Determination of Pulsatilla Saponin D in Pulsatillae Radix from Different Areas

Abstract:

Objective: To establish a determination method for pulsatilla saponin D in Pulsatillae Radix. Method: The content of pulsatilla saponin D was determined by RP-HPLC,chromatographic conditions were as follows:Elite Hypersil ODS₂ C₁₈ column(4.6 mm×250 mm,5 μm),mobile phase of acetonitrile-water-formic acid(38:62:0.1),flow rate 1.0 mL·min^-1,detection wavelength 203 nm,column temperature 25℃.Content difference of pulsatilla saponin D in Pulsatillae Radix from different areas was investigated. Result: Pulsatilla saponin D had good linear relationship in 0.311 2-3.112 μg(r=0.999 9),RSD of instrument precision was 1.36%,RSD of method repeatability was 1.65%,samples solution was stable within 24 h,the average recovery was 98.07%(RSD 0.64%).The content of pulsatilla saponin D in Pulsatillae Radix from Jilin was the highest(0.256%),which was 20 folds higher than Pulsatillae Radix from Guangdong. Conclusion: This method was simple,accurate and repeatable.The content of pulsatilla saponin D in Pulsatillae Radix from different areas had much difference,so materials choice should be careful.