Abstract

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF EPALRESTAT AND METHYLCOBALAMIN IN TABLET DOSAGE FORM

*Mili Patel, Bhakti J. Ladva, Vijay Mahida, Dr. Bhavesh S. Nayak, Hetal K. Patel and Bhumi R. Patel

ABSTRACT

A sensitive, selective and precise RP-HPLC has been developed and validated for the simultaneous determination of Epalrestat and Methylcobalamin tablet formulation. The method utilized spherisorb CNRP C18 column (250 x 4.6 mm id, 5 μm particle size) as the stationary phase while acetonitrile: 0.05 M Potassium Dihydrogen Phosphate buffer (60:40 v/v) pH 4.0 was adjusted using ortho phosphoric acid used as mobile phase. The retention time for Epalrestat and Methylcobalamin were found to be 3.26 and 4.92 minutes, respectively. Analysis was carried out in absorbance mode at 292 nm. Linearity of the developed method followed beer’s law. It found to be linear in range of 10-60 μg/ml for Epalrestat and 5-30 μg/ml for Methylcobalamin with regression co-efficient of 0.9994 and 0.9991 for Epalrestat and Methylcobalamin respectively. %RSD was found to be less than 2 for precision. The LOQ was found to be 0.098 μg/ml for Epalrestat and 0.18 μg/ml for Methylcobalamin and LOD was found to be 0.29 μg/ml for Epalrestat and 0.55 μg/ml for Methylcobalamin. The method was validated as per ICH guideline and it was found to be accurate, precise and robust. Marketed formulation was analyzed successfully.

Keywords: Epalrestat, Methylcobalamin, RP-HPLC, Validation, ICH Guideline.