Validation of Chromatographic Methods of Analysis

Analytical method validation is performed by regulated laboratory, and deals with the testing of significant method characteristics to ensure that under routine use, the analytical method is accurate, precise, specific, reproducible, and rugged over the whole specified range, for which an analyte (s) is determined. The validation of chromatographic methods should be performed before the first routine use of the procedure, and a validation of methods of analysis is crucial in all phases of drug development. The necessity for validation in analytical laboratories is derived from regulations as the current Good Manufacturing Practices (cGMP), Good Laboratory Practices (GLP), and the Good Clinical Practices (GCP). Other regulatory requirements are found in quality and accreditation standards as the International Organization for Standardization (ISO) 9000 series, ISO 17025, the European Norm (EN45001) , United States Pharmacopoeia (USP) , Food and Drug Administration (FDA ) and Environmental Protection Agency (EPA). Analytical procedures are classified as being compendial or non‐compendial in character. Compendial methods are considered to be valid, but their suitability should be verified under actual conditions of use. For non‐compendial procedures, the performance parameters determine specificity/selectivity, linearity, accuracy, precision (repeatability and intermediate precision), detection limit (DL) , quantitation limit (QL) , range, ruggedness, and robustness. In chromatography techniques, selectivity can be proved by the existence of good separation between the analyte and the other components. The accuracy of an analytical method is given by the extent, by which the value obtained deviates from the true value.