Protocol for method validation

Important protocols for method validation in the literature have been derived, amongst others, from the Current Good Manufacturing Practice, Code of Federal Regulations, Food and Drug Administration, National Drug Administration, the United States Pharmacopoeia Convention, the American Public Health Association and the International Conference on Harmonization. 

Figure 8.1 Common European protocol for method validation...

A detailed protocol for laboratory validation of sampling and analytical methods for toxic substances in workplace environments is given in Figure 1. The literature was searched and a method of sampling and analysis was selected. The next step was to evaluate and, if necessary, develop an analytical method that was compatible with the sampling medium. If a satisfactory analytical method became available only then did we undertake generation of a test atmosphere. Then samples were collected with the appropriate collection medium. Both capacity and collection efficiency were evaluated. 

The overall protocol for packaging validation remains the same whether microbial challenge test methods or physical test methods are used. At each stage in the development and production process of a package for a medical device package integrity must be verified. 

As for all other analytical methods, methods for the analysis of products derived from modern biotechnology need to be validated. This validation should follow international standards (e.g. ISO 5725) and comply with, The IUPAC/AOAC/ISO harmonized protocol of method validation [6] or the standards cited above. Validation of testing methods is greatly facilitated by the use and availability of the appropriate reference materials. The following paragraphs specify some important... 

Present-day analytical laboratories are increasingly under pressure to supply objective evidence of their technical competence, of the reliability of their results and performance, and to seek formal certification or accreditation. This pressure may come from the laboratory s customers (e.g., industry and national bodies) but may also be due to scientific considerations. A QM system in place, validation of methods, uncertainty evaluation, the use of primary standards and CRMs, participation in ILCs, and PT, all serve to assure and demonstrate the quality of measurements. Compared to, say, 30 years ago, the stability of the equipment now available is much improved, and a greater range of RMs for method validation and calibration is accessible. Nevertheless, to achieve mutual (international) acceptance of various bodies of evidence for QA activities, a number of protocols have been developed. The most widely recognized protocols used in chemical measurements and testing are the ISO Guide 9000 2000, ISO/IEC 17025 2005, and OECD Guidelines for GLP, as well as its national and sector equivalents. 

Abstract ISO principles of measurement uncertainty estimation are compared with protocols for method development and validation by collaborative trial and concomitant top-down estimation of uncertainty. It is shown that there is substantial commonality between the two procedures. In particular, both require a careful consideration and study of the main effects on the re-... 

We have developed a protocol which describes how data generated from experimental studies commonly undertaken for method validation purposes can be used in measurement uncertainty evaluation. This paper has illustrated the application of the protocol. In the example described, the uncertainty estimate for three analytes in different oil matrices was evaluated from three experimental studies, namely precision, recovery and ruggedness. These studies were required as part of the method validation, but planning the studies with uncertainty evaluation in mind allowed an uncertainty estimate to be calculated with little extra effort. A number of areas were identified where additional experimental work may be required to refine the estimates. However the necessary data could be generated by carrying out additional analyses alongside routine test samples. Again this would minimise the amount of laboratory effort required. 

The third goal is to develop protocols for the validation and harmonisation of chemical and biological methods, to meet EU demands for monitoring of emerging substances. 

M. Kibblewhite, L. Heasman and V. Forster, Development of a validation protocol for methods of... 

The apparatus test includes a protocol about pump, injector and detector functions and includes the requirements listed in Table 19.1 Usually they are easily fulfilled by today s HPLC instrumentation, especially when new. The apparatus test should be performed routinely in adequate time intervals and after each repair. It is a prerequisite for method validation. A proposal can be found in Chapter 24. 

Occupational Safety and Health Administration Analytical Laboratory Precision and Accuracy Data Protocol for Laboratory Validations. In OSHAAnalytical Methods Manual 1st ed. Cincinnati, OH American Conference of Governmental Industrial Hygienists (Pub. No. ISBN 0-936712-66-X), 1985. 

BERM-5 presented the changing outlook of the AOAC International in recognizing the usefulness of incorporating RMs for use in conjunction with their methods validation protocol. 

Within CEN TC 275, a distinct protocol for the conduct of inter-laboratory method validation trials is not selected or required, but it is recommended to apply ISO 5725 or the ISO/International Union for Pure and Applied Chemistry (lUPAC) harmonized protocol. " In the past, results of several national and international interlaboratory trials had been accepted. Nevertheless, the following fundamental aspects of the harmonized protocol for the design of method-performance studies should be fulfilled ... 

The approaches described above give approximate values for the LoD and LoQ. This is sufficient if the analyte levels in test samples are well above the LoD and LoQ. If the detection limits are critical, they should be evaluated by using a more rigorous approach [1, 2, 14]. In addition, the LoD and LoQ sometimes vary with the type of sample and minor variations in measurement conditions. When these parameters are of importance, it is necessary to assess the expected level of change during method validation and build a protocol for checking the parameters, at appropriate intervals, when the method is in routine use. 

Bidigare, R.R., and C.C. Trees. 2000. HPLC phytoplankton pigments Sampling, laboratory methods, and quality assurance procedures. Pp. 154-161 in Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 2, J. Mueller and G. Fargion, eds., NASA Technical Memorandum 2000-209966. 

DQ is performed by the supplier of the equipment or system at the supplier s factory as part of the factory acceptance test (FAT). IQ (based on site acceptance test—SAT), OQ, and PQ are performed on-site at the GMP facility. For a GMP manufacturing facility, the validation activities include the facility design, FTVAC system, environment control, laboratory and production equipment, water system, gases and utilities, cleaning, and analytical methods. Validation protocols (IQ, QQ, and PQ) are prepared for each item, listing all critical steps and acceptance criteria. Deviations are reviewed and resolved before the validation activity proceeds to the next phase. 

A second approach is to prepare validation protocols for each method prior to the commencement of validation activities. This has the advantage of performing method-appropriate experiments while eliminating those portions of validation work that do not apply. It also allows the analysts to set more specific acceptance criteria based on the performance of the individual method instead. A drawback is that the protocol adds to the workload of the analyst since it must be prepared for each validation. This approach also allows for the possibility of greater variance in the approach to validation within the company. 

Once a method has been developed and validated, it should be transferred to each site that intends to use the method. A typical method transfer would take place between the research group that developed and validated the method and the QC group that will use the method to release the finished commercial product, although method transfer may occur at any point where knowledge moves from one group to another. As in the case of method validation, method transfer should be performed under the control of a protocol that details the steps required for the study. 

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