独活寄生汤高效液相色谱指纹图谱的建立和7种有效成分含量的测定

Establishment of HPLC Fingerprint and Determination of 7 Components in Duhuo Jisheng Decoction

目的:建立高效液相色谱(HPLC)指纹图谱法分析独活寄生汤中7种有效成分的方法,为临床独活寄生汤中药处方的质量控制提供参考依据。方法:分析独活寄生汤中药处方现代药理研究文献,筛选HPLC检测主要有效成分,建立HPLC检测上述有效成分的色谱条件,考察精密度、重复性、加样回收率,检测5批次独活寄生汤煎液中独参汤中蛇床子素、二氢欧山芹醇-β-D葡萄糖苷、二氢欧山芹醇、芍药苷、松脂醇二葡萄糖苷、β-蜕皮甾酮、人参皂苷R0含量。HPLC检测独活寄生汤中主要有效成分的色谱条件为:ODSC18柱(4.6 mm×250 mm,5μm)分析柱;1 mL/min;柱温为30℃,进样量10μL,检测方法:设置监测波长0~40 min采用330 nm检测,41~80 min采用230 nm检测;流动相:乙腈-0.08%甲酸(A),水-2.5%异丙醇-0.08%甲酸(B)洗脱程序:0~25 min,10%~26%流动相A,26~30 min,27%~44%流动相A,31~35 min,45%~46%流动相A,41~45 min,47%~48%流动相A;46~65 min,49%~92%流动相A;66~80 min,93%~100%流动相A。结果:蛇床子素、二氢欧山芹醇-β-D葡萄糖苷、二氢欧山芹醇、芍药苷、松脂醇二葡萄糖苷、β-蜕皮甾酮、人参皂苷R0具有较好的线性关系,精密度(日内和日间)RSD值均低于2.5%,7种成分检测重复性RSD值均低于3.5%,7种成分的加样回收率均≥95.0%。5批次独活寄生汤中7种成分含量比较差异有统计学意义(P<0.05)。结论:独活寄生汤主要有效成分可通过HPLC进行定量分析,但不同种含量差异较大,可能与煎药人不同、中药材批次不同有关。

Objective:To establish a HPLC fingerprint method for the analysis of 7 effective components in Duhuo Jisheng decoction,so as to provide reference for the quality control of Chinese medicinal prescription in Duhuo Jisheng Decoction.Methods:The modern pharmacological research literature of Duhuo Jisheng Decoction was analyzed,the main effective components were screened by HPLC,the chromatographic conditions were established for the determination of the above effective components by HPLC,and the precision,repeatability and sample recovery were investigated.The osthol,dihydrocarvacrol-β-D glucoside,dihydrocarvacrol,paeoniflorin and pinoresinol,β-ecdysterone,ginsenoside R0 in 5 batches of Duhuo Jisheng Decoction were detected.The chromatographic conditions for the determination of the main active ingredients in Duhuojisheng Decoction by HPLC were:odsc18 column(4.6 mm×250 mm,5μm);1 mL/min;column temperature was 30℃,injection volume was 10μL;detection method:the detection wavelength was set at 330 nm for 0~40 min and 230 nm for 41~80 min;mobile phase:acetonitrile-0.08%formic acid(a),water-2.5%isopropanol-0.08%formic acid(B)Sequence:0~25 min,10%~26%mobile phase A,26~30 min,27%~44%mobile phase A,31~35 min,45%~46%mobile phase A,41~45 min,47%~48%mobile phase A;46~65 min,49%~92%mobile phase A(2).For 66 to 80 min,93%to 100%mobile phase A.Results:There was a good linear relationship between osthol,dihydrocarinol-β-D glucoside,dihydrocarinol,paeoniflorin,rosinol diglucoside,β-ecdysterone and ginsenoside R0.The RSDS of precision(intra-day and inter-day)were all lower than 2.5%,and the repeatability RSDS of the 7 components were all lower than 3.5%.The recoveries of 7 components were all≥95.0%.There were statistically significant differences in the content of 7 ingredients in weight of 5 batches of Duhuo Jisheng Decoction(P<0.05).Conclusion:the main effective components of Duhuo Jisheng Decoction can be quantitatively analyzed by HPLC,but the contents of different kinds of drugs are quite different,which may be related to different decocting people and different batches of Chinese medicinal.