Review Document Standard EQP LC 01.85 (1)

2014 by Agilent Technologies Enterprise Edition Compliance Services

EQP Name: HPLC Standard EQP

Company Name: _____________________

Customer Name/Title: _____________________

EQP Filename: AgilentRecommendedA.01.12.eqp

Print Date: December 17, 2014 11:57:27 AM

Page 1 / 17 December 17, 2014 11:57:27 AMHPLC Standard EQP


Table of ContentsSection Page



Scope and PurposeOverview

The Equipment Qualification Plan (EQP) documents the activity program that is performed during the qualification services for the

applicable systems. A complete description of the test specifications is provided for the supported services, including setpoints and

acceptance criteria (or limits) for each test. The test specification section of this document is created directly from the EQP file name

listed on the cover.

This document is an abstraction of the EQP file used to perform the service and is generated directly from the electronic Agilent

Equipment Qualification Plan (eEQP) Editor. The purpose of this document is to allow the user to review and record approval of the

EQP that guides the delivery of compliance services provided by the Agilent Automated Compliance Engine.

Statement of Intent

Unless otherwise requested, the qualification is delivered according to the standard test program described in the

Agilent_Recommended EQP. Agilent defines variances as changes to the default recommended values (as stated in the Agilent

Recommended EQP) that fall within a well-defined range. These changes are considered to be within the intended use range of the

system under test.

Customizations are values that (a) subject the system to limits that exceed the typical operational range or (b) additional tests that are

not considered part of the core program required for completion of the selected service. Because custom setpoints and limits may

exceed the operational envelope of the equipment, Agilent reserves the right to warrant conformance only to the closest variance

value. The user is notified of this stipulation at EQP setup time and the qualification report (EQR) will reflect this situation.

A set of ink signature fields, as determined by the creator of this document, can be included at the end of this document. All fields

should be completed or a single set of fields, initialed by an appropriate approver, run through any signature fields that are not to be

used. This is an optional process that allows a paper record of signoff by the appropriate reviewers where a hybrid (electronic/ink)

signature SOP is followed. If this document will be saved electronically and digitally signed in a document management system, it

should be generated without ink signature fields. Signing this document is for the customer's internal documentation purposes, to

assist with the applicable regulatory requirements. It is available for customer review, but it is not a pre-requisite for the delivery. The

delivery of the services is done according to the terms and conditions stated in the corresponding service exhibit. It is recommended

that after approval, this EQP be archived with the electronic EQP file.

Understanding the Test Specification Section in Tabular Review Documents

For Agilent-recommended setpoints and limits, the range of allowable values (L for low, H for high) are included.As applicable,

variances, customizations, and additional setpoints are listed beneath the Agilent recommended values and marked W (within range)

or O (outside of range) in the left margin; values for added setpoint are also marked W or O and displayed after all configurations

values. Dual limits are marked DW or DO.

Optional tests that are enabled are included and marked as such; required tests that are disabled by the customer are included and

marked as such.



Customer Responsibilities

o Safely store and archive this EQP

o Maintain change control and revision history

o Review and optionally sign the EQP, making sure the service delivery is what was approved

o Review and approve any of the following variances from the Agilent recommended:-Within Variance Range: changes to the Agilent recommended that are identified by Agilent as within the operation ranges

determined in our test development

-Outside of Variance Range: changes to the Agilent recommended that Agilent identifies as outside of the operational ranges

determined in our test development. Agilent is not under any obligation to make the instrument pass the more stringent limits that fall

in this range and this detail is called out in the EQP Test Specification

-Optional Tests: additional tests that are available but not part of the core testing suite and cost extra

-Disabled Tests: test for which all possible configurations have been disabled (tests are flagged in the test specification)

Agilent Responsibilities

o Deliver the services following the test programs described in the customer EQP

o Provide a locked and e-signed Qualification Report (EQR) upon completion of the service

o If requested, provide an optional ink-signed EQR CD to the customer

General Statements on the Testing Program

The recommended set of hardware OQ tests described in this EQP derives from Agilent's interpretation of authoritative expert

literature issued by the FDA, USP, GAMP5, ASTM 2500, and others. The OQ test design incorporates both modular and holistic

testing, which is a proven approach, acceptable to regulators. As prescribed by the 4Q qualification methodology for Analytical

Instrumentation Qualification (AIQ), the OQ step is separated from the PQ as recommended by the regulatory guidelines.

Enterprise Edition uses a balanced selection of metrology and chemical tests to directly determine the performance of the systems

without unnecessary reliance on inferred or derived results. For example, direct metrology is used to test pump flow rates and

thermal-controlled column compartment and autosampler modules. Holistic chemical testing is used for the evaluation of the following

critical instrument characteristics: linearity, precision, signal to noise, and carry over.



Enterprise Edition is designed to fit the traditional quality systems used by firms and recognized by regulatory agencies worldwide.

How Enterprise Edition aligns with a traditional, paper-based methodology is described below:

[i] Policy documents dictate the need for validation & qualification of GMP/GLP systems and usually mention the DQ/IQ/OQ/PQ model. The precise procedures for IQ & OQ for each type of equipment are prescribed in an approved SOP, perhaps called SOP #123: Qualification of HPLC Systems. In Enterprise Edition, the EQP has the same role as the traditional Qualification SOP.[ii] The traditional SOP provides lists of tests & limits for the range of system configurations found in the labor department. The EQP follows this concept. The inventory of systems covered by an SOP or EQP changes over time so this is kept as a separate record.[iii] The traditional Qualification SOP typically has blank results forms as attachments to be photocopied for each IQ or OQ event the results recorded in ink with manual calculations. In Enterprise Edition this execution process is streamlined and automated by use of Adobe forms and the Agilent Compliance Engine (ACE) delivery tool. It provides reports with no hand-writing errors; validated calculations; automated pass/fail report; traceability to raw data and a count of number of times a test was run. This automation provides efficiency and enforces compliance to procedure.[iv] The traditional Qualification SOP is approved and released only once replacing need to author individual protocols for each chromatography system. This is the same concept for the EQP. The appropriate tests for each individual configuration are automatically selected by ACE from the list in the approved EQP at time of delivery. The final reports are unique for each system and each qualification event but the single approved EQP can cover a lab, department or as wide a scope as desired.[v] In the traditional qualification methodology there is no convenient provision to record the actual workflow of the tests execution and results. In the event that a test is repeated during the Enterprise Edition delivery, ACE maintains a counter per test which is automatically incremented for GxP compliant work, and the engineer should generate a deviation note within the ACE report.

HOW AGILENT ENTERPRISE EDITION COMPLIANCE SERVICES WORK

Agilent Enterprise Edition Compliance Services



Design Qualification (DQ)Design Qualification (DQ) for commercial lab instruments is recommended by some, but not all, guidances and procedures. Defintions of DQ found in guidances and firm-specific validation procedures vary widely around the world. Some firms require nothing more than a record (such as certificate) from the instrument manufacturer demonstrating that the lab system has been designed for purpose and manufactured to a quality standard. Others treat DQ as the development of a user requirement specification document (URS) which can be matched to the IQ and OQ specifications for a manufacturer. Other firms consider DQ as including the vendor selection activities.

USP Chapter pre-published in USP 31/Supplement defines DQ:

Design qualification (DQ) is the documented collection of activities that define the functional and operational specifications of the instrument and criteria for selection of the vendor, based on the intended purpose of the instrument. Design qualification (DQ) may be performed not only by the instrument developer or manufacturer but also may be performed by the user. The manufacturer is generally responsible for robust design and maintaining information describing how the analytical instrument is manufactured (design specifications, functional requirements, etc.) and tested before shipment to users. Nonetheless, the user should ensure that commercial off-the-shelf (COTS) instruments are suitable for their intended application and that the manufacturer has adopted a quality system that provides for reliable equipment. Users should also determine capability of the manufacturer for support installation, services, and training.

For your reference, Agilent provides the following statements for DQ purposes:

1. All Agilent hardware and software laboratory products including the ACE software used to deliver qualification services, are designed, manufactured, and tested according to Agilent internal Quality Life-Cycle Development Procedures.

2. Certificates of Agilent testing, validation, and conformance to standards are provided with new Agilent instruments and similar certification is provided for ACE software. These documents are checked and recorded in Enterprise Edition Compliance Services IQ.

3. Agilent maintains information describing how products are manufactured and maintains a problem and bug reporting program as required by international software quality guidelines.

4. The OQ specifications in this EQP can be used, as appropriate, by the user to prepare URS. The OQ specifications in this EQP represent the levels of performance acceptable to regulatory agencies for the technique; conform to typical specifications found in Validation literature; are equally suitable for OQ at installation and on-going OQ throughout operational lifetime; are equivalent to the OQ specifications published in the legacy Agilent Classic OQPV protocols; and are suitable for most user requirements.

5. Agilent Technologies is capable of installation, support, preventive maintenance, on-going qualification and re-qualification after repair and user training worldwide.



Installation Qualification (IQ) HardwareHardware IQ checks and tests for Agilent software products include the following:

1. Purchase Order Documents: Allows the customer to verify that the instrument being qualified matches their design requirements (if available) and purchase order.

2. Preparation and Installation Documents: Gathers and records information about preparation and installation documents.

3. System and Installation Documentation: Gathers and records information about reference and user manuals for initial installations.

4. Product Quality Assurance Documents: Collects and records certificates and other forms that verify that the vendor has developed and built the product according to internal standards.

5. Start Up Test: Verifies that all modules start up properly.

6. Instrument Check: Demonstrates that all modules of the instrument are correctly installed and connected. It does not test instrument performance as fully as OQ. This test is not necessary and therefore skipped if an OQ is to be performed by Agilent operator at installation after IQ.

Operational QualificationRefer to the appropriate Test Definitions document for a detailed description of the testing program, set-points and acceptance limits for each system technique, category, and instrument configuration.

Dual-Acceptance LimitsWithin the Equipment Qualification Plan (EQP) of the Agilent Enterprise Edition, each of the tests final result can be compared against two different limits if required. This allows customer-configured OQ to report against a User Limit (limit1) and the Agilent Recommended Limit (limit2) simultaneously.

The Standard_EQP documents have both Limit1 & Limit2 values set the same effectively de-activating this feature. Custom_EQPs can also be prepared on request, making effective use of the Two-Limit feature of the Agilent Compliance Engine (ACE). In those cases, Limit2 will always be the Agilent Recommended limit, and Limit1 will be the limit requested by the user. Agilent will not be under any obligation regarding the OQ testing results against User-requested limits that are more stringent than the Agilent Recommended ones.



Re-Qualification after Repair (RQ) HardwareIn the event of a hardware breakdown followed by an engineering repair of a qualified instrument, it is necessary to re-qualify the system to an appropriate level before release back into operational use.

For some of the instrument techniques, Agilent offers a service contract to repair and re-qualify an instrument during the period between scheduled annual OQs.

The level of re-testing is prescribed in the RQ section of ACE: a form is displayed for the operator showing all types of repair possible and the re-testing required. Part of an example form is shown below.

Re-Qualification After Repair

Pump Strategies

Repair/Replace Strategy Modules OQ/PV Testing

Internal pump head parts, active inlet valve (or AIV cartridge), (parts of) check valves, reference valves, inlet manifold or pump drive, or taking pump head apart to clean (versus repair)

Any pump Flow Accuracy & Precision

Pulse damper, pressure transducer Any pump Flow Accuracy & Precision

Multi-channel gradient valve Quaternary Flow Accuracy & Precision Gradient Composition

The full list of repair and re-test guidance is available for review by customers of the RQ service.

The RQ form in ACE prescribes which tests the operator must perform for each repair circumstance. The test procedure, setpoints, and limits will be an exact repeat of the previous OQ test (a so called regression testing strategy).

Information, descriptions and specifications in this publication are subject to change without notice.

Agilent Technologies, Inc. 2014Published in USA, April 29, 2014

www.agilent.com/chem/enterprise



Standard OQ Test Specifications for Analytical Scale HPLC SystemsThe following tables describe the testing specifications for HPLC systems. Note the different conditions for some tests depending on the intended operating pressure range.

Test Name Setpoints and Parameters Limits

Pump Flow Accuracy and Precision(below 400 bar)

Flow Rate 1 = 0.500 ml/minuteFlow Rate 2 = 5.000 ml/minute*

Accuracy 5.00 % from setpointPrecision 0.50 % RSD

Pump Flow Accuracy and Precision (above 400 bar)

Flow Rate 1 = 0.500 ml/minuteFlow Rate 2 = 5.000 ml/minute*

Accuracy 3.00 % from setpointPrecision 0.50 % RSD

Column Temperature Accuracy and Stability

Temperature 1 = 80.0 CTemperature 2 = 40.0 C**Stability measured at Temperature 2

Diff. from setpoint 3.0 C ( 60 C)Diff. from setpoint 2.0 C (< 60 C)Stability 1.0 C

Wavelength Accuracy (UV-Vis) Wavelength 1 = 205 nm [Maximum]Wavelength 2 = 245 nm [Minimum]Wavelength 3 = 273 nm [Maximum]

Accuracy 2 nm

Wavelength Accuracy (FLD) Wavelength 1 = 350 nm [Maximum]Wavelength 2 = 397 nm [Maximum]

Accuracy 3 nm

Signal Noise and Drift (VWD) ASTM baseline noiseSlope of regression fit for drift

Noise: 0.040 mAUDrift 0.500 mAU/hr

Signal Noise and Drift (DAD, MWD) (below 400 bar)

ASTM baseline noiseSlope of regression fit for drift

Noise: 0.050 mAUDrift 5.000 mAU/hr

Signal Noise and Drift (DAD) (above 400 bar)


G4212A/B Noise: 0.030 mAUG4212A/B Drift: 3.000 mAU/hrOther Noise/Drift: same as < 400 bar

Signal Noise and Drift (MWD)(above 400 bar)


Noise: 0.050 mAUDrift: 5.000 mAU/hr

Signal Noise and Drift (RID) ASTM baseline noiseSlope of regression fit for drift

Noise: 10.000 nRIUDrift 400.000 nRIU/hr

Signal Noise and Drift (ELSD) ASTM baseline noiseSlope of regression fit for drift

Noise: 2.000 mVDrift 5.000 mV/hr

HPLC HARDWARE OPERATIONAL QUALIFICATION

Agilent Enterprise Edition Compliance Services

(*) 2.000 ml/min for G4220B 1290 pump(**) T2 = 60.0 C for 1120 and 1220 systems




Signal Noise and Drift (CD) ASTM baseline noiseSlope of regression fit for drift

Noise: 0.100 uSDrift 10.000 uS/hour

Signal to Noise (UV-Vis) (below 400 bar)

Signal height is divided by ASTM baseline noise for known concentration and known conditions.

Signal to noise 3,000

Signal to Noise (UV-Vis) (above 400 bar)

Signal height is divided by ASTM baseline noise for known concentration and known conditions.

G4212A/B Signal to Noise 10,000Other UV-Vis Signal to noise 3,000

Signal to Noise (RID) Signal height is divided by ASTM baseline noise for known concentration and known conditions.

Signal to noise 2,000

Signal to Noise (FLD) Signal height of Raman peak is divided by noise at different wavelength in flat region of emmision spectrum.

Signal to noise 400

Injection Precision (UV-Vis, RID) (below 400 bar)

Injection volume on column = 20 ul Height RSD 2.00 %Area RSD 1.00 %

Injection Precision (UV-Vis) (above 400 bar)

Injection volume on column = 10 ul Height RSD 2.00 %Area RSD 1.00 %

Injection Precision (ELSD) Injection volume on column = 20 ul Height RSD 3.00 %Area RSD 3.00 %

Injection Precision (FLD) Injection volume on column = 5 ul (2 ul for 1290 w/ CTC samplers)

Height RSD 2.00 %Area RSD 2.00%*

Injection Precision (CD) Injection volume on column = 25 ul Height RSD 2.00 %Area RSD 1.00 %

Injection Carry Over (UV-Vis, RID)

Injection volume on column = 20 ul (2 ul for 1290 w/ CTC samplers)

Height carry over 0.40 %Area carry over 0.20 %

Injection Carry Over (UV-Vis) (above 400 bar)

Injection volume on column = 10 ul (2 ul for 1290 w/ CTC samplers)


Injection Carry Over (FLD) Injection volume on column = 5 ul (2 ul for 1290 w/ CTC samplers)


Injection Carry Over (CD) (below 400 bar)

Injection volume on column = 25 ul Height carry over 1.00 %Area carry over 1.00 %

Response Linearity (UV-Vis) 5 concentrations of certified reference standard

Coefficient of determination (r2) 0.99900R/F precision 5.00 % RSD

Response Linearity (RID, CD)

5 concentrations of certified reference standard

Coefficient of determination (r2) 0.99500R/F precision 10.00 % RSD

Gradient Composition Accuracy (UV-Vis, CD)

20.00 %, 40.00 %, 60.00 %, 80.00 % steps Accuracy 2.00 %

Gradient Composition Noise and Drift (UV-Vis)

20.00%, 40.00%, 60.00%, 80.00% steps Composition noise 2.00%Composition drift 2.00%

Gradient CompositionNoise and Drift (CD)

20.00%, 40.00%, 60.00%, 80.00% steps Composition noise 3.50%Composition drift 3.50%

Standard OQ Test Specifications for Analytical-Scale HPLC Systems (continued)




Gradient Composition Linearity(UV-Vis, CD)

Linear gradient from 100 % to 0 % Coefficient of determination (r2) 0.99900**(at start, 50:50 zone, end)***

Sample Temperature Accuracy Temperature = 4.0 CSamples four vials of water in different tray positions

Diff. from setpoint 2.0 C, 5.0 C

Fraction Collection Select Fraction Collector 1, 2, or 3Select Peak or Time-based collection mode

Peak Presence (Qualitative)

* Area RSD 1.00 % with CTC samplers** r2 0.99000 for CD*** r2 0.997 for 1260 600 bar pump with AIV

Key:Fixed HPLC setpoints/limits Fixed UHPLC setpoints/limits Variance allowed for setpoint(s)

See corresponding attachments for recommended tests definitions for Capillary-Scale and Preparative Scale systems, as well as for Non-Agilent Systems

For multiple-detector systems, only one execution of the Injection Precision & Carry-Over tests will be performed in the standard test program - by default using the UV detector if present. Repeat execution of the test can be added as optional tests for a nominal fee.

End of Section - Standard OQ Test Specifications for Agilent Analytical-Scale HPLC Systems

OQ Test Design and Rationale for Analytical Scale HPLC SystemsMany GMP/GLP enforcement agency inspectors now ask firms to provide a risk assessment of their equipment and computer systems plus a science-based rationale for subsequent validation and qualification testing.

GENERAL RISK STATEMENT: Any HPLC, LCMS, UHPLC, UHPLC_MS, GC, or GCMS system used for raw material testing or final drug product / medical device testing in GMP or used in formal GLP studies will likely fall into a HIGH RISK category. This risk assessment will imply the need for IQ & OQ & on-going qualification. ANY USER SPECIFIC RISK ANALYSIS SUPERCEDES THIS GENERAL RISK STATEMENT.

The rest of this section outlines the science-based rationale for each test in the Agilent hardware OQ plus a brief test design and procedure description.

The recommended set of hardware OQ tests described in this EQP derives from Agilents intepretation of FDA, USP, and GAMP4 guidelines and other authoritative expert literature.

OQ test design incorporates both modular and holistic testing, which is a proven and regulatory acceptable approach. Direct metrology is used to test pump flow rates and thermal-controlled column compartment and autosampler modules. Holistic chemical testing is used for the evaluation of the following critical instrument characteristics: linearity, precision, signal to noise, and carry over.

Certified reference standards and calibrated traceable thermometers and digital flowmeters are used.

Considering the number of setpoints, parameters, and conditions of each recommended OQ test, the proven concepts of worst case, range, and representative have been applied. If a property or characteristic is known to have its worst performance at one end of a range of use, this is the setpoint that should be tested and other setpoints are not required. If a property or characteristic has no known worst case, testing at the high and low points of the range of use is required. If there are too many possible use cases and conditions to realistically test (and none is a worst case), a representative sample for test is the best approach.

Standard OQ Test Specifications for Analytical-Scale HPLC Systems (continued)



OQ Test Design and Rationale for Analytical Scale HPLC Systems (continued)

The test design for HPLC systems covers UV absorbance, fluorescence, evaporative light scattering, refractive index, and conductivity detectors; isocratic, binary, tertiary, and quaternary pumps; most autosampler models; and fraction collectors.

Tests for HPLC Systems (Non-MSD)

1. Pump Flow Accuracy and Precision

Rationale: Accuracy of flow is important for comparability between systems and transferring methods. Flow precision is critical for repeatability of peak height and area.

Procedure: A calibrated digital flowmeter is attached to the waste line of the system flowing pure water at representative back pressure provided by a small guard column. Six readings are taken at each setpoint to determine the flow accuracy and precision. Flow accuracy is calculated as the absolute % difference of the mean of the six flow readings against the setpoint. The precision is calculated as the %RSD of the six flow readings. The two default setpoints (0.5ml/min and 5.0 ml/min) are evaluated in the core test. Extra setpoints and flexible test range are only available in customer-configured EQPs for flow, temperature, and some other tests. The repeat measurements of flow in the flow precision test eliminate the need for measurement of retention time precision (which is an indirect approach to determining flow precision).

2. Column Temperature Accuracy and Stability

Rationale: The thermostat accuracy is important for comparability between systems and transferring methods. Column temperature stability is critical for repeatability of peak height and area.

Procedure: A calibrated digital temperature meter and a proprietary probe are used to measure the temperature of the flowing eluent. With the use of a T-piece, the temperature probe is positioned to be in contact with the heated eluent. A typical column compartment temperature range of use is tested. At the high end of the range, after stabilization, the temperature accuracy is calculated as the absolute difference between what was measured and the setpoint. After completing this measurement at the low end of the range, six readings are taken every four minutes and temperature stability is calculated as the absolute difference between the highest and lowest measured temperatures. The temperature accuracy is calculated as the average of the six readings compared to the setpoint. All readings are reported in Celsius. Both sides of the Agilent column compartment are tested at the same time.

3. Wavelength Accuracy

Rationale: Wavelength accuracy is critical for accuracy of quantitative and qualitative analysis. Wavelength accuracy is also important for comparability between systems and transferring methods.

Procedure for UV absorbance detector (UV, VWD, DAD, PDA, etc.): A traceable caffeine standard is used to determine the wavelength accuracy. In one procedure, for certain models, the caffeine is trapped in the flow cell and a programmable timetable is used to determine the wavelength maxima (205 and 273 nm) and minimum (245 nm). For other models (for example, DAD and PDA), a caffeine injection is made and a spectrum is acquired. The spectral maxima and minimum are determined directly from the scan or the table of scan results. The wavelength accuracy is determined as the absolute difference between the measured and certified wavelength values.

Procedure for fluorescence detector: The detector cell is filled with pure water. Using a programmable timetable, the excitation (350 nm) and Raman band emission (397 nm) wavelengths are determined. The wavelength accuracy is determined as the absolute difference between the measured and theoretical peaks of Raman scattering (in nm).




4. Signal Noise and Drift

Rationale: This test gives an indication of detector sensitivity and stability.

Procedure for UV absorbance detectors: Pumping water at 1 ml/min, the signal is monitored at a specified wavelength over a twenty minute period. The signal noise is calculated based on ASTM E685-93 as the average peak-to-peak noise in a number of signal segments. The drift is calculated as the slope of the linear regression for the signal.

Procedure for evaporative light scattering detectors: With no flow and the inlet to the detector capped, the signal is monitored over a twenty minute period. The signal noise is calculated based on ASTM E685-93 as the average peak-to-peak noise in a number of signal segments. The drift is calculated as the slope of the linear regression for the signal.

Procedure for refractive index detectors: Pumping water at 1 ml/min, the signal is monitored over a twenty minute period. The signal noise is calculated based on ASTM E685-93 as the average peak-to-peak noise in a number of signal segments. The drift is calculated as the slope of the linear regression for the signal.

5. Injection Precision

Rationale: System precision is critical for accuracy of quantitation. Autosampler performance contributes to system precision.

Procedure: A short column is used to separate the evaluation standard from the void volume. Using a traceable standard, six injections from the same standard are made and the height, area, average height, average area, %RSD of height and %RSD of area are determined and calculated.

6. Injection Carry Over

Rationale: Low carry over from a previous injection is critical for accuracy of quantitative and reliability of qualitative analysis. This test challenges the injector system in the HPLC system.

Procedure: Following the six-injection precision test, a blank injection is made. The carry over result is calculated as a ratio of the area of any residual peak found in the blank injection to the area of the previous injection (expressed as a percentage).

7. Signal to Noise

Rationale: Sensitivity is a critical performance feature in quantitative and qualitative analysis. A signal-to-noise value of a representative compound at known concentration provides sensitivity statistics. This measurement is especially critical to establish level of detection.

Procedure for UV absorbance detector and refractive index detector: An evaluation standard is injected and the calculated height, divided by the ASTM noise monitored over a specified range, provides the signal-to- noise result. Procedure for fluorescence detector: Using pure water in the flow cell, the signal is monitored at the emission maximum wavelength of the Raman band of water and then, using a timetable, switched to a no emission wavelength where the noise is monitored. Signal to noise is calculated as the height of the Raman band peak divided by the monitored noise in a spectral region where no scattering is expected.

8. Response Linearity

Rationale: The linearity of a detector is a critical parameter to establish for reliable and accurate quantitative results and is important for comparability between systems and transferring methods.

Procedure: A series of five traceable standards which represent typical concentrations range are injected and evaluated. The response linearity is calculated by determining the coefficient of determination (r2) of the peak areas versus concentration. It is now recognized that regression statistics alone are insufficient and non-sensitive indicators of linearity. Therefore, the % RSD of the response factors for all five peaks is also calculated. In addtion, as an optional extra linearity statistic, ratios of peak areas in the set of five injections can be reported. For example, up to two ratios such as Peak 2 to Peak 1 and Peak 5 to Peak 2 can be selected in the EQP Record of Variances section.



Information, descriptions and specifications in thispublication are subject to change without notice.

www.agilent.com/chem/enterprise


9. Gradient Composition

Rationale: Accuracy and stability of solvent mixing online is critical for consistent and accurate quantitative analysis. Gradient composition is also important for comparability between systems and transferring methods.

Procedure: [Pre-requisite: UV detector is installed.] An acetone tracer is used to determine the solvent gradient composition accuracy, stability, and linearity. The test challenges the system by making compositional changes from 0% to 100% in 20% increments. In addition, a linear ramp down from 100% to 0% is performed where the composition linearity is determined between ranges 95, 75, and 25%. All composition accuracies are calculated as the absolute difference between the mean composition at each setpoint and the theoretical composition. Stability is determined by the noise and drift at each composition step. Linearity is calculated from 95% to 5% in the linear portion of the gradient.

10. Sample Temperature Accuracy

Rationale: The thermostat accuracy is important for comparing systems and transfer methods.

Procedure: Four vials are filled with water and allowed to equilibrate to the temperature setpoint. Similar to the column compartment, the temperature of the water is measured using a traceable digital temperature meter and proprietary probe. Accuracy is determined as the difference between the measured temperature and the setpoint.

11. Fraction Collection (only applicable if collector is installed)

Rationale: It is important to demonstrate that a fraction collector can collect fractions based on peak detection or time.

Procedure: Two injections of a traceable standard are made and fractions are collected in peak-based or time-based mode. This is a qualitative test in which collected fractions are re-injected to prove that they are fractions of the traceable standard.

Protocol revision: LC.01.85

Document Released: October 2014

Protocol Revision: LC.01.86 Document Released: November 2014

Agilent Technologies, Inc. 2014 Published in USA, November 14, 2014



Customer Approval

Name:

Title:

Date:

Signature:

Name:

Title:

Date:

Signature:

Name:

Title:

Date:

Signature:

Name:

Title:

Date:

Signature:



Legal NoticeEnterprise Edition and its primary components (ACE software tool, procedures, test design, metrology tools, chemical reference

standards, and operator training materials) have been designed, tested, validated, and released for commercial use following Agilent's

Life-Cycle Development Quality Assurance methodology.

Date: November 2014

Services R&D Manager: Michael Pope, Santa Clara, California, USA

Services Quality Manager: Julio Hector, Santa Clara, California, USA

Enterprise Edition is endorsed by Dr. Ludwig Huber on behalf of labcompliance.com.

ACE software is patented. Copyright is claimed by this statement for all original work comprising Enterprise Edition. Any unauthorized

use, reproduction, or translation will be prosecuted to the maximum extent possible by law. All customer copies of EQP approval, final

qualification reports, and raw data provided to customer at delivery of the service become the property of the customer.



Lc.01.86 November 2014 Lc.01.85 September 2014

Current Revision Current Revision Release Date Previous Revision Previous Revision Release Date

Protocol Details


toc_item_0005: Protocol Detailstoc_pagenum_0005: 17toc_item_0004: Legal Noticetoc_pagenum_0004: 16toc_item_0003: Customer Approvaltoc_pagenum_0003: 15toc_item_0002: LCOQtoc_pagenum_0002: 9toc_item_0001: What is EEtoc_pagenum_0001: 5toc_item_0000: Scope and Purposetoc_pagenum_0000: 3