Validation of a Developed Analytical Method for Determination of Nateglinide and Metformin HCL in Pure and Pharmaceutical Dosage Form by Reversed-Phase High-Performance Liquid Chromatography and Its Degradation Studies

Abstract

Objective: The objective of the study was to develop a versatile analytical method and validate according to International Council for Harmonization guidelines for simultaneous estimation of nateglinide and metformin HCl by reversed-phase high-performance liquid chromatography (RP-HPLC) in active pharmaceutical ingredient and in tablet dosage form.
 Methods: Analytes, metformin and nateglinide, are separated and eluted from stationary phase luna phenyl hexyl column (150 mm ×4.6 mm, 3.5 μm) (micrometer) using polar mobile phase composed of acetonitrile:1% orthophosphoric acid 30:70 v/v, with flow rate of 1 ml/min for 8 min at ambient column temperature, at 221 nm (nanometer) detection. Acid, base, peroxide, thermal, and photolytic-induced degradation studies were performed on nateglinide and metformin.
 Results: Through isocratic flow, both metformin and nateglinide are detected at retention times of 2.79 min and 5.13 min, respectively, at 221 nm. The linearity and range of analytical method for nateglinide and metformin were 0.61–9.15 μg/ml and 7.5–75.15 μg/ml, respectively. The R2 value for nateglinide was 0.9998 and for metformin HCl was 0.9991. The limit of detection and limit of quantification for nateglinide were 0.21 μg/ml and 0.63 μg/ml and for metformin were 4.8 μg/ml and 14.6 μg/ml, respectively. The % relative standard deviation for method precision was found to be 0.22% and 0.64% for both nateglinide and metformin, respectively. The mean %recovery for nateglinide and metformin was 99.88% and 99.21%, respectively. The %thermal degradation was identified as 17.7% and 17.5% for nateglinide and metformin, respectively.
 Conclusion: The developed chromatographic (RP-HPLC) method was selective, specific, economic, precise, and accurate. Hence, it can be one of the preferred analytical methods of choice for the estimation of nateglinide and metformin by RP-HPLC in pure and in tablet dosage form.