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Calibration Tests of HPLC

Calibration tests of HPLC

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0% found this document useful (0 votes)
90 views16 pages

Calibration Tests of HPLC

Calibration tests of HPLC

Uploaded by

SPPL QAQC
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOC, PDF, TXT or read online on Scribd
You are on page 1/ 16

WAT053374 REV.

12

Performance Main-
tenance Protocol

2690/2695/e2695 Separations Module

Serial Number/
Firmware Version:

Date:

Performer:

©2013 WATERS CORPORATION. ALL RIGHTS RESERVED.

*WAT053374* *REV.12*
WAT053374 REV. 12

Performance maintenance

Goal
This performance maintenance protocol is a guide to performing maintenance tasks on your
system. Packaged as part of the performance maintenance kit, it is intended to ensure optimum
performance and an uninterrupted period of trouble-free operation.

Schedule
Waters recommends that performance maintenance visits be performed once a year. Systems
under heavy use or using aggressive solvents will need more frequent maintenance. For detailed
maintenance schedules, refer to the appropriate operator’s guide(s) located on the Waters web
site. Failure to follow the maintenance schedules as described in the operator’s guides may result
in unreliable operation and inaccurate laboratory results. For some protocols, optional tests can be
performed by Waters field service engineers for added confidence and reliability.

Safety considerations
Some protocols may involve removing instrument covers, exposing the performer to high
voltages, high currents, and moving parts. This presents risks of electric shock and/or damage to
the instrument. To reduce these risks, Waters recommends that these protocols be conducted by
certified Waters field service engineers, who have access to the latest information on Waters
products. Waters does not advise and is not responsible for an unqualified performer initiating
actions that they do not understand. If there is any doubt on the part of the performer, a Waters
certified field service engineer should conduct the performance maintenance activity.

Waters Quality Parts


Waters designs and manufactures Waters Quality Parts ® to the same strict regulatory standards
as your high-performance Waters systems. Waters recommends using Waters Quality Parts for all
maintenance activities. Failure to use Waters Quality Parts may compromise laboratory results
and reliability.

Customer education
Waters offers several operation and maintenance courses for ACQUITY ® UPLC®, Alliance® HPLC
and MS systems as well as courses designed to help you maintain professional competency and
further your knowledge in Chromatography, UPLC, Informatics and Mass Spectrometry. For more
information on available training courses from Waters Educational Services please visit the Waters
web site.

Compliance services
Waters Compliance Services help you achieve and maintain the regulatory compliance of your LC
and MS instruments and software with a combination of expertise, compliant-ready systems and
software, and a portfolio of services to ensure that you have quality data, consistently. To learn
more about these services, please visit the Waters web site.

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 2 OF 16


WAT053374 REV. 12

General tasks

Use the following guidelines when conducting performance maintenance visits. This information is
intended to be a guide only. For example:
 If a particular module has been recently serviced, it may need only testing and
replacement of fewer parts.
 Not all procedures apply to all instruments (specify NA in the Comments section if the
procedure does not apply).
 If the instrument fails to perform adequately during pre-performance maintenance checks,
further service may be required beyond the scope of this protocol. Such service may incur
additional parts and labor charges.
The following table lists the general tasks to be performed during a performance maintenance
visit. Perform and confirm the steps listed for each procedure and record any comments that you
have about the results at the end of each section.
For detailed instructions and a list of the necessary tools, refer to the Operator’s Guide located on
the Waters website.

Post-performance Optional calibration


Inspect Maintenance tasks
maintenance checks tests1

Testing the flow rate


Firmware version2 Filters Initialization diagnostics
accuracy

Testing the Gradient


Fan (inspect from the Solvent management
Static leak test Proportioning Valve
outside of the unit) components
(GPV Test)

Sample management Testing the injection


Valves Compression check
components accuracy

Carousel drive
Lithium battery3
mechanism

PerformancePlus™
degasser

Seal wash system

Fittings and tubing


(leaks)

Error logs

1
Optional tests performed by Waters field service engineers as part of a Total Assurance with Calibration Plan. For
those without the plan, the tests can be performed for an additional charge. Some of these tests may require
additional chemistry kits that are not supplied in the performance maintenance kit.
2
Firmware upgrades, if required, should be performed by a Waters field service engineer prior to beginning
performance maintenance. Firmware upgrades may require re-qualification of the upgraded instrument if it is in a
regulated environment.
3
Batteries are replaced by Waters field service engineers only.

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 3 OF 16


WAT053374 REV. 12

Inspect

Firmware version

Confirm which firmware version is installed and note it on the cover page. Update the firmware if
required.

Firmware check complete.

Yes No N/A

Comments:

Fan

To inspect the fan:

1. Observe the fan on the rear of the instrument.


2. Ensure that the fan grate is clear of debris and dust. The fan should be blowing out-
ward.
3. Clean if necessary.

Fan inspection complete.

Yes No N/A

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 4 OF 16


WAT053374 REV. 12

Valves

Prime/vent valve inspection complete.

Yes No N/A

High pressure valves 1 and 2 inspection complete.

Yes No N/A

Low pressure valve 3 inspection complete.

Yes No N/A

Gradient proportioning valve inspection complete.

Yes No N/A

Comments:

Carousel drive mechanism

Carousel drive mechanism inspection complete.

Yes No N/A

Comments:

PerformancePlus degasser

PerformancePlus degasser inspection complete.

Yes No N/A

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 5 OF 16


WAT053374 REV. 12

Seal wash system

Seal wash system inspection complete.

Yes No N/A

Comments:

Fittings and tubing leak inspection

Fittings and tubing leak inspection complete.

Yes No N/A

Comments:

Error logs

Error log review complete.

Yes No N/A

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 6 OF 16


WAT053374 REV. 12

Maintenance tasks

The following procedures are routinely performed as part of the performance maintenance. Wa-
ters Quality Parts® have been used to replace/rebuild all parts.

 Filters

 Solvent management components

 Sample management components

 Lithium battery

NOTE: Batteries are replaced by Waters field service engineers only.

Yes No N/A

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 7 OF 16


WAT053374 REV. 12

Post-performance maintenance checks

Perform the post-performance maintenance checks in the order listed.

Initialization diagnostics

Turn on power, allow instrument to complete and pass the on-board software checks.

Initialization diagnostics check complete.

Yes No N/A

Comments:

Static leak test

1. Prime the system.


2. Energize the seals.
3. Test the accumulator and primary pump heads.

Static leak test check complete.

Yes No N/A

Pumps PSI/minute % Volume Pass/Fail

Primary Pass Fail

Accumulator Pass Fail

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 8 OF 16


WAT053374 REV. 12

Compression check

1. Prime the system.


2. Purge the injector.
3. Perform the compression check.

Compression check complete.

Yes No N/A

Check Result Pass/Fail

Compression volume µL Pass Fail

Delta decay pressure psi Pass Fail

Final pressure psi Pass Fail

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 9 OF 16


WAT053374 REV. 12

Performance maintenance confirmation

I confirm on
behalf of
Company Name

that this performance maintenance


procedure has been satisfactorily completed on:
Serial No.

Signed: Date:
Customer

Type Name

I certify that the installation has been successfully completed.

Signed: Date:
Waters Engineer

Type Name

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 10 OF 16


WAT053374 REV. 12

Optional calibration tests

Optional calibration tests are performed by Waters field service engineers as part of a Total
Assurance with Calibration Plan. For customers without this plan, these tests can be performed for
an additional charge. Some of these tests may require additional chemistry kits that are not
supplied in the performance maintenance kit.

Optional calibration tests

1. Testing the flow rate accuracy.

2. Testing gradient proportioning.

3. Testing the injection accuracy.

Tools/materials required
• Graduated cylinder,10 mL
• (2) Graduated cylinders, 50 mL or 100 mL
• Stopwatch (calibrated)
• 100% HPLC – grade Methanol
• 100% HPLC -grade water
• 2 mL vial
• Analytical balance (calibrated)

W ALWAYS OBSERVE SAFE LABORATORY PRACTICES WHEN YOU USE THIS EQUIPMENT AND
WHEN YOU WORK WITH SOLVENTS AND TEST SOLUTIONS. KNOW THE CHEMICAL AND
PHYSICAL PROPERTIES OF THE SOLVENTS AND TEST SOLUTIONS YOU USE. SEE THE
MATERIAL SAFETY DATA SHEET FOR EACH SOLVENT AND TEST SOLUTION IN USE.

C NEVER CHANGE DIRECTLY BETWEEN IMMISCIBLE SOLVENTS OR BETWEEN BUFFERED


SOLUTIONS AND ORGANIC SOLVENTS. IMMISCIBLE SOLVENTS FORM EMULSIONS IN THE
FLUID PATHWAYS. BUFFERED SOLUTIONS AND ORGANIC SOLVENTS IN COMBINATION CAN
RESULT IN SALT PRECIPITATION IN THE GRADIENT PROPORTIONING VALVES, PUMP
HEADS, CHECK VALVES, AND OTHER PARTS OF THE SYSTEM. CONFIRM THAT ALL FLUIDS IN
THE SYSTEM ARE MISCIBLE WITH METHANOL.

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 11 OF 16


WAT053374 REV. 12

Testing the flow rate accuracy

1. Obtain a 10 mL graduated cylinder or similar container.

N Ensure that the graduated cylinder is thoroughly dry.


2. Prime the system with fully degassed methanol or water. Set the vacuum degasser (if in-
stalled) to run continuously, or set the sparge rate to 100% A.
3. Prepare the stopwatch to begin timing.
4. Enter the test flow of 1.0 mL/min.
5. When the flow and pressure are stable, insert the outlet tubing into the graduated cylin-
der.
6. When the meniscus reaches the 1.0 mL mark of the graduated cylinder, start the stop-
watch.
7. Stop the timer when the bottom of the meniscus reaches the 4 mL mark on the gradu-
ated cylinder.
8. Record the time required to reach the 4 mL mark.

N If the time is not within the acceptable range, other service adjustments may be
necessary for an additional charge.

Flow rate accuracy test complete.

Yes No N/A

Acceptable time
Measured time Pass/Fail
range to 4 mL mark

2:51 to 3:09 minutes min Pass Fail

Stop watch
serial #:
Calibration
date:

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 12 OF 16


WAT053374 REV. 12

Testing gradient proportioning

1. Place the primed solvent A and B lines from the separations module into two identical 50
or 100 mL graduated cylinders.
2. Fill the cylinders to the 50 mL mark.
3. Set the flow rate to 5 mL/min with 50% solvent A, and 50% solvent B.
4. When the solvent in the A cylinder reaches the 25 mL mark, stop the flow.
5. Record the values for the volumes in each cylinder (Table 2).

N Expected Result: B cylinder volume = A cylinder volume ± 3 mL.

N If the volume is not within the acceptable range, other service adjustments may be
necessary for an additional charge.

Gradient proportioning test complete.

Yes No N/A

Solvent Measured volume Pass/Fail

A mL Pass Fail

B mL Pass Fail

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 13 OF 16


WAT053374 REV. 12

Injection accuracy test

C To ensure the accuracy of test results use forceps, or wear lint-free gloves
to handle the vial used in this test.

Preparing the sample for the injection accuracy test


1. Fill a standard 2 mL vial with degassed HPLC-grade methanol (Method A) or degassed
HPLC-grade water (Method B).
2. Seal the vial with a PTFE (or pre-slit) septum and cap, if available.

N If a PTFE (or pre-slit) septum is not available, use a septum-free cap. Do not use
self-sealing septa (unless pre-slit), as they create a vacuum in the vial during
multiple injections.
3. Zero the analytical balance, and then carefully weigh the vial (using forceps or your
gloved hand to move the vial).
4. Record the weight in grams (W1).
5. Place the weighed vial in position 1 of carousel A, and then place the carousel in the
sample compartment.

Performing the injection accuracy test


1. Program the system to make the appropriate volume and number of injections from the
vial. Use the shortest possible runtime.

Value When Using Value When Using


Parameter
a 250 µL Syringe a 25 µL Syringe

Vial Range 1 1

Injects per Vial 6 16

Run Time 0.1 0.1

Inject Volume 50.0 12.0

2. Remove and reweigh the vial.


3. Record the weight in grams (W2).
4. Use one of the following formulas to calculate the average amount injected per injection:

• For a 250 µL syringe systems (Method A methanol): [(W1– W2)/6] x 1.268 x 1000 =
mg of methanol withdrawn per injection = µL per injection

• For a 25 µL syringe systems (Method A methanol): [(W1– W2)/16] x 1.268 x 1000 =


mg of methanol withdrawn per injection = µL per injection

• For a 250 µL syringe systems (Method B water): [(W1– W2)/6] x 1.0 x 1000 = mg of
water withdrawn per injection = µL per injection

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 14 OF 16


WAT053374 REV. 12

• For a 25 µL syringe systems (Method B water): [(W1– W2)/16] x 1.0 x 1000 = mg of


water withdrawn per injection = µL per injection
5. Record the results.

N If the result is not within the acceptable range, other service adjustments may be
necessary for an additional charge.

Vial Weight Vial Weight Acceptable


Programmed Calculated
Before After 6 or 16 Injection
Injection Injection Pass/Fail
Injection Injections Volume Range
Volume Volume (µL)
(W1) (W2)

g g 50 µL µL 50 µL± 2.0 µL Pass Fail

g g 12 µL µL 12 µL± 1.0 µL Pass Fail

Balance
serial #:
Calibration
date:

Comments:

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 15 OF 16


WAT053374 REV. 12

Calibration confirmation

I confirm on
behalf of
Company Name
that the calibration tests were performed
according to the prescribed procedure, and the
results were accurately recorded on:
Serial No.

Signed: Date:
Customer

Type Name

I certify that the calibration tests were performed according to the


prescribed procedure, and the results were accurately recorded .

Signed: Date:
Waters Engineer

Type Name

Comments:

Waters Corporation
34 Maple Street
Milford, Massachusetts
01757 USA

Waters, MassLynx, The Science of What’s Possible and Waters Quality Parts are registered trademarks of Waters
Corporation. All other trademarks are property of their respective owners.
©2013 Waters Corporation. Printed in the U.S.A. – WAT053374, Rev. 12

WATERS 2690/2695/E2695 PERFORMANCE MAINTENANCE PROTOCOL 16 OF 16

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