HPLC同时测定山豆根中7种生物碱及3种黄酮的含量  

Simultaneous determination of seven alkaloids and three flavonoids in Sophorae Tonkinensis Radix et Rhizoma by HPLC

作　　者：韩馥蔓 王莉鑫 陈影[1] 陈两绵[1] 冯伟红[1] 王锦玉[1] 刘德文[1] 游云[1] 仝燕[1] 

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国中药杂志》2016年第24期4628-4634,共7页China Journal of Chinese Materia Medica

基　　金：中医药行业科研专项(201507004-2);中国中医科学院自主选题(ZZ2014055)

摘　　要：建立HPLC同时测定山豆根中7种生物碱（金雀花碱、氧化苦参碱、氧化槐果碱、N-甲基金雀花碱、槐醇、苦参碱和槐果碱）及3种黄酮（三叶豆紫檀苷、芒柄花素和马卡因）含量的方法。采用Welch XtimateTMC18（4.6 mm×250 mm,5μm）色谱柱;流动相为乙腈（A）-0.01 mol·L-1醋酸铵（氨水调p H 8.0）溶液（B）进行梯度洗脱：0~20 min,4%~14%A;20~30 min,14%~25%A;30~45 min,25%~40%A;45~65 min,40%~55%A;65~75 min,55%A;流速1.0 m L·min-1,柱温30℃,检测波长225 nm。方法学验证结果显示,10种成分的分离度良好,且在各自浓度范围内呈现出良好的线性关系,r≥0.999 7,平均回收率在98.60%~102.6%,RSD为0.60%~3.7%（n=6）,该方法简便、准确,重复性良好。含量测定结果显示,7种生物碱和3种黄酮成分在不同样品中的含量具有显著性差异。由此表明,不同批次的山豆根药材,其内在质量存在较大差异。该法可用于山豆根药材的多成分同步测定,同时可以为山豆根的全面质量评价和质量控制提供科学依据。In this study, an HPLC method was developed for simultaneous determination of seven alkaloids （cytosine, oxymatrine, N-oxysophocarpine, N-methylcytisine, sophoranol, matrine, and sophocarpine） and three flavonoids （ trifolirhizin, fermononetin, and maaekiain） from different samples of Sophorae Tonkinensis Radix et Rhizoma. Samples were analyzed on a Welch XtimateTM C18 col- umn （4. 6 mm × 250 mm, 5 μm） eluted with the mobile phase of acetonitrile （A） and 0. 01 tool · L^-1 ammonium acetate solution （pH 8.0） （B） in a linear gradient mode as follows： 0-20 min,4%-14% A;20-30 min,14%-25% A;30-45 min,25%-40% A;45-65 min,40%-55 % A ;65-75 min,55% A. The flow rate of the mobile phase, the column temperature, and the PDA detector wavelength were set at 1.0 mL · min^- 1 , 30 ℃ , and 225 nm, respectively. For the method validation, these ten compounds showed good separa- tion and satisfactory linearity （r≥0. 999 7 ） within the concentration ranges tested. The mean recoveries were in the range of 98.60% to 102. 6% with the RSD （n =6） between 0. 60% and 3.7%. This method was proved to be simple, accurate and repeatable. The quantitative results showed that there were significant differences in the contents of seven alkaloids and three flavonoids among the dif- ferent samples. This result revealed that the quality of Sophorae Tonkinensis Radix et Rhizoma varied widely. This method could be used for the simultaneous determination of the multi-ingredients from Sophorae Tonkinensis Radix et Rhizoma, which might provide sci- entific evidences to evaluate/control the quality of Sophorae Tonkinensis Radix et Rhizoma, comprehensively.

关 键 词：山豆根 高效液相色谱法 含量测定 生物碱 黄酮 

分 类 号：R284.1[医药卫生—中药学]