EQUIPMENT QUALIFICATION PLAN
(EQP)
Agilent Enterprise Edition Compliance Services
Qualification of Analytical Scale HPLC Systems
Agilent 1200, 1220, 1260, & 1290 Infinity Series (HPLC & RRLC);
Agilent 1120/1100/1090 Series HPLC and Select Non-Agilent HPLC Models,
with operating pressure range below 400 bar
REVIEW DOCUMENT NAME:
Agilent_Recommended_EQP: HPLC
Page 1 of 17
Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
How to Use This Document
This document is an Equipment Qualification Plan (EQP). It covers Design Qualification (DQ), Installation Qualification (IQ),
Operational Qualification (OQ), scheduled repeat OQ, and Re-Qualification after Repair (RQ). It contains information on how
Enterprise Edition Compliance Services work, and also provides a full list of the tests and checks performed as part of Agilent’s
standard Enterprise Edition IQ and OQ services.
The hardware IQ and OQ procedures listed in this document include fixed tests and checks at Agilent recommended criteria and
limits, both for Agilent and non-Agilent systems (see attachment).
All tests in this document exist in all Agilent delivery tools. The tests descriptions for Capillary Scale and Preparative Scale HPLC
are also detailed in dedicated attachments to this document. However, customer-selectable variance to the standard hardware
OQ setpoints is possible to enable testing of chromatography system(s) over their intended range of use. All setpoint menu
selections in the Variance Section are with the validated range of Enterprise Edition. The inventory of systems covered by the
EQP will be maintained as a separate record.
To facilitate the EQP review and approval process, this document is best viewed on-screen using Adobe ®. Also, there are
several pdf file attachments included with this document: (i) Non-Agilent test specifications; (ii) Question and Answer document;
(iii) 21 CFR Part11 Conformance Checklist for the Agilent Compliance Engine (ACE) - the Enterprise Edition delivery tool; (iv) EE
1.76 EQR comparison with previous revisions.
To approve this EQP simply print to paper and sign. To add variances see instructions below. Keep copies for your own records.
Verbal confirmation of approval is sufficient for Agilent service to proceed with scheduling and delivery.
To make variances to the standard hardware OQ setpoints:
[1] Use the pull-down button to select the alternative approval statement “shall follow...the standard specifications with
VARIANCES to OQ setpoints...”; [2] Complete the “EQP Record of Variances to Setpoints from Standard OQ Specifications”
later in this document; [3] Print EQP to paper and [4] ENSURE THE VARIANCE REQUEST IS COMMUNICATED to Agilent service
engineer BEFORE first OQ delivery starts. Do not e-mail/FAX/post copies of your approved EQP to Agilent. BUT CUSTOMER
MUST PROVIDE A COPY OF ANY EQP WITH VARIANCES TO AGILENT OPERATOR ON-SITE TO ENSURE THE VARIANCES ARE
ENTERED INTO DELIVERY TOOL. NO EXTRA FEE TO DELIVER SETPOINT VARIANCES.
For a full process description, click here to go to the EQP Record of Variances section.
Approval of EQP
The undersigned person(s) approve the following:
[1] the use of Enterprise Edition Compliance Services and delivery of the IQ and/or OQ and/or RQ checks and tests appropriate
to the actual configuration, make, and model of those systems covered by the service;
[2] the specifications described in this Standard EQP Review Document where the tests, setpoints, and limits shall follow...
the
FIXEDAgilent
Agilent
recommended
specifications.
the STANDARD
STANDARD FIXED
recommended
specifications.
Name and Role
Signature and Date
[You cannot save form entries with Adobe Reader. Typed entries and menu selections are printed on your official paper copy when you print.]
DO NOT SEND AGILENT A COPY OF YOUR APPROVED EQP. THIS DOCUMENT IS YOUR OWN RECORD OF APPROVAL.
© Agilent Technologies, Inc. 2014
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Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Contents
To go to a section, click on one of the section titles below.
Sections
Page
How Enterprise Edition Compliance Services Work.................................................................................................. 4
Design Qualification (DQ)............................................................................................................................................... 5
Installation Qualification (IQ) Hardware...................................................................................................................... 6
Operational Qualification (OQ) Hardware.................................................................................................................... 7
Standard OQ Test Specifications for Analytical Scale HPLC Systems (*)................................................................. 7
OQ Test Design and Rationale for Analytical Scale HPLC Systems.......................................................................... 9
EQP Record of Variances to Setpoints from Standard OQ Specifications............................................................... 13
Re-Qualification after Repair (RQ) Hardware............................................................................................................ 14
Legal and Endorsement................................................................................................................................................ 15
Revision History............................................................................................................................................................. 15
PDF file attachments to this electronic EQP (open the attachments folder for this document in Adobe):
Why Has Agilent Introduced the New
Compliance Service, Called Enterprise Edition?
Introduction
Table of contents: [click on title for fast navigation]
What Are The High Level Changes In Enterprise Edition
And What Were The Drivers For These Changes?
Any Other Practical Or Process Changes In
Enterprise Edition?
Let’s Dive Into The Details – How Do The Protocols And
Tests In Enterprise Edition Compare To Classic Edition?
List Of Enterprise Edition OQ Tests Versus
Classic OQPV Tests For LC:
What About The Reports, How Are These Different
To OQPV Reports?
What Would I Have To Do If I Wanted To Move My
Annual OQ Service From Classic To Enterprise Edition?
What Are The Main Risks To Migrating To
Enterprise Edition And How To Avoid Them?
Finally, Can You Summarize The High Level
Comparison Of Enterprise Edition Versus
Classic Edition Compliance Services?
Agilent (then we were HP Analytical) introduced
OQPV for our own LC and GC instruments in the early
1990’s and since then we have delivered well over
100,000 OQPV reports to customers around the world.
Despite the undoubted success and acceptance of our
old OQPV (now called Classic Edition to distinguish
from the new Enterprise Edition service) times have
changed. Expectations and requirements of an
OQ have slightly shifted. The number and type of
instruments and software used by our customers has
increased. And of course we are truly in the new world
of computers and electronic media.
So Agilent set out with a team of international experts
3 years ago to create an upgraded compliance service
that would meet the new demands but crucially
maintain the fundamental requirements:
• Always pass FDA and national agency audits
without over-testing or under-testing;
• Challenge the LC or GC system with a scientifically
sound methodology that provides valuable
performance data.
• Meet the quality needs of customers and the spirit &
intention of the GLP & GMP laws.
• Offer this service at a cost-effective price that
makes it more than just worthwhile – we hope it
is the simplest & best qualification choice that a
customer can make.
What Are The High Level Changes In
Enterprise Edition And What Were The
Drivers For These Changes?
The first big driver was the software environment.
A greatly increased number of chromatography data
system (CDS) products are available to control
LC and GC systems. Agilent has ChemStation,
Cerity, EZChrom, OpenLab and some specialist
LCMS/GCMS software. Our customers also use
Empower, Chromeleon, Atlas, Turbochrom and many
others. Classic OQPV was built into ChemStation
software. The Classic OQPV is a miracle of validated
and almost fully automated OQ testing. But these
benefits are therefore limited to Agilent instruments
running on ChemStation. To provide all our customers,
and customers of non-Agilent instruments, a single
OQ solution as good as (or better than) OQPV – it
was clear we had to develop an automation tool
independent of ChemStation and any other CDS.
The Agilent Compliance Engine (ACE) is our new
software tool that manages the workflow and
protocols, calculates results and produces the
reports. Naturally it is fully validated and tested.
Our service engineers carry “ACE laptops” in the
same way as they carry “ChemStation laptops”.
Alternatively our contract customers can have the
ACE software on their own laptops or installed with
Agilent OpenLab networked CDS.
Tests definitions for
Non-Agilent Systems
Q & A: Why Change?
Capillary Scale Tests
Descriptions
Preparative Scale Tests
Descriptions
© Agilent Technologies, Inc. 2014
Part 11 Checklist (ACE)
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EE 1.76 Comparison
Document
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Document Released: April 2014
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How Enterprise Edition Compliance Services Work
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 lab or 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.
Figure 1:
This EQP Review Document is the record of IQ checks and OQ / RQ tests, setpoints, and limits for HPLC systems. The tests already exist in the automation tool called ACE
and are ready to run after the EQP is approved. ACE holds the test forms applicable to the full range of HPLC configurations plus a validated calculation and report generator
engine. At time of delivery, a record of individual system configuration is made by the operator and entered into ACE. The correct test forms are automatically selected by
ACE from its internal catalog of test designs. Each test in the catalog has a blank results template form. The appropriate setpoints and limits for the individual HPLC system
are added by ACE to the forms according to the approved EQP. When each test is run, the results are calculated and forms completed and then collated to make a single
final report called an Equipment Qualification Report (EQR), which is provided in secure PDF format or optional CD disk – printable to paper and stored in a binder and/or
customers’ network storage system.
© Agilent Technologies, Inc. 2014
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Document Released: April 2014
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 <1058> 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 LC, LCMS, GC, GCMS, UV-Vis and Dissolution 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.
© Agilent Technologies, Inc. 2014
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Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Installation Qualification (IQ) Hardware
Hardware 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.
© Agilent Technologies, Inc. 2014
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Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Operational Qualification (OQ) Hardware
Standard OQ Test Specifications for Analytical Scale HPLC Systems
Test Name
Setpoints and Parameters
Limits
Pump Flow Accuracy and Precision
Flow Rate 1 = 0.500 ml/minute
Flow Rate 2 = 5.000 ml/minute*
Accuracy ≤ 5.00 % from setpoint
Precision ≤ 0.50 % RSD
Column Temperature Accuracy and
Stability
Temperature 1 = 80.0 °C
Temperature 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 noise
Slope of regression fit for drift
Noise: ≤ 0.040 mAU
Drift ≤ 0.500 mAU/hr
Signal Noise and Drift (DAD, MWD)
ASTM baseline noise
Slope of regression fit for drift
Noise: ≤ 0.050 mAU
Drift ≤ 5.000 mAU/hr
Signal Noise and Drift (RID)
ASTM baseline noise
Slope of regression fit for drift
Noise: ≤ 10.000 nRIU
Drift ≤ 400.000 nRIU/hr
Signal Noise and Drift (ELSD)
ASTM baseline noise
Slope of regression fit for drift
Noise: ≤ 2.000 mV
Drift ≤ 5.000 mV/hr
Signal Noise and Drift (CD)
ASTM baseline noise
Slope of regression fit for drift
Noise: ≤ 0.100 uS
Drift ≤ 10.000 uS/hour
Signal to Noise (UV-Vis)
Signal height is divided by ASTM baseline
noise for known concentration and known
conditions.
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
Key:
Variance allowed for setpoint(s)
Fixed setpoints/limits
* 2.000 ml/min for G4220B 1290 pump
** T2 = 60.0°C for 1120 and 1220 systems
See attachments for recommended test definitions for Non-Agilent Systems
© Agilent Technologies, Inc. 2014
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Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Operational Qualification (OQ) Hardware (continued)
Standard OQ Test Specifications for Analytical-Scale HPLC Systems (continued)
Test Name
Setpoints and Parameters
Limits
Injection Precision
(UV-Vis, RID)
Injection volume on column = 20 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 (FLD)
Injection volume on column = 5 ul
(2 ul for 1290 w/ CTC samplers)
Height carry over ≤ 0.40 %
Area carry over ≤ 0.20 %
Injection Carry Over (CD)
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) ≥ 0.99900
R/F precision ≤ 5.00 % RSD
Response Linearity
(RID, CD)
5 concentrations of certified reference
standard
Coefficient of determination (r2) ≥ 0.99500
R/F precision ≤ 10.00 % RSD
Gradient Composition
Accuracy (UV-Vis)
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 Composition
Noise and Drift (CD)
20.00 %, 40.00 %, 60.00 %, 80.00 % steps
Composition noise ≤ 3.50 %
Composition drift ≤ 3.50 %
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 °C
Samples four vials of water in different
tray positions
Diff. from setpoint ≥ –2.0 °C, ≤ 5.0 °C (setpoints < 10 °C)
Fraction Collection
Select Fraction Collector 1, 2, or 3
Select Peak or Time-based collection mode
Peak Presence (Qualitative)
* Area RSD ≤ 1.00 % with CTC samplers
** r2 ≥ 0.99000 for CD
Key:
Fixed setpoints/limits
Variance allowed for setpoint(s)
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
© Agilent Technologies, Inc. 2014
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Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Operational Qualification (OQ) Hardware (continued)
OQ Test Design and Rationale for Analytical Scale HPLC Systems
Many 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 Agilent’s 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.
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 (0.5 ml/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. (continues on next page)
© Agilent Technologies, Inc. 2014
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Operational Qualification (OQ) Hardware (continued)
OQ Test Design and Rationale for Analytical Scale HPLC Systems (continued)
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).
© Agilent Technologies, Inc. 2014
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Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Operational Qualification (OQ) Hardware (continued)
OQ Test Design and Rationale for Analytical Scale HPLC Systems (continued)
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.
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.
© Agilent Technologies, Inc. 2014
Page 11 of 17
No reproduction, translation, or use without permission
Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Operational Qualification (OQ) Hardware (continued)
OQ Test Design and Rationale for Analytical Scale HPLC Systems (continued)
The following tests are NOT INCLUDED in the standard OQ for HPLC but can be ordered as EXTRA COST TESTS.
Test Name
Setpoints and Parameters
Limits
Include
Injection Linearity (UV-Vis)
Any choice of 5 injection volumes.
Constant concentration standard is
5 ug/ml caffeine.
Coefficient of determination (r2) ≥ 0.99900
R/F precision ≤ 5.00 % RSD
Injection Response (UV-Vis)
Same setpoint as Injection Precision
test.
Average area ≥ 1,200,000 and ≤ 1,800,000 counts
(For standard cell with 20 ul injection. Result is
corrected for path length and attenuation.)
Wavelength Accuracy
(Extended Test, UV-Vis)
Wavelength 1 = 361 nm [Maximum]
Wavelength 2 = 416 nm [Maximum]
Wavelength 3 = 451 nm [Maximum]
Wavelength 4 = 537 nm [Maximum]
Accuracy ≤ 2 nm
Fixed setpoints/limits
Variance allowed for setpoint(s)
Additional Test 1. Injection Linearity (optional extra test available in custom-configured EQP)
Rationale: Injection linearity of variable volume HPLC injector systems is normally not critical to quantitative or qualitative
analysis. Most HPLC analytical methods use fixed and only nominal injection volumes and do not use variable volume injections
within a single analysis. However, some users may wish to use variable volume injection if the linearity is demonstrated.
Procedure: Five injections of increasing volumes of the same traceable caffeine standard are made. Injection linearity is
calculated from the coefficient of determination (r2) of the peak areas versus injection volume. Also, %RSD of the response
factor for all five peaks is calculated.
Additional Test 2. Injection Response (optional extra test available in custom-configured EQP)
Rationale: The accuracy of the injected volume is normally not critical to quantitative or qualitative analysis. Most HPLC
analytical methods use fixed and only nominal injection volumes and results are not affected by even moderate inaccuracy in
actual injected volume. However, it may be important for comparability between systems and transferring methods, and it is
useful as a diagnostic for establishing that the correct injection syringe/loop/device is installed.
Procedure: A known traceable caffeine standard is injected six times (in the precision tests) and the average response is
calculated. The injection response is the mean of the average areas corrected for sample concentration, cell path length, and
attenuation, and the response within an acceptance window indicates correct volume injected.
Additional Test 3. Wavelength Accuracy, Extended Test: (optional extra test available in custom-configured EQP)
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 holmium oxide standard is used to determine the
wavelength accuracy. In one procedure, for certain models, the holmium oxide is trapped in the flow cell and a programmable
timetable is used to determine the wavelength maxima (241, 278, 287, 361, 416, 451, 537 and/or 641 nm). For other models (for
example, DAD and PDA), a holmium oxide injection is made and a spectrum is acquired. The spectral maxima 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.
© Agilent Technologies, Inc. 2014
Page 12 of 17
No reproduction, translation, or use without permission
Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Operational Qualification (OQ) Hardware (continued)
EQP Record of Variances to Setpoints from Standard OQ Specifications
IGNORE THIS SECTION IF YOU ACCEPT AND APPROVE THE FIXED STANDARD QUALIFICATION TESTS AND SETPOINTS
RECORDED IN THE PRECEDING PAGES OF THIS STANDARD EQP.
EQP with Variance Approval Process: Customer Actions:
1. View in Adobe®; select required setpoint variances below; select the alternative approval statement on page 2.
2. Print to paper to save the selections; sign page 2 of this EQP.
3. Ensure the approved EQP with Variances is provided to Agilent operator on the day of the first delivery before start of OQ.
Counter-sign and date the Agilent operator signature on this page. [End of EQP with Variance approval process. Next step:
wait for your qualification reports.]. Agilent Operator Actions:
1. Enter and save the customer change requests on this page into the ACE tool.
2. Sign and date this page on the customer copy to verify that you made the changes in ACE; return signed copy to customer
for counter-signature.
3. Deliver the qualification by following this EQP and any setpoint variances. (Note: Once the EQP Variances are entered into
ACE these are saved for all future OQ/RQ events where applicable.)
Test Name
Pump Flow Accuracy and Precision
Setpoint
Standard
Variance
Units
Flow Rate 1
0.500
No variance
ml/minute
Flow Rate 2
5.000
No variance
ml/minute
Temperature 1
80.0
No variance
°C
Temperature 2
40.0
No variance
°C
Injection Precision, Carry Over (UV-Vis, RID)
Injection Vol. on Column
20
No variance
ul
Injection Precision, Carry Over (CD)
Injection Vol. on Column
25
No variance
ul
Injection Precision (ELSD)
Injection Vol. on Column
20
No variance
ul
Sample Temperature Accuracy
Temperature
4.0
No variance
°C
Response Linearity
(UV-Vis, RID optional extra statistic)
1st Peak Area Ratio
Not applicable
No variance
2nd Peak Area Ratio
Not applicable
No variance
Wavelength Accuracy (Extended Test, UV-Vis)
Wavelength 1
361
No variance
nm
Wavelength 2
416
No variance
nm
Wavelength 3
451
No variance
nm
Wavelength 4
537
No variance
nm
Injection Volume 1
Not applicable
ul
Injection Volume 2
Not applicable
ul
Injection Volume 3
Not applicable
ul
Injection Volume 4
Not applicable
ul
Injection Volume 5
Not applicable
ul
Injection Volume
Not applicable
ul
Column Temperature Accuracy and Stability
Injection Linearity
Injection Response
Customer:
Agilent Operator (verifies variances are entered into ACE):
Name:
Name:
Signature, Date:
Signature, Date:
For a fully tailored operational qualification program using all the flexibility of Enterprise Edition, contact your local Agilent representative and/or e-mail [email protected]
with your OQ test specification requirements. Fees may apply.
© Agilent Technologies, Inc. 2014
Page 13 of 17
No reproduction, translation, or use without permission
Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Re-Qualification after Repair (RQ) Hardware
In 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.
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).
Dual-Acceptance Limits
Within 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_
EQP’s 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.
© Agilent Technologies, Inc. 2014
Page 14 of 17
No reproduction, translation, or use without permission
Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Legal and Endorsement
Enterprise Edition and its primary components (ACE software tool, procedures, test design, metrology tools, chemical reference
standards, operator training materials) has been designed, developed, tested, validated, and released for commercial use
following Agilent’s Life-Cycle Development Quality Assurance methodology.
Date: April 2014
Services R&D Manager: Michael F. 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.
Revision History
Revision History of HPLC Enterprise Edition Protocols
A.01.84 April 2014
Added support for (1) Agilent G1312C FLD; Ultra-low dispersion kit for G1290 pumps. Removed support
for (1) HP/Agilent 1050 and 1090 Systems; (2) Agilent SFC System.
A.01.83 September 2013
Changed injection volume in the Carry-Over test to 25ul and to 5ul for CD and FLD systems respectively.
A.01.82 April 2013
Added support for (1) G4260A/B and G4621 A/B ELSD Detectors (former Varian 380, 385); (2) G4309A
SFC System; (3) High-Dynamic Range LC Configurations and G4234 column switching valve. Changed
the test scheduling for IP & CO in multi-detector configuration - to run only once in the standard
program. Others can be added as optional tests. Only one instance of the Injection Precision & CarryOver tests will be performed standard for multi-detector systems.
A.01.81 January 2013
Added support for: (1) Agilent 1220 DAD, (2) additional Waters, Dionex and GE ATKA modules and
systems (see Compatibility Matrix for details); (3) Other/Unspecified Modules (General-Purpose
Systems). Updated step 1–4 accuracy label text in Gradient Composition test (analytical and capillary
scales). Calculations remain unchanged. Updated accuracy check calculation in Pump Flow Precision
for Preparative Scale sytems. No regulatory impact. Corrected typo on FLD carry-over test limits.
A.01.80. July 2012
Added support for G42204A 1290 Quaternary pump. Re-instated Injection Precision test for FLD
systems. Updated acquisition method to measure noise on the Raman current in the Signal-to-Noise
test for FLD systems. Improved calculation to separate the artifact from the standard and carry over
peaks. Updated ASTM algorithm for Signal-to-Noise and Noise & Drift tests.
Updated test limits for 1220 system and Flow Accuracy & Precision test.
A.01.79. April 2012
Added support for (1) vial verification testing in IP test, (2) Thermo and Waters additional equipment.
Updated section in Pump flow precision to reflect 10 readings used in calculations for prep pumps.
A.01.78. February 2012
(1) Added support for Waters H-Class pump, injector, column compartment, valve, and detector;
Acquity ELSD. (2) Removed IP/CO tests for FLD detectors — Use Custom EQP as required
A.01.77. December 2011
No changes to LC impacting regulatory approval
A.01.76. August 2011
No changes to LC. Protocol revision made independent from ACE revisions. No regulatory impact.
A.01.75. March 2011
(1) Added support for Agilent 1260 Bio LC. No regulatory impact for previously-approved Standard_EQP
documents.
© Agilent Technologies, Inc. 2014
Page 15 of 17
No reproduction, translation, or use without permission
Agilent_Recommended_EQP
Document Released: April 2014
Enterprise Edition Compliance Services
Revision History (continued)
Revision History of HPLC Enterprise Edition Protocols
A.01.74. September 2010
(1) Added support for Agilent 1220 & 1260 Infinity Systems. (2) Added FLD tests for Injection Precision
& Carry Over. (3) Added Tests definitions for non-Agilent Systems. No regulatory impact for previouslyapproved Standard_EQP documents.
A.01.73. June 2010
(1) Added Additional Tests Selection Boxes. (2) Added Additional Tests Variances tables.
No regulatory impact.
A.01.72. January 2010
(1) Added support for additional Shimadzu and Dionex modules (see Compatibiliti matrix for details);
(2) added support for an extended Wavelength Accuracy test for UV/UV-Vis detectors (NO
REGULATORY IMPACT).
A.01.71. October 2009
(1) Added E-signature fields; (2) changed nomenclature used to list limits (NO REGULATORY IMPACT).
A.01.70. May 2009
(1) Added capillary scale support; (2) added internal/external valve support; (3) reprocess plots for
Injection Precision, Injection Carry Over, Response Linearity, Gradient Composition; (4) updated noise
and drift calculation (NO REGULATORY IMPACT).
A.01.60. SR2, November 2008
(1) Added Agilent Jet Stream Technology support; (2) improved noise calculations for low-noise
detectors.
A.01.60. May 2008
(1) Reset DAD/MWD/VWD noise and drift Test Specification limits to match Classic OQ/PV limits (NO
REGULATORY IMPACT); (2) added second UV detector, second injector, and second and third column
compartment and degasser; (3) added support for ELSD and Dionex conductivity detectors; (4) added
forms: Certificate of System Qualification at end of EQR, Chromatography Report after each applicable
test, Errors and Corrections for operator and customer to record any corrections to EQR, Data Transfer
Audit Log for complete traceability.
A.01.54. January 2008
No changes for HPLC.
A.01.53. August 2007
Added kPa units for pressure tests.
A.01.50. March 2007
Added fraction collector support.
A.01.40. December 2006
No changes for HPLC.
A.01.30. July 2006
(1) Added FLD and RID support; (2) added Signal to Noise test.
A.01.20. March 2006
Added rapid resolution support.
A.01.10. November 2005
Initial HPLC - Analytical Scale - Operational Qualification.
End of EQP Review Document
© Agilent Technologies, Inc. 2014
Page 16 of 17
No reproduction, translation, or use without permission
www.agilent.com/chem/enterprise
Information, descriptions and specifications in this
publication are subject to change without notice.
© Agilent Technologies, Inc. 2014
Published in USA, April 22, 2014
Page 17 of 17