An HPLC method has been developed for routine analysis of negundoside in Vitex negundo Linn. leaf extract. Separation was performed on a C 18 column with 65:35 ( v / v ) buffer solution (containing 0.01 M potassium dihydrogen orthophosphate and 0.01 M heptane sulphonic acid sodium salt; pH 3.0)-methanol as mobile phase. Detection and quantification were achieved by use of UV and photo diode array detectors operated at 251 nm. Response was a linear function of negundoside concentration in the range 1–37.5 μg mL -1 (correlation coefficient 0.9996). The method was validated for system and method precision, accuracy, linearity, and solution stability. The method is sensitive, simple, rapid, accurate, and precise and can therefore be used for routine qualitycontrol analysis of Vitex negundo Linn. leaf powder, including quantitative analysis of negundoside.
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