Analytical Method Validation
- 1. ICH Q 2 GUIDELINES ANALYTICAL METHOD VALIDATION
- 2. The objective of validation of an analytical procedure To demonstrate that it is suitable for its intended purpose. To maintain and assure a higher degree of quality of food and drug products.
- 3. 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
- 4. Definition Validation is the process of establishing documentary evidence demonstrating that a procedure, process, or activity results are accurate & precised. In the pharmaceutical industry, it is very important that in addition to final testing and compliance of products, it is also assured that the process will consistently produce the expected results
- 5. Analytical method validation parameters • Accuracy • Precision Repeatability Intermediate Precision • Specificity • Detection Limit (LOD) • Quantitation Limit (LOQ) • Linearity • Range
- 6. SPECIFICITY • Specificity is the ability to assess unequivocally the analyte in the presence of components which may be expected to be present. • Typically these might include impurities, degradants, etc.
- 7. 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. • This is sometimes termed trueness.
- 8. PRECISION • The precision of an analytical procedure expresses the closeness of agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions. Precision may be considered at three levels: intermediate precision reproducibility. repeatability,
- 9. • Repeatability Repeatability expresses the precision under the same operating conditions over a short interval of time. Repeatability is also termed intra-assay precision . • Intermediate precision Intermediate precision expresses within-laboratories variations: different days, different analysts, different equipment, etc. • Reproducibility Reproducibility expresses the precision between
- 11. DETECTION LIMIT/Limit of Detection /LOD • 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.
- 12. LOQ/LIMIT OF QUANTIFICATION • 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.
- 13. LINEARITY • The linearity 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 • For the establishment of linearity, a minimum of 5 concentrations is recommended. • The correlation coefficient, y-intercept, slope of the regression line should be submitted
- 14. RANGE • The 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.
- 15. ROBUSTNESS • The robustness 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. • typical variations are: - • different columns (different lots and/or suppliers); - temperature; • - flow rate.
- 16. ACCURACY • Recommended Data Accuracy should be assessed using a minimum of 9 determinations over a minimum of 3 concentration levels covering the specified range (e.g., 3 concentrations/3 replicates each of the total analytical procedure). Accuracy was determined by percent recovery study. Recovery study was done by standard addition technique in which pre analysed samples will be spiked with known standard solutions at three different concentrations over the range 50 to 150%. • Calculate the mean, standard deviation, RSD, and percent recovery for all samples. Record results on the datasheet.
- 17. Precision : repeatability, • Repeatability should be assessed using: a) a minimum of 9 determinations covering the specified range for the procedure (e.g., 3 concentrations/3 replicates each); or b) a minimum of 6 determinations at 100% of the test concentration. • Recommended Data The standard deviation, relative standard deviation (coefficient of variation) and confidence interval should be reported for each type of precision investigated.
- 18. DETECTION LIMIT • Based on Visual Evaluation • Based on Signal-to-Noise • Based on the Standard Deviation of the Response and the Slope The detection limit • (DL) may be expressed as: DL = 3.3 σ / S where σ = the standard deviation of the response S = the slope of the calibration curve The slope S may be estimated from the calibration curve of the analyte.
- 19. QUANTITATION LIMIT • Based on Visual Evaluation • Based on Signal-to-Noise Approach • Based on the Standard Deviation of the Response and the Slope • QL = 10 σ /S • where σ = the standard deviation of the response S = the slope of the calibration curve