Qualification of UPLC system IRequest UPLC Community Tips ... · ©2007 Waters Corporation 1 News...

©2007 Waters Corporation 1

News from service 22.05.2012News from service 22.05.2012

� Qualification of UPLC system� IRequest� UPLC Community� Tips & tricks



System QTSystem 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


� 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)

What isWhat is SystemsQTSystemsQT??


� A wizard-based qualification solution

� Simple wizard interface built into Empower 1154 and Empower 2154 - 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.

SystemsQTSystemsQT Key BenefitsKey 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.

SystemsQT Key BenefitsSystemsQT Key Benefits


How is it Installed?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


How is it Installed?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


SystemsQT Project/Documentation SystemsQT Project/Documentation DVDDVD

� Contains the following documents:— Overview— Protocol— Test Descriptions and

Specifications— Amounts— System Set Up— Forms— Manual OQ Procedures— Operator Guides— Maintenance Procedures— Examples Reports


SystemsQT Project/Documentation SystemsQT Project/Documentation DVDDVD

� 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


SystemsQT Screen shotsSystemsQT Screen shots� Enable the option with the

SystemsQT option Disk� Launch SystemsQT from

Run Samples-Qualify System

� Enter Service Password


SystemsQT WizardSystemsQT Wizard

� Introduction � Test Selection


SystemsQT WizardSystemsQT Wizard

� Standard Amounts and Chemistry Details

� Test Selection Summary and Finish


What Systems will it Qualify?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


SQT report example for AquitySQT report example for Aquity--systemsystem


SQT timelineSQT timeline

1.00 0.50 4.00 0.50

0.00 1.00 2.00 3.00 4.00 5.00 6.00

Time in Hours

Systeemin valmisteluTesti-inisoinnitKvalifiointitestitPaperityöt

10.00Perinteinen työkirjakvalifiointi

Käytetty aika tunteina


IQ ja Final ApprovalIQ ja Final Approval--raportitraportit


What Configurations are currently What Configurations are currently available today?available today?

� ACQUITY UPLC— ACQUITY FLR— ACQUITY TQ— ACQUITY SQ— ACQUITY TUV— ACQUITY PDA/2996— ACQUITY ELS

� Alliance/HT— 2489— 2998— 3100— 2487/996/2996/2475/410/

2410/2414


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IRequest will create a call to CIT IRequest will create a call to CIT (customer interaction tracking)(customer interaction tracking)


Call will be reviewed to confrim Call will be reviewed to confrim notifications mail.notifications mail.


Call can be tracked for web site to Call can be tracked for web site to check handling statuscheck handling status


UPLC UPLC communitycommunity

� 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


UPLC communityUPLC community


UPLC UPLC communitycommunity


Tips & TricksTips & Tricks


Recommended Guidelines of Use Recommended Guidelines of Use for the Solvent Managerfor the Solvent Manager

� Keep seal wash primed. — 90-95% 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 24 – 48 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


Controlling Bacterial Growth in Controlling Bacterial Growth in Aqueous Mobile PhasesAqueous 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.


General Recommendations for Sample ManagerGeneral Recommendations for Sample Manager(not applicable for FTN 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


Detector ConsiderationsDetector Considerations� ALWAYS ensure the back pressure regulator is installed if UV

detector is last in line.� Choose a sampling rate to collect 25-50 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.


Effect of Filtering on Peak Shape Effect of Filtering on Peak Shape and Noiseand Noise

Peak shape suffers

Best s/n


Effect of Different Sampling Rate Effect of Different Sampling Rate Settings on Peak Width and Peak Settings on Peak Width and Peak HeigthHeigth

4 sec


System EquilibratedSystem EquilibratedTypical 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


Pump Pressure plot Pump Pressure plot Check Valve ProblemCheck Valve Problem

Primary A

Accumulator A

Accumulator B

Primary B

Inconsistent height of pressure trace


Pressure problemPressure problem


Effect of Contaminated/blocked InEffect of Contaminated/blocked In--line line Filter on Peak Shape/EfficiencyFilter on Peak Shape/Efficiency

Contaminated Frit P = 6400 psiN = 7575

Debris from seal shedding, particulates from buffer,particulates from sample

Sample“Band” Column New Frit

P = 5100 psiN = 9349


Over tightened Finger tight FittingsOver tightened Finger tight Fittings

Deformed Ferrule

Normal Ferrule

Reusable Fitting Installed Correctly

au

0.000

0.010

0.020

0.030

0.040

Overtightened Reusable Fitting

au

0.000

0.005

0.010

0.015

0.020

0.025

Minutes1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40


InIn--line filter installation problemline filter installation problem

N 5sigma = 5025Tailing factor = 1.31

N 5sigma = 6959Tailing factor = 1.17

Wrong position of ferrule

Correct position of ferrule


Column outlet tube, Column outlet tube, wrong position of ferrulewrong position of ferrule


NeedlesNeedlesPart No Model Size

(µl)WettedMaterial

Recommended Applications

700002644 ACQUITY (FL) 30 PEEK™ Default needle

205000362 ACQUITY (FL) 30 SS tubing and tip Robust piercing.Recommended for strongly hydrophobic compounds

205000369 ACQUITY (FL) 30 FEP Teflon™ w.SS tip

Lower cost alternative to SS.Less interaction with hydrophobic compounds compared to PEEK

205000363 ACQUITY (FL) 15 SS tubing and tip Provides lower dispersion then 30ul version

205000507 ACQUITY (FL) 15 Non-deactivatedPEEK-Sil

Strongly hydrophobic peptides

700005179 ACQUITY (FL) 30 Non-deactivatedPEEK-Sil

Small hydrophobic compounds

700005215 H-Class (FTN) 15 Metal Needle 0.007” ID. Use w. all UPLC applications

700005279 H-Class (FTN) 30 Metal Needle 0.01” ID. Use w. HPLC application and larger ext. loops


PlatesPlatesPart No Plate Plate Type186002643 96 round-well, 350 µl

polypropyleneANSI-96well350µl

186002481 96 round-well, 1.0 mlPolypropylene

ANSI-96well1ml

186002482 96 square-well, 2.0 mlpolypropylene

ANSI-96well2ml

186002631 384-well, 100 µlpolypropylene ANSI-384well100µl *

186002632 384-well, 250 µlpolypropylene

ANSI-384well250µl

186000349 96-well w. 700 µl glass insert w. plate

ANSI-96well700µlGlassInsert

186000855 96-well w. 1 ml extended glass insert w. plate

ANSI-96well1mlGlassInsert

186001437 96-well w. 700 µl glass insert,insert only

ANSI-96well700µlGlassInsert

186001436 96-well w. 1 ml extended glass insert, insert only

ANSI-96well1mlGlassInsert

* Not for use with H-Class


VialsVialsPart No. 2 ml screw cap Vial

(12x32mm)ACQUITY SMNeedle depth

offset/ Residual Vol.

H-Class SM FTNNeedle depth

offset/Residual Vol.

186000307C Clear, w. pre-slit PTFE/siliconesepta

2 mm/165µl 0.1 mm

186000847C Amber, w. pre-slit PTFE/siliconesepta

2 mm/165µl 0.1 mm

186000327C Clear, Max Recovery,w. pre-slit PTFE/silicone septa

2 mm/22µl 2.1 mm

186003886C Amber, Max Recovery,w. pre-slit PTFE/silicone septa

2 mm/22µl 2.1 mm

186002636 750 µl PP, w. pre-slit PTFE/siliconesepta

3 mm/70µl

186002639 300 µl PP, w. pre-slit PTFE/siliconesepta

3 mm/20µl

186000385C Clear, Total Recovery,w. pre-slit PTFE/silicone septa

2.5 mm/9µl 0.7 mm


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