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目的建立锦鸡儿中白桦脂酸,齐墩果酸,熊果酸和羽扇豆醇4种萜类化合物定量分析的高效液相色谱方法,并考察了它们的体外α-葡萄糖苷酶活性。方法采用Agilent Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为甲醇-0.1%磷酸水溶液(85∶15),流速为0.6 m L·min~(-1),柱温25℃,检测波长204 nm,对锦鸡儿中的白桦脂酸、齐墩果酸和熊果酸进行含量测定;采用Inert Sustain C18色谱柱(4.6 mm×150 mm,5μm),流动相为乙腈-水(70∶30),流速0.8 m L·min~(-1),柱温30℃,检测波长204 nm,测定锦鸡儿中羽扇豆醇的含量。结果白桦脂酸、齐墩果酸、熊果酸和羽扇豆醇在0.56~5.58、0.42~4.20、0.19~1.92和6.10~61.00μg内线性关系良好,平均加样回收率分别为99.79%、98.51%、98.05%和99.47%,RSD分别为1.53%,1.74%,1.78%和1.94%。结论本试验建立的方法准确可靠,可用于锦鸡儿药材的质量控制;除白桦脂酸外,其他3种化合物均表现出良好的对α-葡萄糖苷酶抑制活性,有望开发成为新的降血糖药物。
OBJECTIVE To establish a method for the quantitative analysis of four terpenoids of betulinic acid, oleanolic acid, ursolic acid and lupeol in Caragana microphylla and to study their in vitro α-glucosidase activity. Methods The mobile phase consisted of methanol-0.1% phosphoric acid solution (85:15) with a flow rate of 0.6 mL · L -1 at an Agilent Eclipse XDB-C18 column (4.6 mm × 250 mm, 5 μm) The content of betulinic acid, oleanolic acid and ursolic acid in Caragana korshinskii were determined by Iodora Sustain C18 column (4.6 mm × 150 mm, 5 μm) at a detection wavelength of 204 nm. The mobile phase consisted of acetonitrile-water (70:30), the flow rate was 0.8 m L · min -1, the column temperature was 30 ℃, the detection wavelength was 204 nm, and the content of lupulonol in Caragana microphylla was determined. Results The results showed that betulinic acid, oleanolic acid, ursolic acid and lupeol had good linearity in the ranges of 0.56 ~ 5.58,0.42 ~ 4.20,0.19 ~ 1.92 and 6.10 ~ 61.00μg with the average recoveries of 99.79%, 98.51 %, 98.05% and 99.47%, respectively. The RSDs were 1.53%, 1.74%, 1.78% and 1.94% respectively. Conclusion The method established in this study is accurate and reliable and can be used for the quality control of Caragana microphylla; besides betulinic acid, the other three compounds all show good α-glucosidase inhibitory activity and are expected to be developed into new hypoglycemic drugs .