Agilent ChemStation Plus Method Validation Pack. User s Guide

Transcription

1 Agilent ChemStation Plus Method Validation Pack User s Guide A

2 Notices copyright Agilent Technologies, Inc. 2001, 2003 and headwork-consulting GmbH ( ) No part of this manual may be reproduced in any form or by any means (including electronic storage and retrieval or translation into a foreign language) without prior agreement and written consent from Agilent Technologies, Inc. as governed by United States and international copyright laws. Microsoft is a U.S. registered trademark of Microsoft Corporation. Manual Part Number G Edition 09/03 Printed in Germany Agilent Technologies, Inc. Hewlett-Packard-Strasse Waldbronn headwork-consulting GmbH 2003 Bonner Talweg Bonn Germany Software Revision This guide is valid for A.01.xx revisions of the Agilent ChemStation Plus Method Validation Pack software, where xx refers to minor revisions of the software that do not affect the technical accuracy of this guide. Warranty The material contained in this document is provided as is, and is subject to being changed, without notice, in future editions. Further, to the maximum extent permitted by applicable law, Agilent disclaims all warranties, either express or implied, with regard to this manual and any information contained herein, including but not limited to the implied warranties of merchantability and fitness for a particular purpose. Agilent shall not be liable for errors or for incidental or consequential damages in connection with the furnishing, use, or performance of this document or of any information contained herein. Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms, the warranty terms in the separate agreement shall control. Technology Licenses The hardware and/or software described in this document are furnished under a license and may be used or copied only in accordance with the terms of such license. Restricted Rights Legend If software is for use in the performance of a U.S. Government prime contract or subcontract, Software is delivered and licensed as Commercial computer software as defined in DFAR (June 1995), or as a commercial item as defined in FAR 2.101(a) or as Restricted computer software as defined in FAR (June 1987) or any equivalent agency regulation or contract clause. Use, duplication or disclosure of Software is subject to Agilent Technologies standard commercial license terms, and non-dod Departments and Agencies of the U.S. Government will receive no greater than Restricted Rights as defined in FAR (c)(1-2) (June 1987). U.S. Government users will receive no greater than Limited Rights as defined in FAR (June 1987) or DFAR (b)(2) (November 1995), as applicable in any technical data. Safety Notices CAUTION A CAUTION notice denotes a hazard. It calls attention to an operating procedure, practice, or the like that, if not correctly performed or adhered to, could result in damage to the product or loss of important data. Do not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met. WARNING A WARNING notice denotes a hazard. It calls attention to an operating procedure, practice, or the like that, if not correctly performed or adhered to, could result in personal injury or death. Do not proceed beyond a WARNING notice until the indicated conditions are fully understood and met. Agilent ChemStation Plus - Method Validation

3 In This Guide This operator manual contains information on how to work with Method Validation Pack Version A or higher. The chapter Installation describes the installation of Method Validation Pack. The chapter Operation describes the general handling of the program including a short description of the main menu items. The chapters Planning and Execution describe the standard steps of a Validation. These are the input of the most important parameters of a Validation (planning) and the input of data necessary for the statistical reduction (execution). In the Utilities chapter, additional options of Method Validation Pack are described, many of which are basic settings which are rarely changed and can be accessed only by authorized persons. Agilent ChemStation Plus - Method Validation Pack 3

4 4 Agilent ChemStation Plus - Method Validation

5 Contents 1 Overview 11 Preface to Version A Validation 14 Validation Definition 16 Validation and QM requirements 17 2 Installation 23 Before the Installation 24 Start of Installation 26 Installation of additional components 26 Users of VALIDAT versions (Pre-MVP installations) 27 Requalification 28 Network 28 Licences 29 Support for SQL-Servers 30 Client/Server 30 DMS (Document Management System) 30 Viewing the program log file 35 Viewing the database log file 36 Configuring VALIDAT.CFG 37 Database sizes 41 Starting the program 43 3 Operation 45 Security Concept 46 Levels of Entry 46 Agilent ChemStation Plus - Method Validation Pack 5

6 Contents Auditing 47 Locking a Validation 47 Operation 49 The Validation assistant 50 Main window 52 About DMS (Document Management System) 59 DMS History 62 4 Planning 63 Checkpoints and validation extend 64 Planning a Validation 67 Checkpoints 70 Configuration data 72 External documents 73 Store Method 73 Validation type 73 Checkpoints 75 Precision 75 Ruggedness/Robustness 77 Linearity 82 Calibration function 83 Detection and Determination limit 85 Accuracy 87 Selectivity / Specificity 90 Ring experiment 91 Output settings 93 5 Execution 103 Input Agilent ChemStation Plus - Method Validation Pack

7 Contents Data input window 106 Robustness 110 Lab capability 110 Calibration function/linearity 110 Detection and Determination Limit 111 Accuracy 111 Selectivity/Specificity 112 Ring experiment 112 Report 113 Additional Report Options 118 Graphics Utilities 121 Configuration 122 Levels of significance 122 Other variables 124 Archive 126 A Formulae 127 Precision 128 Relative Standard Deviation CV 128 Dixon test for outliers 128 Grubbs outlier test 128 Neumann Trend Test 129 Repeatability Limit r 129 Error of Result u 129 Confidence Interval 129 Ruggedness 130 Dixon 130 Grubbs 130 Neumann Trend Test 131 Agilent ChemStation Plus - Method Validation Pack 7

8 Contents Cochran test for variance homogeneity 131 Bartlett test for variance homogeneity 132 F-test for variance homogeneity 133 Repeatability standard deviation 133 Standard deviation mean value 133 Reproduce Standard Deviation 133 F-test for Variance Homogeneity 134 t-test for Ruggedness 134 Horwitz-test 135 Linearity 137 Scaling factor C 137 Result of F-test 139 Lab Capability 141 Short term deviation sst and long term deviation slt 141 Method effectiveness index 142 Detection and Determination Limit 144 Limit of Detection / Blank method 144 Limit of Detection / Calibration line method 144 Accuracy 146 Nominal Value Comparison 146 Wilcoxon-test (signed rank test) 146 Comparison of two samples 147 Recovery 148 Selectivity 150 Standard Addition Method 150 Extended Spiking Method 151 Chromatography 152 Ring Experiment Agilent ChemStation Plus - Method Validation Pack

9 Contents B Qualification Tests 155 File Verification/Installation Qualification (IQ) 156 Performance Qualification (OQ/PV) - Support Services 157 C Error Messages 159 D Known Problems 161 E Glossary 163 F Interfacing ChemStation Plus and Method Validation Pack 167 Validation, components and checkpoints 168 Step 2: Data input or import 175 ChemStation Plus modules and their functionality in the method validation process 176 General configuration 177 Extended planning 179 Minimizing the sample number by using one run for Multiple checkpoints 181 Managing MVP data as ChemStore custom fields 186 Data completion wizard 187 User management 189 ChemStation 190 Index 193 G Literature 195 Agilent ChemStation Plus - Method Validation Pack 9

10 Contents 10 Agilent ChemStation Plus - Method Validation Pack

11 Agilent ChemStation Plus Method Validation Pack User s Guide 1 Overview Preface to Version A Validation 14 Validation Definition 16 The program Method Validation Pack supports Validation of methods in accordance with various guidelines such as DIN, USP and ICH guidelines. It runs under Microsoft Windows NT 4.0 and Microsoft Windows ChemStation Plus Method Validation Pack ( MVP ) is a compound-specific software permitting the validation of methods with different compound types such as main compounds, secondary compounds and trace compounds. It offers a list of tests ( checkpoints ) for result calculation and result evaluation based on advanced statistical calculations. It combines results, statistics and related information in a final validation report. MVP offers an all-inclusive list of checkpoints allowing the calculation of validation reports in accordance with all major method validation guidelines. It supports the requirements of 21 CFR part 11 for electronic records and electronic signatures and can be used in regulated environments. The following checkpoints necessary for the Validation can be planned and executed: Precision Ruggedness/Robustness Lab capability Linearity Calibration function Detection and Determination Limit Accuracy Agilent Technologies 11

12 1 Overview Selectivity/Specificity Ring Experiment Method Validation Pack analyses the data input statistically and gives as a result the characteristics corresponding to a specific checkpoint, e.g. standard deviation, confidence interval, statements on linearity, homogeneity, systematic and/or proportional errors etc. Starting with the hierarchical planning of a Validation, all steps up to the printed report are supported. The report, including graphics and lists of content, can then be exported to an external word processing program where it can be worked upon. 12 Agilent ChemStation Plus - Method Validation Pack

13 Overview 1 Preface to Version A Preface to Version A Version A of Method Validation Pack contains many new or extended functions: IQ Extended Installation Qualification OQ/PV Extended Operation Qualification/Performance Verification (Support Services) ORACLE SQL Database Document Management System stores revision of all important parts of a Validation to track the complete life cycle of a validation (full History Management) New top-level navigation bar for easier function access Electronic signature for 21 CFR part 11 support Better support for manual integration (batch review) Support for GC back-injektor Multi-instrument installations supported Modifications of already exported validations possible Import wizard can directly connect to XLS data sources Agilent ChemStation Plus - Method Validation Pack 13

14 1 Overview Validation Validation Validation is the formal and systematic proof that an analytical method permits an evaluation of its quality characteristics with adequate and defined reliability. These characteristics are defined in various regulations (ICH, DIN, ISO). Validated methods are a mandatory requirement for successful audits in regulated environments (e.g. FDA). Validation Analytical methods do not usually determine all details of executions that have influence on the deviation of results. Random errors of test substances may influence the results to some degree. Lab capability Lab capability refers to the routine use of checking methods in a laboratory. Routine use means the use of an analytical method as it develops through frequent (routine) use during quality checks. For more detail, see Checkpoints on page 75 Documentation Results, in the form of printouts, and all raw data, must be maintained in accordance with documentation rules. An analytical method becomes valid through the signature of the head of the institution or his/her deputy. Validity of an analytical method Validation is valid for an approved analytical method, a defined product and for the analytical laboratory in which the method is used. An additional validation is required in the case of Substantial changes concerning the quality of the product Changes in the analytical method which influence the result Changed customer requirements, e.g. requirements of an authority 14 Agilent ChemStation Plus - Method Validation Pack

15 Overview 1 Validation The scope of the additional validation can vary from a validation modification, e.g. an additional or updated checkpoint for one or multiple compounds, to a full revalidation. Extent of Validation The extent of the validation has to be modified to suit the application sector and the type of analytical method used. It must be authorized by the head of the testing institution or his/her deputy. In larger laboratories, the head of department must authorize the extent of Validation Agilent ChemStation Plus - Method Validation Pack 15

16 1 Overview Validation Definition Validation Definition According to accepted international quality standards, Validation means: Confirmation on the basis of a test plus the proof that particular requirements for a specific, intended use are met. Therefore, a validation is possible only if a specific, intended use is established. For an analytical method, this means that the range of application including working range and matrix definition must be determined. It is obligatory for an analytical method to be recorded. It also means that a validation can be executed only on the basis of sufficient expert knowledge because only then can the specific, intended use be judged. The above general definition can be transferred to the validation of analytical methods: Validation is the formal and systematic proof that an analytical method gives results for a defined range of application with adequate and defined reliability. Statistical data have to be processed for a validation. They must permit the judgment of the suitability of a method as well as the judgment of the results as processed under routine conditions with the method. See Appendix A for details. Verification demands that stipulated requirements are met, instead of particular requirements for a specific, intended use (see the above definition). We speak of verification if, for example, specific data are supplied in a standard or a specification and the adherence to these data must be checked. Validation, on the other hand, means that the analyst gives a proof of for his/her specific case. The definition of the American Food and Drug Administration FDA is of great importance: Establishing documented evidence which provides a high degree of assurance that a specific process will consistently produce a product meeting its predetermined specifications The FDA requirements thus refer to processes to be validated during fabrication of products of a specific and continuous quality. This concept of validation can be transferred to different industries. 16 Agilent ChemStation Plus - Method Validation Pack

17 Overview 1 Validation Definition Validation and QM requirements Different validation requirements may be demanded according to the QM system used. The validation requirements according to ISO and GLP are summarized here as examples. ISO Reliability and comparability of analytical methods are in the foreground here. When ISO is used, the result uncertainty has a special meaning. All data processed during a validation are summarized into a degree of uncertainty which determines the respective analytical method. The customer must be informed if he/she wishes it. Validation of procedures Validation is the confirmation by a test plus the proof that particular requirements for a specific, intended use are met. When do users have to validate their methods? They must validate all methods that are not described in any regulatory publication as, for example, a pharmacopoeia release or other guidance document, to confirm that the method is applicable to the intended use. In particular, these are: methods which are not determined in normative documents, self-developed methods, methods in accordance with normative documents that are used outside their intended range of application extensions of methods in accordance with normative documents The validation must be exercised on a scale which is necessary to fulfill the requirements of the intended use. The laboratory must record the results and the methods used for the validation. It must determine whether the method is applicable for the intended use. NOTE Validation can contain methods for sampling, use and transport. Agilent ChemStation Plus - Method Validation Pack 17

18 1 Overview Validation Definition NOTE One of the following methods, or a combination of them, should be used to determine the method characteristics: calibration with reference norms or reference materials; comparison with results gained by other methods; comparison between laboratories; systematic assessment of factors which influence the result; assessment of precision on the grounds of an understanding of the theoretical basics of the method and practice. NOTE Should changes be made to the validated but not standardized methods, the influence of such changes should be recorded, and a new validation should be done if necessary. The range and the exactness of the values determined by the validation method (e.g. precision, detection limit, selectivity, linearity, repeatability limit or comparison limit, ruggedness against external influences) must meet customer requirements. NOTE Validation includes description of requirements, determination of procedure characteristics, a test to make sure the requirements by using a methods are met, and a statement of validity. NOTE Regular tests are necessary if a method is continuously developed to make sure customer requirements are met. Any change of requirements which require modification of the development program must be tested and authorized. NOTE Costs, risks and technical possibilities must be considered carefully. In many cases the range and uncertainties of values (e.g. for precision, detection limit, selectivity, linearity, reproducibility, repeatability, ruggedness, sensitivity) cannot be precisely determined because of a lack of information. 18 Agilent ChemStation Plus - Method Validation Pack

19 Overview 1 Validation Definition GLP General GLP requirements at the high level of the OECD are more concerned with data recording and the validation of computerized systems than with method validation. This is expected by the authorities. There are many regulations which cannot be explained here (see bibliography at the end of the book). Here are some general GLP principles of validation, describing the general responsibilities and requirements of quality work. They are mandatory prerequisites for any work procedures that require validation, thus including method validation. Management responsibilities of test organization Management must at least introduce methods to make sure that computerized systems are suitable for intended use. They must be validated, used and cared for in accordance with the principles of Good Laboratory Practice. Responsibilities of the test manager The test manager must make sure that the computerized systems used during tests are validated. Apparatus, materials and test solutions Apparatus used for the acquisition, recording and reproduction of data (including computerized systems) and for the control of environmental conditions important for testing must be suitably housed. They must be suitably constructed and equipped with enough power. Standard work instructions Standard work instructions must be available at least for the following areas: Apparatus, materials and test solutions... Computerized systems Validation, operation, maintenance, safety, change control, back-up Recording, storing and tracing Validation documents for computerized systems must be kept for fifteen years in an archive Agilent ChemStation Plus - Method Validation Pack 19

20 1 Overview Validation Definition Validation expenditure The greatest challenge for a laboratory is the determination of an acceptable validation expenditure and the extent of a validation. Commercial interests stand in the way of a complete validation. The following rules may serve as a guideline: Procedures published in standards and official method collections may be taken as validated. Laboratory-specific deviations, however, must be validated. One well-known example of a published standard is the use of the Microsoft Windows PC operating systems. It is typically not expected that users validate this software, because it has been validated through the use of many millions of users. Self-developed methods ( house methods ) must be completely validated. The higher the health or environmental risk in the case of wrong results, the higher the validation expenditure must be. The higher the commercial risk is (product liability, loss of customer), the higher the validation expenditure must be. The less the practical experience available with a method is, the more extensive the validation must be. The more important for a company a product or a method is, the more extensive the validation must be. The better the guaranteed reliability is, by control charts or ring experiments, the less extensive the validation must be. The higher the regulation expectations (GLP/GMP) are, the more conscientious the validation, and especially the documentation, must be. The more complex a method is, the more complex the validation (e.g. trace analysis); the easier the method, the more a calibration of apparatus may be sufficient. The more extensive a validation supplied by the apparatus manufacturer is, the less validation the user has to do. The higher the customer requirements with respect to reliability and exactness is (e.g. product analysis with narrow specification limits), the more expressive a validation must be. 20 Agilent ChemStation Plus - Method Validation Pack

21 Overview 1 Validation Definition Errors and problems During audits and inspections, certain sensitive points in a validation often come to light. Just observing them and trying to avoid them improves the quality of a validation considerably. Validation often occurs under unreal conditions: the best employee using the newly calibrated apparatus. This often results in unrealistically good values, which cannot be upheld during routine operation. So, one key recommendation in validation is: Use realistic specifications in your validation plan (claim by Ludwig Huber, personal note). Wrong results are just not expected, which of course is wrong in itself. Misoperation and faulty machines cannot be excluded. Validation should offer a measure of security as high as possible but with a reasonable effort. Absolute security can neither be obtained nor paid for. Too much emphasis is placed on precision and not enough on accuracy. Precision may be obtained easily by repeated tests, accuracy however always needs a reference (such as reference material or reference methods, occasionally reference laboratories). Too much emphasis is placed on coincidental deviations and possible errors during execution and calibration, and too little during sample taking. During a report, the results are presented with an exactness inconsistent with validation results. Anxiety in employees is underestimated (is my method as good as I claim it to be?). Validation turns into an impediment to innovation: you are reluctant to change methods accepted by the customer and the organizations. You want to avoid questioning, although you are sure to have found a better method to work with. Validation is done by chance: testing is done on a random basis, and no planning or proper final report are done. The Validation has been executed but raw data and documentation is completely confused. The procedure cannot be duplicated. Method Validation Pack effectively overcomes the last two points by providing a validation plan and a validation report at the end. We recommend that these documents are stored together with the raw data. Agilent ChemStation Plus - Method Validation Pack 21

22 1 Overview Validation Definition Conclusion: MVP therefore allows all documents related to the method validation to be stored in its DMS system and also makes revalidation easy by storing and managing all Method Validation Pack revisions in its document management system etc. Responsibility Validation responsibilities must be determined in a quality management system: Who determines which methods are to be validated? Who determines the validation extent and releases the validation plan? Who executes the validation in the laboratory? Who prepares the validation report and releases it? Who determines revalidation steps? Usually a validation plan and its release is done by the laboratory head, but different options are of course possible. It is best to summarize all validation rules into a validation manual. Corresponding regulations have to be determined for the development, change and release of analytical methods. It should be made clear whether validations are already necessary for the release of analytical methods. If this is the case, it should also be determined which, if any, validation results are included into the analytical methods. Revalidation A validation cannot be valid for an unlimited amount of time. A revalidation is necessary if changes are made concerning the analytical method (e.g. a different solution, different extraction conditions) the analytical apparatus (e.g. a different detector) the sample (e.g. a different matrix, different product composition) customer requirements (e.g. new rules and regulations) In most cases, the effort for a revalidation is lower than that for the original validation. Only checkpoints that are affected by changes have to be validated. 22 Agilent ChemStation Plus - Method Validation Pack

23 Agilent ChemStation Plus Method Validation Pack User s Guide 2 Installation Before the Installation 24 Start of Installation 26 Support for SQL-Servers 30 This chapter describes the installation of Method Validation Pack. Agilent Technologies 23

24 2 Installation Before the Installation Before the Installation Hardware requirements To run Method Validation Pack, you need a computer with either the operating system Microsoft Windows NT 4.0 or Microsoft Windows Physical memory required is 96 MB RAM for Windows NT, and 128 MB RAM for Windows 2000 respectively. License Number When Method Validation Pack is started for the first time, it asks for a license number. You can find the license number on the license agreement. This license number is case sensitive. Service packs Service pack 6a or later is required for Microsoft Windows NT 4.0; or SP 2/3 for Microsoft Windows 2000 respectively. Year 2000 Use only versions of operating systems and hardware that are classified as year 2000-compliant to avoid Y2K problems. Method Validation Pack itself (the program and its components) uses only documented system calls for time and date access. Internet Explorer Method Validation Pack needs Microsoft Internet Explorer 5.5 or higher to show the online help and the online manual. Netscape Browser It is possible to use Netscape Browser. However, it restricts the convenient use of the online help because the browser window cannot be embedded in Method Validation Pack. 24 Agilent ChemStation Plus - Method Validation Pack

25 Installation 2 Before the Installation Printer Method Validation Pack needs an installed printer to show the reports on screen. If no printer is installed, the program cannot be started. Whether the printer is a local printer or a network printer is irrelevant. Hard disk space Method Validation Pack needs approximately 40 MB hard disk space for installation. The Validation files typically need KB per file. This depends on the size of embedded graphics. If possible, use vector graphics (WMF-format) rather than bitmap graphics (BMP) to reduce validation file size. Using large bitmap graphics in validation comments also has an impact on the DMS size. Hard disk requirements for the Oracle database are discussed later in this chapter. Agilent ChemStation Plus - Method Validation Pack 25

26 2 Installation Start of Installation Start of Installation Before installation, make sure that all user programs except the desktop are closed, otherwise the installation may fail. Begin the installation of Method Validation Pack by starting the program SETUP.EXE on the Installation CD. If your CD drive is configured to autostart, the insertion of the CD is sufficient. Installation of additional components Method Validation Pack needs the following components for correct installation and operation. These components are not automatically installed. Ensure that this has been done prior to Method Validation Pack installation. ChemStation/ChemStore ChemStation Plus Rev A or higher with the following modules: ChemStation Data Acquisition software Rev. A or higher (optional) ChemStore C/S Rev B (mandatory) Security pack Rev B (optional) ADO 2.5 ADO 2.5 (Microsoft Data Access Components 2.5) (installed by ChemStore) or higher Adobe Acrobat Reader 4.0 (part of standard bundle) The Microsoft Data Access Components installs a layer to access local databases or central SQL servers. Method Validation Pack uses Microsoft JET database to save local data (i.e. MS Access format). Microsoft Windows 2000 already contains the 2.5 version of ADO. This version is fully compatible with Method Validation Pack. Adobe Acrobat Reader 4.0 or 5.0 is required to display the online manual. 26 Agilent ChemStation Plus - Method Validation Pack

27 Installation 2 Start of Installation Users of VALIDAT versions (Pre-MVP installations) If you are using VALIDAT for Windows 1.31 you can import your files into Method Validation Pack A Files are automatically converted and saved in the new format. If you are using VALIDAT 2000 your files are also be converted during import. Files of VALIDAT version are converted only after query. Until they have been converted, the extended audit functions of version 2 cannot be used. Administrator NOTE If you want to start MVP from the programs menu, use this User-ID Password combination for the first logon. It is recommended to start MVP from the ChemStation Plus ChemStore module. During installation, the administrator admin is installed with the password admin. Use this login to identify yourself the first time you start the program. You should then change the password immediately and install further users. VALIDAT.CFG contains the path for the configuration files under ConfigPath=. The default is. (i.e. the installation directory). If the installation directory is write-protected, a different path can be installed under ConfigPath=. The files VALUSER.BDB, VALBASE.VD, V2kAUDIT.BDB and, v2kimport.bdb must then be copied into the new directory (see below). Installation requirements You need the following for the operation of Method Validation Pack: Microsoft Windows NT 4.0 SP 6a, or Microsoft Windows 2000 SP 2/3 Internet Explorer 4.01/5.00 or Netscape Browser 4.x Microsoft Data Access Components 2.5 Adobe Acrobat Reader 4.0 Agilent ChemStation Plus - Method Validation Pack 27

28 2 Installation Start of Installation Requalification Method Validation Pack can carry out installation qualification (File qualification). Operational qualification of the ChemStation Plus Method Validation Pack software requires checking interfaces to ChemStation Data Acquisition and to ChemStore database as well as a regression testing of the MVP software. MVP OQ is therefore possible only with Agilent Validation protocols and can thus only be executed by an Agilent-certified engineer. Network Validation files can be opened by one user only at a time. A network installation is possible insofar as Method Validation Pack can be installed on a network drive. It then saves its configuration and Validation files on the network drive. The configuration files and the installation files can be separated by using the file VALIDAT.CFG. VALIDAT.CFG Two paths can be defined in VALIDAT.CFG: ConfigPath defines the saving of VALUSER.BDB, V2KAUDIT.BDB and VALBASE.VD, HelpFiles defines the saving of online documentation. The information for the HelpFiles must be a valid URL (e.g. on your Intranet server). If you are using the Client/Server or DMS option of Method Validation Pack, then you can setup VALIDAT.CFG with the DB-Installer utility. Example ConfigPath=\\server\Val2000Config HelpFiles= Use the following syntax if you want to use a path instead of an http-address for your HelpFiles: HelpFiles=file:///\\server\VALIDAT_2000_help 28 Agilent ChemStation Plus - Method Validation Pack

29 Installation 2 Start of Installation Licences Method Validation Pack is licensed per machine. It may be installed on one computer only at a time. It may not be used as a concurrent licence or a network licence. Contact our support if you have questions concerning the licensing of Method Validation Pack. Agilent ChemStation Plus - Method Validation Pack 29

30 2 Installation Support for SQL-Servers Support for SQL-Servers Client/Server Optionally, Method Validation Pack can store its data on central SQL-Servers. The following SQL-server is currently supported: ORACLE Server DMS (Document Management System) Method Validation Pack can save document revisions in its optional Document Management System. This DMS is able to hold the history of all important elements of validation, i.e. validation database, reports, IQ/OQ results etc. The DMS helps users to support GLP requirements for data integrity and data security. It stores all validation raw- and meta-data, and offers data revisioning for full versioning. 30 Agilent ChemStation Plus - Method Validation Pack

31 Installation 2 Support for SQL-Servers Installation of a SQL-Server with DB-Installer Method Validation Pack provides an installation and database maintenance utility for easier setup of the SQL-server. Figure 1 DB-Installer main screen The basic tasks of this program are: Installation of the database (for both validation files and DMS) Viewing the program log file Viewing the database installation log file Configuring VALIDAT.CFG DMS archiving Agilent ChemStation Plus - Method Validation Pack 31

32 2 Installation Support for SQL-Servers Figure 2 Database installation screen Installation of the database generates the tablespaces and databases for Method Validation Pack and the DMS. Existing logins can be deleted. There are three size schemes for installation: small, medium and large. Choose in accordance with the case studies explained in the manual. Take care when selecting the server type and the DB path: although DB-Installer can be started from any client computer, the DB-path must be valid for the server and not for any existing client drive mapping. Server: Name of the server. This corresponds to the net alias name under Oracle SQL Server installations. The net alias must be configured with the Net8 configuration assistant before running DB-Setup (normally done during ChemStore setup). 32 Agilent ChemStation Plus - Method Validation Pack

33 Installation 2 Support for SQL-Servers Delete existing: Existing VALIDAT/DMS databases and VALIDAT/DMS logins are deleted. This affects only MVP and not ChemStore. Take care when selecting this options, as you might overwrite an existing database with important data. DB-Installer displays a warning message if these options are selected. In any case, an Oracle DBA is needed for creating and deleting databases/logins on the server. DB-Path: The DB-Path on the server. Take care when selecting this path, since the installation program will not (and cannot) check the validity of this path. Be sure that there is sufficient free space on the specified drive. The directories specified must exist, since they are not created by DB-installer. For advanced installation, you might specify separate paths for the tablespaces. This could significantly increase performance. Server type: The server type is important for correctly customizing the database installation script. All these servers require different scripts. Hence, specifying the incorrect type results in error messages and an unstable database. For MVP the only valid selection is Oracle Database size: The database size can be chosen from small, medium and large schemes. See the detailed size scheme in section Database size: on page 33. The small scheme should be used for a demo or notebook installation only. The medium scheme is intended for normal usage. The large scheme is for heavy usage of the DMS part of the database especially as it occurs e.g. with a 21 CFR part 11 validation type. Log file: The log file lists all messages received from the server and additional information about what will be set up. This log file is stored in the installation directory of MVP. Agilent ChemStation Plus - Method Validation Pack 33

34 2 Installation Support for SQL-Servers Command: This window shows the command currently in execution. Be aware that certain commands take a lot of time (e.g. creation of the database file). Create DB: This commands starts the database installation. You are prompted for an administrative account of your SQL server. Extended configuration: You can select separate directories for each of the eight tablespaces. This could improve performance significantly when these directories are located on separate drives. In normal installations, there is no need for this extended configuration, and same path for all is recommended (this is the default). Figure 3 Extended Database path configuration. 34 Agilent ChemStation Plus - Method Validation Pack

35 Installation 2 Support for SQL-Servers Viewing the program log file Figure 4 Program log file screen The error log file contains all error reports of Method Validation Pack. Detailed tracing information is included. Information: All error reports contain information about the error type, the code module, tracing information and the date/time of the error. Support: If there are any unusual or frequent entries in the log file, contact Agilent Technologies. Try to be as specific as possible when talking to the support in order to present a clear picture of the problems. Agilent ChemStation Plus - Method Validation Pack 35

36 2 Installation Support for SQL-Servers Viewing the database log file Figure 5 Database log file screen The database log file contains information about DB-Setup runs. Information: All DB-Setup and DMS archive runs are logged to this database log file. If the database is not running properly or not running at all, check this file and contact Agilent Technologies. 36 Agilent ChemStation Plus - Method Validation Pack

37 Installation 2 Support for SQL-Servers Configuring VALIDAT.CFG Figure 6 Configuring validat.cfg This file contains basic information for Method Validation Pack about configuration paths and database connects. ConfigPath=: Path for configuration files. May be in UNC format. Default is. (i.e. installation directory). HelpFiles=: Path for all help and documentation files. May be in UNC or URL format. Default is. (i.e. installation directory). Agilent ChemStation Plus - Method Validation Pack 37

38 2 Installation Support for SQL-Servers TempDBFiles=: Path for temporary database files. May be in UNC format. Default is <installation dir>\tempdb. HomeServer=: Name of the installation's home server. The home server is used for account checking, audit and validation default settings. Must be identical to the configured Net8 alias. Server type: If MVP is installed as C/S application, the server type must be specified as Oracle. Defaults: This sets the above information to the described defaults: ConfigPath= and HelpFiles= is set to. (i.e. current directory = installation dir), TempDBFIle= is set to the <installation directory>\tempdb SERVER= is set to the server specified in DB-Setup. Update window: This updates the VALIDAT.CFG window to the information specified above. Load configuration: This reads the VALIDAT.CFG file from disk. Save configuration: This stores the VALIDAT.CFG window to disk. VALIDAT.CFG: Contains all VALIDAT.CFG information. 38 Agilent ChemStation Plus - Method Validation Pack

39 Installation 2 Support for SQL-Servers DMS archiving Figure 7 DMS archiving The DMS size can be reduced by archiving old entries to an external file system and deleting them in the DB. All entries are still accessible but do not consume space on the database server. All entries can still be accessed because the archive process is fully transparent to the DMS client, i.e. a user will see no difference when the DMS has been archived. During an archive process, the DMS Binary Large Objects (BLOBs) are moved from the Oracle server to the file system. This move is completely transparent for all MVP clients as long as there is access to the target directory of this move ( read permission necessary). Be sure to specify a directory that is accessible from all MVP clients. It is recommended to use UNC-paths (no drive mappings) or a DFS (distributed file system) so that the same access is available from all clients. Agilent ChemStation Plus - Method Validation Pack 39

40 2 Installation Support for SQL-Servers The archive process can run simultaneously with standard MVP operation, but it might slow down database responses. Archive all BLOBs: All BLOBs are archived. Archive old BLOBs: Only BLOBs older than the specified number of days are archived. Archive until 50% is reached: This archives BLOBs until 50% free space is reached in the DMS. Flat file structure: This generates a flat directory structure during the archiving process. Normally, a descriptive directory structure is generated. Generic names: Generic names do not contain any information about the contents of the BLOBs, they are subsequently numbered. Save BLOBs under: The archiving path for the BLOBs. Ensure that there is enough free space to hold all files. Since this path is accessed only once in writing mode, you could use WORM (write one read multiple) media for DMS archiving storage. This path may or may not be online afterwards. If it is offline, DMS gives a warning message during BLOB access. Start archiving: This starts the archiving process. Depending on the requested operation, this may need significant time. Log file: The log file contains a short overview of the archiving process. 40 Agilent ChemStation Plus - Method Validation Pack

41 Installation 2 Support for SQL-Servers Database sizes Method Validation Pack supplies three pre-defined schemes for database installation: small, medium and large. The databases are separated into multiple tablespaces for better performance and administration. In particular, the validation/comment-table and the DMS/BLOB (binary large objects) tables reside in special (large) tablespaces. SMALL VDB Database MB Initial Size 30 Maximal Size 300 File Growth 15 Comments Initial Size 100 Maximal Size 700 File Growth 40 DMS Database MB Initial Size 20 Maximal Size 200 File Growth 5 BLOBs Initial Size 180 Maximal Size 1800 File Growth 45 Agilent ChemStation Plus - Method Validation Pack 41

42 2 Installation Support for SQL-Servers MEDIUM VDB Database MB Initial Size 600 Maximal Size 1200 File Growth 30 Comments Initial Size 1400 Maximal Size 2800 File Growth 70 DMS Database MB Initial Size 500 Maximal Size 1000 File Growth 25 BLOBs Initial Size 4500 Maximal Size 9000 File Growth 225 LARGE VDB Database MB Initial Size 1200 Maximal Size 2400 File Growth 60 Comments Initial Size 2800 Maximal Size 5600 File Growth Agilent ChemStation Plus - Method Validation Pack

43 Installation 2 Support for SQL-Servers DMS Database MB Initial Size 2000 Maximal Size 4000 File Growth 100 BLOBs Initial Size Maximal Size File Growth 900 Database parameters have been chosen for unattended operation; tablespaces and datafiles auto-extend until their maximum file size has been reached. Nevertheless, you should keep in mind that frequent DB administration may be necessary for correct and secure operation (backup tasks, size checks etc.). Your database administrator may manually expand your database if necessary. There is no limitation in DB sizes in Method Validation Pack. If you have any questions regarding database layout, database installation or performance or size issues, contact our support. Starting the program When you are starting the program for the first time, it initializes all databases and writes important default entries. Figure 8 Starting the init-process Agilent ChemStation Plus - Method Validation Pack 43

44 2 Installation Support for SQL-Servers Figure 9 Init process finished - program will restart The default login is username admin with password admin. NOTE Since MVP uses ChemStore user management, be sure to add method validation permission before logging in. 44 Agilent ChemStation Plus - Method Validation Pack

45 Agilent ChemStation Plus Method Validation Pack User s Guide 3 Operation Security Concept 46 Operation 49 Welcome screen 50 Main window 52 Main menu 53 DMS (Document Management System) 59 DMS History 62 This chapter describes the security concept of Method Validation Pack, program operation using mouse and keyboard, and the main menu options. Agilent Technologies 45

46 3 Operation Security Concept Security Concept Method Validation Pack uses five levels of entry to guarantee consistency of data necessary for Validation.These levels are configured in the ChemStation Plus user administration, for details see Chapter 12 Interfacing ChemStation Plus and Method Validation Pack on page 167. Figure 10 Login dialogs Levels of Entry Method Validation Pack has five user levels. You log into the program by entering your user name and the matching password. Report level Logging into the system with a name and a password of level 1 gives you access to report output only. At this level, data changes are impossible. Input level Logging into the system with a name and a password of level 2 gives you access to a Validation (data input and graphics/report output). 46 Agilent ChemStation Plus - Method Validation Pack

47 Operation 3 Security Concept Planning level/ Configuration level Logging into the system with a name and a password of level 3 gives you access to planning. Level 4 gives you access to configuration rights. You may change the configuration values of each Validation (reaction of statistical tests). Administration level Logging into the system with a name and a password of level 5 gives you access to all functions of Method Validation Pack including program configuration (global settings). Auditing Auditing allows the actions of Method Validation Pack to be monitored. Program operations, including data such as time, user and action can be registered. The Audit is divided into program auditing, default Validation auditing and Validation auditing. For further information, see chapter Utilities on page 121. Locking a Validation A validation can be locked so that data cannot be changed. This leaves reports and graphics as output and export options. You can lock validations, components or checkpoints via the context menu (the right mouse button). You can use locking to protect your data after the data input has been finalized. Unlocking a validation can be performed only by saving the validation under a new file name. This can only be done by an administrator. Agilent ChemStation Plus - Method Validation Pack 47

48 3 Operation Security Concept Both locking an unlocking a validation requires an electronic signature to be applied. Figure 11 Electronic signature dialog 48 Agilent ChemStation Plus - Method Validation Pack

49 Operation 3 Operation Operation Method Validation Pack is a Microsoft Windows program and can be operated via mouse clicks and keyboard in accordance with the Microsoft Windows standard. If you have already used other Microsoft Windows programs, you should have no problems with Method Validation Pack. Context menu During operation of Method Validation Pack, the right mouse button opens the context menu of your current program option. The context menu can consist of functions concerning a selected component, or the attributes of a graphic or a report. All functions of the context menu are also accessible via the main menu bar. Adjustment to user All settings that you change in the menus or tool bars are recorded for you at the end of a session. They are automatically loaded during your next login. Navigation bar On the left of the screen, the navigation bar presents the most important top-level functions of Method Validation Pack. You can use this bar for directly selecting Auditprogram, default validation and validation audit DMS Document Management System ExportExport dialog ImportImport wizard for templates New validationnew validation wizard PlanningPlanning wizard ReportReport settings (output formats etc.) SecuritySecurity settings Agilent ChemStation Plus - Method Validation Pack 49

50 3 Operation Operation Welcome screen The assistant is displayed at the start of Method Validation Pack except when you have disabled it under Utilities/options.The assistant helps you by speeding up operation of the software, and offers a quick and easy way to resume your work. Figure 12 Welcome screen The assistant offers the following options: Creation of a new Validation Opening an existing Validation Opening the last Validation (per user) The Validation assistant The Validation assistant serves to create a new Validation in a very simple manner. It can also be used to add a new component. During program start, the assistant can be used only to create a new Validation. Within the program Method Validation Pack, the assistant can be opened via the Validation menu or the help menu. 50 Agilent ChemStation Plus - Method Validation Pack

51 Operation 3 Operation It guides you step by step through all important configuration and planning steps of a validation. It is recommended to use the assistant for all users, although advanced users of MVP may prefer to configure their validations manually. Figure 13 The validation assistant Follow the instructions of the assistant to create a new Validation or component. You can choose between a Development and a 21 CFR part 11 validation. A development validation is less strict in its security settings, i.e. less DMS entries are made and the audit level can be lower than everything. Agilent ChemStation Plus - Method Validation Pack 51

52 3 Operation Operation A part 11 validation complies to the strict regulations of the 21 CFR part 11, i.e. the audit level is everything and all important steps of a validation are tracked in the DMS (validation file, overall and component report and MVS files) and some actions are not possible. You can switch from a part 11 validation to a development validation but not vice-versa. Main window Figure 14 Main window 52 Agilent ChemStation Plus - Method Validation Pack

53 Operation 3 Operation The main window is displayed after you have started the program, entered a password and selected an existing Validation or created a new one. At the left of the main window is the navigation tree of the program. It displays the open Validations, components and checkpoints. Fast selection and execution of functions is thus possible. Depending on what you have selected, different functions can be selected via the context menu (right mouse button) or the main menu. Main menu All program functions of Method Validation Pack can be executed via the main menu. Note that, depending on what you are currently doing in the program, not all options may be accessible. The particular functions that you can select depend on your user rights. Should a function be inaccessible, check whether your current selection and your user rights allow this function. File Menu Select this option to create, save and print a new Validation file. You can also save Validations as templates as in MS Word 97 and use them as a basis for new Validations. The print preview creates a double-page preview of the current report window. You can print and export a Validation via the preview. Agilent ChemStation Plus - Method Validation Pack 53

54 3 Operation Operation The option send creates a mail to send your Validation files without having to start the mail program. Validation list The file menu also contains a list of the five last Validation files you worked on. If an entry is displayed but cannot be selected, it has been deleted. Edit Menu Select this option to use the clipboard (copy, cut and paste). It also offers Undo/Redo to revoke actions. Note that it is impossible to change the content of the report windows as the reports are write-protected. You can change only the format. Validation Menu Select this option to add or remove components, create reports and change the settings of a Validation. The settings include the configuration values of the statistical tests, the output formats and the headers. 54 Agilent ChemStation Plus - Method Validation Pack

55 Operation 3 Operation Validations, components and checkpoints can be locked so that data cannot be changed. Unlocking a Validation can be performed only by saving the Validation under a new file name. This can be done only by an administrator. You can also open the Validation assistant. It helps you to create a new Validation or component. A Validation report can be created via the function Validation planning report. Security Menu The option Auditing shows you all actions as recorded by the program (who did what, when and with what). The entries of the Audit logs can be sorted by clicking on a header. If you click, for example, on the header date the corresponding entries are sorted either ascending or descending. Alternatively, they can be grouped into the empty space above. This is done by dragging a single header into the empty space above the header line. Grouping can be done several times on different levels so that recorded data are finally structured in a tree. Auditing is divided into program auditing, default Validation auditing and Validation auditing. Furthermore, all changes in data input can now be recorded. This option can be selected under Validation/Properties/Configuration. If the validation type is development, for part 11 auditing the audit is preconfigured and cannot be changed by any user. Note that, as long as the auditing window stays open while you work with different functions of Method Validation Pack (e.g. call of a report), the auditing window is updated and may slow down the program considerably. Report Menu Select this option to open the entire Validation report and export it. Agilent ChemStation Plus - Method Validation Pack 55

56 3 Operation Operation The export formats available are DOC (Word 97/2000 format) and HTML (without graphics), RTF (including graphics) and TXT. RTF is supported by all standard word processors and includes all formatting information and graphics displayed in Method Validation Pack reports. View Menu Select this option to fade in the status bar, to update the display and to open the Internet browser. The options Zoom in/zoom out allow you to change the size of the displayed report in steps of 10%. The minimum is 10%, the maximum 400% of the original size. Zoom in/zoom out can also be done via the shortcut Ctrl-F3/F2. Zoom to fit aligns the width of a report window to match the window size. Utilities Menu The options Properties for new Validations and Output settings serve to adjust basic settings for new Validations. Options serves to make a personal adjustment of the program. You can determine the font, the color and the size of graphics and reports. You can also determine general settings for the start of the program and data saving. 56 Agilent ChemStation Plus - Method Validation Pack

57 Operation 3 Operation Window Menu Select this option to change windows settings (align horizontally, align vertically and cascade). Agilent ChemStation Plus - Method Validation Pack 57

58 3 Operation Operation Help Menu Select this option to call up the help function or the help assistant. The help was done in HTML format. You can navigate via links within the help function as well as access the online manual (also HTML format). You can put the help on a Web server of your Intranet to maintain it centrally. This can be configured via the file VALIDAT.CFG and the entry HelpFiles ( Configuring VALIDAT.CFG on page 37). About... You can display program information such as serial number, program version and so on. Have information data ready when you call our support. You can call detailed information about installation via the system info if the MSINFO component of Microsoft is installed on your computer. See appendix B on page 155 for further information about File Verification and Support Services. Support Services can be performed only by Agilent Technologies Support 58 Agilent ChemStation Plus - Method Validation Pack

59 DMS (Document Management System) Operation 3 DMS (Document Management System) Method Validation Pack contains a complete document management system for secure data storage in a database by tracking revisions of validation, documents and other important files. DMS is automatically started along with the program and can be displayed by clicking the DMS-button in the navigation bar, or by selecting the DMS-button in the task bar. Figure 15 DMS window For full data security and data integrity, DMS entries are either automatically invoked when actions are triggered that require DMS tracking or called manually using the toolbar entry. Agilent ChemStation Plus - Method Validation Pack 59

60 3 Operation DMS (Document Management System) DMS stores important information, which is validation raw data, such as configuration and planning, along with the analytical results and the meta data. Along with this information, DMS stores all important information (properties) along with the entry: Name of entry Original path Label Purpose of entry ( manual revision, automatic revision ) Version Document type Size Checksum Status (normal, checked-out and finalized) User domain and user computer Real name of user (display name) Reason for check-in Check-in date Date of parent entry (i.e. base node entry) All DMS entries are displayed in a hierarchical list. Entries can be selected to view all properties. The state of a document (normal, checked-out and finalized) can be changed by double-clicking to the status field. This requires sufficient permissions (at least configuration level). You can expand/collapse the DMS tree. With Rebuild, you regenerate the tree with the latest DMS contents. You can select to display the DMS tree with all available information ( include all documents ). When you click a version child node once, it displays its properties. When you double-click a validation (VDB-file), or click on its selection field, it asks to restore the selected version. When you select a document file, it displays a 60 Agilent ChemStation Plus - Method Validation Pack

61 Operation 3 DMS (Document Management System) preview window allowing zoom, print, export or copy of the selected document. The actions depends on the type of entry: documents are displayed in a preview window, validations are stored in the file system. The DMS is always aligned to the current validation, i.e. it contacts the same database as the validation itself. Hence, you will notice a short delay when you activate the DMS for the first time the connection must be established. If you are using a local DMS database, you are prompted to add a new database when you have allocated more than 700MB. The old DMS databases can still be selected and retrieved with the server selection in the DMS window. Click on the combo box and DMS displays all existing local DMS databases. Select one and DMS asks you to connect it. You can go back to the current DB by selecting Current DMS or by selecting a validation in the MVP window and go back to the DMS window. Agilent ChemStation Plus - Method Validation Pack 61

62 3 Operation DMS History DMS History When you open validation files from the Oracle server, you can select history. This opens a list of all available revisions of the selected validation. When you select an old revision, it is loaded. When you save it, it becomes the current ( latest ) version of the validation history. Nevertheless, you have no data loss, since the validation you have overwritten still exists as revision n-1. Figure 16 Validation History A validation from history is always opened as a locked validation to avoid accidentally working on an older revision. 62 Agilent ChemStation Plus - Method Validation Pack

63 Agilent ChemStation Plus Method Validation Pack User s Guide 4 Planning Checkpoints and validation extend 64 Planning a Validation 67 Checkpoints 75 Output settings 93 Validation planning defines the important basic data for a Validation, such as the product, the technical equipment used, the product components and, for each component, the checkpoints. Agilent Technologies 63

64 4 Planning Checkpoints and validation extend Checkpoints and validation extend A validation is composed of different checkpoints. These are often called validation elements or validation parameters. Results are occasionally called method data. A checkpoint is the key element in MVP. A checkpoint defines one set of experiments (typically at least 6 runs) that are executed to receive results for calculating the quality of a specific requirement. These results allow the proof that the specific requirements can be met with the actual analytical method. Typically, multiple checkpoints must be calculated for each compound in order to prove that all key requirements for a analytical method validation can be met. The applicability of each checkpoint for a certain compound is defined in various regulatory guidelines, and it depends on the nature of the compound (Main, side, trace compound etc.). Different names are used for checkpoints. This is a compilation: accuracy trueness, accuracy of the mean, bias precision repeatability intermediate precision reproducibility calibration (curve) linearity range Comprises systematic and random deviations, is quantitatively often referred to as uncertainty of result Only systematic deviations, is based on comparison with a reference (sample, method) Only random deviations, can be described as standard deviation Laboratory internal: one person in charge of an apparatus, can be described as repeat standard deviation or repeatability limit Can be described as a laboratory internal comparison Laboratory-to-laboratory: ring experiment conditions, can be described as reproduce standard deviation or reproduce limit Connection signal/concentration, e.g. first, second or third order, weighted or unweighted, ground or matrix calibration (un-weighted or weighted) Linear correlation between signal and concentration Concentration range which is covered with acceptable accuracy, ideally variance homogeneity can be shown at its ends 64 Agilent ChemStation Plus - Method Validation Pack

65 Planning 4 Checkpoints and validation extend recovery specificity, selectivity ruggedness, robustness stability detection limit quantitation limit, determination limit Statement about losses because of sample preparation steps or matrix influence, if dependent on concentration, the recovery function must be determined To exclude impairment through accompanying substances To see impairment through changed parameter Samples and calibration substances and their solutions, mostly checked during ruggedness The smallest detectable amount Smallest quantifiable amount There is often more than one possibility to judge and categorize checkpoints. For example, an analyte adsorption at the sample matrix may touch the checkpoints accuracy, specificity, calibration function, recovery and determination limit. In this case it is optional which checkpoint you select. Agilent ChemStation Plus - Method Validation Pack 65

66 4 Planning Checkpoints and validation extend To judge the amount of time needed for a validation, the following ICH (International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human use) table is helpful: Table 1 ICH - Table to judge need of time Purpose of analytical test Identity Purity Purity Assay Checkpoints qualitative yes/no quantitative threshold value traces qualitative Accuracy Precision repeatability quantitative content intermediate precision - + (1) - + (1) reproducibility - - (3) - - (3) Specificity (2) (2) Detection Limit - - (3) + - Quantitation Limit Linearity Range not necessary as a rule (1) intermediate precision can be ignored as soon as reproducibility is determined + necessary as a rule (2) faulty specificity of one procedure can be compensated by different analytical methods (3) can be necessary in isolated cases Ruggedness can be checked during an advanced phase of development. 66 Agilent ChemStation Plus - Method Validation Pack

67 Planning 4 Planning a Validation Planning a Validation NOTE Planning a validation requires at least planning permission. New Validation A new Validation is created from the File menu. You then add components to the new Validation. Finally, the range of checkpoints is defined. The checkpoint names can be changed in the table of contents. Double-click twice slowly on the respective entry. Changed names are adopted in both the checkpoint reports and in the table of contents of the validation. Method Validation Pack offers a selection of predefined checkpoints ranges. You can either select them, or enter your own selection of checkpoints. The checkpoints have to be planned in detail. This could mean the selection of a sub-method for a checkpoint, the entry of header data, number of measured values and dimensions. Example: A validation in Method Validation Pack is made up of components and checkpoints. Components versus compounds A component is similar to a ChemStation compound but it can be more. A standard validation may be setup as follows: You can add checkpoints to the components Agilent ChemStation Plus - Method Validation Pack 67

68 4 Planning Planning a Validation and complete the planning by filling out the planning dialogs of each checkpoint. Data is entered (either manually or via study import, see below) and you can generate graphics and reports. Now you want to make some comparisons; for example, you want to check precision and linearity with more data. To do this, you can make a copy of Component 1 by dragging component 1 to the Validation 1" line. MVP asks you whether to move the component, to copy its planning or to make a complete copy of it (including all data): To make a comparison based on more data, you select Copy planning. MVP creates a new component named Copy of component 1": You can rename all entries of this hierarchical list in the same way as in the Microsoft Windows Explorer, i.e. click the entry, it switches to edit mode and you can rename it: 68 Agilent ChemStation Plus - Method Validation Pack

69 Planning 4 Planning a Validation Enter the planning of the checkpoints of Component 1 with more data and adjust the number of values (e.g. 8). Assuming that you have already entered your data in the precision of component 1, you can open both data windows to copy the existing data: Agilent ChemStation Plus - Method Validation Pack 69

70 4 Planning Planning a Validation You can copy data by using the clipboard or by dragging selected data directly with the mouse. To do so, select the source data. Multiple cells can be marked as in Excel (including clicking on the header cells). Now move the selection with the right mouse key while pressing the shift-key of the keyboard. Release the mouse on the first cell of the target area. MVP copies all data. Now you can add additional values to the data grid to complete precision input for Component 1 with more data. You can generate reports for both checkpoints and/or components to compare results. That is why MVP does not use the term compound but component. A component is more than an analytical compound. Furthermore, you can rearrange the order of your validation in the same way as you copied the planning. You can select checkpoints and/or components and move them to a different node in the validation. You can even move or copy data between validations to make overall validations out of data and results from simple validations. Checkpoints A checkpoint is the implementation of a calculation method in MVP. It contains the planning (specification of the data structure), the data itself (either manually entered or automatically imported) and some options (e.g. graphical settings). The following checkpoints are supported by MVP: Precision Ruggedness/Robustness Lab capability Linearity Ring Experiment Calibration function Detection and Determination Limit Accuracy Selectivity/Specificity Some of these checkpoints contain more than one calculation method (e.g. accuracy). Clicking on Options in the planning dialog of a checkpoint displays both the additional settings for MVP integration and a short help for selecting the correct calculation method. 70 Agilent ChemStation Plus - Method Validation Pack

71 Planning 4 Planning a Validation Depending on the checkpoint, you can define the data structure as follows: Number of data points (samples) Definition of calculation method (where applicable) Selection of multiple injections and/or multiple determinations (where applicable) Dimension of data Header data for informational purpose Additional data (nominal values etc.) Some checkpoints have a more complex structure. Ring Experiment, Robustness and Lab Capability use more than one data column (lab column, series) for data input. Hence, you must define the number of values for each of these columns. Agilent ChemStation Plus - Method Validation Pack 71

72 4 Planning Planning a Validation After you have defined the checkpoints, you can enter data input (see above) or setup MVP for data transfer with Chemstation/ChemStore (see below). Configuration data Figure 17 Properties of a validation When you are creating a new validation, the configuration values and output formats are determined by these settings. They can be changed only by the administrator. The values for a specific validation can be changed by a user with extended planning rights (see Chapter 3, Operation ). A component report or overall report can be generated to show the current configuration values. All values are listed on one page. You can specify the statistical reliabilities behind their respective calculations. 72 Agilent ChemStation Plus - Method Validation Pack

73 Planning 4 Planning a Validation External documents You can specify a list of external document to be printed with your validation document. The attachments have the same header and footer as the validation and are included in the page numbering. Thus, all important information (data, SOPs, method description, graphics, calculations, reports) is contained in a single document. Figure 18 External documents Store Method With this function, you can directly add a method file to the DMS. This is for archiving or documentation purpose. It is recommended to store the completely developed method before starting the validation. If the method consists of several files, create an archive first and store the archive. NOTE If you want to add the analytical method from ChemStation Plus data acquisition as a text file, use ChemStation s print Method command to create a method in text format. For details, refer to ChemStation online help and the ChemStation manual. Validation type You can select whether your validation is a Development validation or a 21 CFR part 11 validation. Agilent ChemStation Plus - Method Validation Pack 73

74 4 Planning Planning a Validation A development validation has lower security settings, i.e. DMS entries are written less frequently, audit logging is less strict. A part 11 validation on the other hand has the maximum security enabled and writes DMS entries and audit-log entries frequently (level everything ). You can switch from a part 11 validation to a development validation but not vice-versa. The final validation of a method must always be performed under the part 11 setting. The same applies to validation work in a regulated environment. 74 Agilent ChemStation Plus - Method Validation Pack

75 Planning 4 Checkpoints Checkpoints In this section, the checkpoints used are described. The mathematical procedures and formulae used can be found in appendix B. Precision Figure 19 Precision planning Agilent ChemStation Plus - Method Validation Pack 75

76 4 Planning Checkpoints Precision is a qualitative concept. It describes the extent of conformity between results obtained during repeated use of a set analytical method under recurrent and comparable conditions. Monitoring the precision records random errors. Usually, the repeatability or reproducibility limit is given as a quantitative measurement for precision. Precision describes in a quantitative way how close analytical results are to the expected value or the right value. Precision is influenced by the systematic error and by random errors of a result. Usually, the confidence range determined by repeatability or reproducibility conditions is given as a quantitative measurement for precision. Precision can be planned as precision in the true sense, or as repeatability from linearity. In both cases, the nominal values of the variation coefficient can be entered. Precision can be performed with multiple injections. 76 Agilent ChemStation Plus - Method Validation Pack

77 Planning 4 Checkpoints Ruggedness/Robustness Figure 20 Ruggedness/Robustness planning Ruggedness Ruggedness is defined as the independence of an analytical result from changes in other parameters, which could influence the result. Small susceptibility to malfunctioning or ruggedness is the prerequisite for transferring this method to other laboratories. The following factors may contribute to the malfunctioning of an analytical method: Factors depending on persons Technical equipment Instrument settings Aging and quality deviations of separation material and detectors Agilent ChemStation Plus - Method Validation Pack 77

78 4 Planning Checkpoints ADDENDUM: Quality deviation of reagents Stability of sample solutions and/or calibration solutions In order to setup a good quality of ruggedness experiments, it is recommended to first make a judgement on which parameters have the biggest potential of accidental (or deliberate) modification. For example, if you do not have air conditioning in your lab, don't use a TCC and work next to a couple of GCs, it might be a good idea to test your separation at different ambient temperatures. After working out these critical factors/changes, you can design your planning of this checkpoint accordingly, and run the corresponding experiments. After a successful validation of your method, and transfer of the method to routine use, it might be very useful to monitor the potential factors of error as defined in the ruggedness testing on a day-to-day basis. One easy implementation of this monitoring is to check these factors during daily system suitability testing in the routine lab and include the test results in a daily or weekly report. Comparative standard deviation Many cases of malfunctioning can be eliminated by standardization or more precise description of working procedures. As a measure for ruggedness, the comparative standard deviation is calculated and listed. The comparative standard deviation describes the extent of random deviations of the results that can be expected from one sample under different conditions (different technical equipment, different laboratories, different people testing, different times). To determine the comparative standard deviation, you need at least two testers from two laboratories. The two laboratories could be two locations within a single department. Each tester must analyze the samples at least six times under repeatable conditions. Ruggedness of an analytical method is given if the deviation of laboratory mean values is not significantly different from the deviation of all measured values. Simplified ruggedness method Ruggedness should show the reliability of an analysis with respect to the influence of external parameters. Proper ruggedness tests include carrying out a series of analyses over a specific period of time, the influence of the ingredients of the mobile phase, including ph deviations, different columns, 78 Agilent ChemStation Plus - Method Validation Pack

79 Planning 4 Checkpoints temperatures and flow rates. Ruggedness of a method is distinguished by the fact that change of parameters within a reasonable range has no significant influence on the result. The F-test and the t-test can be applied during a statistical test. Furthermore, the results can be compared with a given nominal value for deviation. As a rule, the nominal value of deviation is taken over from the result of precision. The practical significance for the result is more important than the numerical comparison. This is mainly the case for methods to quantify amounts near the limit of determination. The stability of the method with respect to different test persons and different technical equipment over a longer period of time is illustrated under Intermediate Precision. Agilent ChemStation Plus - Method Validation Pack 79

80 4 Planning Checkpoints Lab capability NOTE This is not a pharma test criterion and is used mainly in governmental and screening labs. Figure 21 Lab Capability planning Knowledge of lab capability is necessary for accurate estimates of process capability (see DIN parts 33 and 11). Lab capability is concerned with the relationship between the dispersion of values caused by the measuring process and the requirement, such as the specification. Lab capability occupies a special position among Validation concepts because it does not exclusively refer to the analytical method. For lab capability, the indices Cm and Pm are defined analogous to Cp. Cmk and Pmk are defined analogous to Cpk: 80 Agilent ChemStation Plus - Method Validation Pack

81 Planning 4 Checkpoints Lab capability index Cm Corrected Lab capability index Cmk or Lab performance index Pm Corrected Lab performance index Pmk or Cm = ( OSG USG) 6sst Cmk = min( OSG Xb, Xb USG) 3sst Cmk = amount( SG Xb) 3sst Pm = ( OSG USG) 6slt Pmk = min( OSG Xb, Xb USG) 3slt Pmk = amount( SG Xb) 3slt Impact index Imp = This index is the reciprocal of the lab performance index. Meanings: 1 Pm 100% OSG = upper specification limit USG = lower specification limit sst = Short term standard deviation Xb = reference value for the characteristic value, for example, analytically obtained value of a sample judged as typical, mean value of various results, obtained from the quality control of various samples, value planned by the producer for the checkpoint of the product. Lab capability can be carried out by multiple injections. Agilent ChemStation Plus - Method Validation Pack 81

82 4 Planning Checkpoints Linearity Figure 22 Linearity planning Linearity calculates a linear regression using the least squares error for the model y=a+bx. Linearity can be carried out by multiple injections and multiple determinations. Weighting A 1/x or 1/x 2 weighting can be chosen. Proportionality Proportionality can be calculated. To do this, a level is determined on which further measured values can be entered. 82 Agilent ChemStation Plus - Method Validation Pack

83 Planning 4 Checkpoints Calibration function Figure 23 Calibration Function planning The calibration function is the correlation between the expected value of the test characteristic, e.g. the extinction (see DIN part 13), and the content, e.g. a mass concentration. Calibration Calibration is the process of analysis of calibrator solutions, which are solid or gaseous standards of known content. It serves to define the calibration function. The dependence of the signal y on the unknown variable x can be described by the general curve equation y = a + bx Agilent ChemStation Plus - Method Validation Pack 83

84 4 Planning Checkpoints for the linear case. This curve equation contains two parameters: first the method blank value a, determined by the measuring method, as the ordinate segment, and secondly the sensitivity b as defined by the slope of the curve. The values for a and b must be determined by calibration. As well as the linear function, a fit for a 2nd order calibration is also made, which generally leads to a reduction of the residual variance. Linearity is given if the decrease of the residual variance is shown not to be significant on the basis of an F-test. The calibration function can be carried out by multiple injections and multiple determinations. You can choose the maximal degree of order of the calibration curve. 1st order is standard and 3rd order is the maximum (cubic curve). 84 Agilent ChemStation Plus - Method Validation Pack

85 Planning 4 Checkpoints Detection and Determination limit Gleiches Bild wie oben Figure 24 Detection and Determination Limit planning The detection limit is the smallest amount of substance that can be detected qualitatively during one analysis with a defined statistical certainty. The determination limit is the smallest amount of a substance that can be detected quantitatively during one analysis with a statistical certainty to be determined. The determination limit is higher than the detection limit. You need the relative error of result to calculate the determination limit. Both the detection limit and the determination limit can be calculated by the standard deviation of blank values (blank value method) or the residual standard deviation of regression data (calibration curve method). Only one of the two methods may be suitable for practical purposes. Both methods, however, are almost equal with respect to the detection limit. Agilent ChemStation Plus - Method Validation Pack 85

86 4 Planning Checkpoints The calibration curve method is suitable for determining the determination limit. Options The calibration curve method and the blank value method can both be selected as determination methods. If you select the calibration curve method, you must enter the concentration and the measured values during execution. If you select the blank value method, you must enter the measured values and the slope of the calibration function. ICH You can also use the calculation method as described by ICH. S/N ratio No calculations are done for signal-to-noise ratio. Instead, a path name of the chromatogram can be specified. It is imported and displayed for the checkpoint report. Formats of graphics The formats for import are BMP (Microsoft Windows Bitmap) WMF (Microsoft Windows Metafile) 86 Agilent ChemStation Plus - Method Validation Pack

87 Planning 4 Checkpoints Accuracy Figure 25 Accuracy planning Accuracy, in contrast to the quantitative concept of precision, is a qualitative concept. It describes the extent of correspondence between the expected value and the conventional value. Accuracy is influenced by the systematic error. The systematic error is divided into a constant systematic error and a proportional systematic error. A constant systematic error is not influenced by the concentration of the substance to be determined. If the error is influenced by the concentration, it is a proportional systematic error. A qualitative measurement for accuracy is the systematic deviation of the result. Agilent ChemStation Plus - Method Validation Pack 87

88 4 Planning Checkpoints Comparison with the nominal value There are two tests to compare the results of the analysis with the nominal value: the nominal value t-test and the Wilcoxon-Matched-Pairs-Signed-Rank-Test. The t-test tells you whether the nominal value lies within the range of confidence of an analytical result. The Wilcoxon test checks whether the distribution of values of one measuring row is symmetrical around a nominal value in a way which does not contradict the assumption that the nominal value and the mean value are identical. The approximate value for the nominal value may be a result which is known with a higher exactness than may be expected by the analytical method (for example, in samples with certified result). The nominal value may also be the known and defined amount of a substance added to a solution which did not contain the substance before (placebo method). Comparing two independent samples Where the content values of various samples with different contents have been determined by two different methods, the results can be compared with the difference t-test. In order to do so, you must create the test quantity t. The result can then be compared with the t-value listed for the desired probability. Where the result is greater than the listed value, the systematic difference between the tested method and the pre-defined probability is proven. Extended spiking method The extended spiking method allows constant and proportional systematic errors to be determined, even when there are no samples with known content values. It is suitable for analytical methods which consist of weighing, diluting and measuring steps. The analysis value thus determined must be proportional to the weigh-in value, and the analyte must be such that it can be added to the sample in defined amounts. Standard addition method The standard addition method is usually needed in trace analysis. 88 Agilent ChemStation Plus - Method Validation Pack

89 Planning 4 Checkpoints The slope and linearity of the calibration function that was created with matrix-free samples may be changed by the sample matrix. This is often the case in trace analysis. Such effects can be recognized by addition of standards of the analyte to the true sample containing the matrix. Constant systematic errors cannot be recognized in this way. Accuracy by recovery According to the ICH guideline, accuracy can be proven by recovery experiments. This is guaranteed by comparing the results of one analyte against an analyte sample in the presence of the matrix (substance: other impurities, compound: auxiliary material). It is helpful in this context to guarantee recovery for different concentrations. As the rule, three levels of the expected value (70%, 100% and 130%) are sufficient. Method comparison According to the ICH guideline, accuracy can be proven by comparison to a second, already validated method of proven accuracy. Conformity of mean values and variance with the F-test and the t-test is examined. Recovery The nominal/actual comparison serves to find constant and proportional systematic errors. There must be a number of samples with different contents and linearity must be given. The content values have to be known to a more exact degree than expected by the analytical method. Options You can choose between comparison with a nominal value and comparison of two independent samples. If you choose the comparison with a nominal value, you have to enter a nominal value during planning. If you choose the comparison between two independent values, you have to enter the two measuring rows to be compared during execution. Other methods used may be the extended spiking method or the standard addition method. Selection of the extended spiking method requires that you enter the mixing ratio and the spiking amount during planning. During execution, three measuring rows must be entered. Selection of the standard addition method requires that you enter the number of values with matrix Agilent ChemStation Plus - Method Validation Pack 89

90 4 Planning Checkpoints during planning. You must also enter the content and the measured value without matrix as well as the spiking amount and the measured value with matrix during execution. Nominal value A nominal value for the variation coefficient VC can be entered for method comparison and for accuracy by recovery. NOTE Some checkpoints can use results from the identical runs. If applicable, this multiple data usage saves time and increases efficiency of the method validation. For more details, see Appendix F Minimizing the sample number by using one run for Multiple checkpoints on page 181 Selectivity / Specificity Figure 26 Selectivity / Specificity planning 90 Agilent ChemStation Plus - Method Validation Pack

91 Planning 4 Checkpoints Selectivity means that an analytical method can distinguish the substance to be determined from other substances in the sample. Selectivity in chromatography Selectivity is important in chromatography because this technique is used mainly with mixtures. Selectivity can be described by the relative retention of analyte and interfering substance(s). Selectivity thus defined describes the separation ability of the chromatographic system for the components 1 and 2. Nominal value A nominal value can be given for the dilution R. Ring experiment Figure 27 Ring Experiment planning Agilent ChemStation Plus - Method Validation Pack 91

92 4 Planning Checkpoints Ring experiments are used to prove that a method can be transported to an entirely different location (other site, company etc.) and that it still delivers good results. A ring experiment can be seen as a more general robustness. A robust method should also perform well during a ring experiment and vice versa. Nevertheless, the requirements are different: while robustness checks day-to-day influences (different ph, temperature, column etc.), a ring experiment reflects a shift in the whole environment and should be more difficult to meet. The same basic requirements as for ruggedness apply to the ring experiment. When calculating the repeatability limit and the reproducibility limit, the factor 2.8 is used instead of t * 2. In contrast to ruggedness, at least two measured values can be entered per series. In this case, however, not all calculations are possible (not enough values). This is the case, for example, for trend test and tests for outliers. 92 Agilent ChemStation Plus - Method Validation Pack

93 Planning 4 Output settings Output settings The output settings dialog contains various settings related to output formatting, i.e. extent of report, data precision, additional texts etc. Figure 28 Output formats Data format Data precision for all checkpoints can be determined here. Calculations are always done with the maximum precision. Agilent ChemStation Plus - Method Validation Pack 93

94 4 Planning Output settings Figure 29 Data formats The default option is automatic. Thus the number of significant digits after the decimal separator of the input are automatically determined during calculation. If the most precise input data has 6 digits after the decimal separator, all input data are displayed with 6 digits after the decimal separator. For derived data, an extra digit is displayed. This setting is recommended to maintain clarity of arrangement during operation of Method Validation Pack. Alternatively, you can deactivate automatic calculation by clicking the check box. You can then enter the number of digits you want to have as a basis for calculation. Extend of report You can determine which index numbers should be included in the report. 94 Agilent ChemStation Plus - Method Validation Pack

95 Planning 4 Output settings Precision Figure 30 Output formats (Precision) Repeatability limit Tabled t-value Error of result Confidence interval Neumann trend test Dixon or Grubbs test for outliers Agilent ChemStation Plus - Method Validation Pack 95

96 4 Planning Output settings Ruggedness Figure 31 Output formats (Ruggedness/Robustness) Neumann trend test Dixon or Grubbs test for outliers Variance homogeneity Repeatability limit Reproducibility limit Error of result Range of confidence (repeatability conditions) Range of confidence (reproducibility conditions) Lab capability In the case of lab capability, all data are listed and displayed in the report design window for completeness. 96 Agilent ChemStation Plus - Method Validation Pack

97 Planning 4 Output settings Linearity/Calibration function Figure 32 Output formats (Linearity) Figure 33 Output formats (Calibration Function) Vector y=ax+b - curve equation in the case of linearity Square sum of the residuals (only calibration function) Agilent ChemStation Plus - Method Validation Pack 97

98 4 Planning Output settings Residual standard deviation Mean value and standard deviation of y Multiple correlation coefficient Results of F- and t-tests Method standard deviation Detection limit Figure 34 Output formats (Detection Limit) Standard deviation Procedure standard deviation Critical value y k Residual standard deviation (additional for calibration curve method) 98 Agilent ChemStation Plus - Method Validation Pack

99 Planning 4 Output settings Accuracy general Figure 35 Output formats (Accuracy) Result t-test Joined samples Difference of value pairs Mean value and standard deviation of differences Nominal value comparison Result of Wilcoxon test (additional for nominal value comparison) Extended spiking method Test quantity for a and b Threshold quantity t division Standard additional method Result F-test Result t-test Agilent ChemStation Plus - Method Validation Pack 99

100 4 Planning Output settings Recovery Standard deviation for a,b Residual standard deviation Method standard deviation Test quantities ta and tb t-distribution Ring experiment Figure 36 Output formats (Ring Experiment) Neumann trend test Dixon or Grubbs test for outliers Variance homogeneity Repeatability limit Reproducibility limit Error of result Range of confidence (repeatability conditions) Range of confidence (Reproducibility conditions) 100 Agilent ChemStation Plus - Method Validation Pack

101 Planning 4 Output settings Summary All above settings are preconfigured during installation to useful defaults. As long as you do not have the need to enable special settings, there is no need to change anything. The only exception is the text section, where you can find all relevant texts of your validation report (title, comment page, header and footer, default comments), which should always be edited to produce presentable and nicely customized reports. Figure 37 Configurable texts Agilent ChemStation Plus - Method Validation Pack 101

102 4 Planning Output settings 102 Agilent ChemStation Plus - Method Validation Pack

103 Agilent ChemStation Plus Method Validation Pack User s Guide 5 Execution Input 105 Report 113 After checkpoint planning is completed, data can be entered. You select data input by the context menu of your checkpoints and start entering data. You have three methods for entering data: 1 using the data grids (either manually or via clipboard) 2 using the import wizard (semi-automatic data import) 3 automatic via data import from ChemStore study (preferable), Chapter F on page 168 It is not always possible to use the automatic way of importing data. During Robustness or Ring Experiments, you might need to use systems not controlled by ChemStation/ChemStore. (maybe even without any PC-connection). The data input spreadsheet of Method Validation Pack is modelled on MS Excel and supports clipboard functions, Undo/Redo and Drag and Drop (moving with the help of the mouse). You press the Shift or Ctrl-key at the same time as moving a data marker (more than one cell) to its destination. During this action the mouse curser changes its appearance. Unwanted values You can cancel the marking of an unwanted data value by pressing Ctrl at the same time as clicking the left mouse button. The value is displayed as crossed out during data input as well as during reporting. It is not used for calculations. Agilent Technologies 103

104 5 Execution Unwanted values and missing values could result from problems during data acquisition (injector problems, sample vial problems etc.). They are documented (part 11) but are not part of the calculations. NOTE FIn both cases, you must electronically sign the change. Missing values You can enter the measured value MV if no value exits. This may be the case if the measuring system could not supply a value. NOTE In both cases you must electronically sign then change. Figure 38 Electronic signature dialog Zero values and negative values In Validation/Settings/Configuration you can determine whether the input of zero values or negative values is possible. The use of negative values excludes the use of unwanted values. 104 Agilent ChemStation Plus - Method Validation Pack

105 Execution 5 Input Input This section describes the options the user has during manual data input. Figure 39 Data input Multiple injections All checkpoint data entries can be configured for multiple injections. The multiple injections are averaged before further calculations. They do not change the population of the checkpoint. Single values and their mean values are displayed in the report. Additionally, the single values are displayed graphically. Multiple determinations Some checkpoints offer multiple determinations per concentration level. Multiple determinations enter into the calculation as independent values. They therefore increase the population of the checkpoint. Agilent ChemStation Plus - Method Validation Pack 105

106 5 Execution Input Multiple determinations can be combined with multiple injections. Data input window The data input window shows the following buttons: Comment Opens the checkpoint execution comment. Figure 40 Comment dialog with integrated word-processor Import Opens the import assistant, which offers you data transfer from external result files. Later versions will offer an online connection to data systems. 106 Agilent ChemStation Plus - Method Validation Pack

107 Execution 5 Input Figure 41 Import assistant The import assistant (or import wizard) supports a way to get data out of external summary reports. Normally, all major data systems should be able to provide customizable summary reports. The import assistant displays a definition dialog enabling MVP to automatically load this summary data to its data input grids. The following definitions can be made: Name and description of the import mask Source type (file extension) and path Field separator (delimiter) Row options Data positions (with optional transposition of the data) Column information (X/Y-data, information for multiple injections and determinations) After all settings have been made, you can enter a test mode ( apply & test ) to verify the correctness of your import mask definition. You can select a summary report and see how the import wizard reads the data using your mask. Agilent ChemStation Plus - Method Validation Pack 107

108 5 Execution Input Figure 42 Import wizard Report OK Cancel Confirms data input, closes the data window and creates the respective report. The Validation file is saved if the option automatic saving after data changes is activated. Confirms data input and closes the data input window. The Validation file is saved if the option automatic saving after data changes is activated. Interrupts data input and closes the data input window. 108 Agilent ChemStation Plus - Method Validation Pack

109 Execution 5 Input Precision You must enter the number of measured values as determined during planning. Precision can be executed either as precision or as repeatability by linearity. Treatment of outliers Method Validation Pack recognizes outliers from the data entered. Figure 43 Outlier detection It offers three solutions (this applies only to single injections): The outlier can be kept in the list of measured values. Click on the keep button. The outlier remains with the entered data and is registered as an outlier in data output. The outlier is eliminated so that the number of results in the list of measured values is reduced as if, for example, only 5 instead of 6 values were entered. Data output is done accordingly. The planning is corrected automatically so that the number of values planned for the row is reduced by one. Click on the eliminate button to choose this option. Outliers may be eliminated only if the minimum amount of values defined in the configuration is not reduced by more than one value (see Chapter 6, Utilities ) This means that, generally, just one outlier may be eliminated in any row. Agilent ChemStation Plus - Method Validation Pack 109

110 5 Execution Input The outlier can be replaced by three new values. You must enter the three new values in the fields reserved for them and then click on the button replace by three new values. Then the results of the list of measured values are calculated as if, for example, 8 instead of 6 values were entered. Data output is made accordingly. The planning is corrected automatically so that the number of values planned for the row is enlarged by 2. The first of the three new values replaces the outlier and the remaining two are added at the end. Robustness You must enter the number of rows and measured values per row as determined during planning. The number of measured values may be different for each row. Treatment of outliers The treatment of outliers is described in the checkpoint precision/accuracy. Lab capability You must enter the number of samples and measured values per sample as defined during planning. The number of measured values may be different for each sample. Calibration function/linearity You must enter the number of concentrations and measured values as defined during planning. The number of measured values and concentrations are the same. 110 Agilent ChemStation Plus - Method Validation Pack

111 Execution 5 Input Detection and Determination Limit During planning, either the calibration curve method or the blank value method has been chosen. Calibration curve method You must enter the number of concentrations and measured values as defined during planning. The number of measured values and concentrations are the same. Blank value method You must enter the number of measured values as defined during planning and also the slope b of the linear calibration function in the top line of data input. Accuracy During planning, either the comparison with the nominal value, the comparison of two independent values, the extended spiking method or the standard addition method has been chosen. Comparison with the nominal value You must enter the number of measured values as defined during planning. The nominal value is determined by the planning. Comparison of two independent samples You must enter the two rows of measured values to be compared as defined during planning. The number of measured values and the values of the independent sample are the same. Extended spiking method You must enter the three measuring rows, three weights and the spiking amount as defined during planning. The number of values in all three columns is the same. Agilent ChemStation Plus - Method Validation Pack 111

112 5 Execution Input Standard addition method You must enter the contents and the measured values without matrix as well as the spiking amount and measured values with matrix according to the number of measured values defined during planning. The number of values for content and measured value without matrix as well as spiking amount and measured value with matrix are the same. The number of measured values with and without matrix may be different. Recovery You must enter the number of concentrations and measured values as defined during planning. The number of concentrations and measured values is the same. Selectivity/Specificity Selectivity in Chromatography You must enter the retention times, the peak width of the analyte concerned and the interfering substances as well as the dead times according to the number of rows and measured values per row as defined during planning. Ring experiment The measured values must be entered according to the number of rows and measured values per row determined during planning. The number of measured values may differ in each row. Treatment of outliers The treatment of outliers is described in the precision checkpoint section. 112 Agilent ChemStation Plus - Method Validation Pack

113 Execution 5 Report Report The report of method validation experiments, statistics and the final result is the ultimate goal in each validation experiment and validation documentation. Due to its importance, the reporting tool in MVP is very flexible and comprehensive. It allows for easy configuration of reports and uses an MS Word processing reporting engine offering a generic and well-known report format. For full traceability, each validation is stored as a result version in the DMS system (overall and component report for 21 CFR part 11 type of validations). Reports are available on three major levels: a report can be created for a single checkpoint or a component. It can also be an overall report. A list of contents is placed in front in the case of a component report or an overall report. A configuration page with statistical parameter is placed at the end. The report can be printed, exported (RTF or HTML format) or displayed in the double page print preview. Agilent ChemStation Plus - Method Validation Pack 113

114 5 Execution Report Figure 44 Validation report 114 Agilent ChemStation Plus - Method Validation Pack

115 Execution 5 Report Report generation The report contains the following parts: Page header (default header line) Page footer Table of contents Header data (defined per checkpoint) Planning comment (entered per checkpoint) Agilent ChemStation Plus - Method Validation Pack 115

116 5 Execution Report Measured values Graphics 116 Agilent ChemStation Plus - Method Validation Pack

117 Execution 5 Report Analysis (Execution comment) The position of the execution comment is optional. It can be determined under Utilities/Options. DMS signature info These parts can be configured under validation/properties/output settings (texts) or directly in the related dialog (planning, data input, graphics display). Agilent ChemStation Plus - Method Validation Pack 117

118 5 Execution Report Additional Report Options Report options Figure 45 Additional report options under Utilities-options User-specific settings can be determined under Utilities/Options : Font for report Background color Graphic size (usually 75% of the side width) Graphic settings The following options can be determined per graphic. They are saved with the Validation file: Line color and symbols for graphics Display of simultaneous confidence interval Display of residual graphic 118 Agilent ChemStation Plus - Method Validation Pack

119 Execution 5 Report Scaling of X/Y axis and fineness of ticks on the axis (the default setting is auto ) Formatting of the report You can format all elements of the report. Method Validation Pack has a complete word-processor integrated That gives you full access to many style settings. Figure 46 Paragraph settings Graphics Whenever possible, validation checkpoints have a graphics type to illustrate data and results. Agilent ChemStation Plus - Method Validation Pack 119

120 5 Execution Report Figure 47 Graphic window You can format a graphic by right-clicking it and selecting and changing options. These options are stored with the validation. Figure 48 Graphics settings 120 Agilent ChemStation Plus - Method Validation Pack

121 Agilent ChemStation Plus Method Validation Pack User s Guide 6 Utilities Configuration 122 Archive 126 The Utilities menu determines the settings and output formats for new Validations. This configuration can be carried out only by the administrator. Agilent Technologies 121

122 6 Utilities Configuration Configuration Basic system settings for new validations are determined here. In particular, this means the significance levels for various tests and several other variables including the standard random sampling requirement, which may not be reduced during planning and execution. Levels of significance Figure 49 Configuration settings The main formulae used for calculation are described in Appendix A: Formulae. You can configure the following levels of significance: 122 Agilent ChemStation Plus - Method Validation Pack

123 Utilities 6 Configuration Dixon and Grubbs outlier test (precision, ruggedness, ring experiment). The system default is 5% one-sided. Neumann trend test (precision, ruggedness, ring experiment). The default is 5% one-sided. T-values for repeatability limit r, error of result u and the confidence intervall (precision). The system default is 5% two-sided. Threshold value for the partial F-test (linearity). The system default is 5% one-sided. T-value for the critical value yk and for the auxiliary value D (detection and determination limit). The system defaults are 1% one-sided and 1% two-sided respectively. T-value for the t-test and the Wilson test (comparison of nominal and actual value). The system default is 5% two-sided. t-test for connected random sample (method comparison). The system default is 5% two-sided. Cochran, Bartlett homogeneity test and F-test (ruggedness, ring experiment). The system default is 5% one-sided. F-test and t-test for ruggedness (ruggedness, ring experiment). The system defaults are 5% one-sided and 5% two-sided respectively. T-values for repeatability limit r and reproducibility limit R and for error of results ur and ur (ruggedness, ring experiment). The default is 5% two-sided. t-values for t-test selectivity (recovery). The system default is 5% two-sided. t-values for t-test extended spiking method (selectivity). The system default is 5% two-sided. Homogeneity of variance with F-test, slope difference with t-test (selectivity/standard addition). The system defaults are 5% one-sided and 5% two-sided respectively. Agilent ChemStation Plus - Method Validation Pack 123

124 6 Utilities Configuration Other variables Figure 50 Other variables The variables are as follows: Standard random sampling requirement The standard random sampling requirement defined here may not be reduced during planning the relevant checkpoint. This is the default for Sample Sizes. Example: If you set the minimum standard sample number to 6 in other variables, under Sample sizes you can find the default value 6 for all checkpoints. This value can be changed in the sample sizes menu, but it not be changed on the checkpoint planning level (which is one access level lower that the configuration level). 124 Agilent ChemStation Plus - Method Validation Pack

125 Utilities 6 Configuration Figure 51 Sample sizes Ring experiment During calculation of the repeatability limit r and the reproducibility limit R, the factor is replaced by 2.8 in the DIN ISO The confidence interval and the values depending on it change accordingly. Otherwise, the calculations of the ring experiment are the same as for ruggedness. Agilent ChemStation Plus - Method Validation Pack 125

126 6 Utilities Archive Archive Method Validation Pack contains an archive function. This function permits the saving of a Validation file, the report, the results of file verification and if available support services in a Method Validation Pack archive (*.VAR). This archive can be used for freezing and transporting a validation including its current state (validation file and report) and the system state (file verification and support services). It may be used for sending (mail) or archiving. This type of archiving cannot to used to reduce the size of the server DMS database. To reduce DMS database size, please see DMS archiving in the installation chapter. Figure 52 Archive manager The archive manager allows the display of archive elements and their preview, print and export. The content of an archive can be saved again. 126 Agilent ChemStation Plus - Method Validation Pack

127 Agilent ChemStation Plus Method Validation Pack User s Guide Appendix A Formulae Precision 128 Ruggedness 130 Linearity 137 Lab Capability 141 Detection and Determination Limit 144 Accuracy 146 Recovery 148 Selectivity 150 Ring Experiment 153 This chapter describes the formulae used for calculations. The levels of significance and the limits are the default values of the primary program installation. The levels of significance and limits used for reductions and results are the settings determined in Utilities/Configuration. Mean Value x n x i = i = 1 n Standard Deviation s = n ( x i x) i = 1 n 1 Agilent Technologies 127

128 A Formulae Precision Precision Relative Standard Deviation CV CV = s - 100% x Dixon test for outliers Sort all values in ascending order x ( 1) x ( 2)... x ( n 1) x ( x) Q Q x ( n) x = ( n 1) for n 6 x ( n) x ( 1) x ( n) x = ( n 1) for 7 n 12 x ( n) x ( 2) Result If Q > Q ( Pn, ) Outlier detected ( x ( 1) or x ) ( n) Grubbs outlier test Sort all values in ascending order x ( 1) x ( 2)... x ( n 1) x ( n) max{ x r ( n) x, x x ( 1) } m = s 128 Agilent ChemStation Plus - Method Validation Pack

129 Formulae Precision A Result If r m > r m Outlier detected ( x ( Pn, ) ( 1) or x ) ( n) Neumann Trend Test W = n ( n 1) s 2 ( x i x i 1 ) 2 i = 1 Result If W > Trend detected W m( P, n) Repeatability Limit r r = t 95%, f = n 1 2 s Error of Result u u = s t 95%, f = n n Confidence Interval upper limit = x + u lower limit = x u Agilent ChemStation Plus - Method Validation Pack 129

130 A Formulae Ruggedness Ruggedness The Grubbs and Dixon outlier tests and the Neumann trend test are made as for precision per row. Dixon Sort all values in ascending order x ( 1) x ( 2) x ( n 1) x n Q Q x ( n), j x = ( n 1), j for n 6 x ( n), j x ( 1), j x ( n), j x = ( n 1), j for 7 n 12 x ( n), j x ( 2), j Result If Q > Q Outlier detected x ( P, n) ( ( 1 ), j or x ) ( n), j Grubbs Sort all values in ascending order x ( 1) x ( 2) x ( n 1) x n r m = max { x ( n), j x j, x j x ( 1), j } s j Result If r > mj, r Outlier detected x m 1 ( P, n) ( ( ), j or x ) ( n), j 130 Agilent ChemStation Plus - Method Validation Pack

131 Formulae Ruggedness A Neumann Trend Test Applied for all series Mean Value x j = n j x i, j i = 1 n j 1 W j = ( x i, j x i + 1, j ) 2 ( n j 1) s j i = 1 Result If W j > W m Trend detected ( P, n) Standard Deviation n j s j = n j ( x i, j x j ) 2 i = 1 n j 1 Cochran test for variance homogeneity Test for Cochran variance homogeneity for the same number of random sample n per row, number of rows (k > 2). Agilent ChemStation Plus - Method Validation Pack 131

132 A Formulae Ruggedness C = 2 s max k 2 s j j = 1 Result If C > C no variance homogeneity ( P, n) Bartlett test for variance homogeneity Test for Bartlett variance homogeneity for the same or different number of random samples per row, number of rows k > 2. k χ 2 = 1 -- c ( N k) logs 2 2 ( n j 1) logs j j = 1 N c Result k = n j j = 1 k n j 1 N k = j = 1 + 3( k 1) 1 k ( 2 )s j s 2 = j N k If χ 2 2 > χ ( P, k 1) no variance homogeneity 132 Agilent ChemStation Plus - Method Validation Pack

133 Formulae Ruggedness A F-test for variance homogeneity Test for variance homogeneity with F-test for the same or a different number of samples per row, number of rows= 2. F = max 2 s ---- s 2 1 2, s 2 s 1 Result If F > F no variance homogeneity ( α, nm, ) n = degree of freedom in dividend, m = dof in divisor Repeatability standard deviation k 1 s r N ( k n 1 2 = )s j j j = 1 Standard deviation mean value k 1 s z = n k 1 j ( x j x) 2 j = 1 Reproduce Standard Deviation Comparison standard deviation Agilent ChemStation Plus - Method Validation Pack 133

134 A Formulae Ruggedness s R = s n 1 z n 0 n 0 = 1 k 1 k N --- n 2 j j = 1 F-test for Variance Homogeneity Test for variance homogeneity with the F-test for the same or a different number of samples per row, number of rows = 2. F-Test for k > 2, α = 1% : F = 2 s ---- z 2 s r Result Ruggedness OK, if F F α k 1,, N k t-test for Ruggedness t-test for ruggedness with same or different number of samples per row, number of rows = 2. t-test for k = 2, α = 1% : x 1 x 2 t = ( n 1 1)s 1 + ( n 2 1)s ( n 1 + n 2 2) n 1 n n 1 + n Agilent ChemStation Plus - Method Validation Pack

135 Formulae Ruggedness A Result Ruggedness OK, if t t α k 1,, N k Horwitz-test The Horwitz-test for ruggedness is performed. Result If the relative standard deviation is smaller than the maximal relative standard deviation defined by Horwitz then the procedure is robust. The limit for the Horwitz.test is reduced by considering the input values to be %, i.e. if the mean value of the input values is 90.3, the mean value is used in the formula for the Horwitz limit. Horwitz-test reproducibility limit R R = t 95%, f = N k 2 s R Repeatability limit r r = t 95%, f = N k 2 s r Insecurity of result Error of result (repeatability) u r = s r t 95%, f = N k N Error of result (reproducibility) s R u R = t 95%, f = N k N Agilent ChemStation Plus - Method Validation Pack 135

136 A Formulae Ruggedness Mean value of all mean values x = k n j x ji j = 1i = 1 N Confidence intervals Repeatability: x ± u r Reproducibility: x ± u R 136 Agilent ChemStation Plus - Method Validation Pack

137 Formulae Linearity A Linearity For better numerical stability, the X and Y values are transformed to x and y values. MAX MIN = = maximum of X minimum of X Scaling factor C y = g k, 0 + g k, 1 x + g k, 2 x g kk, x k A = MAX MIN MAX MIN B = Matrix X X = x 1 x 1 x1 x x 2 x 2 x2 x x 3 x 3 x3 x x n x n xn xn Agilent ChemStation Plus - Method Validation Pack 137

138 A Formulae Linearity Vector y y = y 1 y 2 y 3 y n Reversed matrices K k K k = t ( X kxk ) 1 where t is the transposition of the matrix, Polynomial coefficients g for degree k k = 1, 2, 3, 4 g k = g k, 1 g k, 1 g k, 2 g k, n Sum of the squares of the residuals t t SSDR k = y'y g kxky, k = 1, 2, 3, 4 Residual standard deviation s yk = SSDR k n k 1 s 0 2 = square of residual standard deviation 138 Agilent ChemStation Plus - Method Validation Pack

139 Formulae Linearity A Variance polynomial coefficients (diagonal elements of matrices) 2 t VAR( g k ) = s yk ( X kxk ) 1, k = 1, 2, 3, 4 Difference of variances S k 2 = SSDR k SSDR k 1, k = 123,, Test value PG k PG k ( n k 2) SSDR k SSDR k 1 = , k = 123,, SSDR k + 1 Threshold of F-distribution 2 PG 1 tab = t 95% ( n k 2), k = 123,, Result of F-test PG 1 PG 1tab ( ) and r k 95% : function is linear PG 1 PG 1tab ( ) and r k < 95% : function is conditionally linear PG 1 > PG 1tab ( ) and PG 2 PG 2tab ( ) : function is quadratic PG i > PG i( tab) for i = 1, 2 and PG 3 PG 3tab ( ) : function is cubic PG i > PG i( tab) for i = 1, 2, 3: function is higher than cubic Method Standard Deviation s s y, 1 x0 = A g 11, Agilent ChemStation Plus - Method Validation Pack 139

140 A Formulae Linearity Relative method standard deviation V x0 = s x % x 140 Agilent ChemStation Plus - Method Validation Pack

141 Formulae Lab Capability A Lab Capability If calculate Xb is checked during planning, Xb is reduced as the mean value of all entered values. Short term deviation s st and long term deviation s lt m=1, direct calculation 1 s st or s lt = n 1 1 ( x x ) 2 i1 1 i = 1 number of series = 1, with moving ranges n j x i1 x i 1 s st or s lt = s i = 2 AMR = ( n 1 1) multiple measurements per series (m>1) m j 1 s j = x n 1 1 ( ij x j ) 2 i = 1 n j m 1 s st or s lt N ( m n 1 2 = )s j j j = 1 m N = j = 1 n j Agilent ChemStation Plus - Method Validation Pack 141

142 A Formulae Lab Capability Lab Capability Index Lab capability index C m (double sided) USL LSL C m = s st Corrected Lab capability index C mk (single sided USL) USL X C b mk = s st Corrected Lab capability index C mk (single sided LSL) X C b LSL mk = s st Method effectiveness index Method effectiveness index P m (double sided) USL LSL P m = s lt Corrected Method effectiveness index P mk (single sided USL) USL X P b mk = s lt Corrected Method effectiveness index P mk (single sided LSL) X P b LSL mk = s lt Result If R C m and C mk S method is not reliable If S C m and C mk T method is relatively reliable 142 Agilent ChemStation Plus - Method Validation Pack

143 Formulae Lab Capability A If T C m and C m lab capability is given Agilent ChemStation Plus - Method Validation Pack 143

144 A Formulae Detection and Determination Limit Detection and Determination Limit Formulae applied from DIN Limit of Detection / Blank method Critical value n 1 y k = y b + n ( 1 y 2 ) ny 2 i i = 1 Limit of detection y x k y N = b 1 ( ) --- m t 99% n n Limit of Detection / Calibration line method Critical value 2 s y1 2 y k = a+ t 99% ( n 2) s m a (with m=interval of prediction=1) Limit of detection y x k a n = b 144 Agilent ChemStation Plus - Method Validation Pack

145 Formulae Detection and Determination Limit A Limit of quantitation x B = 1 1 ( h x h s x0 t 99% ( n 2) N x) m n n ( x i x) i = 1 with s x0 = method standard deviation s x0 n ( y i ŷ) 2 i = 1 n 2 1 = and h = b relative error of result Agilent ChemStation Plus - Method Validation Pack 145

146 A Formulae Accuracy Accuracy Nominal Value Comparison Mean value Standard deviation of differences t-test x = x µ n 1 2 s x = x n 1 i i = 1 n( x) 2 t = x n s x Result If t t α( n 1) nominal value is observed If t > t nominal value is not observed α( n 1) Wilcoxon-test (signed rank test) T + = c i R i c i = 1 for x > 0 i = 1 c x 0 where i = 0 for x Agilent ChemStation Plus - Method Validation Pack

147 Formulae Accuracy A Result If W α T + W α n k, --- n k, systematic error. nominal value is observed otherwise: Comparison of two samples Differences d i = x i y i for i = 1, 2,, n Mean value and standard deviation d t-test n 1 = -- d n i i = 1 n 1 2 s d = d n 1 i i = 1 nd ( ) 2 t = d n s d Result If t t α( n 1) nominal value is observed If t t α( n 1) nominal value is not observed Agilent ChemStation Plus - Method Validation Pack 147

148 A Formulae Recovery Recovery Matrix calculation equal to linearity. Slope b = A g , C Intercept a = g 11, AB g , C Standard deviation 1 s a --- C 2 Vg AB = k, C 2 Vg k, 2 AB C 2 CVg k, 01 s b = A --- C 2 Vg k, 00 Method standard deviation s x0 = s y1 C s x relative: V a x0 = % x t-test t a = a t s b = b a s b 148 Agilent ChemStation Plus - Method Validation Pack

149 Formulae Recovery A Result If t a t α( n 3) no constant systematic error If t b t α( n 3) no proportional systematic error Agilent ChemStation Plus - Method Validation Pack 149

150 A Formulae Selectivity Selectivity Standard Addition Method Linearity without matrix: y = A + Bx with matrix: y' = A' + B'z Check standard deviations of B and B with F-test F = max s B s B', s B' s B If F F α, ( n 2, m 2) variance homoegeneity t-test for slopes t = B B' ( n 2)s y1 + ( m 2)s y'1 s B s B' m + n s y1 s y'1 n m n + m Result If t t α, ( n + m 4) no difference Content A' x P = B' 150 Agilent ChemStation Plus - Method Validation Pack

151 Formulae Selectivity A Content standard deviation s xp = s y'1 1 B' 1 ( A' y' ) m m B' 2 ( z i z) 2 i = 1 Extended Spiking Method Calculation a and b a i = 2x 2i x 1i b i = x 3i x i z Mean values a n a i = i = 1 b n = n b i i = 1 n Standard deviation s a n 2 n 2 a n i b n i 2 i = 1 2 i = 1 a i b n i n = i = 1 s i = 1 n 1 b = n 1 t-values t a a b = n t s b = n a s b Agilent ChemStation Plus - Method Validation Pack 151

152 A Formulae Selectivity Result If t a t α, ( n 1) no constant systematic error If t b t α, ( n 1) no proportional systematic error Chromatography Relative retention time t α R2 t 0 12, = t R1 t 0 Resolution ( R 12, 2 t R2 t 0 ) ( t R1 t 0 ) = w 1 + w 2 Result If R 1.5 Selectivity is okay 152 Agilent ChemStation Plus - Method Validation Pack

153 Formulae Ring Experiment A Ring Experiment The reductions for the ring experiment are identical to those for ruggedness. The following exception applies: during standard configuration the factor is replaced by 2.8 for the reduction of the repeatability limit r and the reproducibility limit R. Agilent ChemStation Plus - Method Validation Pack 153

154 A Formulae Ring Experiment 154 Agilent ChemStation Plus - Method Validation Pack

155 Agilent ChemStation Plus Method Validation Pack User s Guide Appendix B Qualification Tests Method Validation Pack can carry out the following Qualification Procedures to make safe program operation possible. Figure 53 About dialog with IQ and OQ/PV functions Agilent Technologies 155

156 B Qualification Tests File Verification/Installation Qualification (IQ) File Verification/Installation Qualification (IQ) Installation Qualification makes sure that the program is correctly installed. This means that all program components are there in their correct versions. To guarantee correct installation, the following files are checked for exact correspondence with respect to time/date and size. If there is no exact correspondence, the installation is terminated. Method Validation Pack.exe ValKonv.dll MVClient.exe ValMathGer.dll V2KUtilities.dll MV.macro ValMathEng.dll DMS.ocx MVSetup.exe ValMathGerS.dll V2KAddon.ocx ValMathEngS.dll MVPatch.exe The following files are also checked; non-exact correspondence, however, leads to a warning only: ValMathGer.dll prg.ico TX_htm32.dll ValMathEng.dll splash.bmp or default.bmp TX_rtf32.dll ValMathGerS.dll Auto_req_eng.org TX_bmp32.flt ValMathEngS.dll Auto_req_ger.org TX_tif32.flt ValKonv.dll VSFlex7.ocx TX_wmf32.flt V2KUtilities.dll ActBar.ocx TX_word.dll DMS.ocx TX4OLE.ocx olch2x32.ocx V2KAddon.ocx TX32.dll olch3x32.ocx An incorrect version of the above components does not necessarily lead to an incorrect program function or a false calculation. Performance Qualification must always be executed to guarantee validity of results. 156 Agilent ChemStation Plus - Method Validation Pack

157 Qualification Tests File Verification/Installation Qualification (IQ) B Performance Qualification (OQ/PV) - Support Services The full OQ/PV of the MVP software program covers the MVP functionality as well its interaction with the ChemStore database and the ChemStation for data acquisition. Performance qualification of the Agilent ChemStation Plus Method Validation Pack must therefore be executed by an Agilent service representative. Performance Qualification guarantees that the program supplies the same results after installation as during program Validation. A special Validation is opened, a report is generated and compared to a recorded report. The following tolerances are permitted Date deviations Deviations of content (depending on the printer) Deviations of nomenclature (specificity/selectivity) Other deviations are not allowed. They lead to a failed Performance Qualification. If this happens, you should call Method Validation Pack support to analyze the problem. Performance Qualification can check separately for configurations with an English report with a point as decimal separator and a German report with a comma as decimal separator. Should only one of the four above mentioned configurations be used with Method Validation Pack, Performance Qualification needs to be executed for this one configuration only. Agilent ChemStation Plus - Method Validation Pack 157

158 B Qualification Tests File Verification/Installation Qualification (IQ) 158 Agilent ChemStation Plus - Method Validation Pack

159 Agilent ChemStation Plus Method Validation Pack User s Guide Appendix C Error Messages Method Validation Pack recognizes the suitability of used formulae and procedures for data. It intercepts errors resulting from these, and displays an appropriate warning. As a rule, you do not see error messages displayed because the formulae and procedures are suitable for all data. Error messages for the mathematical function library If the number of measured values for some checkpoints is extraordinarily high, or the configuration is unusual, the table values necessary for a result may not exist for the tests incurred. Method Validation Pack displays the following message in the report window: The following error occurred during calculation: Name of error: no table value Error type 10 Very rarely, the number or the structure of input data is incompatible with the formulae or procedures. Method Validation Pack recognizes this and displays the message: The following error occurred during calculation: Name of error: error in mathematical library Agilent Technologies 159

160 C Error Messages During calculation the following errors may occur: 0 - Division by Logarithm of a negative number 2 - Root of a negative number 3 - Valid range of values is exceeded (underflow) 4 - Valid range of values is exceeded (overflow) 5 - Other errors 6 - Loss of computer exactness 7 - Error buffer full 8 - Singular matrix could not be inverted 9 - Memory allocation not possible (low on memory) 10 - No table value In the case of a user interface problem, Method Validation Pack writes login information to method validation pack.log. You can view this file by using an editor (i.e. notepad) or by using the DB-Installer program. Keep this file available when you contact the support, since it contains valuable information about the error, its description and hopefully the reason why it occurred. 160 Agilent ChemStation Plus - Method Validation Pack

161 Agilent ChemStation Plus Method Validation Pack User s Guide Appendix D Known Problems Here are some points of the current version which may cause difficulties. The Internet Explorer 4.01/5.00 is necessary for program operation for the time being. It therefore must be installed. Problems with the installation of Microsoft Data Access Components 2.5 (Setup errors). The ADO 2.5 installer was written by Microsoft and not headwork-consulting. If problems occur, contact us with a description as detailed as possible. We will send you a new installation. During startup, Method Validation Pack checks whether or not a printer is installed. If it is not, the program is terminated. The reason for this is the word processor in Method Validation Pack. It needs information from the attached printer to format correctly. It is not important whether the printer is a local one or a network printer. Should problems occur during conversion of Method Validation Pack for Windows 1.31 files, send us the files. We will then adapt the converter. An exact 1:1 copy of old VAL files is impossible because the content structure of the VDB Validation files is different. In particular, old faulty Validation files may not be converted correctly. You should therefore check the new Validation file after importing it. During import of an old Validation, the old configuration values are not read. Instead, the current global configuration is used for the converted Validation. Therefore, check the configuration values of your converted Validations. Method Validation Pack installs a number of components on your computer which may be used (partly) by other programs. During installation, Method Validation Pack observes the usual conventions such as check of version number and date and so on. If a program no longer works after the installation of Method Validation Pack, an up-to-date component may be the reason. Contact the respective manufacturer of the program. Agilent Technologies 161

162 D Known Problems If Method Validation Pack itself no longer works after updating another program, call us. It would be helpful to receive information on which files were updated by the program. Also contact us if a problem occurs during use of the Method Validation Pack program. It is not important whether it is a real error, a slight inconsistency or your special wishes for a new version. Note that, as long as the auditing window stays open while you work with different functions of Method Validation Pack (e.g. call of a report), the auditing window is updated and may slow down the program considerably. Display of a log file may take some time if it is a large one. Therefore, only the last entries are displayed. If multiple determinations and multiple injections are combined, input grids and reports may become somewhat confusing. This, however, is not an error, as it is unavoidable. Report formatting is an ongoing process and will be optimized in future versions of Method Validation Pack. 162 Agilent ChemStation Plus - Method Validation Pack

163 Agilent ChemStation Plus Method Validation Pack User s Guide Appendix E Glossary Term quantitation limit, determination limit Dixon Test extended spiking method accuracy Grubbs-Test calibration function Definition The determination limit is the smallest amount of a substance that can be detected quantitatively during one analysis with a statistical certainty to be determined. According to the ICH guideline, the detection and determination limits can be calculated based on the standard deviation of the signal and the calibration curve (slope). See outlier test The extended spiking method allows constant and proportional systematic errors to be determined, even when there are no samples with known content values. It is suitable for analytical methods that consist of weighing, diluting and measuring steps. The analysis value thus determined must be proportional to the weigh-in value, and the analyte must be such that it can be added to the sample in defined amounts. Accuracy is a qualitative concept. It describes the extent of correspondence between the expected value and the conventional value. Accuracy is influenced by the systematic error. The systematic error is divided into a constant systematic error and a proportional systematic error. A constant systematic error is not influenced by the concentration of the substance to be determined. If the error is influenced by the concentration it is a proportional systematic error.a quantitative measurement for accuracy is the systematic deviation of the result. See outlier test The calibration function is the correlation between the expected value of the test characteristic, e.g. the extinction (see DIN part 13), and the content, e.g. a mass concentration. Agilent Technologies 163

164 E Glossary Term Calibration curve method Blank value method linearity Lab capability Method comparison detection limit Neumann-Test precision Proprotional systematic deviation Ring Experiment ruggedness, (robustness) Definition For the calculation of these two values, the concentration and the slope must be entered. If you select the blank value method, you must enter the measured values and the slope of the calibration function. Linearity calculates a linear regression using the least squares error for the model y=a+bx. The optional test on proportionality checks whether a one-point calibration is valid. For this, the influence of the deviation on the determination of the content by a one-point calibration is revised at a given concentration by means of real multiple determinations. Lab capability is concerned with the relationship between the dispersion of values caused by the measuring process and the requirement, such as the specification. Lab capability occupies a special position among validation concepts because it does not exclusively refer to the analytical method. According to the ICH guideline, accuracy can be proven by comparison to a second, already validated method. Conformity of mean values and variance with the F-test and the t-test are examined. The detection limit is the smallest amount of substance that can be detected qualitatively during one analysis with a defined statistical certainty. According to the ICH-guideline, the detection and determination limits can be calculated based on the standard deviation of the signal and the calibration curve (slope). See trend test It describes the extent of conformity between results obtained during repeated use of a set analytical method under recurrent and comparable conditions. Monitoring the precision records random errors. See accuracy The ring experiment tests whether the deviation of the values from the lab mean values is coincidental, and therefore there is a possible delimitation between coincident deviations and non-coincident deviations. Result comparison: Ruggedness is defined as the independence of an analytical result from changes in other parameters, which could influence the result. Ruggedness is given if the spread of the lab mean values has no significant influence on the total spread of all measured values. Comparison to a reference: Here the ruggedness is tested with the help of t-test, F-test and the nominal value standard deviation (typically the result of precision) 164 Agilent ChemStation Plus - Method Validation Pack

165 Glossary E Term selectivity Signal-/Noise-ratio Standard addition method Validation True value Recovery (rate) Repeatability Definition Selectivity means that an analytical method can distinguish the substance to be determined from other substances in the sample. Selectivity in chromatography can be described by the relative retention of analyte and interfering substance(s). Optionally, a chromatogram (BMP /WMF format) can be imported). According to the ICH-guideline, the detection and determination limits can be calculated by visual evaluation based on the signal/noise ratio. Therefore, it is necessary to import a chromatogram here (BMP/WMF format). Slope and linearity of the calibration function that was created with matrix-free samples may be changed by the sample matrix. This is often the case in trace analysis. Such effects can be recognized by addition of standards of the analyte to the true sample containing the matrix. Constant systematic errors cannot be recognized in this way. Establishing documented evidence which provides a high degree of assurance that a specific process will consistently produce a product meeting its predetermined specifications (FDA) The true value is the unknown but correct value. The nominal/actual comparison serves to find constant and proportional systematic deviations. There must be a number of samples with different contents, and linearity must be given. The content values have to be known to a more exact degree than that expected by the analytical method. Repeatability from linearity: For this checkpoint, multiple injections are needed; a judgement of the results is made with the help of the relative standard deviation VC. Agilent ChemStation Plus - Method Validation Pack 165

166 E Glossary 166 Agilent ChemStation Plus - Method Validation Pack

167 Agilent ChemStation Plus Method Validation Pack User s Guide Appendix F Interfacing ChemStation Plus and Method Validation Pack Validation, components and checkpoints 168 ChemStation Plus modules and their functionality in the method validation process 176 General configuration 177 Extended planning 179 Study creation & Creation of a method validation sequence template 181 Managing MVP data as ChemStore custom fields 186 Data completion wizard 187 User management 189 ChemStation 190 Method Validation Pack is smoothly integrated into ChemStation/ChemStore by supplying a unique add-on module. This control supplies functionality for: Extended configuration Extended planning Study creation Creation of a method validation sequence template Automatic import of study data Data completion wizard Agilent Technologies 167

168 F Interfacing ChemStation Plus and Method Validation Pack Validation, components and checkpoints NOTE Validation is the formal and systematic proof that an analytical method permits an evaluation of its quality characteristics with adequate and defined reliability. These characteristics are defined in various regulations (ICH, DIN, ISO). Having validated methods is a mandatory requirement for successful audits in regulated environments (e.g. FDA). A validation in Method Validation Pack is made of components and checkpoints. Components versus compounds For data upload from ChemStation Plus, the MVP client compares the component name in MVP with the ChemStation Plus compound name taken from calibration. If there is no match of calibrated compound name and the MVP component, you are prompted to assign a peak from the chromatogram (regardless of calibrated or uncalibrated peaks) to the MVP component. The following example illustrates the differences between a ChemStation Plus compound and an MVP component: A component is similar to a ChemStation compound but it can be more. A standard validation may be setup as follows: Figure 54 Basic validation structure with two components You can add checkpoints to the components Agilent ChemStation Plus - Method Validation Pack

169 Interfacing ChemStation Plus and Method Validation Pack F Figure 55 Complete validation structure with checkpoints and complete the planning by filling out the planning dialogs of each checkpoint. For details on adding components and planning, on how to execute, see Planning a Validation on page 67. Data is entered (either manually for non-agilent ChemStation Plus systems or non-pc systems, through file import or via study import, see below) and you can generate graphics and reports. After you have completed data input for the initial validation and report generation, you might find in some cases that the current set of results is not satisfactory. In order to check if the reason for these unsatisfactory results resides in the method or in the checkpoint planning of the compounds, you might want to make some comparisons; for example, you might want to check precision and linearity with more data. To do this, you can make a copy of Component 1 by dragging it to the Validation 1 line. MVP asks you whether to move the component, to copy its planning or to make a complete copy of it (including all data): Figure 56 Popup dialog for manipulation of validation structure To make a comparison based on more data you select Copy planning. MVP creates new component named Copy of component 1 : Agilent ChemStation Plus - Method Validation Pack 169

170 F Interfacing ChemStation Plus and Method Validation Pack Figure 57 Component 1 duplicated You can rename all entries of this hierarchical list in the same way as in the Microsoft Windows Explorer, i.e. click the entry, it switches to edit mode and you can rename it: Figure 58 Renamed copy of Component 1 Display the planning dialogs of the Component 1 with more data checkpoints and adjust the number of values (e.g. 8). Assuming that you have already entered your data in the precision of component 1, you can open both data windows to copy the existing data: 170 Agilent ChemStation Plus - Method Validation Pack

171 Interfacing ChemStation Plus and Method Validation Pack F Figure 59 Data input windows for direct data transfer You can copy data by using the clipboard or by dragging selected data directly with the mouse. To do so, select the source data (important: to make a multi-line selection, you release the mouse key outside the cells). Now move the selection with the right mouse key while pressing the shift key of the keyboard. Release the mouse on the first cell of the target area. MVP copies all data. Agilent ChemStation Plus - Method Validation Pack 171

172 F Interfacing ChemStation Plus and Method Validation Pack Now you can add additional values to the data grid to complete precision input for Component 1 with more data. You can generate reports for both checkpoints and/or components to compare results. That is why MVP does not use the term compound but component. A component is more than an analytical compound. The benefit of the component principle of MVP is that it offers the tool to check and improve your method validation experiments and results while offering full traceability and data integrity. Another application of MVP s smart copy functionality is the rearrangement of the order of your validation in the same way as you copied the planning. You can select checkpoints and/or components and move them to a different node in the validation. You can even move or copy data between validations to make overall validations out of data and results from simple validations. Executing method validation Step-by-step. Step 1 Step 2 Step 3 checkpoints selection and checkpoint planning data input or import result creation = report creation 172 Agilent ChemStation Plus - Method Validation Pack

173 Interfacing ChemStation Plus and Method Validation Pack F Step 1: Checkpoints selection and checkpoint planning A checkpoint is the implementation of a calculation method in MVP. It contains the planning (specification of the data structure), the data itself (either manually entered or automatically imported) and some options (e.g. graphical settings). The following checkpoints are supported by MVP: Precision Ruggedness/Robustness Lab capability Linearity Calibration function Detection and Determination Limit Accuracy Selectivity/Specificity Ring Experiment Some of these checkpoints (in accordance with the different requirements in ICH, USP and other regulatory guidelines) contain more than one calculation method (e.g. accuracy). The planning dialog of a checkpoint displays both the additional settings for MVP integration and a short help for selecting the correct calculation method. Agilent ChemStation Plus - Method Validation Pack 173

174 F Interfacing ChemStation Plus and Method Validation Pack Figure 60 Planning dialog (accuracy) Depending on the checkpoint, you can define the data structure as follows: Number of data points (samples) Definition of calculation method (where applicable) Selection of multiple injections (for example, in order to use an averaged result from two or more runs for each data point) and/or multiple determination (where applicable) Dimension of data Header data for informational purpose Additional data (nominal values etc.) 174 Agilent ChemStation Plus - Method Validation Pack

175 Interfacing ChemStation Plus and Method Validation Pack F Some checkpoints have a more complex structure. Ring Experiment, Robustness and Lab Capability use more than one data column (lab column, series) for data input. Hence, you must define the number of values for each of these columns. Figure 61 Planning dialog example (Robustness) Step 2: Data input or import After you have defined the checkpoints, you can enter data input (see above) or set up MVP for data transfer with the Chemstation/ChemStore (see below). Agilent ChemStation Plus - Method Validation Pack 175

176 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process ChemStation Plus modules and their functionality in the method validation process The validation workflow of the Method Validation Pack with ChemStation Plus is as follows: Method Validation Pack Start-up of Method Validation Pack (either stand-alone or from Review Client). Be sure to use the correct login a local login allows MVS export to a local ChemStore DB, a server login (Oracle) allows the export to the server ChemStore DB Creation of a new validation (planning), saving of initial DMS revision (for details on validation planning see Planning a Validation on page 67). Automatic generation of sequence template and study ChemStation Entering Method Validation mode Loading MVP-generated sequence Refine sequence flow (standards, calibration etc.) it is valid to duplicate or delete runs created by the sequence template Start data acquisition in Chemstation Method Validation Pack Load validation Load study data and run data completion Display data forms (where necessary) to complete data input (e.g. concentrations) Generate overall report Generate DMS revision Print validation report, display graphics, export reports 176 Agilent ChemStation Plus - Method Validation Pack

177 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F General configuration Under validation properties you can find the standard set of configuration options for your validation: Configuration values Output settings Default header data External documents Methods Validation type (development validation and part 11 validation, see below) Validation type You can select whether your validation is a Development validation or a 21 CFR part 11 validation. A development validation has lower security settings, i.e. DMS entries are written less frequently, audit logging is less strict. A part 11 validation on the other hand has the maximum security enabled and writes DMS entries and audit-log entries frequently. You can switch from a part 11 validation to a development validation but not vice-versa. The final validation of a method must always be performed under the part 11 setting. The same applies to validation work in a regulated environment. Store Method You can store a method description file in the document management system. Since ChemStation uses directories for its methods, you cannot directly store these methods. Use a one-file method description, either as a method text export from ChemStation or as a user-created document containing a detailed analytical method description. Agilent ChemStation Plus - Method Validation Pack 177

178 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Figure 62 Validation Properties dialog Extended configuration With the extended configuration under Utilities options C/S Study Configuration, you can specify special integration parameters, e.g. study parameters. 178 Agilent ChemStation Plus - Method Validation Pack

179 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F Figure 63 Program options These settings are automatically set for a part 11 compliant validation. Furthermore, under Utilities option C/S General, you can select the behavior for data handling (MV=missing values). If you change any parameter under Utilities option C/S, you must click on save afterwards. The settings are valid for new exports of validations only, and not for existing ones. Extended planning Checkpoint Planning is extended by several combo boxes directly interfacing into ChemStation Plus result management for defining The data type used for data import Additional header info that is directly retrieved from the study Agilent ChemStation Plus - Method Validation Pack 179

180 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Figure 64 Additional header info Figure 65 Extended planning dialog 180 Agilent ChemStation Plus - Method Validation Pack

181 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F Study creation & Creation of a method validation sequence template After you have finalized the planning, you create the ChemStore study and create a method validation sequence template for ChemStation by pressing Create Study and MVS for ChemStore. Figure 66 Study creation (MVS Status tab) Minimizing the sample number by using one run for Multiple checkpoints If you have defined checkpoints with identical data structure (e.g. linearity and calibration), you can choose Multiple Data Usage. Multiple Data Usage means that acquisition runs are used for more than one checkpoint to reduce time and cost. Agilent ChemStation Plus - Method Validation Pack 181

182 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Figure 67 Multiple data usage definition You can choose whether to create one sequence template per validation component or one template for the whole validation. Using one sequence template for the whole validation allows unattended creation of all required data in one run. Using one template per component allows smaller (and faster) sequences and hence faster partial results in MVP. Figure 68 Validation export as Method Validation Sequence After all definitions have been made, the method validation sequence template is stored in the Document Management System. 182 Agilent ChemStation Plus - Method Validation Pack

183 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F Figure 69 Study successfully created or updated Figure 70 DMS-dialog for file check-in You can now see your validation and method validation sequence in the DMS window. If you are searching for a specific entry in the DMS, use the search dialog. It gives you powerful options to define your query. Figure 71 DMS-directory If you want to check the contents of the sequence template double-click the DMS entry. You can zoom, export or print the displayed file. Agilent ChemStation Plus - Method Validation Pack 183

184 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Figure 72 Preview of MVS-file in DMS After the study has been created you can see it in ChemStore. A method validation study is marked with (MV): 184 Agilent ChemStation Plus - Method Validation Pack

185 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F Figure 73 Assign study dialog in Review Client The method validation template can then be loaded from ChemStation (see below, Method Validation mode activated) and can be refined to match analytical needs (calibration runs, standards etc.). NOTE If you use the ChemStore data management module, make sure that you have enabled transfer data after each analysis in the transfer settings of the ChemStore setup in the ChemStation (see also ChemStore Concepts guide). With ChemStation Plus Security Pack this transfer is mandatory and cannot be disabled. During acquisition, the data is transferred to the study as usual. The data is marked with a unique identifier the run-id for Method Validation. These run-ids are used for the automatic data import afterwards. The run-ids are stored as ChemStore custom field data. They can be reviewed and they are traceable for data integrity in the ChemStore database. For more information on MVP custom fields in ChemStation Plus, see the next section Agilent ChemStation Plus - Method Validation Pack 185

186 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Managing MVP data as ChemStore custom fields After the acquisition has finished, you can import the validation data from the ChemStore database. The data is identified by the run-id generated when the method validation sequence template was generated and stored to the ChemStore study. It always imports only the latest result version from ChemStore. One point needs careful consideration: with ChemStation Plus, you are able to reanalyze (e.g. reintegrate) your results that were generated with the Method Validation sequence prior to uploading the data in the MVP client. If this happens, the evidence of the method being validated can be in danger because a change of integration events changes key parameters of an analytical method. For a proper documentation of any change that might occur in the ChemStation Plus, it is recommended to report the ChemStore run-id along with the analytical results in the MVP report. The ChemStore run-id is available under planning options as additional header information. MVP transfers method validation data to ChemStation Plus to uniquely map and match method validation data with the ChemStation data acquisition and result management system. This data is stored in ChemStore custom fields along with each acquisition run. Entries in these custom fields can be generated only though the MVP client. All MVP Custom fields in ChemStore are read-only through the application software. MVP exclusively manages the following ChemStore custom fields: MV_checkpoint storing the MVP checkpoint information (which test was used with this run) MV-Component the name of the MVP component in the validation comparable to the Chemstation Plus compound, see also Components versus compounds on page 168. MV_runID storing a unique run identifier that allows full traceability from MVP results to ChemStation Plus result management. This is particularly useful for example, if you notify an outlier result in your method validation. In order to check and/or reanalyze this run, you have only to query for the run-id to immediately receive the run with its result that is under investigation.. NOTE This custom field data is, in contrast to all other custom fields, not accessible in the ChemStation data acquisition module under Sequence > ChemStore setup > Custom field values. They are displayed for each run in the Sequence table sample information window. 186 Agilent ChemStation Plus - Method Validation Pack

187 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F Figure 74 Load study (Data Wizard tab) Data completion wizard Not all data for checkpoint evaluation and reporting are available during data acquisition, because some information (such as concentration for linearity results) can only be added after data acquisition. MVP therefore has an integrated data completion wizard As long as a checkpoint is not complete, the related report cannot be generated (maybe partially) and you see an invalid data entry in the table of contents. Use Run Data Wizard in the Data Wizard panel to display all incomplete checkpoints. With a double-click, you can open their data grids. Input fields with dark background are locked and intended for automatic fill from a study. If you see any missing data here, it is caused by incomplete study data. Input fields with normal background are for manual completion. Agilent ChemStation Plus - Method Validation Pack 187

188 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Figure 75 Wizard for data completion You can switch the behavior for incomplete data under Validation > Properties > Configuration > Data specification/audit. Settings are available for 0 and negative value usage. Reporting You can start reporting (graphics or text) via the menu or the context menus of components, checkpoints and validations. In the MVP integration toolbar is an additional control for the standard functions Overall report, Generate Revision and Export. Figure 76 Wizard for reporting (Reporting tab) 188 Agilent ChemStation Plus - Method Validation Pack

189 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F User management User management of the Method Validation Pack is done via the ChemStore user management, i.e. all user administration is done within ChemStore. Standard ChemStore users have no method validation permission. For more details on the MVP user levels, refer to section Security Concept on page 46. If Method Validation Pack is started directly from the Review Client or stand-alone without any method validation permission, it displays a warning dialog and terminates. Figure 77 ChemStore user manager (Review Client) Agilent ChemStation Plus - Method Validation Pack 189

190 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process ChemStation The ChemStation has a new Method Validation entry in the ChemStore menu to activate the new mode. The toolbar is extended by new buttons to activate the new mode, and to open the browser for Method Validation Sequence Templates (i.e. MVS-files). These new icons are displayed only in sequence mode of ChemStation. This browser is connected to the DMS, and displays all open jobs for the current user (i.e. all validations planned and exported to a study and a sequence template). Changes to the ChemStation functionality in the method validation mode The method validation mode changes the ChemStation functionality to fully support the method validation concept: 4 It denies access to the standard sequence load and save tools and corresponding smart icons. 5 It generates a default generic name mthval.s for the method validation sequence. All sequence table information of the method validation sequence is available in the DMS system of the MVP application 6 It denies access to the ChemStore setup menu items in the sequence and ChemStore menu of the ChemStation 7 It manages jobs from MVP as pending sequences in a new window, accessible though the sequence import smart icon and from the ChemStore menu Method Validation Sequence Browser This browser is connected to the DMS and displays all open jobs for the current user (i.e. all validations planned and exported to a study and a sequence template). From the ChemStation method validation import interface, you can select a job for loading by clicking on it in the list of pending jobs and pressing the select button. This marks the job as active and it cannot be used by a different user. When it is done, it is marked as finalized. When started, the browser displays all jobs of the current user and jobs intended for all users. Uncheck Jobs for all users to see only your jobs. 190 Agilent ChemStation Plus - Method Validation Pack

191 Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process F Show only latest version displays only current versions of jobs and does not display the full history. Pausing and resuming method validation sequences When you want to interrupt working with the MVS file, or when you terminate the ChemStation with a method validation sequence loaded, the MVS file is abandoned and is again available for selection. If an MVS file needs to be deleted e.g because of accidental creation, an administrator can mark this job as finalized. It then disappears from the list of pending MVS sequences. Figure 78 MVS browser connected to DMS The MVS browser can sort the displayed columns when you click on the corresponding header cells. Agilent ChemStation Plus - Method Validation Pack 191

192 F Interfacing ChemStation Plus and Method Validation Pack ChemStation Plus modules and their functionality in the method validation process Figure 79 Graphical user interface buttons and menu items to enable/disable the method validation mode in ChemStation 192 Agilent ChemStation Plus - Method Validation Pack