Qualification of UPLC system IRequest UPLC Community Tips & tricks Waters Corporation 1

Transcription

1 News from service Qualification of UPLC system IRequest UPLC Community Tips & tricks 2007 Waters Corporation 1

2 2007 Waters Corporation 2

3 System QT What is SystemsQT? Key benefits How is it installed? Overview of the SystemsQT Documentation DVD Overview of the SystemsQT wizard screens Timeline and report examples What configurations are available today 2007 Waters Corporation 3

4 What is SystemsQT? SystemsQT is the new Waters System Qualification Tool It is a Replacement product for ConnectionsAQT for Waters ALLIANCE Systems and ACQUITY UPLC SYSTEMS (collectively known as System AQT) 2007 Waters Corporation 4

5 SystemsQT Key Benefits A wizard-based qualification solution Simple wizard interface built into Empower 1154 and Empower using Run Samples The SystemsQT wizard displays the qualification tests based on the empower project, for system qualifications all tests are selected To run just specific tests simply deselect the tests you do not require Waters Corporation 5

6 SystemsQT Key Benefits A separate Empower Project which contains all the methods required to qualify a specific system configuration Simply restore the project, set-up the system, insert your samples, equilibrate and run - its that easy! All the SystemsQT qualification documentation can be saved to a customer specific location for easy access, review and retrieval All SystemsQT project calculations are automated Waters Corporation 6

7 How is it Installed? SystemsQT is integrated within Empower For Empower 1154, Service Pack L or greater must be loaded For Empower 2154, Feature Release 4 or greater must be loaded 2007 Waters Corporation 7

8 How is it Installed? The Waters Compliance Specialist must restore the appropriate SystemsQT project for the system to be qualified The qualification project and documentation are located on the SystemsQT Documentation DVD The projects names on the DVD are in the format: SQT_SystemName_DetectorName Localized projects will be available in Japanese, Chinese & Korean 2007 Waters Corporation 8

9 SystemsQT Project/Documentation DVD Contains the following documents: Overview Protocol Test Descriptions and Specifications Amounts System Set Up Forms Manual OQ Procedures Operator Guides Maintenance Procedures Examples Reports 2007 Waters Corporation 9

10 SystemsQT Project/Documentation DVD The Waters Compliance Specialist copies the appropriate documents to a user-specified location The SystemsQT Project/Documentation DVD is the property of Waters and is not left with the customer 2007 Waters Corporation 10

11 SystemsQT Screen shots Enable the option with the SystemsQT option Disk Launch SystemsQT from Run Samples-Qualify System Enter Service Password 2007 Waters Corporation 11

12 SystemsQT Wizard Introduction Test Selection 2007 Waters Corporation 12

13 SystemsQT Wizard Standard Amounts and Chemistry Details Test Selection Summary and Finish 2007 Waters Corporation 13

14 What Systems will it Qualify? All new Instruments under Empower and Empower 2 control will be introduced with SystemsQT as the qualification tool All systems currently qualified by the ConnectionsAQT products will eventually be added to SystemsQT 2007 Waters Corporation 14

15 SQT report example for Aquity- system 2007 Waters Corporation 15

16 SQT timeline Time in Hours Käytetty aika tunteina Systeemin valmistelu Testi-inisoinnit Kvalifiointitestit Paperityöt Perinteinen työkirjakvalifiointi Waters Corporation 16

17 IQ ja Final Approval-raportitraportit 2007 Waters Corporation 17

18 What Configurations are currently available today? ACQUITY UPLC ACQUITY FLR ACQUITY TQ ACQUITY SQ ACQUITY TUV ACQUITY PDA/2996 ACQUITY ELS Alliance/HT /996/2996/2475/410/ 2410/ Waters Corporation 18

19 irequest 2007 Waters Corporation 19

20 IRequest 2007 Waters Corporation 20

21 IRequest 2007 Waters Corporation 21

22 irequest 2007 Waters Corporation 22

23 IRequest will create a call to CIT (customer interaction tracking) 2007 Waters Corporation 23

24 Call will be reviewed to confrim notifications mail Waters Corporation 24

25 Call can be tracked for web site to check handling status 2007 Waters Corporation 25

26 UPLC community Share, ask, and interact with ACQUITY UPLC experts and scientists in dynamic discussions Access ACQUITY UPLC publications and user experiences from around the globe Review exclusive FAQ's, tips & tricks, and tutorials Explore latest cutting edge ACQUITY UPLC applications and information Access: Web site home ->ACADEMIC -> Acquity users community 2007 Waters Corporation 26

27 UPLC community 2007 Waters Corporation 27

28 UPLC community 2007 Waters Corporation 28

29 Tips & Tricks 2007 Waters Corporation 29

30 Recommended Guidelines of Use for the Solvent Manager Keep seal wash primed % water Re-prime solvent lines before starting Sys Prep or Start-up Use 100 µl mixer for TFA/ACN gradients at low wavelengths (for BSM only) Run gradients starting with at least some organic 0.1% Provides more consistent and predictable gradient formation than from starting at 0% Change aqueous mobile phases often every hr Remember: UPLC flow rates are much lower, thus mobile phases are consumed at a lower rate and last longer Do not top of mobile phase but prepare new 2007 Waters Corporation 30

31 Controlling Bacterial Growth in Aqueous Mobile Phases ACQUITY UPLC columns are packed with 1.7 µm particles and thus require 0.2 µm end frits ACQUITY UPLC columns are very fine and efficient filters UPLC column frits can plug due to bacterial growth in highly aqueous mobile phases Bacteria is the most common cause of UPLC column plugging and can easily be avoided by filtering through a 0.22 µm filter ph 7 phosphate buffer is VERY susceptible to bacterial growth Electron micrograph of Rod-shaped bacteria on the bed side of the inlet frit removed from column Waters Corporation 31

32 General Recommendations for Sample Manager (not applicable for FTN Sample Manager) Use Default Conditions 10 µl loop and PLUNO (partial loop uses needle overfill) injection will give good peak shape, good recovery, good linearity for range of injection volumes appropriate to 2.1 x 50 and 2.1 x 100 mm ACQUITY columns Stay within the recommended limits for each injection mode and loop size Use Load Ahead when cycle time is an issue Use Full Loop when best precision and accuracy is critical Use Partial Loop (pressure assisted) when sample consumption is critical Characterize needle and loop volumes when changing weak wash solvents as well as loops and needles Use steel tip needles for capmat covers on well plates Always use the column stabilizer tube even if temperature is not used 2007 Waters Corporation 32

33 Detector Considerations ALWAYS ensure the back pressure regulator is installed if UV detector is last in line. Choose a sampling rate to collect points across the peak. Choose the initial digital filter time constant according to the requirements of the method. If speed is most important start with FAST. If sensitivity is most important start with NORMAL. Optimize digital filter time constants once method is developed to balance sensitivity and resolution Waters Corporation 33

34 Effect of Filtering on Peak Shape and Noise Peak shape suffers Best s/n 2007 Waters Corporation 34

35 Effect of Different Sampling Rate Settings on Peak Width and Peak Heigth 4 sec 2007 Waters Corporation 35

36 System Equilibrated Typical pressure trace indicating that the system is equilibrated and ready for injections Note the pressure ripple in psi for the past minute = 18. This should normally be < 30 psi indicating that the solvent manager is operating well. Same distance between pump strokes Same height of pump stroke 2007 Waters Corporation 36

37 Pump Pressure plot Check Valve Problem Inconsistent height of pressure trace Primary A Accumulator A Primary B Accumulator B 2007 Waters Corporation 37

38 Pressure problem 2007 Waters Corporation 38

39 Effect of Contaminated/blocked In-line Filter on Peak Shape/Efficiency Debris from seal shedding, particulates from buffer, particulates from sample Contaminated Frit P = 6400 psi N = 7575 Sample Band Column New Frit P = 5100 psi N = Waters Corporation 39

40 Over tightened Finger tight Fittings Reusable Fitting Installed Correctly au Normal Ferrule Overtightened Reusable Fitting au Deformed Ferrule Minutes 2007 Waters Corporation 40

41 In-line filter installation problem N 5sigma = 5025 Tailing factor = 1.31 Wrong position of ferrule N 5sigma = 6959 Tailing factor = 1.17 Correct position of ferrule 2007 Waters Corporation 41

42 Column outlet tube, wrong position of ferrule 2007 Waters Corporation 42

43 Needles Part No Model Size (µl) Wetted Material Recommended Applications ACQUITY (FL) 30 PEEK Default needle ACQUITY (FL) 30 SS tubing and tip Robust piercing. Recommended for strongly hydrophobic compounds ACQUITY (FL) 30 FEP Teflon w. SS tip Lower cost alternative to SS. Less interaction with hydrophobic compounds compared to PEEK ACQUITY (FL) 15 SS tubing and tip Provides lower dispersion then 30ul version ACQUITY (FL) 15 Non-deactivated PEEK-Sil ACQUITY (FL) 30 Non-deactivated PEEK-Sil Strongly hydrophobic peptides Small hydrophobic compounds H-Class (FTN) 15 Metal Needle ID. Use w. all UPLC applications H-Class (FTN) 30 Metal Needle 0.01 ID. Use w. HPLC application and larger ext. loops 2007 Waters Corporation 43

44 Plates Part No Plate Plate Type round-well, 350 µl polypropylene round-well, 1.0 ml Polypropylene square-well, 2.0 ml polypropylene well, 100 µl polypropylene well, 250 µl polypropylene well w. 700 µl glass insert w. plate well w. 1 ml extended glass insert w. plate well w. 700 µl glass insert, insert only well w. 1 ml extended glass insert, insert only ANSI-96well350µl ANSI-96well1ml ANSI-96well2ml ANSI-384well100µl * ANSI-384well250µl ANSI-96well700µlGlassInsert ANSI-96well1mlGlassInsert ANSI-96well700µlGlassInsert ANSI-96well1mlGlassInsert * Not for use with H-Class 2007 Waters Corporation 44

45 Vials Part No C 2 ml screw cap Vial (12x32mm) Clear, w. pre-slit PTFE/silicone septa ACQUITY SM Needle depth offset/ Residual Vol. H-Class SM FTN Needle depth offset/ Residual Vol. 2 mm/165µl 0.1 mm C Amber, w. pre-slit PTFE/silicone septa 2 mm/165µl 0.1 mm C Clear, Max Recovery, w. pre-slit PTFE/silicone septa 2 mm/22µl 2.1 mm C Amber, Max Recovery, w. pre-slit PTFE/silicone septa 2 mm/22µl 2.1 mm µl PP, w. pre-slit PTFE/silicone septa 3 mm/70µl µl PP, w. pre-slit PTFE/silicone septa 3 mm/20µl C Clear, Total Recovery, w. pre-slit PTFE/silicone septa 2.5 mm/9µl 0.7 mm 2007 Waters Corporation 45

46 Kysymyksiä? 2007 Waters Corporation 46