Simultaneous Estimation of Pregabalin and Methylcobalamine in Pharmaceutical Formulation by RP-HPLC Method

Abstract

A simple, precise, rapid, selective, and economic reversed phase high-performance liquid chromatography (RP-HPLC) method has been established for simultaneous analysis of PRG and MC. A Phenomenex C18 (250×4.6 mm i.d) chromatographic column equilibrated with mobile phase water: methanol (60:40 v/v) adjusted to pH 6.5 with Triethylamine (1% v/v) was used. Mobile phase flow rate was maintained at 1 ml/min and effluents were monitored at 218 nm. The sample was injected using a 20 μl fixed loop, and the total run time was 10 min. Experimental conditions such as pH of mobile phase, column saturation time, selection of wavelength, etc. were critically studied and the optimum conditions were selected. The calibration curve 50-300 μg/ml for PRG and 0.5-2.0 μg/ml for MC. The limit of quantification was 24.10 μg/ml for PRG and 0.40 μg/ml for MC respectively. This HPLC procedure is economic, sensitive, and less time consuming than other chromatographic procedures. It is an importance tool for analysis of combined dosage form.