Development of a Validated Stability Indicating Method for Quantification of Amoxicillin, Clarithromycin and Lansoprazole in Bulk and Pharmaceutical Dosage form by RP-HPLC
P. Sushma
Department of Pharmaceutical Analysis & Quality Assurance, A. U. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, AP 530003, India.
A. K. M. Pawar *
Department of Pharmaceutical Analysis & Quality Assurance, A. U. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, AP 530003, India.
M. Divya
Department of Pharmaceutical Analysis & Quality Assurance, A. U. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, AP 530003, India.
*Author to whom correspondence should be addressed.
Abstract
Objective: The main objective of the present work is to develop an efficient, unique, reliable Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method for the simultaneous quantification of Amoxicillin (AMX), Clarithromycin (CTM) and Lansoprazole (LPZ) in bulk and pharmaceutical formulations.
Methods: The chromatographic separation was achieved by using Kinetex column C18 (100 x 4.6 mm, 2.6 µm) with Buffer (2.5 g of hexane sulphonic acid and 1ml of Triethylamine which are added to 1000 ml of HPLC water and adjusted its pH at 5.0 with Ortho phosphoric acid) and acetonitrile in the ratio of 70: 30 (%v/v) as a mobile phase at flow rate of 1.0 ml/min. The column effluents were monitored by a photodiode array detector at wavelength predetermined at 240 nm.
Results: The method produced reliable results at optimized chromatographic conditions. The method was linear at concentration range of 15-225 µg/ml of AMX, 15-225 µg/ml of CTM and 0.9-13.5 µg/ml of LPZ with regression coefficients of 0.9999, 0.9999, and 0.9999 respectively. The retention times of AMX, CTM, LPZ were obtained as 1.513, 3.124, 3.770 min respectively. Results obtained for system suitability, precision, LOD and LOQ were in acceptable range and were validated according to the guidelines of the International Council for Harmonization (ICH).
Conclusion: The proposed method was validated in accordance with ICH and all the obtained results were found satisfactory and were successfully applicable to the analysis of the bulk and the pharmaceutical formulations.
Keywords: Reverse phase high performance liquid chromatography (RP-HPLC), amoxicillin, clarithromycin, lansaprazole, validation