A Validated Stability Indicating LC Method for Nateglinide

ABSTRACT

A simple, isocratic, rapid and accurate reverse phase high-performance liquid chromatography (RP-HPLC) method was developed for the quantitative determination of Nateglinide. The developed method is also applicable for determination of related substance in bulk drugs. The chromatographic separation was achieved on a Hypersil C18 (250 × 4.6 mm 5 μm) column using aqueous mixture of 0.025 M potassium hydrogen phosphate and 0.1% triethyl amine, v/v (pH 3.0 with dilute phosphoric acid)—methanol (25:75, v/v) as a mobile phase. Solution concentrations were measured on a weight basis to avoid the use of an internal standard. The chromatographic resolutions between Nateglinide and its potential impurities A and B were found to be greater than four. Forced degradation studies were performed for Nateglinide using acid (0.5 N hydrochloric acid), base (0.5 N sodium hydroxide), oxidation (3% hydrogen peroxide) heat (60°C) and UV light (254 nm). The limit of detection and limit of quantification of Nateglinide, impurities A and B were found to be 0.05 and 0.15 μg /mL, respectively for 20 μL injection volume. The percentage recovery of Nateglinide was ranged from 98.4 to 100.9. The percentage recovery of impurities in Nateglinide sample was ranged from 96.8 to 103.5. The developed RP-HPLC method was validated with respect to linearity, accuracy, precision, robustness, and forced degradation studies prove the stability indicating power of the method.