HPLC法同时测定山豆根中4种生物碱及其含量

Simultaneous content determination of four kinds of alkaloids in subprostrate sophora by HPLC

目的:对不同来源的山豆根提取物中苦参碱类生物碱成分进行定量测定。方法:采用HPLC法对山豆根提取物中苦参碱、氧化苦参碱、槐果碱和N-甲基野靛碱4种成分含量进行测定。色谱柱:Agilent C_(18)柱(4.6 mm×250 mm,5μm);流动相:甲醇-磷酸盐缓冲液[pH=6.3](20:80);检测波长:220 nm;流速:1.0 mL·min^(-1);柱温:37℃。结果:方法学验证结果显示,4种成分分离度良好,在各自浓度范围内呈现出良好的线性关系,r≥0.999 1,平均回收率在99.17%~100.78%,RSD为1.61%~2.20%(n=6)。含量测定结果显示,不同样品中生物碱成分含量具有显著性差异。广西省的4份山豆根样品中苦参碱类生物碱成分含量分别为39.375、59.247、40.398、53.13 mg·g^(-1),高于其他省份,四川省的山豆根样品中苦参碱类生物碱成分含量较低。结论:不同批次山豆根药材,内在质量存在较大差异,HPLC法可用于山豆根药材的多种成分同步测定,该方法简便、快捷、准确、重复性好,为山豆根质量评价和控制提供科学依据。

Objective:To establish the HPLC method to simultaneously determinate the content of 4 kinds of matrine alkaloids in subprostrate sophora samples from different sources.Methods:HPLC method was adopted to simultaneously determinate 4 kinds of alkaloids(matrine,oxymatrine,sophocarpine,and N-methylcytisine)from different samples of subprostrate sophora.Chromatographic conditions was as follows:Agilent C18 column(4.6 mm×250 mm,5μm);mobile phase:methanolphosphate buffer solution[pH 6.3](20:80);UV detection wavelength:220 nm;flow rate:1.0 mL·min-1;column temperature:37℃.Results:Results showed that four compounds had good resolution and linear relation(r≥0.999 1)in the tested concentration ranges.The mean recoveries were in the range of 99.17%to 100.78%and the RSD(n=6)was in the range of 1.61%to 2.20%(n=6).The quantitative results showed that there were significant differences in the contents of four alkaloids from different samples.The content of matrine alkaloids in four samples from Guangxi province were 39.375,59.247,40.398 and 53.13 mg·g-1 respectively,which were higher than those from other provinces.The content of alkaloids in samples from Sichuan is relatively low.Conclusion:The quality of subprostrate sophora from different province varied widely.HPLC method could be used for the simultaneous determination of the multi-ingredients from subprostrate sophora,this method was simple,accurate and repeatable,which could provide evidences for quality evaluation and control of subprostrate sophora.