Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF MELATONIN AND PYRIDOXINE IN TABLET DOSAGE FORM BY HPLC

Sangeeta Rathod*, Ankita Bhavsar, Bhagirath Patel

ABSTRACT

A simple, rapid and accurate High performance Liquid Chromatography method was developed and validated for simultaneous estimation of Melatonin and Pyridoxine in tablet dosage form. The High performance Liquid Chromatography method has shown adequate separation of Melatonin and Pyridoxine in its tablet Dosage form. The separation was achieved BDS hypersil C18, with an isocratic system of 0.2M KH2PO4 buffer (pH - 4): methanol: TEA in the ratio of 70:30:0.1 v/v. The mobile phase at a flow rate of 1.0 ml/min, Injection volume 20μl and wavelength of detection used was 244 nm. The retention time for Melatonin and Pyridoxine were obtained as 3.760 min and 6.107 min, respectively. The linearity of the proposed method was investigated in the range of 3-9 μg/ml and 10-30μg/ml for Melatonin and Pyridoxine respectively. Correlation coefficient was 0.9939 and 0.9995 for Melatonin and Pyridoxine respectively. The developed method was validated as per International conference Harmonization (ICH) guideline for its accuracy, precision, Limit of detection and Limit of quantitation.

Keywords: Melatonin, Pyridoxine, HPLC method development, Validation.