DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR QUANTITATION OF AMOXICILLIN TRIHYDRATE IN BULK AND PHARMACEUTICAL FORMULATIONS USING BOX-BEHNKEN EXPERIMENTAL DESIGN

Abstract

A rapid and sensitive novel reversed phase-HPLC method with UV detection at 229 nm was developed for routine analysis of amoxicillin trihydrate in bulk drugs and pharmaceutical formulations (Damoxy and Amoxil). Chromatographic separation was performed on a Capacel Pak C₁₈ type MG column using isocratic elution of mobile phase containing mixture of phosphate buffer and methanol (50:50, %v/v) with flow rate at 1.0 ml/min and pH 3.0. The method was validated for linearity (r² = 0.999), accuracy, precision, sensitivity, system suitability, selectivity, and robustness testing. A 3²-factorial design was employed utilizing box-behnken experimental design for the validation of robustness using three factors such as composition of mobile phase (X₁), flow rate (X₂), pH (X₃); while peak area (Y₁) and retention time (Y₂) were used as responses. System showed minor change in the mobile phase composition and flow rate affects the responses produced, while pH has no significant effect on the responses. Finally the developed experimental design was validated by statistical analysis using ANOVA showed that the proposed method was simple, sensitive, and highly robust for routine analysis of the amoxicillin trihydrate.

Keywords:

• amoxicillin trihydrate
• experimental design
• HPLC
• linearity
• robustness
• validation

ACKNOWLEDGMENT

The authors want to express their thanks and gratitude to Ranbaxy Lab. Ltd., Gurgaon, India, Aurobindo Pharma Ltd., (Hyderabad, India) for providing the gift sample of drugs and Central Instrumental Facility (CIF) Laboratory, Jamia Hamdard, New Delhi, India for carry out the present work.