双酯型乌头生物碱对照提取物的制备及其在川乌 - 中国药品标准杂志.doc

双酯型乌头生物碱对照提取物的制备及其应用研究 朱月1,2，王瑞忠1, 3，张聿梅1，鲁静1 (1.中国食品药品检定研究院，北京 100050；2. 北京泰德制药股份有限公司，北京100176；3. 山西中医学院，太原030619) 摘要 目的：以定量用双酯型乌头生物碱对照提取物替代单体对照品，修订川乌含量测定方法。方法：制备定量用对照提取物，标定和协作标定,对其中的乌头碱、次乌头碱和新乌头碱含量进行赋值。采用HPLC法，流动相为乙腈和0.2%冰醋酸溶液（三乙胺调pH至6.2），梯度洗脱， 检测波长235nm；并比较使用单体对照品和对照提取物的含量测定结果。结论：定量用对照提取物可替代乌头碱等单体对照品，用于川乌药材中双酯型生物碱的含量测定。关键词：双酯型乌头生物碱对照提取物；乌头碱；新乌头碱；次乌头碱；川乌 Studies on Diester Aconitum Alkaloid Reference Extract Preparation and Its Application Zhu Yue1, 2, Wang Ruizhong1, 3, Zhang Yumei1, Lu Jing1 (1. National Institutes for Food and Drug Control, Beijing 100050; 2. Beijing Tide Pharmaceutical Co.,Ltd., Beijing 100176; 3. Shanxi University of Traditional Chinese Medicine, Taiyuan 030619) Abstract Objective: To modify the assay method of Aconiti Radix by using diester aconitum alkaloid reference extract as the reference substance to substitute the single reference substance. Method: Prepared the quantitative reference extract , standardized and calibrated the contents of aconitine, hypaconitine and mesaconitine by HPLC. The mobile phase was acetonitrile and 0.2% solution of acetic acid adjusted pH to 6.2 with triethylamine, gradient elution, and the detection wavelength was 235 nm. The contents between calculated with the single reference substance and the reference extract were compared. Conclusion: The diester aconitum alkaloid reference extract could substitute the single reference substance in the assay of diester aconitum alkaloid of Aconiti Radix. Key words: diester aconitum alkaloid reference extract; aconitine; mesaconitine; hypaconitine; Aconiti Radix 川乌、草乌等乌头类药材中主要有效成分及毒性成分为双酯型二萜生物碱类,其中主要成分为乌头碱、次乌头碱和新乌头碱。《中国药典》2015年版一部川乌、草乌项下收载了乌头碱、次乌头碱和新乌头碱三种双酯型二萜生物碱的含量测定,作为质量控制指标。由于单体对照品的制备工艺复杂，耗费大量人力、物力资源，一直被认为是中药质量控制的瓶颈。而定量用对照提取物是解决对照品来源的途径之一，在国内外药典中已有应用[1-2]。本文报道双酯型乌头生物碱对照提取物的制备，对照提取物中乌头碱、次乌头碱、新乌头碱的含量定值，以及川乌药材中以对照提取物为对照的含量测定方法的建立，并比较了使用对照提取物与单体对照品含量测定的结果，两种方法的标准偏差均小于3.5%。 1仪器、试剂与材料 Waters 2695型高效液相色谱仪，2996型DAD检测器；AB-8大孔树脂（国药集团化学试剂有限公司）；硅胶GF254（青岛海洋化工厂）。 乌头碱、次