基于5种生物碱定量分析的马钱子总碱提取工艺优化

Extraction Process Optimization of Total Alkaloids Based on Simultaneous Determination of Five Alkaloids in Strychnos nux-vomica L.

首先以乙腈-水为流动相,梯度洗脱,体积流量1 mL/min,柱温35℃,检测波长260 nm,建立同时测定5种马钱子生物碱（士的宁、马钱子碱、士的宁氮氧化物、马钱子碱氮氧化物与番木鳖次碱）含量的反相HPLC法;然后以上述5种生物碱的总量为指标,采用L9（33）正交实验法,对马钱子总碱的回流提取工艺条件（溶剂用量、质量浓度和提取次数）进行优化.结果表明,上述5种马钱子生物碱质量浓度线性范围依次为14.64～468.48、11.625～372.0、0.555～17.76、0.891～28.512μg/mL与2.5～80.0 μg/mL,回收率分别为100.3％、101.7％、97.7％、98.8％与99.6％.优化后马钱子总碱的回流提取工艺条件为：加入20倍量体积分数70％的酸性乙醇,回流提取4次,每次1h.在此提取条件下,马钱子提取物中5种生物碱总质量分数达3.633％.反相HPLC法同时测定5种马钱子生物碱含量的方法简便、准确;以此作为马钱子总碱提取工艺优化的指标更为全面、可靠.

The RP-HPLC analysis of was performed on a Merck C18 column （4.6×250 mm,5 μm） for simultaneous determination of the contents of five alkaloids （strychnine,brucine,strychnine Noxide,brucine N-oxide and vomicine） in Strychnos nux-vomica L..The mobile phase was a mixture of acetonitrile （A） and aqueous （B）,gradient elution,the flow rate was 1.0 mL/min,the detection wavelength was at 260 rm,and the column temperature was at 35 C.The linear range of strychnine,brucine,strychnine N-oxide,brucine N-oxide and vomicine was 14.64～468.48 μg/mL,11.625～372.0 μg/mL,0.555～17.76 μg/mL,0.891～28.512 μg/mL and 2.5～80.0 μg/mL,respectively.The average recovery of these five alkaloids was 100.3％,101.7％,97.7％,98.8％ and 99.6％,respectively.With the total content of five alkaloids as the indexes,the extraction conditions of total alkaloids from S.nux-vomica,such as solvent amounts,concentration and extraction times,were optimized by L9 （33） orthogonal experimental design.The optimal extraction process was heat reflux extraction with 20-fold,70％ acidic alcohol for 4 times,one hour for each time.Under this optimum condition,the total content of the five alkaloids of the extraction was 3.633％.The RP-HPLC method was simple,accurate and suitable for simultaneous determination of strychnine,brucine,strychnine N-oxide,brucine N-oxide and vomicine of S.nux-vomica.The optimization of the extraction process of the total alkaloids was more comprehensive and reliable because of the total content of the five alkaloids as the indicator.