超高效液相色谱全波长扫描法同时测定倍生颗粒中5种成分含量

Simultaneous determination of five constituents in Beisheng granules by UPLC-full wavelength scanning method

目的采用超高效液相色谱全波长扫描法同时测定倍生颗粒中人参皂苷Rb1、毛蕊异黄酮葡萄糖苷、2,3,5,4'-四羟基二苯乙烯-2-O-β-D葡萄糖苷(以下简称二苯乙烯苷)、阿魏酸、大黄酸5种活性成分含量。方法采用Waters-ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8μm),以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.3 ml/min;全波长扫描(190~400 nm);柱温30℃;进样量3µl。结果人参皂苷Rb1、毛蕊异黄酮葡萄糖苷、二苯乙烯苷、阿魏酸、大黄酸5种活性成分分别在203、260、320、316、254 nm波长段分离良好,线性范围分别为0.013~0.210μg(r=0.9994)、0.027~0.432μg(r=0.9999)、0.038~0.600μg(r=0.9999)、0.005~0.084μg(r=0.9999)、0.003~0.048μg(r=0.9992);精密度试验、重复性试验、稳定性试验的RSD分别为0.17%~1.73%、0.40%~1.49%、0.21%~1.98%(n=6);加样回收率分别为92.71%、92.44%、96.83%、105.71%、102.61%(n=6)。结论该检测方法简便、快捷、结果准确,可为倍生颗粒的质量控制提供参考。

Objective To establish the method for simultaneous determination of ginsenoside Rb1,calycosin7-O-β-D-glucopyranoside,2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside(stilbene glucoside),ferulic acid,and rhein in Beisheng granules by UPLC-full wavelength scanning method.Methods The Waters-ACQUITY UPLC HSS T3(2.1 mm×100 mm,1.8μm)chromatographic column was used.The mobile phase was acetonitrile and 0.1%formic acid-water solute,a linear gradient,the flow rate was 0.3 ml/min and full-wavelength scanning with detection wavelength of 190-400 nm.The column temperature was 30℃,and sample volumn was 3μl.Results Ginsenoside Rb1,calycosin7-O-β-D-glucopyranoside,stilbene glucoside,ferulic acid and rhein were well separated at 203,260,320,316 and 254 nm wavelength,respectively.The linear ranges were 0.013-0.210μg(r=0.9994),0.027-0.432μg(r=0.9999),0.038-0.600μg(r=0.9999),0.005-0.084μg(r=0.9999),0.003-0.048μg(r=0.9992).The RSDs of precision,repeatability and stability tests were 0.17%-1.73%,0.40%-1.49%,0.21%-1.98%(n=6),respectively.The recovery rates were 92.71%,92.44%,96.83%,105.71%and 102.61%(n=6),respectively.Conclusions The method is simple,rapid and accurate,and can be used as a reference for quality control of Beisheng granules.