藏药白花龙胆质量标准研究

Quality standard study on Tibetan medicine Gentianae Szechenyii Flos

为了对藏药白花龙胆药材进行有效的质量控制,该研究建立了白花龙胆药材的质量控制方法与标准。参照《中国药典》2010年版附录相关方法,对白花龙胆药材的水分、灰分及醇溶性浸出物进行测定;采用薄层色谱法,以硅胶G为薄层板,乙酸乙酯-甲醇-水-甲酸（7∶1.5∶1∶0.2）为展开剂,以大花龙胆及乌奴龙胆苷A为对照物质,建立其薄层鉴别方法;采用HPLC建立主要成分乌奴龙胆苷A的含量测定方法：采用Ultimate XB-C18色谱柱,以甲醇-0.02%磷酸水（52∶48）为流动相,流速1.0 m L·min-1,柱温25℃;检测波长240 nm。结果以三氯化铝试液为显色剂,白花龙胆中各成分在紫外光灯（366 nm）下检视,能得到较好的分离,在与对照药材和对照品的相应位置上均显示相同的颜色荧光斑点;含量测定方法学研究结果表明,乌奴龙胆苷A在10.01~400.32 mg·L-1呈良好的线性关系,回归方程为Y=1 539.5X-33.339（r=0.999 7）,平均回收率为99.68%（RSD 1.92%）。19批白花龙胆药材样品的测定结果表明,乌奴龙胆苷A的质量分数为14.48~31.51 mg·g-1;醇溶性浸出物的结果为24.21%~31.60%,水分为11.25%~12.74%;10批代表性样品总灰分为4.64%~6.12%。以上结果表明建立的标准具有较好的专属性和准确性,可用于白花龙胆药材的质量控制。

In order to efficiently control the quality of the Tibetan medicine Gentianae Szechenyii Flos, the quality standard was established in this study. The tests of water content, total ash and ethanol-soluble extractives of the crude drugs were carried out based on the methods recorded in appendix of Chinese Pharmacopeia（2010 edition, volume 1 ）. The TLC method was established by using reference drug and gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid （7： 1.5： 1 ： 0. 2） as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on a Ultimate XB-C18 （4. 6 mm ×250 mm,5 μm） column, using methanol-water（0.02% phosphoric acid） （52：48） as the mobile phase at a flow rate of 1.0 mL · min^-1. The column temperature is 25 ℃ and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light（366 nm）. The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 10. 01-400. 32 mg · L^-1 with the regression equation of Y= 1 539.5X -33. 339（r =0. 999 7）, and the average recovery was 99. 68% （RSD 1.92% ）. The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 19 batches samples were varied in the ranges of 14.48-31.51 mg· g^-1, 11.25%-12. 74% and 24. 21%-31.60% , respectively, and total ash was 4. 64% -6. 12% detected from 10 batches samples. The recommended standards of quantitative indexes are tlaat the mass fractions of gentiournoside A and extractives are not less than 15.0 mg · g^-1 （ 1.5% ） and 21.0% , respectively; the water and total ash are not more than 13.0% and 6.0%, respectively.