钩藤总生物碱提取工艺的优化  

Optimization of the extraction technology of total alkaloids from Uncaria rhynchophylla

作　　者：王明阳 唐颖 刘芳 陈彩霞 石宝生 袁晓艳 Wang Mingyang;Tang Ying;Liu Fang;Chen Caixia;Shi Baosheng;Yuan Xiaoyan(Department of Analytical Chemistry,School of Pharmacy,Zunyi Medical University,Zunyi Guizhou 563099,China)

机构地区：[1]遵义医科大学药学院分析化学教研室,贵州遵义563099

出　　处：《遵义医科大学学报》2020年第5期656-661,共6页Journal of Zunyi Medical University

基　　金：贵州省教育厅青年科技人才成长项目(NO:黔教合KY字[2016]211);贵州省中医药管理局中医药、民族医药科学技术研究课题(NO:ZYY-2018-096);遵义医科大学大学生创新创业项目(NO:XYDC-2018-096)。

摘　　要：目的获取中药钩藤总生物碱的最优化提取工艺条件。方法采用邻苯二甲酸氢钾-盐酸缓冲液-溴甲酚绿法测定钩藤总生物碱含量,以钩藤总生物碱得率为评价指标,考察乙醇体积分数、提取时间、液料比和温度四个因素对超声提取钩藤中总生物碱含量的影响,按照均匀设计法优化提取工艺。同时,采用高效液相色谱法,以乙腈-0.1%三乙胺磷酸盐溶液(pH 7)进行梯度洗脱,对钩藤中钩藤碱、异钩藤碱、去氢钩藤碱、异去氢钩藤碱进行含量测定。结果钩藤超声浸提最佳条件为:40%乙醇作为提取溶剂,超声时间1 h,液料比25∶1(mL/g),温度30℃,总生物碱得率0.40%。在高效液相色谱法的条件下,4种生物碱分离良好,钩藤中四种生物碱的总含量0.34%,占总生物碱含量的85%。方法学验证结果显示,4种成分在各自范围内线性关系良好,精密度、稳定性、重复性良好,RSD值均≤3%,平均加样回收率在88.2%~91.5%。结论采用均匀设计法优选最佳的钩藤总生物碱的提取工艺,稳定合理。本研究为钩藤资源的充分利用提供了理论和方法上的参考,为进一步研发钩藤产品奠定基础。Objective To obtain the optimal extraction conditions of the total alkaloids from Uncaria rhynchophylla(U.rhynchophylla).Methods Determination of total alkaloids from U.rhynchophylla was performed by potassium hydrogen phthalate-hydrochloric acid buffer solution-bromocresol green method.Taking the total alkaloids of U.rhynchophylla as an evaluation index,four factors,such as the ethanol-concentration,the extraction time,the liquid-to-material ratio and the temperature,were investigated to optimize the extraction process according to the uniform design method.Meanwhile,high performance liquid chromatography(HPLC)was used to determine the contents of 4 alkaloids in U.rhynchophylla by gradient elution with acetonitrile-0.1%triethylamine phosphate solution(pH=7).Results The optimal conditions for cold water extraction of U.rhynchophylla were:40%ethanol as extraction solvent,ultrasonic extraction time of 1h,liquid-to-material ratio of 25:1(mL/g),temperature of 30℃and total alkaloids of 0.40%.Under the conditions of HPLC,the four alkaloids were well separated and the total content of the four alkaloids from U.rhynchophylla were 0.34%,which accounted for 85%of the total alkaloids.Methodological verification results showed that the four components had a good linear relationship in their respective ranges with good precision,stability and repeatability.RSD values were≤3%and the average sample recovery was 88.2%~91.5%.Conclusion This study showed that the method of uniform design was stable and feasible and could optimize the extraction process of total alkaloids from U.rhynchophylla.This study further provided theoretical and methodological references for the full utilization of U.rhynchophylla and laid the foundation for future development of U.rhynchophylla.

关 键 词：钩藤 生物碱类化合物 均匀设计 高效液相色谱法 定量分析 

分 类 号：R917[医药卫生—药物分析学]