DEVELOPMENT, STANDARDIZATION AND VALIDATION OF HPLC METHOD FOR ESTIMATION OF βSITOSTEROL IN VITEX NEGUNDO SEED EXTRACT
Journal Title: Int J of Pharm Rev& Res - Year 2015, Vol 5, Issue 2
Abstract
A simple, precise and accurate high-performance liquid chromatographic method has been developed for determination of β-sitosterol in the hydroalcoholic extact of Vitex negundo Linn. A Reverse phase HPLC column CrestPack RP C18 (Make: Jasco; 250 mm x 4.6 mm I.D; particle size 5μm) was used as stationary phase with a mobile phase comprising of acetonitrile : methanol (70:30) v/v at a flow rate of 1.0 mL min-1 and UV detection at 210nm with a run time of 6 mins. The proposed method was validated for linearity, accuracy, precision, LOD and LOQ. LOD for beta-sitosterol was found to be 0.5μg/mL whereas LOQ for the same was found to be 1.5μg/mL. The method was found to be linear over the range 1 μg/mL to 5μg/mL with coefficient of regression 0.9732 for betasitosterol. Intra-day and inter-day precision studies showed % RSD to be less than 2 and hence the method was precise. Stock solution stability studies were carried out for beta-sitosterol. Stability samples in methanol after a week revealed that all beta-sitosterol in solution form were stable. The validated HPLC method can be used for a routine quality control analysis and quantitation of β-sitosterol from Vitex negundo Linn.
Authors and Affiliations
Divya K Shetty , Swati S Patil
Keywords
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