柿叶总三萜的质量标准研究

Study on the Quality Standard of Total Triterpenoids in Diospyros kaki Leaves

目的：建立柿叶总三萜的质量标准。方法：采用原位预处理-薄层色谱（TLC）法对样品中总三萜成分（齐墩果酸、熊果酸）进行定性鉴别。以熊果酸为对照品,采用分光光度法测定总三萜的含量。采用高效液相色谱法测定齐墩果酸、熊果酸的含量：色谱柱为COSMOSIL5 C18,流动相为甲醇-0.2%磷酸（82∶18,V/V）,流速为1.0 ml/min,检测波长为210 nm,柱温为25℃,进样量为10μl。结果：齐墩果酸、熊果酸的TLC图斑点清晰,分离度好。总三萜检测质量浓度线性范围为7.52-45.12μg/ml。齐墩果酸、熊果酸检测质量浓度线性范围分别为18.4-184、18.8-188μg/ml（r=0.9999）;精密度、稳定性、重复性试验的RSD〈2%;加样回收率分别为98.83%-102.11%（RSD=1.21%,n=6）、99.00%-103.80%（RSD=1.82%,n=6）。结论：所建标准可用于柿叶总三萜的质量控制。

OBJECTIVE：To establish the quality standard of total triterpenoids in Diospyros kaki leaves. METHODS： In situ pretreatment-TLC was used for the qualitative identification of total triterpenoids （oleanolic acid and ursolic acid）. With the reference of ursolic acid, spectrophotometry was used to determine the content of total triterpenoids. HPLC was used to determine the contents of oleanolic acid and ursolic acid; the column was COSMOSIL5 C18 with mobile phase of methanol -0.2% phosphoric acid （82 ： 18, V/V） at a flow rate of 1.0 ml/min, the detection wavelength was 210 nm, the column temperature was 25 ℃, and the injection volume was 10 μl. RESULTS： TLC of oleanolic acid and ursolic acid showed clear spots and good separation. The linear range of total triterpenoids was 7.52-45.12 μg/ml. The linear range was 18.4-184 μg/ml for oleanolic acid（r=0.999 9） and 18.8-188μg/ml for ursolic acid （r=0.999 9） ; RSDs of precision, stability and reproducibility tests were lower than 2% ; recoveries were 98.83%-102.11% （RSD= 1.21%, n=6） and 99.00% -103.80% （RSD= 1.82 %, n=6）, respectively. CONCLUSIONS ： The standard can be used for the quality control of total triterpenoids in D. kaki leaves.