FDA方法学验证

1、 Guidance for IndustryQ2B Validation of AnalyticalProcedures: MethodologyNovember 1996ICHGuidance for IndustryQ2B Validation of AnalyticalProcedures: MethodologyAdditional copies are available from:the Drug Information Branch (HFD-210,Center for Drug Evaluation and Research (CDER,5600 Fishers Lane,

2、Rockville, MD 20857 (Tel 301-827-4573/cder/guidance/index.htmorOffice of Communication,Training, and Manufacturers Assistance (HFM-40Center for Biologics Evaluation and Research (CBER1401 Rockville Pike, Rockville, MD 20852-1448,/cber/guidelines.htm(Fax 888-CBERFAX

3、or 301-827-3844(Voice Information 800-835-4709 or 301-827-1800U.S. Department of Health and Human ServicesFood and Drug AdministrationCenter for Drug Evaluation and Research (CDERCenter for Biologics Evaluation and Research (CBERNovember 1996ICHTable of ContentsI.II. INTRODUCTION . . . . . . . . . .

4、 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1SPECIFICITY (1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2A. Identification (1.1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

5、 . . . . . . . . . . . . 2B. Assay and Impurity Test(s (1.2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2LINEARITY (2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3RANGE (3. . . . . . . . . . . . . . . . . . .

6、 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4ACCURACY (4. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5A. Assay (4.1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

7、 . . . . . . . . . 5B. Impurities (Quantitation (4.2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6C. Recommended Data (4.3. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6PRECISION (5. . . . . . . . . . . . . . . . . . . . . . . . .

8、. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6A. Repeatability (5.1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7B. Intermediate Precision (5.2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7C. Reproduci

9、bility (5.3. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7D. Recommended Data (5.4. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7DETECTION LIMIT (6. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

10、 . . . . . . . . . . . . . 7A. Based on Visual Evaluation (6.1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7B. Based on Signal-to-Noise (6.2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8C. Based on the Standard Deviation of the Response and t

11、he Slope (6.3. . . . . . . . 8D. Recommended Data (6.4. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8QUANTITATION LIMIT (7. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8A. Based on Visual Evaluation (7.1. . . . . . . . .

12、 . . . . . . . . . . . . . . . . . . . . . . . . . . . 9B. Based on Signal-to-Noise (7.2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9C. Based on the Standard Deviation of the Response and the Slope (7.3. . . . . . . . 9D. Recommended Data (7.4. . . . . . . . . . . . .

13、 . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10ROBUSTNESS (8. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10SYSTEM SUITABILITY TESTING (9. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10III. IV. V. VI. VII. VI

14、II. IX. X.iGUIDANCE FOR INDUSTRY1Q2B Validation of AnalyticalProcedures: MethodologyI. INTRODUCTIONThis document is complementary to the ICH guidance entitled Text on Validation of AnalyticalProcedures (ICH Q2A, which presents a discussion of the characteristics that should beconsidered during the v

15、alidation of analytical procedures. Its purpose is to provide someguidance and recommendations on how to consider the various validation characteristics for eachanalytical procedure. In some cases (for example, demonstration of specificity, the overallcapabilities of a number of analytical procedure

16、s in combination may be investigated in order toensure the quality of the drug substance or drug product. In addition, the document provides anindication of the data that should be presented in a new drug application.All relevant data collected during validation and formulae used for calculating val

17、idationcharacteristics should be submitted and discussed as appropriate.Approaches other than those set forth in this guidance may be applicable and acceptable. It is theresponsibility of the applicant to choose the validation procedure and protocol most suitable fortheir product. However, it is imp

18、ortant to remember that the main objective of validation of ananalytical procedure is to demonstrate that the procedure is suitable for its intended purpose. Dueto their complex nature, analytical procedures for biological and biotechnological products insome cases may be approached differently than

19、 in this document.Well-characterized reference materials, with documented purity, should be used throughout thevalidation study. The degree of purity necessary depends on the intended use.In accordance with the parent document, and for the sake of clarity, this document considers thevarious validati

20、on characteristics in distinct sections. The arrangement of these sections reflectsthe process by which an analytical procedure may be developed and evaluated.This guidance was developed within the Expert Working Group (Quality of the International Conference onHarmonisation of Technical Requirement

21、s for Registration of Pharmaceuticals for Human Use (ICH and has beensubject to consultation by the regulatory parties, in accordance with the ICH process. This document has been endorsedby the ICH Steering Committee at Step 4 of the ICH process, November 1996. At Step 4 of the process, the final dr

22、aft isrecommended for adoption to the regulatory bodies of the European Union, Japan and the United States. This guidancewas published in the Federal Register on May 19, 1997 (62 FR 27464, and is applicable to drug and biologicalproducts. This guidance represents the Agencys current thinking on the

23、validation of analytical procedures. It does notcreate or confer any rights for or on any person and does not operate to bind FDA or the public. An alternative approachmay be used if such approach satisfies the requirements of the applicable statute, regulations, or both.1In practice, it is usually

24、possible to design the experimental work such that the appropriatevalidation characteristics can be considered simultaneously to provide a sound, overall knowledgeof the capabilities of the analytical procedure, for instance: Specificity, linearity, range, accuracy,and precision.II. SPECIFICITY (1An

25、 investigation of specificity should be conducted during the validation of identification tests, thedetermination of impurities, and the assay. The procedures used to demonstrate specificity willdepend on the intended objective of the analytical procedure.It is not always possible to demonstrate tha

26、t an analytical procedure is specific for a particularanalyte (complete discrimination. In this case, a combination of two or more analyticalprocedures is recommended to achieve the necessary level of discrimination.A. Identification (1.1Suitable identification tests should be able to discriminate b

27、etween compounds of closelyrelated structures which are likely to be present. The discrimination of a procedure maybe confirmed by obtaining positive results (perhaps by comparison with a known referencematerial from samples containing the analyte, coupled with negative results from sampleswhich do

28、not contain the analyte. In addition, the identification test may be applied tomaterials structurally similar to or closely related to the analyte to confirm that a positiveresponse is not obtained. The choice of such potentially interfering materials should bebased on sensible scientific judgment w

29、ith a consideration of the interferences that couldoccur.B. Assay and Impurity Test(s (1.2For chromatographic procedures, representative chromatograms should be used todemonstrate specificity, and individual components should be appropriately labeled.Similar considerations should be given to other s

30、eparation techniques.Critical separations in chromatography should be investigated at an appropriate level. Forcritical separations, specificity can be demonstrated by the resolution of the twocomponents which elute closest to each other.In cases where a nonspecific assay is used, other supporting a

31、nalytical procedures shouldbe used to demonstrate overall specificity. For example, where a titration is adopted toassay the drug substance for release, the combination of the assay and a suitable test forimpurities can be used.2The approach is similar for both assay and impurity tests:1. Impurities

32、 are available (1.2.1For the assay, this should involve demonstration of the discrimination of theanalyte in the presence of impurities and/or excipients; practically, this can be doneby spiking pure substances (drug substance or drug product with appropriatelevels of impurities and/or excipients an

33、d demonstrating that the assay result isunaffected by the presence of these materials (by comparison with the assay resultobtained on unspiked samples. For the impurity test, the discrimination may beestablished by spiking drug substance or drug product with appropriate levels ofimpurities and demon

34、strating the separation of these impurities individually and/orfrom other components in the sample matrix.2. Impurities are not available (1.2.2If impurity or degradation product standards are unavailable, specificity may bedemonstrated by comparing the test results of samples containing impurities

35、ordegradation products to a second well-characterized procedure, e.g.,pharmacopoeial method or other validated analytical procedure (independentprocedure. As appropriate, this should include samples stored under relevantstress conditions: Light, heat, humidity, acid/base hydrolysis, and oxidation. F

36、or the assay, the two results should be compared. For the impurity tests, the impurity profiles should be compared.Peak purity tests may be useful to show that the analyte chromatographic peak isnot attributable to more than one component (e.g., diode array, massspectrometry.III. LINEARITY (2A linea

37、r relationship should be evaluated across the range (see section 3 of the analyticalprocedure. It may be demonstrated directly on the drug substance (by dilution of a standard stocksolution and/or separate weighings of synthetic mixtures of the drug product components, usingthe proposed procedure. T

38、he latter aspect can be studied during investigation of the range.Linearity should be evaluated by visual inspection of a plot of signals as a function of analyteconcentration or content. If there is a linear relationship, test results should be evaluated byappropriate statistical methods, for examp

39、le, by calculation of a regression line by the method ofleast squares. In some cases, to obtain linearity between assays and sample concentrations, thetest data may have to be subjected to a mathematical transformation prior to the regression3analysis. Data from the regression line itself may be hel

40、pful to provide mathematical estimates ofthe degree of linearity.The correlation coefficient, y-intercept, slope of the regression line, and residual sum of squaresshould be submitted. A plot of the data should be included. In addition, an analysis of thedeviation of the actual data points from the

41、regression line may also be helpful for evaluatinglinearity.Some analytical procedures, such as immunoassays, do not demonstrate linearity after anytransformation. In this case, the analytical response should be described by an appropriatefunction of the concentration (amount of an analyte in a samp

42、le.For the establishment of linearity, a minimum of five concentrations is recommended. Otherapproaches should be justified.IV. RANGE (3The specified range is normally derived from linearity studies and depends on the intendedapplication of the procedure. It is established by confirming that the ana

43、lytical procedure providesan acceptable degree of linearity, accuracy, and precision when applied to samples containingamounts of analyte within or at the extremes of the specified range of the analytical procedure.The following minimum specified ranges should be considered.For the assay of a drug s

44、ubstance or a finished (drug product: Normally from 80to 120 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 (e.g., metered dose inhalers, is just

45、ified;For dissolution testing: +/-20 percent over the specified range; e.g., if thespecifications for a controlled released product cover a region from 20 percent,after 1 hour, up to 90 percent, after 24 hours, the validated range would be 0-110percent of the label claim;For the determination of an

46、impurity: From the reporting level of an impurity2 to120 percent of the specification;See sections on "Reporting Impurity Content of Batches" of the corresponding ICH Q3A guidance entitled "Impurities in New Drug Substances" (61 FR 372; January 4, 1996 and ICH Q3B draft guidance

47、"Impurities in NewDrug Products" (61 FR 11268; March 19, 1996.24For 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 controlled.Note: For v

48、alidation of impurity test procedures carried out during development,it may 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 percentstandard is used, linearity should cover the range from the reporting level o

49、f theimpurities 3 to 120 percent of the assay specification.V. ACCURACY (4Accuracy should be established across the specified range of the analytical procedure.A. Assay (4.11. Drug substance (4.1.1Several methods of determining accuracy are available.(a Application of an analytical procedure to an a

50、nalyte of known purity (e.g.,reference material;(b Comparison of the results of the proposed analytical procedure with those of asecond well-characterized procedure, the accuracy of which is stated and/ordefined (independent procedure, see section 1.2.;(c Accuracy may be inferred once precision, lin

51、earity, and specificity have beenestablished.2. Drug product (4.1.2Several 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 beanalyzed have been added;(b In

52、 cases where it is impossible to obtain samples of all drug productcomponents, it may be acceptable either to add known quantities of the analyte to3 Ibid .5the drug product or to compare the results obtained from a second, well-characterized procedure, the accuracy of which is stated and/or defined

53、(independent procedure, see section 1.2;(c Accuracy may be inferred once precision, linearity, and specificity have beenestablished.B. Impurities (Quantitation (4.2Accuracy should be assessed on samples (drug substance/drug product spiked withknown amounts of impurities.In cases where it is impossib

54、le to obtain samples of certain impurities and/or degradationproducts, it is considered acceptable to compare results obtained by an independentprocedure (see section 1.2. The response factor of the drug substance can be used.It should be clear how the individual or total impurities are to be determ

55、ined, e.g.,weight/weight or area percent, in all cases with respect to the major analyte.C. Recommended Data (4.3Accuracy should be assessed using a minimum of 9 determinations over a minimum of 3concentration levels covering the specified range (e.g., 3 concentrations/3 replicates eachof the total

56、analytical procedure.Accuracy should be reported as percent recovery by the assay of known added amount ofanalyte in the sample or as the difference between the mean and the accepted true valuetogether with the confidence intervals.VI. PRECISION (5Validation of tests for assay and for quantitative d

57、etermination of impurities includes aninvestigation of precision.A. Repeatability (5.1Repeatability should be assessed using:(1 A minimum of 9 determinations covering the specified range for the procedure(e.g., 3 concentrations/3 replicates each; or(2 A minimum of 6 determinations at 100 percent of

58、the test concentration.6B. Intermediate Precision (5.2The extent to which intermediate precision should be established depends on thecircumstances under which the procedure is intended to be used. The applicant shouldestablish the effects of random events on the precision of the analytical procedure. Typicalvariations to be studied include days, analysts, equipment, etc. It is not necessary to studythese effects individually. The use of an experimental design (matrix is encouraged.C. Reproducibility (5.3Reproducibility is assessed by means of an interl