增液汤中麦冬皂苷D的纯化制备和HPLC含量测定研究

Purification preparation and determination of ophiopogonin D in Zengye decoction by using HPLC method

目的建立增液汤提取物中麦冬皂苷D的提取纯化制备方法,并建立增液汤中麦冬皂苷D含量的HPLC-UV测定方法。方法采用大孔树脂柱和硅胶柱色谱纯化制备麦冬皂苷D;采用Agilent Zorbax SB-C18柱(4.6×250 mm,5μm),柱温30℃,流动相为有机相(乙腈-甲醇3∶1)-水(56∶44),流速为1.0 ml/min,检测波长为203 nm。结果制得的麦冬皂苷D纯度大于98%,可用于含量测定研究;麦冬皂苷D在进样量0.364~5.46μg范围内与峰面积呈现良好的线性关系,其相关系数为0.999 9,平均加样回收率为98.38%,RSD值为1.28%。应用所建立的方法对3批增液汤提取物及麦冬药材中的麦冬皂苷D进行了含量测定。结论此方法准确可靠,可为增液汤提取物的质量控制提供参考。

Objective To purify and prepare Ophiopogonin D and establish the content determination method in Zengye Decoction. Methods Macroporous resin and silica column chromatography were used to purify and prepare Ophiopogonin D. The HPLC separation was achieved on an Agilent Zorbax SB-C18 analytical column（ 250 mm × 4. 6 mm,5μm） with a gradient elution of acetonitrile/methanol（ 3∶ 1） and water（ 56∶ 44） at a flow rate of 1. 0 m L/min. The detection wavelength was 203 nm and the column temperature was 30 ℃. Results The purity of Ophiopogonin D obtained was no less than 98%. The calibration curve showed good linearity between concentration of Ophiopogonin D and peak area with r value of 0. 9999. The mean recovery was 98. 38% with RSD value of 1. 28%. The developed method was successfully applied for the analysis of Ophiopogonin D in three batches of Zengye decoction. Conclusion The developed method is accurate and reliable. It can be helpful for determination of Ophiopogonin D and quality control of Zengye decoction.