Method Development and Validation for Simultaneous Estimation of Teneligliptin and Pioglitazone by UHPLC Method

Objectives

To develop and validate a sensitive, accurate, simple, precise and cost-effective UHPLC method for the simultaneous determination of Teneligliptin and Pioglitazone in pure and its tablet formulation form and validating this developed method as per Guidelines of ICH (Figure 1).

Materials and Methods

The chromatographic separation was done by using column Agilent C18 (2.5 μm; 4.6x100 mm ID), isocratic mobile phase consists of Methanol: 0.1% TEA (PH-6 WITH OPA) 60: 40%v/v. The flow rate of mobile phase is 0.9 mL/min. The separation was carried out at 241 nm wavelength. The current method for accuracy, precision, linearity, specificity, robustness and ruggedness was validated as per ICH guidelines.

Results

Teneligliptin and Pioglitazone retention time observed at 2.382 and 3.315 min respectively. The graphs showing peak area against concentration demonstrated linear between 2-10 μg/mL for Teneligliptin and 1.5-7.5 μg/mL for Pioglitazone. This relationship exhibited a high level of linearity with a Regression coefficient (R²) of 0.999. The determined limit of detection is 0.0843 and 0.0084 μg/mL while the limit of quantification was found to be 0.255 and 0.025 μg/mL for Teneligliptin and Pioglitazone respectively. The assay percentage of the available formulation was found to be 99.72 and 100.51 for Teneligliptin and Pioglitazone.

Conclusion

The Validation parameters indicate the effective separation of the drug substance from their degradants effectively. This developed method shows the` suitability for the routine quantitative analysis of Teneligliptin and Pioglitazone in its pure and their available pharmaceutical formulations for quality control purpose.