A Comprehensive Review of RP-HPLC In Bioanalytical Method Validation and Sample Preparation

Bioanalytical method development is essential for the accurate quantification of drugs and their metabolites in biological matrices such as plasma, serum, or urine Techniques like liquid-liquid extraction (LLE), solid phase extraction (SPE), and protein precipitation are commonly used to isolate analytes from complex biological samples. Among various analytical tools, HPLC is widely preferred due to its speed, specificity, accuracy, and precision. HPLC is especially suitable for analysing low dose drugs and multicomponent formulations, making it a critical component in pharmaceutical research and development. Method development and validation are vital at all stages of drug discovery and manufacturing to ensure that analytical procedures are fit for their intended purpose. Regulatory bodies require validated methods for various stages of drug development, including Investigational New Drug (IND) applications, New Drug Applications (NDA), and Abbreviated New Drug Applications (ANDAs). Validated HPLC methods support pharmacokinetic and toxicokinetic studies, which are essential for assessing drug safety and efficacy. This article focusing on parameters such as selectivity, sensitivity, linearity, precision, accuracy, and stability. The development of robust HPLC methods contributes significantly to the success of new drug applications and ongoing pharmaceutical research.