中国中药杂志

2018, v.43(02) 363-368

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HPLC-DAD双波长法同时测定柴胡中5种皂苷的含量
Simultaneous determination of five saponins in Bupleuri Radix by HPLC-DAD dual wavelength method

刘玉梅;周安;俞年军;韩荣春;张威;朱月健;曹勇;李翔宇;彭代银;
LIU Yu-mei;ZHOU An;YU Nian-jun;HAN Rong-chun;ZHANG Wei;ZHU Yue-jian;CAO Yong;LI Xiang-yu;PENG Dai-yin;The Experimental Research Center,School of Pharmacy,Anhui University of Chinese Medicine;Anhui Puren Prepared Slices of Chinese Crude;Anhui Jiren Phaumaceutical Co.;

摘要(Abstract):

环氧醚型和异环双烯型皂苷为中药柴胡中的两类主要皂苷成分,但由于其紫外光谱存在差异,目前缺少针对2类皂苷成分同时进行定量的测定方法。为此,该文建立一种同时测定中药柴胡中环氧醚型(柴胡皂苷a,c,d)和异环双烯型(柴胡皂苷b_1,b_2)5种皂苷成分含量的双波长高效液相色谱分析方法。采用Welch Topsil-C_(18)色谱柱(4.6 mm×250 mm,5μm)柱温30℃,流动相为乙腈-水(0.1%磷酸)梯度洗脱,流速为1.0 m L·min-1,检测波长分别为:208 nm(柴胡皂苷a,c,d)、254 nm(柴胡皂苷b_1,b_2)。结果表明5种柴胡皂苷分离度良好,线性范围分别为:9.70-C_(18)色谱柱(4.6 mm×250 mm,5μm)柱温30℃,流动相为乙腈-水(0.1%磷酸)梯度洗脱,流速为1.0 m L·min-1,检测波长分别为:208 nm(柴胡皂苷a,c,d)、254 nm(柴胡皂苷b_1,b_2)。结果表明5种柴胡皂苷分离度良好,线性范围分别为:9.701 935.00(r=0.999 4),8.201 935.00(r=0.999 4),8.201 380.00(r=0.999 3),6.901 380.00(r=0.999 3),6.901 640.00(r=0.999 0),5.251 640.00(r=0.999 0),5.25630.00(r=0.999 4),5.15630.00(r=0.999 4),5.15618.00 mg·L618.00 mg·L(-1)(r=0.999 5),线性关系良好,平均加样回收率在97.7%(-1)(r=0.999 5),线性关系良好,平均加样回收率在97.7%100.2%,RSD均小于3%(n=6)。所建立的方法简便、快捷、重复性好,可用于中药柴胡中5种柴胡皂苷类成分的含量测定。
Epoxy ether type and isophthalene type saponin are the main saponins of Bupleurum chinense. However,due to the difference of their UV spectrum,there is no quantitative method for simultaneous determination of these two kinds of saponins. In this paper,a dual-wavelength high performance liquid chromatography( HPLC) was developed for simultaneous determination of five saponins in epoxidized ether( saikosaponin a,c,d) and isosorbide type( saikosaponin b_1,b_2). The mobile phase was eluted with acetonitrile-water(0. 1% phosphoric acid) gradient at a column temperature of 30 ℃ and a flow rate of 1. 0 m L·min-1. The detection wavelengths were208 nm for saikosaponins a,c,and d,and 254 nm for saikosaponins b_1 and b_2. The results showed that the separation of five kinds of saikosaponin was good,with the linear range of 9. 70-1 935. 00( r = 0. 999 4),8. 20-1 380. 00( r = 0. 999 3),6. 90-1 640. 00(r = 0. 999 0),5. 25-630. 00( r = 0. 999 4),and 5. 15-618. 00 mg·L100.2%,RSD均小于3%(n=6)。所建立的方法简便、快捷、重复性好,可用于中药柴胡中5种柴胡皂苷类成分的含量测定。
Epoxy ether type and isophthalene type saponin are the main saponins of Bupleurum chinense. However,due to the difference of their UV spectrum,there is no quantitative method for simultaneous determination of these two kinds of saponins. In this paper,a dual-wavelength high performance liquid chromatography( HPLC) was developed for simultaneous determination of five saponins in epoxidized ether( saikosaponin a,c,d) and isosorbide type( saikosaponin b_1,b_2). The mobile phase was eluted with acetonitrile-water(0. 1% phosphoric acid) gradient at a column temperature of 30 ℃ and a flow rate of 1. 0 m L·min-1. The detection wavelengths were208 nm for saikosaponins a,c,and d,and 254 nm for saikosaponins b_1 and b_2. The results showed that the separation of five kinds of saikosaponin was good,with the linear range of 9. 70-1 935. 00( r = 0. 999 4),8. 20-1 380. 00( r = 0. 999 3),6. 90-1 640. 00(r = 0. 999 0),5. 25-630. 00( r = 0. 999 4),and 5. 15-618. 00 mg·L(-1)( r = 0. 999 5),respectively. The average recoveries were 97. 70 %-100. 2 % and the RSD was less than 3 %( n = 6). The method is simple,rapid and reproducible. It can be used for the determination of five kinds of saikosaponins in B. chinense.

关键词(KeyWords): 柴胡;柴胡皂苷;双波长法;高效液相色谱法-二极管阵列检测器
Bupleurum;saikosaponin;dual wavelength method;HPLC-DAD

Abstract:

Keywords:

基金项目(Foundation): 国家中药标准化项目(ZYBZH-C-AH-02);; 安徽高校科研创新平台团队项目(皖教秘科)

作者(Authors): 刘玉梅;周安;俞年军;韩荣春;张威;朱月健;曹勇;李翔宇;彭代银;
LIU Yu-mei;ZHOU An;YU Nian-jun;HAN Rong-chun;ZHANG Wei;ZHU Yue-jian;CAO Yong;LI Xiang-yu;PENG Dai-yin;The Experimental Research Center,School of Pharmacy,Anhui University of Chinese Medicine;Anhui Puren Prepared Slices of Chinese Crude;Anhui Jiren Phaumaceutical Co.;

DOI: 10.19540/j.cnki.cjcmm.20171027.001

参考文献(References):

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