Abstract

METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LINAGLIPTIN, GLIMEPIRIDE AND METFORMIN HYDROCHLORIDE IN SYNTHETIC MIXTURE BY RP-HPLC

Manali Ram*, Dr. Priyanka Yadav and Tirthraj Rupavatiya

ABSTRACT

A specific, sensitive, accurate, precise and robust Reverse Phase High Performance Liquid Chromatography method was developed and validated for the simultaneous estimation of Linagliptin, Glimepiride and Metformin Hydrochloride in a synthetic mixture. The separation was completed with Agilent Zorbax SB-CN column which operated at 25 °C column temperature. In this method development mobile phase comprised Buffer and Acetonitrile, utilized in a gradient program to ensure effective separation of the three drugs. Flow rate was 1.0 mL/min and 10 μL injection volume of was used for sample analysis. The UV detection wavelength was optimized at 225 nm, with a total run time of 25 minutes. Linearity was performed in the concentration ranges of 500–1500 μg/mL for Metformin, 1.25–3.75 μg/mL forLinagliptin and 1.0-3.0 μg/mL for Glimepiride, with give excellent correlation coefficients (r² = 0.9998 for Metformin, 0.9999 for Linagliptin and 0.9987 for Glimepiride). The developed method was validated as per the current ICH Q2 (R2) standards guidelines for accuracy/ specificity, precision, linearity, robustness, limit of detection and limit of quantification. The precision and robustness of the method were confirmed with RSD less than 2%, which indicate that the developed method was found reliable with reproducible.

Keywords: Linagliptin, Glimepiride, Metformin, HPLC, Validation.