the original method from the drop down list see Figure 2 on page 23 The drop down list shows the module number of the pump and the revision of its characterization The current revision is V1 0 Future revisions of ISET might provide updated characterizations To determine a pump s name by its specific module number please refer to List of supported ISET modules on page 12 7 Ifthe solvent system required by the method includes only water methanol or acetonitrile you need make no changes to the solvent model In ISET v1 0 water methanol and acetonitrile are fully characterized for gradient compositions 8 If solvents other than water methanol or ace
7. T 6 The solution for instrument to instrument method transfer 1290 Infinity LC with ISET 9 2 Installing and Configuring ISET 15 Installing and Configuring ISET 16 ISET Removal and Reinstallation 18 3 Setting Up ISET Parameters 19 Preparing the 1290 Infinity System for emulation 20 Setting up the basic ISET parameters 21 Fine tuning the emulation 26 4 Understanding ISET Functionality 29 Understanding ISET Functionality 30 4 1290 Infinity ISET P 1290 Infinity ISET e09 ee 1 0790 What is ISET O O O o a o a What is Intelligent System Emulation Technology ISET 6 The solution for instrument to instrument method transfer 1290 Infinity LC with ISET 9 This chapter gives a definition and brief overview of ISET and its intended use Agilent Technologies 1 Whatis ISET What is Intelligent System Emulation Technology ISET Instrument to instrument method transferability is an important topic for all laboratories throughout all industries where HPLC and UHPLC methods are transferred between different departments and locations with different LC instruments In the pharmaceutical industry the transfer of analytical methodology between R amp D contract research organizations and manufacturing is an essential part in the development of a new pharmaceutical product Several hundred observations from the FDA and a proposal for a new chapter in USP 1224 Transfer of analytical procedures emphasize the actuality an
8. amplers G1313A G1329A G1329B G1367A 100 uL syringe G1367B 100 uL syringe G1367C 100 uL syringe G1367D 100 uL 40 pL syringe G1367E 100 uL 40 uL syringe G1377A G5667A 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 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 Future revisions of ISET will also cover LC systems from other vendors 12 1290 Infinity ISET WhatisISET 1 Specifications Table 1 Retention Time RT Deviation RT 5 for RT gt 6 min 0 3 min for RT lt 6 min ART 5 for ART gt 2 min 0 1 min for ART lt 2 min A RT RT ART gt Demo Trial A 700 hour demonstration trial version of ISET is available with G4
9. at ISET has calculated This gradient time table will be used to emulate the original method Check that the stoptime of the original method and the emulation method are the same 13 When all method settings are finished click OK to close the method setup screen The ISET emulation method will then be prepared During the run of an ISET method it is not possible to change the method parameters 1290 Infinity ISET 25 3 Setting Up ISET Parameters Fine tuning the emulation 26 When the original system and the ISET system both use standard configurations no offset setting should be required As a rule of thumb only the excess delay volume of the system with respect to the standard configuration contributes to the offset The total delay volume offset setting is then composed of the original system s contribution minus the ISET system s contribution If the emulated method was already run under ISET the results of those previous runs may then be used to accomplish a perfect match of the emulated method with respect to the original if there are still remaining retention time differences In critical applications where the gradient is too early or reaches the column head later than expected the separation and selectivity are changed and influence the resolution and retention times For fine tuning the delay volume may be adjusted to improve the emulation 1 Mark the Enable manual fine tuning check box to display the fine tunin
10. ations in retention times and often also in selectivity The ISET emulation considers the delay volume of both pump and autosampler The delay volume from the autosampler through the column compartment to the detector is around 5 ul when red capillaries with 0 12 mm ID are installed and 14 ul when green capillaries with 0 17 mm ID are installed These differences in volume are usually minor they are not part of ISET and can be neglected in most cases If the volume is large enough to have an effect the Delay Volume Offset function is available in ISET to compensate for the additional volume The gradient delay resulting from geometric volume is shown in Figure 3 on page 31 Volume Figure 3 Step gradients are used to evaluate system delay volume and composition offset The system delay volume is determined at 10 of the step height transition volume is from the 10 height to the 90 height and the system dwell volume is determined at 50 of the step height 31 4 32 Understanding ISET Functionality Composition differences The primary and most visible aspect of compressibility settings is reduction of pressure ripple but for predictable and accurate pumping the flow accuracy is more critical Additionally with the high pressure blending concept of binary pumps this flow accuracy translates into compositional accuracy The compressibility of the solvents in use affect retention time stability and predictability espec
11. d importance of this topic Which parameters affect method transfer 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 Therefore identical LC methods used on different LC instrumentation could result in different retention time and chromatographic resolution 6 1290 Infinity ISET Whatis ISET 1 What is Intelligent System Emulation Technology ISET Delay volume Retention Time Gradient mixing behaviour Resolution E Pressure x flow rate 3 gt Delay volume Retention Time a Extra column volume Resolution e Injection volume Sensitivity o Retention Time Retention Time 3 Resolution Resolution 2 Data rate Resolution o Extra column volume Retention Time S Path length Sensitivity uw A wide range of instrument parameters have a direct impact on the retention times resolution and sensitivity of a separation The impact of delay volume and gradient mixing 1290 Infinity ISET The delay volume of an LC system determines how fast the gradient reaches the column Further the mixing behavior influences the gradient profile Both these factors delay volume and mixing behavior are determined by the instrument design and the consequences for method transfer are differences in retention times and in resolution What is ISET What is In
12. d to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated condi tions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly performed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indi cated conditions are fully under stood and met 1290 Infinity ISET In this book 1290 Infinity ISET In this book The manual describes the Agilent 1290 Infinity Intelligent System Emulation Technology ISET V1 0 Revision It contains the following What is ISET This chapter gives a definition and brief overview of ISET and its intended use Installing and Configuring ISET This chapter gives detailed step by step instructions for installing a new system and upgrading an existing system Setting Up ISET Parameters This chapter gives detailed step by step instructions for setting up the standard ISET parameters and for setting up and using a verification method to confirm that ISET is functioning as expected It also contains explanations of the advanced ISET parameters with step by step instructions for setting them up Understanding ISET Functionality This chapter explains in detail how ISET works Contents Contents 1 WhatisISET 5 What is Intelligent System Emulation Technology ISE
13. for example to plot the profile of the pump as it delivers a stepwise gradient in the proportion of the solvents In practice there is always a delay and the step gradient is smoothed and transformed into a curve that is characteristic of the pump Every model of pump has its own characteristic profile and this leads to slight differences in nominally similar gradients on different pumps Often these are sufficient to cause problems in the transfer of some methods and the challenge is to measure the characteristics of different pumps and reproduce them on the Agilent 1290 Infinity LC system pump This is the heart of the ISET system however the accurate performance of the 1290 Infinity is still needed to execute the settings ISET reveals and translates those parameters to provide an accurate method transfer from older Agilent systems to the 1290 Infinity system and vice versa 1290 Infinity ISET 1290 Infinity ISET Understanding ISET Functionality 4 System delay or dwell volume The system delay or dwell volume includes the volume of the system flow path from the point of mixing of the mobile phase components to the top 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 delay volume before any change made at the pump arrives at the head of the column Any differences in delay or dwell volume cause vari
14. g parameters 2 If necessary enter a Typical Operating Pressure During the formation of gradients using Water Methanol or Water Acetonitrile the system pressure changes This change in pressure affects the volume of the damper of the pump which results in a change in volume The Typical Operating Pressure can be used to compensate for this change in volume 3 If necessary enter a Delay Volume Offset The Delay Volume Offset can be used to compensate for a difference in delay volume between the original instrument and the ISET system caused for example by wider ID capillaries or an additional mixer that is non standard system configurations Positive Delay Volume Offsets shift the gradient to a later position negative Delay Volume Offsets shift the gradient to an earlier position 4 You can click View Emulation Set to display the modifications to the gradient that ISET has calculated and will be used to emulate the original method 1290 Infinity ISET Setting Up ISET Parameters Fine tuning the emulation 5 When all method settings are finished click OK to close the method setup screen The ISET emulation method will then be prepared NOTE During the run of an ISET method it is not possible to change the method parameters 3 1290 Infinity ISET 27 3 Setting Up ISET Parameters Fine tuning the emulation 28 1290 Infinity ISET 1290 Infinity ISET ria ISET Functionality Understanding ISET Functionality 3
15. ially when the back pressure in the system changes for example ageing of column To minimize this effect the pump provides a compressibility compensation feature that optimizes the stability of the flow accuracy according to the solvent type The compressibility compensation is set to a default value and can be changed through the user interface Without compressibility compensation the following happens during a stroke of the first plunger e The pressure in the plunger chamber increases and the volume in the chamber is compressed depending on back pressure and solvent type e When dispensing a more compressible solvent against pressure the displacement rate of the piston is reduced to compensate for the expansion of the solvent while it travels down the column When a compressibility value is set the processor calculates a compensation volume that is dependent on the back pressure in the system and the selected compressibility This compensation volume is added to the normal stroke volume and compensates for the loss of volume during the delivery stroke of the first plunger Composition differences are generated by the HPLC by imperfect compensation of the solvent compressibility for example due to mismatch of the compressibility settings and the actual solvent compressibility Accurate blending of mobile phase composition is vital for predictable retention While historically the equipment was well known for its reproducibility
16. in terms of accuracy of the mobile phase composition offsets may have been allowed However in order to emulate the historic behavior these systematic offsets characteristic of the individual instrument classes need to be taken into consideration As an example consider volume contraction When mixing water in an organic solvent the basic pump concept if it is low pressure proportioning or high pressure dispensing may introduce a significant offset This may even be different for various solvents and may even change with runnning conditions 1290 Infinity ISET 1290 Infinity ISET Understanding ISET Functionality 4 such as B or pressure and of course the compressibility settings that the original pump compensated for The composition differences generated by a 1100 quaternary pump for different settings of solvent compressibility are shown in Figure 4 on page 33 Figure 4 Composition offset of an Agilent 1100 quaternary pump for acetonitrile water at different compressibility settings When a gradient timetable is executed on an U HPLC instrument the shape of the resulting gradient formed at the top of the column depends mainly on e the system volume and geometry between the point of mixing usually in the pump and the column head e the accuracy with which the programmed composition is delivered to the column When an HPLC system is characterized with regard to gradient shape it is important to separate the effec
17. le has a counter that tracks the number of times it has been used You can install and reactivate ISET a maximum of five times after this the ISET USB dongle is no longer usable 1290 Infinity ISET P 1290 Infinity ISET e09 0Q0 gt 3 Setting Up ISET Parameters O O O o a e d Preparing the 1290 Infinity System for emulation 20 Setting up the basic ISET parameters 21 Fine tuning the emulation 26 This chapter gives detailed step by step instructions for setting up the standard ISET parameters and for setting up and using a verification method to confirm that ISET is functioning as expected It also contains explanations of the advanced ISET parameters with step by step instructions for setting them up gg Agilent Technologies 19 3 Setting Up ISET Parameters Preparing the 1290 Infinity System for emulation 1 Set up the 1290 Infinity system with the correct column and mobile phases for the method to be transferred If possible transfer the column from the original system The differences in selectivity between similar types of column for example C18 from different manufacturers is likely to be greater than differences caused to the separation by parameters that ISET controls 2 Download the method parameters for all modules from the original method OR Enter the method parameters into the new 1290 Infinity method in the normal way Ensure that all values are correctly transferred and that appropriate values a
18. ment method transfer 1290 Infinity LC with ISET CI DADI A Sig 245 4 Ref 380 100 Z CHEM32 T_5 95_4 10MIN_F000 800 2011 03 24 10 04 20PEST__210000018 0 mU 120 A 100 4 g mrin 1 2 3 O DADI A Sig 245 4 Ref 260 100 Z CHEM32 OE_X 95_4 16_F600 800 201 1 03 29 09 02 00 PEST2_210000042 D 1 2 3 4 O DADI A Sig 245 4 Ref 380 100 Z CHEM32 DE_X 95_4 16_F000 800 2011 03 28 16 32 47 PEST_2110000018 D 1 2 2 4 5 mrin Figure 1 Chromatograms of a gradient separation of a pesticide mixture run on A 1100 Series Quaternary Pump G1311A Autosampler G1367A B 1290 Infinity LC without emulation C 1290 Infinity LC in emulation mode The result similar retention times and similar resolution without modifying the instrument or the original method NOTE The sample and method conditions in the figure above are used to display a critical transfer situation The method is not optimized and not meant to be a good example for best resolution For ISET it is key to always reproduce the original resolution It delivers a good separation in C if the separation in A is good and a compromised separation in C if the separation in A is compromised ISET truly reproduces the original separation pattern 1290 Infinity ISET 11 1 Whatis ISET List of supported ISET modules Revision 1 of ISET supports the emulation of the following modules Pumps G1311A G1311B G1311C G1312A G1312B G1312C G5611A Autos
19. pressure 26 U USB dongle 16 UV detection wavelength 30 1290 Infinity ISET Index 1290 Infinity ISET 37 Index 38 1290 Infinity ISET Index 1290 Infinity ISET 39 www agilent com In This Book The manual describes the Agilent Intelligent System Emulation Technology ISET It contains the following Instructions for installing and configuring ISET e Details on setting up the ISET parameters e Detailed information about how ISET works Agilent Technologies 2011 Printed in Germany 09 11 G4220 90310 EE Agilent Technologies
20. r Empty Cut Copy Paste Shift Times min 1ser al e 2 Expand the Advanced section of the pump method parameters and ensure that the following check boxes are marked Synchronized in the Minimum Stroke section Use Solvent Types in the Compressibility section Advanced Minimum Stroke Channel A Channel B Automatic Automat V Synchronized Compressibility Y Use Solvent Types For ISET to work correctly it is vital to ensure best accuracy of both flow and composition of the mobile phase Critical performance factors are the stroke settings and the solvent correction Only when the built in solvent libraries are used is the operation accurate enough to expect correct results uL 22 1290 Infinity ISET 1290 Infinity ISET Setting Up ISET Parameters 3 3 Verify that the following method parameters for the pump show the correct values as given in your original method solvent flow rate solvents A and B and solvent composition pressure limits stop and post times gradient timetable 4 Expand the ISET section of the method parameters ISET T Emulation 5 Mark the Enable ISET check box Currently only one version is available in the Enable ISET field Future versions may come with updates that have either more aspects to emulate or a wider range of supported instrument configurations The Model Parameter section of the method setup is displayed ISET a Emulation Enable
21. re provided for the data collection rate detector PeakWidth setting 3 Allow the system to equilibrate 20 1290 Infinity ISET Setting Up ISET Parameters 3 Setting up the basic IS parameters Setting up the basic ISET parameters In this section you will transfer your original method to the 1290 Infinity system and activate ISET by selecting the original hardware configuration All known differences in the behavior between the original LC instrument and the current 1290 Infinity target system will be addressed by ISET e ISET is installed see Installing and Configuring ISET on page 15 TE When ISET is installed but not enabled the grey icon is displayed in the Pump Dashboard sone panel the orange icon indicates that ISET is enabled ISET icon OOO PEELE EEE Dr Trimm 2 1290 Infinity ISET 21 3 Setting Up ISET Parameters 1 Right click in the Pump Dashboard panel and select Method from the context menu The method setup dialog box is displayed Flow Advanced Timetable 1 100 events a 1 000 mimin function centric view SE Time min AR Bro max e a i 1 o 100 0 Water VOZ 0 00 5000 5000 1000 1000 00 A 2 100 0 Water V 02 X O o m m 1 oa Acetonitrle vo e v 2 100 0 Acetonitrile V 02 Pressure Limits Min 0 00 gt bar Max 1 000 00 bar Stoptime Posttime As Injector No Limit o of min min f Add Remove ClearAll Clea
22. technology is based on two components the 1290 Infinity performance specification and the ISET emulation algorithm Agilent 1290 Infinity Performance specification The 1290 Infinity LC with its broad power range unmatched flow and composition accuracy ultra low delay volume superior sensitivity delivers the key requirement for the implementation of ISET technology 1200 Infinity Series Power Range 1290 Infinity LC 0 1 2 3 4 5 mL min The wide power range of the 1290 Infinity LC ensures that both HPLC and UHPLC methods from narrow bore to standard bore columns can be executed 1290 Infinity ISET 9 What is ISET The solution for instrument to instrument method transfer 1290 Infinity LC with ISET The ISET emulation algorithm With the detailed knowledge about the system behavior of the target LC instrument and the high accuracy of the 1290 Infinity LC ISET is able to create an emulation function which delivers similar gradient conditions as the selected instrument The results are similar retention times and similar chromatographic resolution Response 90 Gradient slope Mixing behaviour Programmed gradient step Typical UHPLC system 10 1100 Series Binary system Delay volume Time The 1290 Infinity LC with ISET delivers a gradient very similar to the 1100 Series Binary LC emulating the delay volume and mixing behavior 1290 Infinity ISET Whatis ISET 1 The solution for instrument to instru
23. telligent System Emulation Technology ISET Response 90 Gradient slope Mixing behaviour Programmed gradient step Typical UHPLC system 10 1100 Series Binary system Delay volume Time Comparison of delay volumes and gradient mixing behaviors between a typical UHPLC system and an 1100 Series Binary LC using a tracer experiment On a typical UHPLC system the mixed solvents reach the column much earlier and the set composition is also achieved earlier due to the steeper gradient slope Absorbance mAU 800 700 600 500 400 Typical UHPLC system with low delay volume 1100 Series Binary LC 1 1 5 2 2 5 3 35 Time min Different solvent compositions at the column due to different delay volumes and gradient mixing behaviors result in different retention times and resolutions Column Poroshell 120 3 0 x 50 mm 2 7 ul Flow rate 0 85 mL min Mobile Phase Water Acetonitrile Gradient 0 min 10 Acetonitrile 3 min 90 Acetonitrile 1290 Infinity ISET Whatis ISET 1 The solution for instrument to instrument method transfer 1290 Infinity LC with ISET The solution for instrument to instrument method transfer 1290 Infinity LC with ISET The Intelligent System Emulation Technology enables the 1290 Infinity LC to execute other HPLC and UHPLC methods and deliver nearly the same chromatographic results without any change of the instrument or the original method all by simple mouse click This
24. tonitrile are required by the method mark the manually select ISET solvent model check box and click the down arrow and select an appropriate solvent system from the drop down list select Generic when buffers or solvent mixtures are used select Aqueous Channel A Organic Channel B or Organic Channel A Aqueous Channel B for pure solvents as appropriate You can also select Acetonitrile Water or Methanol Water if the solvents used are similar to these 24 1290 Infinity ISET Setting Up ISET Parameters 3 9 Mark the manually set check box for compressibility and enter the compressibility settings that were used in the original method This is especially important if the compressibility values in the original method are different from the default values The parameters you set up to this step are those that ISET uses to correct for the behavior of the original pump module 10 Click the Emulated Sampler down arrow and select the sampler and configuration from the drop down list ISET calculates delay volumes using the factory installed values of capillaries syringes and needle seats If other capillaries are installed fine tune the delay volume see Fine tuning the emulation on page 26 11 If a needle seat for higher injection volumes is installed enter the correct needle seat volume in the Seat field 12 You can click View emulation set if you want to display the modifications to the gradient time table th
25. trument driver to generate a virtual timetable that is executed by the 1290 Infinity pump to emulate another HPLC system When the emulation mode is enabled the 1290 Infinity emulating the original pump and auto sampler can run the original gradient method and achieve a very similar separation to that produced on the emulated system see Figure 1 on page 11 1290 Infinity ISET 35 Index Index A autosamplers 12 Cc column temperature 30 column 20 compressibility 25 32 D delay volume offset 26 31 delay volume 30 demonstration version 13 drivers revision 16 E emulated pump 24 emulated sampler 25 enable ISET 23 extra column volume 30 F firmware update 18 firmware revision 16 flow accuracy 32 flow path 30 flow rate 23 30 G gradient timetable 30 gradient composition 24 gradient timetable 23 36 injection volume 30 instrument parameters 30 ISET icon 21 M main board replacement 18 method setup 22 method parameters 20 minimum stroke 22 mixing behavior 30 mobile phase composition 30 mobile phase 20 N needle seat 25 0 offset 26 ompressibility 22 OpenLABCDS 16 P peakwidth 20 posttime 23 pressure limits 23 pressure ripple 32 pump configuration 17 pumps 12 reinstallation 18 removal 18 ripple 32 S solvent types 22 solvent composition 23 specifications 13 stoptime 23 synchronized 22 system delay volume 30 31 T trial version 13 typical operating
26. ts of the geometric volumes from those of static composition errors produced by the pump Figure 4 on page 33 shows the composition offset generated by a 1100 quaternary pump for different settings of solvent compressibility 33 4 Understanding ISET Functionality ISET functionality The physical relationship between a programmed timetable and the system response can be described by a transfer function Figure 5 on page 34 Amn O Timetable System response ro A it 7 E 4 Timetable System response Input x transfer function complex Output Figure 5 Different gradient shapes resulting from the same time table for different instrument characteristics For system B with a considerably lower delay volume than system A it is possible to compute a virtual timetable from the transfer functions of the two instruments that corresponds to the programmed timetable of system A and that generates a gradient response on system B that is equivalent to that of system A Figure 6 on page 35 34 1290 Infinity ISET Understanding ISET Functionality 4 Understanding ISET Functionality Method Timetable a Input A 1813 Nas Timetable b lim ir 4 Output B eo Figure 6 Operation of the1290 Infinity system in a mode that emulates the gradient response of a larger delay volume system The transfer functions generated from thorough system characterizations are used by ISET the ins

Agilent 1290 Infinity with ISET

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