Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SELECTED ANTI INSOMNIA DRUG IN BULK AND FORMULATED PRODUCT

Ms. Supriya Vijay Kamble*, Dr. Mahesh J. Patil, Dr. Mahesh J. Patil and Ms. Sonal Nalkar

Abstract

A simple precise, rapid and accurate UV-Visible Spectrophotometer and High Performance Liquid Chromatography Method was developed for simultaneous estimation of Melatonin and L- Theanine, in pharmaceutical dosage form. This method was carried out on Inertsil- ODS C18 (250x4.6 mm, 5μ) column with a mobile phase consisting of Methanol: Buffer (60:40v/v). me of The retention time of Melatonin and L-Theanine is 4.25min and 5.99 min respectively. The flow rate was 1.0 ml/min with UV detection at 278 nm. The linear regression data for the linearity plot showed good relationship with correlation coefficient value for Melatonin and L-Theanine were r2= 0.999 and r2= 0.999. The stock solution equivalent to20ppm to 80ppm with respect to both drugs are prepared in combination of Melatonin and L-Theanine. The relative standard deviation for intra-day precision has been found to lower than 2.0%.0 The method is validated According to ICH guidelines. The development was validated in terms of specificity, estimated by accuracy, linearity, limit of detection, limit of quantification and solution stability. Sensitivity was estimated by determination of LOD and LOQ and values are found to be 0.25 and 0.77 for Melatonin and 0.34 and 1.05 for L-Theanine. The purposed method can be successfully applied for the determination of these drugs in combined dosage forms.

Keywords: melatonin, L-Theanine, UV Visible Spectrophotometer, High Performance Liquid Chromatography, Method Validation, ICH guidelines.