OBJECTIVE

PUBLISHED GUIDANCES

TYPES OF ANALYTICAL METHOD TO BE VALIDATED

CONSIDERATIONS PRIOR TO METHOD VALIDATION

TYPICAL ANALYTICAL PERFORMANCE

CHARACTERISTICS USED IN METHOD VALIDATION

REVALIDATION

POSSIBLE QUESTIONS

REFERENCES

PUBLISHED GUIDANCES

ICH-Q2A “Text on Validation of Analytical Procedure:(1994)

ICH-Q2B “Validation of Analytical Procedures: Methodology: (1995)

CDER “Reviewer Guidance: Validation of Chromatographic Method” (1994)

CDER “Submitting Samples and Analytical Data for Method Validations” (1987)

CDER Draft “Analytical Procedures and Method Validation” (2000)

CDER “Bioanalytical Method Validation for Human Studies” (1999)

USP<1225> “Validation of Compendial Methods” (current revision)

SUBMISSION TO THE COMPENDIARATIONALE

PROPOSED ANALYTICAL PROCEDURE

DATA ELEMENTS

TYPES OF ANALYTICAL PROCEDURES TO BE VALIDATEDIdentification tests.Quantitative tests for impurities' content.Limit tests for the control of impurities.Quantitative tests of the active moiety in

samples of drug.substance or drug product or other

selected component(s) in the drug product.

CONSIDERATIONS PRIOR TO METHOD VALIDATION

Suitability of InstrumentStatus of Qualification and Calibration

Suitability of MaterialsStatus of Reference Standards, Reagents, etc.

Suitability of AnalystStatus of Training and Qualification Records

Suitability of DocumentationWritten analytical procedure and proper approved protocol with pre-established acceptance criteria.

EXAMPLES OF METHODS THAT REQUIRE VALIDATION DOCUMENTATION

CHROMATOGRAPHIC METHODS

SPECTROPHOTOMETRIC METHODS

CAPILLARY ELECTROPHORESIS METHODS

PARTICLE SIZE ANALYSIS METHODS

DISSOLUTION METHODS

TITRATION METHODS

AUTOMATED ANALYTICAL METHODS

ANALYTICAL METHOD VALIDATION

Validation of an analytical method is the process by which it is established, by laboratory studies, that the performance characteristics of the method meet the requirements for the intended analytical applications.

TYPICAL ANALYTICAL PERFORMANCE CHARACTERISTICS USED IN METHOD VALIDATION

Specificity (Selectivity)LinearityRangeAccuracyPrecisionDetection LimitQuantitation LimitRobustnessSystem Suitability

Testing

SPECIFICITY

SPECIFICITY is the ability to assess unequivocally the analyte in presence of components which may be expected to be present.

DETERMINATION

IDENTIFICATION TESTS

ASSAY AND IMPURITY TEST(S) Impurities are available Impurities are not available

LINEARITYLINEARITY of an analytical procedure is its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample.

DETERMINATION- Linearity should be evaluated by visual inspection of a plot of signals as a function of analyte concentration or content.

NOTEFor the establishment of linearity, a minimum of five concentrations is recommended.

RANGE

RANGE of an analytical procedure is the interval between the upper and lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity.

DETERMINATION-The specified range is normally derived from linearity studies and depends on the intended application of the procedure.

ACCURACYACCURACY of an analytical method is the closeness of

test results obtained by that method to the true value.

DETERMINATION-Accuracy should be established across

the specified range of the analytical procedure.ASSAY

Drug SubstanceDrug Product

IMPURITIES (QUANTITATION)

NOTE

Accuracy should be assessed using a minimum of 9 determinations over a minimum of 3 concentration levels covering the specified range (i.e., three concentrations and three replicates of each).

PRECISION

PRECISION of an analytical method is the degree of agreement among

individual test results when the method is applied repeatedly to multiple

samplings of a homogenous sample.

DETERMINATION- A sufficient number of aliquots of a

homogeneous sample are assayed to be able to calculate statistically valid

estimates of standard deviation or relative standard deviation.

Repeatability

Intermediate precision

Reproducibilty

DETECTION LIMITDETECTION LIMIT of an individual analytical procedure is the lowest amount of analyte in a sample which can be detected but not necessarily quantitated, under the stated experimental conditions.

DETERMINATION- Several approaches for determining the detection limit are possible, depending on whether the procedure is a non-instrumental or instrumental.

BASED ON VISUAL EXAMINATION

BASED ON SIGNAL TO NOISE RATIO

QUANTITATION LIMIT

QUANTITATION LIMIT of an individual analytical procedure is the lowest amount of analyte in a sample which can be quantitatively determined with suitable precision and accuracy.

DETERMINATION- Several approaches for determining the detection limit are possible, depending on whether the procedure is a non-instrumental or instrumental.

BASED ON VISUAL EXAMINATION

BASED ON SIGNAL TO NOISE RATIO

LOQ, LOD and SNRLimit of Quantitation Limit of DetectionSignal to Noise Ratio

noise

Peak ALOD

Peak BLOQ

Baseline

RUGGEDNESS

NOTEIncluded in

but not in

RUGGEDNESS of an analytical method is the degree of reproducibility of test results obtained by the analysis of the same samples under a variety of conditions, such as different laboratories different analyst, different instruments, different lots of reagent, different elapsed assay times, different assay temperatures, different days, etc.

ROBUSTNESSROBUSTNESS of an analytical procedure is a measure of its capacity to remain unaffected by small, but deliberate variations in method parameters and provides an indication of its reliability during normal usage.

DETERMINATION- The evaluation of robustness should be considered during the development phase and depends on the type of procedure under study.

SYSTEM SUITABILITY TESTINGSYSTEM SUITABILITY TESTING is an integral part of many analytical procedures. The tests are based on the concept that the equipment, electronics, analytical operations and samples to be analyzed constitute an integral system that can be evaluated as such.

Recommended Validation Characteristics of the Various Types of Tests

REVALIDATION MAY BE NECESSARY IN THE FOLLOWING CIRCUMSTANCES:

changes in the synthesis of the drug substance; changes in the composition of the finished product;changes in the analytical procedure;The degree of revalidation required depends on the nature of the changes. Certain other changes may require validation as well.

The United State Pharmacopoeia 24; The National

Formulary 19; 2000: [1225] VALIDATION OF

COMPENDIAL METHODS.

www.labcompliance.com/methods/meth_va

htm#introduction

http://www.fda.gov/cder/guidance/2396dft.htm

www.fda.gov/ohrms/dockets/

ac/02/slides/3841s1_07_lachman.PPT

http://www.fda.gov/cder/guidance/ameth.htm

http://www.ich.orghttp://www.fda.gov/cder/guidance/4252fnl.htmhttp://www.pharmtech.com/pharmtech/

dat a/articlestandard/pharmtech/102003/48314/article.pdfhttp://www.ivstandards.com/tech/

reliability/part17.asphttp://www.aoac.org/