METHOD DEVELOPMENT AND VALIDATION OF AMOXICILLIN TRIHYDRATE BY HPTLC IN BULK AND PHARMACEUTICAL DOSAGE FORM

Amoxicillin Trihydrate is an extensively used antibiotic, and the accurate determination of its content in bulk and pharmaceutical dosage forms is of greatest importance. This thesis focuses on the development and validation of a High-Performance Thin-Layer Chromatography (HPTLC) method for the analysis of Amoxicillin Trihydrate. The study begins with an inclusive review of the existing analytical methods for Amoxicillin Trihydrate, highlighting the advantages and limitations of each technique. HPTLC is selected as the method of choice due to its simplicity, cost-effectiveness, and potential for routine analysis in pharmaceutical laboratories. The Chromatographic separation was carried out on Merck, Germany, TLC plates of silica gel 60 F_254, 10 cm x10 cm using n-Hexane: ethyl acetate in the ratio of 7:3 (% v/v) as a mobile phase followed by densitometric measurements at 230 nm. The following method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantification, and specificity according to the ICH (International Conference on Harmonization) guidelines. The calibration curve and results were found to be linear with regression equation of y = 1.2832x + 3341, between 2000 to 12000 ng/ band for Amoxicillin trihydrate respectively, with significantly high value of regression coefficient is (r²) > 0.9936 with linear and homoscedastic residuals. The limit of quantification and limit of detection was found to be 1107.217 ng/band and 365.382 ng/ band respectively. The validated HPTLC method was successfully applied for the determination of Amoxicillin trihydrate in the commercial formulation.