A Stability Indicating RP-HPLC Method Validation for Simultaneous Estimation of Linagliptin and Empagliflozin in Pharmaceutical Dosage Form

Abstract

Aims: Linagliptin and Empagliflozin is a new drug combination for the treatment of Diabetes Mellitus which is one of the oldest and lethal diseases of the mankind. Aim of the research work was to develop and validate novel, rapid, sensitive, specific, robust stability indicating analytical method for the simultaneous estimation of Linagliptin and Empagliflozin in the pharmaceutical dosage form as fixed dose formulation. Study Design: Method development and validation was performed as recommended in ICH guideline “Validation of analytical procedures: Test and Methodology Q2(R1)”. Methodology: Method develop with chromatographic parameters as Phenyl column (150mm×4.6 mm, 3.5 mm particle size), Column temperature was 30°C, HPLC system with PDA detector and mobile phase contained a degassed mixture 0.1% Perchloric Acid: Acetonitrile (75:25 v/v). The flow rate was set to 1.2 ml/min with responses measured at 226 nm, injection volume was 20 µl, and run time of 22 mins. Results: The retention time of Linagliptin and Empagliflozin was 10.2 min and 15.7 min respectively. Linearity was established in the range of 10-30 µg/ml for Linagliptin and 20-60 µg/ml for Empagliflozin with correlation coefficients more than 0.999. The percentage recoveries were between (98.4-100.5%) and (99.2-100.4%) for Linagliptin and Empagliflozin respectively. Validation parameters were evaluated according to the International Conference on Harmonization (ICH) Q2 R1 guidelines. The forced degradation studies were performed by using HCl, NaOH, H2O2, thermal and UV radiation. The developed method was successfully applied for the quantification and hyphenated instrumental analysis. Conclusion: Significance of developed method is that it can be utilize for routine or unknown sample analysis of assay of Linagliptin and Empagliflozin in pharmaceutical dosage form developed by various Pharmaceutical Industry.