藏药二十一味寒水石丸中的总鞣质、没食子酸和鞣花酸的含量测定

Determination of total tannins, gallic acid and ellagic acid in Ershiyiwei Hanshuishi pill

目的：建立二十一味寒水石丸中总鞣质、没食子酸和鞣花酸的含量测定方法.方法：以干酪素作为吸附剂,碱性条件下显色,以磷钼钨酸为氧化剂,用紫外可见分光光度法测定鞣质含量;采用HPLC法同时测定没食子酸和鞣花酸的含量.使用迪马Diamonsil-2 C18（4.6 mm×250mm,5μm）色谱柱,以乙腈-0.1磷酸水梯度洗脱,体积流量分别为1.0mL/min,柱温为30℃;检测波长270nm,测定.结果：以没食子酸计的鞣质含量在0.5～ 5μg·ml-1的范围内线性关系良好（r=0.9991）,平均回收率为95.77％,RSD为0.70％（n=6）;没食子酸在0.32～1.6μg范围内线性关系良好（r=0.9999）,平均回收率为98.70％,RSD为1.01％（n=9）,鞣花酸在0.372～.1.86μg范围内线性关系良好（r=0.9999）,平均回收率为99.63％,RSD为0.75％（n=9）.结论：本方法准确、可靠简便,具有较强专属性,可用于常用藏药二十一味寒水石丸中总鞣质、没食子酸和鞣花酸的含量测定.

Objective： To establish a stable method for determination of total tannins, gallic acid and ellagic acid in Ershiyiwei Hanshuishi pill. Methods： The UV-vis method was used to determine the content of total tannins of Ershiyiwei Hanshuishi pill with casein as sorbent, phosphorus molybdenum-tungsten acid as oxidant. Gallic acid and ellagic acid were determined by HPLC. The Diamonsil-2 Cls column（4.6 mmx250 mm, 5μm） column was used with gradient elution. Mobile phase was composed of acetonitrile-0.1% phosphoric acid in solution, and the flow rate was 1 mL/min. The column temperature was kept at 25 ℃.The detection wavelength was set at 270 nm. Results： The linear relationship of standard curve was good within the range of 0.5-5μg/mL, r = 0.9991,and the average recovery was 95.77% with RSD of 0.70% （n=6）. The linear range of gallic acid was 0.32-1.6μg（r=0.9999）, and the average recovery rate was 98.70%, with RSD of 1.01%. The linear range of ellagic acid was 0.372-1.86txg（r=0.9999）, the average recovery rate was 99.63% with RSD of 0.75% respectively. Conclusion： The method is accurate, reliable, specific and can be used to determine the total tannins, gallic acid and ellagic acid in Ershiyiwei Hanshuishi pill.