analytical method validation

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ANALYTICAL METHOD VALIDATION : 

BY: P. VANAJA ANALYTICAL METHOD VALIDATION WWW.PHARMACYGRADUATES.ORG

INTRODUCTION : 

INTRODUCTION Method validation is the process used to confirm that the analytical procedure employed for a specific test is suitable for its intended use. Results from method validation can be used to judge the quality, reliability and consistency of analytical results. WWW.PHARMACYGRADUATES.ORG

TYPES OF ANALYTICAL PROCEDURES TO BE VALIDATED : 

TYPES OF ANALYTICAL PROCEDURES TO BE VALIDATED Identification 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. WWW.PHARMACYGRADUATES.ORG

Analytical methods need to be validated or revalidated : 

Analytical methods need to be validated or revalidated Before their introduction into routine use Whenever the conditions change for which the method has been validated Whenever the method is changed WWW.PHARMACYGRADUATES.ORG

Laboratory Analytical Method Flow During the Drug Development Process : 

Laboratory Analytical Method Flow During the Drug Development Process WWW.PHARMACYGRADUATES.ORG

STEPS IN METHOD VALIDATION : 

STEPS IN METHOD VALIDATION Develop a validation protocol or operating procedure for the validation. Define the application, purpose, and scope of the method. Define the performance parameters and acceptance criteria. Define validation experiments. Verify relevant peformance characteristics of equipment. Qualify materials WWW.PHARMACYGRADUATES.ORG

Slide 7: 

Define criteria for revalidation. Define type and frequency of system suitability tests and/or analytical quality control(AQC) checks. Document validation experiments and results in the validation report. WWW.PHARMACYGRADUATES.ORG

Slide 8: 

Perform pre-validation experiments. Adjust method parameters or acceptance criteria if necessary. Perform full internal (and external) validation experiments. Develop SOPs for executing the method in the routine. WWW.PHARMACYGRADUATES.ORG

PARAMETERS FOR METHOD VALIDATION : 

PARAMETERS FOR METHOD VALIDATION SPECIFICITY LINEARITY RANGE ACCURACY WWW.PHARMACYGRADUATES.ORG

Slide 10: 

ROBUSTNESS SYSTEM SUITABILITY TESTING PRECISION DETECTION LIMIT QUANTITATION LIMIT WWW.PHARMACYGRADUATES.ORG

SPECIFICITY : 

SPECIFICITY The ability to assess unequivocally the analyte in the presence of components which may be expected to be present. An investigation of specificity should be conducted during the validation of identification tests, the determination of impurities, and the assay. WWW.PHARMACYGRADUATES.ORG

IDENTIFICATION : 

IDENTIFICATION Suitable identification tests should be able to discriminate between compounds of closely related structures which are likely to be present. WWW.PHARMACYGRADUATES.ORG

ASSAY AND IMPURITY TEST(S) : 

ASSAY AND IMPURITY TEST(S) For chromatographic procedures, representative chromatograms should be used to demonstrate specificity, and individual components should be appropriately labeled. For critical separations, specificity can be demonstrated by the resolution of the two components which elute closest to each other. WWW.PHARMACYGRADUATES.ORG

Slide 14: 

In cases where a nonspecific assay is used, For ex: where a titration is adopted to assay the drug substance for release, the combination of the assay and a suitable test for impurities can be used. WWW.PHARMACYGRADUATES.ORG

Slide 15: 

For 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 is not attributable to more than one component (e.g., diode array, mass spectrometry). WWW.PHARMACYGRADUATES.ORG

LINEARITY : 

LINEARITY Ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample. WWW.PHARMACYGRADUATES.ORG

Slide 17: 

Linearity should be evaluated by visual inspection of a plot of signals as a function of analyte concentration or content. To obtain linearity between assays and sample concentrations, the test data may have to be subjected to a mathematical transformation prior to the regression analysis. WWW.PHARMACYGRADUATES.ORG

Slide 18: 

Some analytical procedures, such as immunoassays, do not demonstrate linearity after any transformation. In this case, the analytical response should be described by an appropriate function of the concentration (amount) of an analyte in a sample. For the establishment of linearity, a minimum of five concentrations is recommended. WWW.PHARMACYGRADUATES.ORG

RANGE : 

RANGE The interval between the upper and lower concentration of analyte in the sample. WWW.PHARMACYGRADUATES.ORG

Slide 20: 

For the assay of a drug substance or a finished (drug) product: Normally from 80 to 120 percent of the test concentration; For the determination of an impurity: From the reporting level of an impurity 2 to 120 percent of the specification; Note: For validation of impurity test procedures carried out during development, it may be necessary to consider the range around a suggested (probable) limit. WWW.PHARMACYGRADUATES.ORG

ACCURACY : 

ACCURACY The accuracy of an analytical procedure expresses the closeness of agreement between the value which is accepted either as a conventional true value or an accepted reference value and the value found. WWW.PHARMACYGRADUATES.ORG

ASSAY : 

ASSAY Drug substance: (a)Application of an analytical procedure to an analyte of known purity (e.g.,reference material); (b)Comparison of the results of the proposed analytical procedure with those of a second well-characterized procedure, the accuracy of which is stated and/or defined (c)Accuracy may be inferred once precision, linearity, and specificity have been established. WWW.PHARMACYGRADUATES.ORG

Slide 23: 

2. Drug product: (a) Application of the analytical procedure to synthetic mixtures of the drug product components to which known quantities of the drug substance to be analyzed have been added; (b) Accuracy may be inferred once precision, linearity, and specificity have been established. WWW.PHARMACYGRADUATES.ORG

IMPURITIES : 

IMPURITIES Accuracy should be assessed on samples (drug substance/drug product) spiked with known amounts of impurities. It should be clear how the individual or total impurities are to be determined, e.g. weight/weight or area percent, in all cases with respect to the major analyte. WWW.PHARMACYGRADUATES.ORG

RECOMMENDED DATA : 

RECOMMENDED DATA Accuracy should be assessed using a minimum of 9 determinations over a minimum of 3 concentration levels covering the specified range Reported as percent recovery by the assay of known added amount of analyte in the sample or as the difference between the mean and the accepted true value together with the confidence intervals. WWW.PHARMACYGRADUATES.ORG

ROBUSTNESS : 

ROBUSTNESS 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. WWW.PHARMACYGRADUATES.ORG

Slide 27: 

It should show the reliability of an analysis with respect to deliberate variations in method parameters. Examples of typical variations are: • Stability of analytical solutions • Extraction time WWW.PHARMACYGRADUATES.ORG

Slide 28: 

In the case of liquid chromatography, Influence of variations of pH in a mobile phase Influence of variations in mobile phase composition Different columns (different lots and/or suppliers) Temperature Flow rate In the case of gas-chromatography, Different columns Temperature Flow rate WWW.PHARMACYGRADUATES.ORG

PRECISION : 

PRECISION The precision of an analytical procedure expresses the closeness of agreement between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions. WWW.PHARMACYGRADUATES.ORG

Slide 30: 

WWW.PHARMACYGRADUATES.ORG

SYSTEM SUITABILITY TESTING : 

SYSTEM SUITABILITY TESTING 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. System suitability test parameters to be established for a particular procedure depend on the type of procedure being validated. WWW.PHARMACYGRADUATES.ORG

DETECTION LIMIT : 

DETECTION LIMIT The detection limit of an individual analytical procedure is the lowest amount of analyte in a sample which can be detected but not necessarily quantitated as an exact value. WWW.PHARMACYGRADUATES.ORG

BASED ON VISUAL EVALUATION : 

BASED ON VISUAL EVALUATION The detection limit is determined by the analysis of samples with known concentrations of analyte and by establishing the minimum level at which the analyte can be reliably detected. WWW.PHARMACYGRADUATES.ORG

BASED ON SIGNAL-TO-NOISE : 

BASED ON SIGNAL-TO-NOISE Determination of the signal-to-noise ratio is performed by comparing measured signals from samples with known low concentrations of analyte with those of blank samples and establishing the minimum concentration at which the analyte can be reliably detected. A signal-to-noise ratio between 3 or 2:1 is generally considered acceptable for estimating the detection limit. WWW.PHARMACYGRADUATES.ORG

BASED ON THE STANDARD DEVIATION OF THE RESPONSE AND THE SLOPE : 

BASED ON THE STANDARD DEVIATION OF THE RESPONSE AND THE SLOPE The detection limit (DL) may be expressed as: 3.3 σ DL= ____________ S σ = the standard deviation of the response S = the slope of the calibration curve WWW.PHARMACYGRADUATES.ORG

QUANTITATION LIMIT : 

QUANTITATION LIMIT The 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. The determination of impurities and/or degradation products. WWW.PHARMACYGRADUATES.ORG

BASED ON VISUAL EVALUATION : 

BASED ON VISUAL EVALUATION The quantitation limit is generally determined by the analysis of samples with known concentrations of analyte and by establishing the minimum level at which the analyte can be quantified with acceptable accuracy and precision. WWW.PHARMACYGRADUATES.ORG

BASED ON SIGNAL-TO-NOISE : 

BASED ON SIGNAL-TO-NOISE Determination of the signal-to-noise ratio is performed by comparing measured signals from samples with known low concentrations of analyte with those of blank samples and by establishing the minimum concentration at which the analyte can be reliably quantified. A typical signal-to-noise ratio is 10:1. WWW.PHARMACYGRADUATES.ORG

BASED ON THE STANDARD DEVIATION OF THE RESPONSE AND THE SLOPE : 

BASED ON THE STANDARD DEVIATION OF THE RESPONSE AND THE SLOPE The quantitation limit (QL) may be expressed as: 10 σ QL= _______________ S σ = the standard deviation of responses S = the slope of the calibration curve WWW.PHARMACYGRADUATES.ORG

REFERENCES : 

REFERENCES www.ich.org Pharmaceutical process validation: an international third addition http://www.fda.gov/cder/guidance/ichq2a.pdf http://www.nihs.go.jp/drug/validation/q2bwww.html WWW.PHARMACYGRADUATES.ORG

THANQ : 

THANQ WWW.PHARMACYGRADUATES.ORG