Development and Validation of HPTLC Method for Simultaneous Estimation of Alogliptin Benzoate and Pioglitazone Hydrochloride in Bulk Drugs and Combined Dosage Forms

Abstract

A new, simple, precise, accurate and selective high performance thin-layer chromatographic (HPTLC) method has been developed and validated for the simultaneous estimation of Alogliptin benzoate (ALG) and Pioglitazone hydrochloride (PIO) in bulk and tablet dosage form. Chromatographic separation was carried out on Merck HPTLC aluminium sheets of silica gel 60F254 using Acetonitrile: 1% ammonium acetate in Methanol (4.5:5.5 v/v) as mobile phase followed by densitometry analysis at 254 nm. The reliability of the method was assessed by evaluation of linearity (500-3000 ng/spot for Alogliptin benzoate as well as for Pioglitazone). The accuracy of method was assessed by recovery studies and was found to be within range of 98-102% for both Alogliptin benzoate and Pioglitazone. The developed method was validated with respect to linearity, accuracy (recovery), and precision. The results were validated statistically as per ICH Q2 R1 guidelines and were found to be satisfactory. Due to non-availability of product, the simulation was done by using Piosys® tablets (Pioglitazone 30mg) and API of Alogliptin benzoate. The proposed method was successfully applied for the determination of Alogliptin benzoate and Pioglitazone in the mixture.