Method validation is a process that demonstrates that a method will successfully meet or exceed the minimum standards recommended in the Food and Drug Administration (FDA) guidance for accuracy, precision, selectivity, sensitivity, reproducibility, and stability. This article discusses the validation of bioanalytical methods for small molecules with emphasis on chromatographic techniques. We present current thinking on validation requirements as described in the current FDA Guidance and subsequent 2006 Bioanalytical Methods Validation Workshop white paper.K EYWORDS: bioanalytical , validation , precision , accuracy , sensitivity , selectivity , reproducibility
INTRODUCTIONBioanalytical methods are used for the quantitation of drugs and their metabolites in biological matrices. In today ' s drug development environment, highly sensitive and selective methods are required to quantify drugs in matrices such as blood, plasma, serum, or urine. Chromatographic methods (high-performance liquid chromatography [HPLC] or gas chromatography [GC]) have been widely used for the bioanalysis of small molecules, with liquid chromatography coupled to triple quadrupole mass spectrometry (LC/MS/ MS) being the single most commonly used technology. After developing a method with desired attributes, the method is validated to establish that it will continue to provide accurate, precise, and reproducible data during study-sample analysis. Method validation is a process that demonstrates that the method will successfully meet or exceed the minimum standards recommended in the Food and Drug Administration (FDA) Guidance 1 for accuracy, precision, selectivity, sensitivity, reproducibility, and stability. The validation is performed using a control matrix spiked with the compounds to be quantifi ed. This article discusses the validation of bioanalytical methods for small molecules with emphasis on chromatographic techniques. We present current thinking on validation requirements as described in the current FDA Guidance 1 and subsequent 2006 Bioanalytical Methods Validation Workshop white paper. 2
VALIDATION PARAMETERSBioanalytical methods can be developed in the laboratory conducting the validation or obtained from another laboratory or literature. The results from a method validation can be no better than the quality of the method that was developed. Thus, before beginning the method validation, it is important that the method is set up and tested in the laboratory. For methods obtained externally, modifi cations may be necessary to achieve the desired performance of the method relative to how it was developed originally. This process will help ensure that when validation begins, chances for its successful completion (and more important, successful sample analysis) are high. During method validation, values for validation parameters are obtained. The essential parameters required according to the FDA Guidance 1 are selectivity, sensitivity, accuracy, precision, reproducibility, and stability. While obtaining these parameters, other param...