瓦布贝母中生物碱的含量测定  

Determination of alkaloids in Fritillaria wabuensis

作　　者：黎萍 李敏 蒋静 吴雪婷 付春梅[1] 刘秀秀[1] 晁若冰[1] LI Ping;LI Min;JIANG Jing;WU Xueting;FU Chunmei;LIU Xiuxiu;CHAO Ruobing(West China School of Pharmacy,Sichuan University,Chengdu,Sichuan,610041 P.R.China;Sichuan Guoqing Chuanbeimu Biotechnology Co.,Ltd.,Chengdu,Sichuan,624000 P.R.China)

机构地区：[1]四川大学华西药学院,四川成都610041 [2]四川国青川贝母生物科技股份有限公司,四川成都624000

出　　处：《华西药学杂志》2020年第6期655-659,共5页West China Journal of Pharmaceutical Sciences

基　　金：道地药材川贝母、川芎、附子规模化种植及精准扶贫示范研究项目(2017YFC1701800)。

摘　　要：目的测定瓦布贝母中3种特征性生物碱西贝碱苷、西贝碱和贝母辛以及总生物碱的含量,并评价瓦布贝母栽培品的质量。方法采用HPLC-ELSD法测定3种特征性生物碱的含量,色谱柱为Inertsil ODS-SP柱,流动相(A)为0.1%三氟乙酸水溶液,流动相(B)为乙腈,梯度洗脱,ELSD漂移管温度为98℃,气体流速3.0 L·min^-1,不分流进样。参考2015年版《中国药典》中川贝母项下含量测定的方法,采用酸性染料比色法测定总生物碱的含量;将样品中生物碱苷类成分经稀硫酸于90℃水浴中,加热2 h水解为其苷元,同时对测定方法进行了改进和优化。结果HPLC-ELSD法专属好,待测的3种生物碱均可以完全分离;西贝碱苷、西贝碱和贝母辛进样量分别为0.59~5.90、0.96~9.60、0.54~5.40μg,峰面积对数与进样量对数的线性关系良好(r>0.9999);进样精密度、日内精密度和日间精密度峰面积对数值的RSD均小于0.5%;方法回收率分别为97.8%、99.9%、100.3%。优化后的酸性染料比色法能够测定生物碱及其苷的总量,线性关系良好(r>0.9999),重复性试验的RSD=2.26%,回收率为100.7%。结论所用方法专属、简便、准确,适用于测定瓦布贝母中特征性生物碱和总生物碱的含量,可为瓦布贝母栽培品的质量评价提供依据。OBJECTIVE To determine three characteristic alkaloids of imperialine-3-β-D-glucoside,imperialine,peimisine and total alkaloids in cultivated Fritillaria wabuensis,and to evaluate its quality.METHODS An HPLC-ELSD method was developed to determine the three characteristic alkaloids.It was conducted on an Inertsil ODS-SP column with 0.1%TFA aqueous solution(mobile phase A)and acetonitrile(mobile phase B)eluting in a gradient program.The temperature of drift tube of ELSD was 98℃,and the gas flow rate was 3.0 L·min-1 with splitless injection.Referring to the assay method of F.cirrhosa in the Chinese Pharmacopoeia(2015),the acid dye colorimetry was used for determination of the total alkaloids in F.wabuensis.Alkaloid glycosides in sample were hydrolyzed into aglycones by heating in 90℃water bath for two hours with dilute sulfuric acid,and the assay method was improved and optimized.RESULTS The HPLC-ELSD method was specific.Linear range of imperialine-3-β-D-glucoside,imperialine,peimisine were respectively within 0.59-5.90μg,0.96-9.60μg and 0.54-5.40μg.Their correlation coefficients of the linear regression equations of logarithm of peak area and logarithm of sample size were greater than 0.9999.The RSD of injection precision,intra day precision and inter day precision of the logarithm of peak area were all less than 0.5%,and the recoveries were 97.8%,99.9%and 100.3%,respectively.The alkaloids and glycosides in sample were both determined by the optimized acid dye colorimetry.It had a good linear relationship(r>0.9999).The RSD of repeatability test was 2.26%,and the recovery rate was 100.7%.CONCLUSION The methods are specific,simple and accurate,which are suitable for determination of the characteristic alkaloids and total alkaloids in cultivated F.wabuensis and provide the basis for its quality evaluation.

关 键 词：瓦布贝母 西贝碱苷 西贝碱 贝母辛 总生物碱 含量测定 HPLC-ELSD 酸性染料比色法 质量评价 

分 类 号：R917[医药卫生—药物分析学]