超高效液相色谱法检测二仙汤中淫羊藿黄酮类成分的含量  

Study on the Ultra-High Performance Liquid Chromatography(UPLC) Method Determination the Content of Flavonoids from Epimedii Folium in Erxian Tang

作　　者：郭红霞[1] 腊贵晓[1] 代丹丹[1] 理向阳[1] 杨铁钢[1] GUO Hongxia, LA Guixiao, DAI Dandan, LI Xiangyang, YANG Tiegang(Economic Crop Research Institute, Henan Academy of Agricultural Sciences/Henan Provincial Engineering Technology Research Center of Economic Crop Seedling, Zhengzhou Henan China 45000)

机构地区：[1]河南省农业科学院经济作物研究所/河南省经济作物育苗工程技术研究中心,河南郑州450008

出　　处：《中医学报》2018年第2期286-290,共5页Acta Chinese Medicine

基　　金：河南省农业科学院科研发展专项资金项目{豫财预[2017]76-2}

摘　　要：目的:建立超高效液相色谱法同时测定二仙汤中淫羊藿主要黄酮类成分的检测方法。方法:采用ZORBAX Eclipse Plus-C18色谱柱(3.0 mm×150 mm,3.5μm);以体积分数0.1%甲酸和浓度5 mmol·L^(-1)乙酸铵水溶液(A相)和甲醇(B相)为流动相进行线性梯度洗脱;梯度洗脱程序:0.0~0.5 min,A相的比例保持95%,0.5~1.5 min,A相的比例由95%降到70%,1.5~3.5 min,A相的比例由70%降到50%,3.5~6.0 min,A相的比例由50%降到30%,6~12 min,A相的比例由30%降到10%,12~15 min,A相的比例由10%降到5%;流速为0.45 mL·min^(-1);进样量:1μl;柱温:40℃;检测波长为270 nm。测定对照品混合液、二仙汤水提取液和体积分数50%乙醇提取液中朝藿定B、朝藿定C、淫羊藿苷以及淫羊藿次苷Ⅱ的含量。结果:超高效液相色谱法检测朝藿定B、朝藿定C、淫羊藿苷以及淫羊藿次苷Ⅱ的日间精密度RSD为0.21%~0.40%,日内精密度RSD为0.54%~0.89%,稳定性RSD为0.26%~0.90%,回收率为104.8%~116.7%。同一提取剂的二仙汤提取液中,不同主要黄酮类成分存在明显的差异,表现为朝藿定C>淫羊藿苷>朝藿定B>淫羊藿次苷Ⅱ含量;二仙汤体积分数50%乙醇提取液中检测的主要黄酮类成分均显著高于二仙汤水提取液,差异均有统计学意义(P<0.01)。结论:超高效液相色谱法具有十分良好的精密度、稳定性以及回收率,符合方法学的要求,可用于二仙汤中淫羊藿主要黄酮类成分的含量测定,为二仙汤的质量控制提供依据。Objective：To establish a ultra-high performance liquid chromatography（UPLC） method determination the content of flavonoids from Epimedii folium in Erxian Tang.Methods：Chromatographic column：ZORBAX Eclipse Plus-C18 column（3.0 mm × 150 mm,3.5 μm）;0.1% formic acid and 5 mmol·L^（-1） ammonium acetic acid solution（A phase） and methanol（B phase） were used as mobile phase to perform linear gradient elution;Gradient elution program：0.0-0.5 min with keeping 95%A phase,0.5-1.5 min with A phase ratio from 95% to 70%,1.5-3.5 min with A phase ratio from 70% to 50%,3.5-6 min with A phase ratio from 50% to 30%,6-12 min with A phase ratio from 30% to 10%,12-15 min with A phase ratio from 10%to 5%.The flow rate was 0.45 mL·min^（-1）,the injection volume was 1 μL,column temperature was 40 ℃.Detection wavelength was at 270 nm.The epimedin B,epimedin C,icariin and icariside Ⅱ in the control product mixture,water of Erxian Tang and 50%ethanol extracting solution.Results：In the determinations of epimedin B,epimedin C,icariin and icariside Ⅱ with the UPLC method,the RSD of interday and intraday precision were respectively 0.21%-0.40% and 0.54%-0.89%,the RSD of stability and average recovery rate were respectively 0.26%-0.90% and 104.8%-116.7%.The main flavonoids from Epimedii folium in the same extracting solution of Erxian Tang were obviously different and ranked for epimedin Cicariinepimedin BicarisideⅡ;the main flavonoids from Epimedii folium in the 50% ethanol extracting solution were significantly higher than those in the water extracting solution,their differences were statistically significant（all P0.01）.Conclusion：The UPLC method has very good precision,stability and recovery,which could meet the requirements of method validation to be used for the content determination of the main flavonoids from Epimedii folium in Erxian Tang and provide the foundations for quality control of Erxian Tang.Reference citation：GUO Hongxia,LA Guixiao,DAI Dandan,et al.Study on the Ultra-High Perf

关 键 词：淫羊藿 二仙汤 超高效液相色谱法 朝藿定B 朝藿定C 淫羊藿苷 淫羊藿次苷Ⅱ 含量测定 

分 类 号：R284.2[医药卫生—中药学]