Method development and validation for estimation of melatonin in tablet by using RP-HPLC

Jadhav Khanderao Rajaram*, Sonawane Vijayraj Narendra, Jadhav Shivraj Popat, Somvanshi Deepak Balasaheb, Patil, Chandrashekhar Dinkar

Sheeshakti Shaikshanik Sanstha’s Divine College of Pharmacy, Satana, Nashik, Maharashtra, India

ABSTRACT

In the present work new, Simple Reversed-Phase High Performance Liquid Chromatographic method was developed and validated for the estimation of Melatonin in tablets. A sharp peak was obtained 7.125 min using Methanol: Phosphate buffer (PH 3.0) in the ratio of 40:60 as mobile phase at a flow rate of 1.0 ml/min, on HPLC Waters 515 system containing UV detector with Empower software. The separation was achieved on the C18 analytical column (250 mm × 4.6 mm, 5.0 μm). The detection was carried out at 223 nm. The method gave good resolution and suitable retention time. The results of the analysis in this method were validated in terms of specificity, accuracy, precision, linearity, and range. The method was found to be reliable, reproducible, rapid, and economic. The RP-HPLC method developed in this study showed specificity and selectivity with linearity in the working range and good precision and accuracy, making it very suitable for the estimation of Melatonin in tablets.

Keywords:

Melatonin, Assay, RP-HPLC, Method validation