高效液相色谱一测多评法同时测定木丹颗粒中9种成分

Simultaneous Determination of Nine Components in Mudan Granules by HPLC-QAMS

目的建立高效液相色谱一测多评法(HPLC-QAMS)同时测定木丹颗粒中巴西苏木素、(±)原苏木素B、四氢非洲防己碱、四氢黄连碱、延胡索乙素、去氢紫堇碱、毛蕊异黄酮葡萄糖苷、毛蕊异黄酮和芒柄花素的含量。方法采用Agilent Zorbax SB-C_(18)色谱柱(250 mm×4.6 mm,5μm),柱温30℃;流动相为乙腈-0.2%甲酸溶液,梯度洗脱,流速1.0 mL·min^(-1);检测波长分别为280 nm[检测巴西苏木素、(±)原苏木素B、四氢非洲防己碱、四氢黄连碱、延胡索乙素和去氢紫堇碱]和254 nm(检测毛蕊异黄酮葡萄糖苷、毛蕊异黄酮和芒柄花素)。以延胡索乙素为内参物,建立其他8种成分的相对校正因子,测定含有量。结果巴西苏木素、(±)原苏木素B、四氢非洲防己碱、四氢黄连碱、延胡索乙素、去氢紫堇碱、毛蕊异黄酮葡萄糖苷、毛蕊异黄酮和芒柄花素分别在4.91~98.20,3.57~71.40,1.03~20.60,2.39~47.80,1.62~32.40,1.79~35.80,1.06~21.20,0.58~11.60,1.86~37.20μg·mL^(-1)范围内线性关系良好(r≥0.9991),平均加样回收率(RSD)分别为100.01%(0.73%)、99.17%(0.59%)、97.93%(0.78%)、98.81%(0.98%)、98.78%(0.43%)、97.74%(0.60%)、96.99%(0.62%)、96.96%(0.46%)和99.28%(0.37%),一测多评法所得结果与外标法结果无明显差异。结论该方法操作便捷、结果准确,重复性好,可用于同时测定木丹颗粒中巴西苏木素、(±)原苏木素B、四氢非洲防己碱、四氢黄连碱、延胡索乙素、去氢紫堇碱、毛蕊异黄酮葡萄糖苷、毛蕊异黄酮和芒柄花素含量。

Objective To establish a high performance liquid chromatography-quantitative analysis of multi-components by single-marker(HPLC-QAMS)method for determination of brazilin,(±)protosappanin B,tetrahydrocolumbamine,tetrahydrocoptisine,tetrahydropalmatine,dehydrocorydaline,calycosin7-O-β-D-glucopyranoside,calycosin and formononetin in Mudan granules.Methods The analysis was performed on an Agilent Zorbax SB-C_(18) column(250 mm×4.6 mm,5μm),mobile phase of acetonitrile-0.2%formic acid aqueous solution in a linear gradient mode.The flow rate was 1.0 mL·min^(-1).The detection wavelengths were set at 280 nm for brazilin,(±)protosappanin B,tetrahydrocolumbamine,tetrahydrocoptisine,tetrahydropalmatine and dehydrocorydaline;and 254 nm was used for calycosin7-O-β-D-glucopyranoside,calycosin and formononetin.The column temperature was 30℃.Tetrahydropalmatine was used as the internal standard.The relative correction factors(RCF)between the tetrahydropalmatine and other eight constituents were established and calculate the quantitation.Results Brazilin,(±)protosappanin B,tetrahydrocolumbamine,tetrahydrocoptisine,tetrahydropalmatine,dehydrocorydaline,calycosin7-O-β-D-glucopyranoside,calycosin and formononetin showed good linear relationships within the ranges of 4.91-98.20,3.57-71.40,1.03-20.60,2.39-47.80,1.62-32.40,1.79-35.80,1.06-21.20,0.58-11.60,1.86-37.20μg·mL^(-1)(r≥0.9991),respectively.The average recoveries(RSD)were 100.01%(0.73%),99.17%(0.59%),97.93%(0.78%),98.81%(0.98%),98.78%(0.43%),97.74%(0.60%),96.99%(0.62%),96.96%(0.46%)and 99.28%(0.37%),respectively.There was no significant difference between the calculated values of HPLC-QAMS method and the measured values of the external standard method.Conclusion The HPLC-QAMS method is simple,accurate and reproducible,which can be used for simultaneous determination of brazilin,(±)protosappanin B,tetrahydrocolumbamine,tetrahydrocoptisine,tetrahydropalmatine,dehydrocorydaline,calycosin7-O-β-D-glucopyranoside,calycosin and formononetin in Mudan granules.