VICH Topic GL2 (Validation: Methodology)

1 The European Agency for the Evaluation of Medicinal ProductsVeterinary Medicines Evaluation Unit7 Westferry Circus, Canary Wharf, London E14 4HB, UKSwitchboard: (+44-171) 418 8400 Fax: (+44-171) 418 8447E_Mail: 1998 Reproduction and/or distribution of this document is authorised for non-commercial purposes only provided the EMEA isacknowledgedCVMP/ vich /591/98-FINALL ondon, 10 December1998 vich Topic GL2(Validation: Methodology) Step 7 Consensus GuidelineGUIDELINE ON VALIDATION OFANALYTICAL PROCEDURES:METHODOLOGYTRANSMISSION TO CVMPD ecember 1997 TRANSMISSION TO INTERESTED PARTIESJ anuary 1998 COMMENTS REQUESTED BEFOREA pril 1998 FINAL APPROVAL BY CVMPD ecember 1998 DATE FOR COMING INTO OPERATION BYOctober 1999 FOR IMPLEMENTATION AT STEP 7 - 22/10/1998 Page 1 of 10 EMEA 1998 vich GL2 (VALIDATIONMETHODOLOGY)October 1998 For implementation Step 7 VVVVALIDATION OF AAAANALYTICALPPPPROCEDURES::::MMMMETHODO LOGYR ecommended for Implementationat Step 7 of the vich Processon 22 October 1998by the vich Steering CommitteeTHISGUIDELINE HAS BEEN DEVELOPED BY THE APPROPRIATEVICH EXPERTWORKINGGROUP ONTHE BASIS OF THEICHGUIDELINES ON THE SAME SUBJECT AND HAS BEEN SUBJECT TOCONSULTATION BY THE PARTIES,IN ACCORDANCE W ITH THEVICH THEPROCESS THE FINAL DRAFT IS RECOMMENDED FOR ADOPTION TO THE REGULATORY BODIES OF THEEUROPEANUNION,JAPAN IMPLEMENTATION AT STEP 7 - 22/10/1998 Page 2 of 10 EMEA 1998 VALIDATION OF ANALYTICAL PROCEDURES:METHODOLOGYTABLE OF Assay and Impurity Test(s).

2 42. Linearity ..43. Impurities (Quantitation).. Recommended Intermediate Recommended DETECTION Based on Visual Based on Based on the Standard Deviation of the Response and the Recommended QUANTITATION LIMIT .. Based on Visual Based on Signal-to-Noise Based on the Standard Deviation of the Response and the Recommended ROBUSTNESS ..99. SYSTEM SUITABILITY IMPLEMENTATION AT STEP 7 - 22/10/1998 Page 3 of 10 EMEA 1998 VALIDATION OF ANALYTICAL PROCEDURES:METHODOLOGYINTRODUCTIONThis document is complementary to the parent document which presents a discussion ofthe characteristics that should be considered during the validation of analytical purpose is to provide some guidance and recommendations on how to consider thevarious validation characteristics for each analytical some cases (forexample, demonstration of specificity), the overall capabilities of a number ofanalyticalprocedures in combination may be investigated in order to ensure the quality of the drugsubstance or drug product.

3 In addition, the document provides an indication of the datawhich should be presented in a registration application .All relevant data collected during validation and formulae used for calculating validationcharacteristics should be submitted and discussed as other than those set forth in this guideline may be applicable and acceptable. Itis the responsibility of the applicant to choose the validation procedure and protocol mostsuitable for their product. However it is important to remember that the main objective ofvalidation of an analytical procedure is to demonstrate that the procedure is suitable for itsintended purpose. Due to their complex nature, analytical procedures for biological andbiotechnological products in some cases may be approached differently than in reference materials, with documented purity, should be used throughoutthe validation study. The degree of purity necessary depends on the intended accordance with the parent document, and for the sake of clarity, this documentconsiders the various validation characteristics in distinct sections.

4 The arrangement ofthese sections reflects the process by which an analytical procedure may be developed practice, it is usually possible to design the experimental work such that the appropriatevalidation characteristics can be considered simultaneously to provide a sound, overallknowledge of the capabilities of theanalytical procedure, for instance: specificity, linearity,range, accuracy and investigation of specificity should be conducted during the validation of identificationtests, the determination of impurities and the assay. The procedures used todemonstrate specificity will depend on the intended objective of the is not always possible to demonstrate that an analytical procedure is specific for aparticular analyte (complete discrimination). In this case a combination of two or moreanalytical procedures is recommended to achieve the necessary level identification tests should be able to discriminate between compoundsof closely related structures which are likely to be present.

5 The discrimination ofa procedure may be confirmed by obtaining positive results (perhaps bycomparison with a known reference material) from samples containing theanalyte, coupled with negative results from samples which do not contain theanalyte. In addition, the identification test may be applied to materialsstructurally similar to or closely related to the analyte to confirm that a positiveFOR IMPLEMENTATION AT STEP 7 - 22/10/1998 Page 4 of 10 EMEA 1998response is not obtained. The choice of such potentially interfering materialsshould be based on sound scientific judgement with a consideration of theinterferences that could and Impurity Test(s)For chromatographic procedures, representative chromatograms should beused to demonstrate specificity and individual components should beappropriately labelled. Similar considerations should be given to otherseparation separations in chromatography should be investigated at anappropriate level.

6 For critical separations, specificity can be demonstrated bythe resolution of the two components which elute closest to each cases where a non-specific assay is used, other supporting analyticalprocedures should be used to demonstrate overall specificity. For example,where a titration is adopted to assay the drug substance for release, thecombination of the assay and a suitable test for impurities can be approach is similar for both assay and impurity Impurities are availableFor the assay, this should involve demonstration of the discrimination of theanalyte in the presence of impurities and/or excipients; practically, this can bedone by spiking pure substances (drug substance or drug product) withappropriate levels of impurities and/or excipients and demonstrating that theassay result is unaffected by the presence of these materials (by comparisonwith the assay result obtained on unspiked samples).For the impurity test, the discrimination may be established by spiking drugsubstance or drug product with appropriate levels of impurities anddemonstrating the separation of these impurities individually and/or from othercomponents in the sample Impurities are not availableIf impurity or degradation product standards are unavailable, specificity may bedemonstrated by comparing the test results of samples containing impurities ordegradation products to a second well-characterized procedure :pharmacopoeial method or other validated analytical procedure (independentprocedure).

7 As appropriate, this should include samples stored under relevantstress conditions: light, heat, humidity, acid/base hydrolysis and for the assay, the two results should be for the impurity tests, the impurity profiles should be purity tests may be useful to show that the analyte chromatographic peakis not attributable to more than one component ( , diode array, massspectrometry). linear relationship should be evaluated across the range (see section 3) of theanalytical procedure. It may be demonstrated directly on the drug substance (bydilution of a standard stock solution) and/or separate weighings of synthetic mixtures ofthe drug product components, using the proposed procedure. The latter aspect can bestudied during investigation of the should be evaluated by visual inspection of a plot of signals as a function ofanalyte concentration or content. If there is a linear relationship, test results should beFOR IMPLEMENTATION AT STEP 7 - 22/10/1998 Page 5 of 10 EMEA 1998evaluated by appropriate statistical methods, for example, by calculation of aregression line by the method of least squares.

8 In some cases, to obtain linearitybetween assays and sample concentrations, the test data may need to be subjected toa mathematical transformation prior to the regression analysis. Data from theregression line itself may be helpful to provide mathematical estimates of the degree correlation coefficient, y-intercept, slope of the regression line and residual sum ofsquares should be submitted. A plot of the data should be included. In addition, ananalysis of the deviation of the actual data points from the regression line may also behelpful for evaluating analytical procedures, such as immunoassays, do not demonstrate linearity afterany transformation. In this case, the analytical response should be described by anappropriate function of the concentration (amount) of an analyte in a the establishment of linearity, a minimum of 5 concentrations is approaches should be specified range is normally derived from linearity studies and depends on theintended application of the procedure.

9 It is established by confirming that the analyticalprocedure provides an acceptable degree of linearity, accuracy and precision whenapplied to samples containing amounts of analyte within or at the extremes of thespecified range of the analytical following minimum specified ranges should be considered:- for the assay of a drug substance or a finished (drug) product: normally from 80 to120 percent of the test concentration;- for content uniformity, covering a minimum of 70 to 130 percent of the testconcentration, unless a wider more appropriate range, based on the nature of thedosage form, is justified;- for dissolution testing: +/-20 % over the specified range; , if the specifications fora controlled released product cover a region from 20%, after 1 hour, up to 90%,after 24 hours, the validated range would be 0-110% of the label for the determination of an impurity: from the reporting level of an impurity1to 120%of the specification;for impurities known to be unusually potent or to produce toxic or unexpectedpharmacological effects, the detection/quantitation limit should be commensuratewith the level at which the impurities must be :for validation of impurity test procedures carried out during development, itmay be necessary to consider the range around a suggested (probable) limit;- if assay and purity are performed together as one test and only a 100% standard isused, linearity should cover the range from the reporting level of the impurities1to120% of the assay specification.

10 Should be established across the specified range of the analytical chapters Reporting Impurity Content of Batches of the corresponding vich -Guidelines: Impurities in New Drug Substances and Impurities in New Drug Products FOR IMPLEMENTATION AT STEP 7 - 22/10/1998 Page 6 of 10 EMEA Drug SubstanceSeveral methods of determining accuracy are available:a) application of an analytical procedure to an analyte of known purity ( material);b) comparison of the results of the proposed analytical procedure with those ofa second well-characterized procedure, the accuracy of which is statedand/or defined (independent procedure, see );c) accuracy may be inferred once precision, linearity and specificity have Drug ProductSeveral methods for determining accuracy are available:a) application of the analytical procedure to synthetic mixtures of the drugproduct components to which known quantities of the drug substance to beanalysed have been added;b) in cases where it is impossible to obtain samples of all drug productcomponents, it may be acceptable either to add known quantities of theanalyte to the drug product or to compare the results obtained from a secondwell-characterized procedure, the accuracy of which is stated and/or defined(independent procedure, see ).