UV AND HPLC METHOD DEVELOPMENT OF AZADIRACHTIN AND GYMNEMIC ACID IN POLYHERBAL CHURNA AND ITS VALIDATION

Dr. Vijay R. Salunkhe* and Dr. C. S. Magdum

ABSTRACT

The present research work is associated with UV and HPLC method development of Azadirachtin and Gymnemic acid in polyherbal churna and its validation .UV method for simultaneous estimation of Azadirachtin and Gymnemic acid was developed using 95% methanol as a solvent. By scanning, the each solution was in the range of 200-400 nm. 210 nm was selected as a wavelength for Azadirachtin while 217 nm for Gymnemic acid. Method was validated by linearity ,range ,accuracy, precision (intraday and interday),LOD LOQ. HPLC method for simultaneous estimation of Azadirachtin and Gymnemic acid was developed using HPLC system of JASCO UV -2075 with C18Intresil, 4.6(i.d.) x 263 nm columns. Chromatogram for marker was developed using mobile phase methanol :acetonitrile in the ratio of 60:40 v/v. Separation was achieved with good resolution as 6.3 , Retention time as 1.9417 ,3.2083,asymmetry 1.15,0.87 and theoretical plates 1180,7955 for Azadirachtin and Gymnemic acid respectively. Method was validated by parameters as linearity, range, accuracy, precision (intraday and interday), LOD LOQ ,robustnees . Churna was analyzed in comparision with standard Azadirachtin and Gymnemic acid. The quantification of Azadirachtin and Gymnemic acid in churna chromatogram was done by comparing peak areas from chromatogram of standard Azadirachtin and Gymnemic acid.

Keywords: UV, HPLC, method development, Azadirachtin, Gymnemic acid, polyherbal churna, validation.