Abstract

BIOANALYTICAL VALIDATION OF ANTIMICROBIAL DRUGS IN RABBIT PLASMA USING REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY: A REVIEW

Dantinapalli V. L. Sagar*, Sanjay Saxena

Abstract

Bioanalytical method development and validation are essential for the reliable quantification of antimicrobial drugs in biological matrices and for generating dependable pharmacokinetic and toxicokinetic data. Reverse-phase highperformance liquid chromatography (RP-HPLC) is widely used for bioanalytical investigations because of its reproducibility, versatility, sensitivity, and suitability for quantitative analysis. Rabbit plasma is frequently employed in preclinical studies, although endogenous proteins, lipids, salts, and metabolites can interfere with chromatographic determination and therefore require appropriate sample preparation. This review discusses the principles involved in developing and validating RP-HPLC methods for the determination of antimicrobial drugs in rabbit plasma. Important aspects include plasma sample preparation, chromatographic optimization, calibration characteristics, selectivity, specificity, accuracy, precision, recovery, matrix effects, carryover, dilution integrity, and stability. Previous literature has demonstrated the importance of systematic bioanalytical method development for pharmacokinetic, bioavailability, bioequivalence, and toxicokinetic studies (Tiwari & Tiwari, 2010; Nunsavathu & Thangabalan, 2024). Current regulatory recommendations, particularly ICH M10, provide a standardized framework for demonstrating the reliability of bioanalytical procedures (ICH, 2022). Properly developed and validated RP-HPLC methods can therefore provide reliable analytical support for preclinical evaluation of antimicrobial drugs.

Keywords: Antimicrobial drugs, rabbit plasma, bioanalytical validation, RP-HPLC, pharmacokinetics, sample preparation, chromatography.