人参皂甙Rb_3分离纯化的研究
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摘要
摘 要
本文研究了三七叶甙中的特有有效成分人参皂甙Rb3的分离纯化工艺，以期
得到含量满足不同新药要求的产品，为新药开发奠定物质基础。
以三七叶甙粗提物为原料，在反相半制备液相色谱实验的基础上，直接应用
反相制备液相色谱分离纯化单体皂甙Rb3，分别考察了上样量、流动相甲醇-水的
组成及流动相流速对分离过程的影响，最终确定了适宜的制备色谱条件，得到
Rb3含量为 65%~70%的样品，得率能达到 20%左右，达到中药二类原料药含量要
求。
为了得到更高含量的Rb3产品，采用二次上样的方法，所得产品中Rb3含量最
高能达到 92.06%。考虑到得率因素，将流动相体系改为乙腈-水溶液，选择粒径
更小的填料，并改变色谱柱的尺寸，从而提高了柱效和分离效果。初步建立了制
备色谱条件，所得人参皂甙Rb3 含量为 86.2%，有望达到中药一类原料药含量要
求。同期制得Rc的纯度在 95%以上。
采用多组分普遍化速率模型对三七叶甙的反相半制备型和制备型高效液相
色谱的分离纯化过程进行了模拟，模拟结果与实验数据的规律性吻合较好，对过
程操作具有一定的指导意义。
本文所建立的人参皂甙Rb3的分离纯化工艺简便、高效，对人参皂甙Rb3单体
的药理研究及新药开发具有重要价值。
ABSTRACT
This paper dealt with the isolation and purification of the pharmaceutical
component-ginsenoside Rb3 from the traditional Chinese medicine-Panax notoginseng,
hoping to get the product of which the content can meet the need of the different new
drug and settling the material foundation for the new drug development.
The notoginseng folium saponins were used as raw material and directly separated
by reversed phase preparative liquid chromatography. The effects of sample volume,
construction and flow rate of the mobile phase on the separation process were
invested. The suitable conditions were determined respectively. The content and the
receive efficiency of ginsenoside Rb3 in the crude product were about 65%~70% and
20% respectively.
In order to get the product (content>90%), the crude product was used as raw
material and separated again, through which the content of Rb3 in the product reached
about 92.06%. To raise the yield, a semi-preparative chromatography was used which
is constituted of the following: the mix solution of methl cyanide and water as the
mobile phase, the C18 of 10μm particle diameter as packing φ40×300mm column.
The primary preparative conditions were determined respectively. The content of the
product Rb3 is about 86.2%, and which of Rc is above 95%.
The general multi-component rate model of preparative chromatography was
established. The result of simulation showed good agreement with experimental data
in regularity. The general multi-component rat model is very useful for guiding the
preparative process.
The process for isolation and purification of ginsenoside Rb3 described here is
simple and effective, and it will be very valuable to pharmacological research and
new drug development.
本文研究了三七叶甙中的特有有效成分人参皂甙Rb3的分离纯化工艺，以期
得到含量满足不同新药要求的产品，为新药开发奠定物质基础。
以三七叶甙粗提物为原料，在反相半制备液相色谱实验的基础上，直接应用
反相制备液相色谱分离纯化单体皂甙Rb3，分别考察了上样量、流动相甲醇-水的
组成及流动相流速对分离过程的影响，最终确定了适宜的制备色谱条件，得到
Rb3含量为 65%~70%的样品，得率能达到 20%左右，达到中药二类原料药含量要
求。
为了得到更高含量的Rb3产品，采用二次上样的方法，所得产品中Rb3含量最
高能达到 92.06%。考虑到得率因素，将流动相体系改为乙腈-水溶液，选择粒径
更小的填料，并改变色谱柱的尺寸，从而提高了柱效和分离效果。初步建立了制
备色谱条件，所得人参皂甙Rb3 含量为 86.2%，有望达到中药一类原料药含量要
求。同期制得Rc的纯度在 95%以上。
采用多组分普遍化速率模型对三七叶甙的反相半制备型和制备型高效液相
色谱的分离纯化过程进行了模拟，模拟结果与实验数据的规律性吻合较好，对过
程操作具有一定的指导意义。
本文所建立的人参皂甙Rb3的分离纯化工艺简便、高效，对人参皂甙Rb3单体
的药理研究及新药开发具有重要价值。
ABSTRACT
This paper dealt with the isolation and purification of the pharmaceutical
component-ginsenoside Rb3 from the traditional Chinese medicine-Panax notoginseng,
hoping to get the product of which the content can meet the need of the different new
drug and settling the material foundation for the new drug development.
The notoginseng folium saponins were used as raw material and directly separated
by reversed phase preparative liquid chromatography. The effects of sample volume,
construction and flow rate of the mobile phase on the separation process were
invested. The suitable conditions were determined respectively. The content and the
receive efficiency of ginsenoside Rb3 in the crude product were about 65%~70% and
20% respectively.
In order to get the product (content>90%), the crude product was used as raw
material and separated again, through which the content of Rb3 in the product reached
about 92.06%. To raise the yield, a semi-preparative chromatography was used which
is constituted of the following: the mix solution of methl cyanide and water as the
mobile phase, the C18 of 10μm particle diameter as packing φ40×300mm column.
The primary preparative conditions were determined respectively. The content of the
product Rb3 is about 86.2%, and which of Rc is above 95%.
The general multi-component rate model of preparative chromatography was
established. The result of simulation showed good agreement with experimental data
in regularity. The general multi-component rat model is very useful for guiding the
preparative process.
The process for isolation and purification of ginsenoside Rb3 described here is
simple and effective, and it will be very valuable to pharmacological research and
new drug development.
引文
参 考 文 献
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31~33
[3]Kosuge, Takuo, Yokota, et.al.Yakugaku Zasshi,1981,101(6)：501~506
[4] 宋烈昌，刘杰，张毅，三七巴布剂中透皮吸收促进剂的选择，药学通报，1982，
17（2）：67~62
[5] 郝朝庆，杨帆，三七皂甙—对大鼠系膜血管微循环的影响，药学学报，1986，
7（3）：252~254
[6] 国家医药管理局，全国人参科技资料汇编，1988，26~29
[7] 唐第光，中成药，1990，12（8）：5~6
[8] Wu,Chieh-His, Tsai, Byi-Rong, Hsieh, Wen-Tsong.The preventive effects of G115
on balloon injury-induced meointima formation in rats.Life
Sciences,2001,669~679
[9] Carai, Mauro A.M,Agabio,Roberta, Bombardelli, Ezio. Potential use of medicinal
plants in the treatment of alcoholism. Fitoterapia,2000,38~42
[10]张新郁，双波长薄层扫描法测定三七片中三七皂甙R1人参皂甙Rb1，色谱，
1986，4（3）：183~189
[11] Vanhaelen-Fastré,RenéJ.,Faes, High-performance thin-layer chromatographic
determination of six major ginsenosides in Panax ginseng.Journal of
chromatography A,2000：269~276
[12]黄永焯，王宁生，三七及制剂中化学成分分析方法概述，中药新药与临床药
理，2002，13（3）：194~197
[13]章观德，中草药，1981，12（11）：23
[14]叶基荣，叶美琪，导数光谱法鉴别片仔癀，中成药，1990，(8)：9~11
[15]刘润民，张建平，三七种仁油的化学分研究，中草药，1990，21（6）：2~5
[16]张云华，李修禄等，超临界流体色普法测定三七及云南白药中人参二醇和人
参三醇的含量，药学学报，1991, 100：764~767
[17]Yamaguchi H.Thin-layer chromatographic determination of panaxadiol and
panaxatriol by ultroviolet derivatization. Chem. Pharm. Bull, 1988, 36(9)：34~68
[18]于世林，高效液相色谱法及应用，北京：化学工业出版社，1999，351~352
55
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[19]Snyder LR, Kirkland J.J., Introduction to modern liquid chromatography, 2nd ed.
John Wiley, New York, 1979
[20]王俊德编，高效液相色谱法，中国石化出版社，1992，121~123
[21]张晓彤主编，液相色谱检测方法，北京：化学工业出版社，2001，19~20
[22]美国 SP 紫外检测器说明书
[23]蒸发光散射 ELSD 检测器说明书
[24]于世林，高效液相色谱法及应用，北京：化学工业出版社，1999，351~352
[25]ESA 库仑列阵电化学检测器说明书
[26]He H., Edeki T., Wood A.J.J.mDetermination of low plasma timolo concentrations
following topical application of timolo eye drops in humans by HPLC with
electrochemical detection, J. Chromatogr.B, 1994, 661: 351~356
[27]孙守成，气相色谱和高压液相色谱及其质谱联用技术发展近况，现代仪器，
2003，3，36~39
[28]张玉奎，张庆合，张维冰等，中国色谱学的近期（1998-1999）发展，色谱，
2001，19（1），37~46
[29]XIE Jian-jun. Chinese Journal of Chromatography, 1999,17(3): 232~234
[30]ZHANG Yu-qing, FENG Lin-xian, FAN Zhi-qiang. Chinese Journal of
Chromatography, 1999,17(1): 35~37
[31]Lin L, Wu C, Yan Z, et al. Chromatographia, 1998,47: 689~694
[32]LIU Ying, CHEN Yuan-yin, ZENG Zhao-niu, et al. Chinese Journal of
Chromatography, 1998,16(5): 393~395
[33]Zeng Z R, Liu M. Chromatographia, 1998,48:817~821
[34]WU Jian-qiang, ZHU Dao-qian, WANG Qing-hai, et al. Chinese Journal of
Analytical Chemistry, 1999,27(2): 174~177
[35]JIANG Yan-jie, ZHAO Chun-xia, WANG Qing-hai, et al. Chinese Journal of
Analytical Chemistry, 1999,27(9): 1 022~1 025
[36]YUAN Li-ming FU Ruo-nong, DAI Rong-ji, et al. Chemical Journal of Chinese
University, 1998,19(2): 207~209
[37]Zhang H, Yuan X, Fu R, et al. J Chromatogr A, 1998,809:65~73
[38]LIANG Li, WANG Jia-liang. Chinese Journal of Analytical Chemistry,
1999,27(3): 271~275
[39]ZHOU Dong-mei, ZOU Han-fa, YANG Li, et al. Chinese Journal of Analytical
Chemistry, 1999,27(12): 1 370~1 375
[40]SHANG Zhen-hua, ZHOU Dong-mei, GUO Wei, et al. Journal of Instrumental
Analysis, 1998,17(1): 1~4
[41]SHANG Zhen-hua,ZHOU Dong-mei, GUO Wei, et al. Journal of Instrumental
Analysis, 1998,17(2):8~12
56
参考文献
[42]WANG Jin-fang, MENG, Zi-hui, ZHOU Liang-mo, et al. Chinese Journal of
Analytical Chemistry, 1999,27(7): 745~748
[43]ZHANG Jin-hui, JIANG Zhong-hua, CHEN Hui-peng, et al. Chiese Journal of
Chromatograph, 1999,17(3): 253~256
[44]LIU Tong, GENG Xin-du. Chiese Journal of Chromatograph, 1998,16(1): 30~34
[45]朱良漪，分析仪器手册，北京：化学工业出版社，1997，841
[46]Bulingame A L,Boyd R K,Gaskell S J.Mass spectrometry [J].Anal Chem,
1994,66(12):634~683
[47]于良涛，三七叶甙中人参皂甙单体Rb3分离纯化的研究：[硕士学位论文]，天
津；天津大学，2002
[48]李超生，潘书洋等，高效液相色谱法测定红参中人参皂甙 Rg1 含量，特定研
究，1998，1：13~15
[49]王旭，周富荣，HPLC 法测定西洋参中人参皂甙 Rb1 的含量，中国中药杂志，
1997，24（4）：227~228
[50]秦桂莲，徐飞等，RP-HPLC 测定洋参醇皂甙注射液中人参皂甙 Rb3 的含量，
中成药，2000，22（5）：338~339
[51]李毅，三七叶甙类新药生产工艺研究：[硕士学位论文]，天津；天津大学，
2004
[52]徐智秀，肖红斌等，高效液相色谱－质谱－质谱法分析人参皂甙，色谱，2000，
18（6）：521~524
[53]Nowakowski R. Computer simulation of non-line preparative chromatography
processed[J]. Chromatographia,1989,28(5/6):293~302
[54]J. C. Gidding, Dynamis of Chromatography, M. Dekker, New Youk,NY.1965
[55]B. C. Lin, Z. Ma, S. Golshan－Shirazi, G. Guiochon, Optimizing mobile phase
composition its flow rate and column temperature in HPLC using taboo search. J.
Chromatography., 1990,185:500~511
[56]Helfferich F, Klein G. Multicomponent chromatography-theory of interference.
Marcel Dekker, New York, 1970,35~45
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Wiley,1980, 84~94
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1980,35:2366~2377
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19~23
[1] 郑虎占，董泽宏，佘靖，中药现代研究与应用，北京：学苑出版社，1997，
23~25
[2] 魏均娴，杜元冲，三七—现代科学研究及应用，云南：科技出版社，1992，
31~33
[3]Kosuge, Takuo, Yokota, et.al.Yakugaku Zasshi,1981,101(6)：501~506
[4] 宋烈昌，刘杰，张毅，三七巴布剂中透皮吸收促进剂的选择，药学通报，1982，
17（2）：67~62
[5] 郝朝庆，杨帆，三七皂甙—对大鼠系膜血管微循环的影响，药学学报，1986，
7（3）：252~254
[6] 国家医药管理局，全国人参科技资料汇编，1988，26~29
[7] 唐第光，中成药，1990，12（8）：5~6
[8] Wu,Chieh-His, Tsai, Byi-Rong, Hsieh, Wen-Tsong.The preventive effects of G115
on balloon injury-induced meointima formation in rats.Life
Sciences,2001,669~679
[9] Carai, Mauro A.M,Agabio,Roberta, Bombardelli, Ezio. Potential use of medicinal
plants in the treatment of alcoholism. Fitoterapia,2000,38~42
[10]张新郁，双波长薄层扫描法测定三七片中三七皂甙R1人参皂甙Rb1，色谱，
1986，4（3）：183~189
[11] Vanhaelen-Fastré,RenéJ.,Faes, High-performance thin-layer chromatographic
determination of six major ginsenosides in Panax ginseng.Journal of
chromatography A,2000：269~276
[12]黄永焯，王宁生，三七及制剂中化学成分分析方法概述，中药新药与临床药
理，2002，13（3）：194~197
[13]章观德，中草药，1981，12（11）：23
[14]叶基荣，叶美琪，导数光谱法鉴别片仔癀，中成药，1990，(8)：9~11
[15]刘润民，张建平，三七种仁油的化学分研究，中草药，1990，21（6）：2~5
[16]张云华，李修禄等，超临界流体色普法测定三七及云南白药中人参二醇和人
参三醇的含量，药学学报，1991, 100：764~767
[17]Yamaguchi H.Thin-layer chromatographic determination of panaxadiol and
panaxatriol by ultroviolet derivatization. Chem. Pharm. Bull, 1988, 36(9)：34~68
[18]于世林，高效液相色谱法及应用，北京：化学工业出版社，1999，351~352
55
参考文献
[19]Snyder LR, Kirkland J.J., Introduction to modern liquid chromatography, 2nd ed.
John Wiley, New York, 1979
[20]王俊德编，高效液相色谱法，中国石化出版社，1992，121~123
[21]张晓彤主编，液相色谱检测方法，北京：化学工业出版社，2001，19~20
[22]美国 SP 紫外检测器说明书
[23]蒸发光散射 ELSD 检测器说明书
[24]于世林，高效液相色谱法及应用，北京：化学工业出版社，1999，351~352
[25]ESA 库仑列阵电化学检测器说明书
[26]He H., Edeki T., Wood A.J.J.mDetermination of low plasma timolo concentrations
following topical application of timolo eye drops in humans by HPLC with
electrochemical detection, J. Chromatogr.B, 1994, 661: 351~356
[27]孙守成，气相色谱和高压液相色谱及其质谱联用技术发展近况，现代仪器，
2003，3，36~39
[28]张玉奎，张庆合，张维冰等，中国色谱学的近期（1998-1999）发展，色谱，
2001，19（1），37~46
[29]XIE Jian-jun. Chinese Journal of Chromatography, 1999,17(3): 232~234
[30]ZHANG Yu-qing, FENG Lin-xian, FAN Zhi-qiang. Chinese Journal of
Chromatography, 1999,17(1): 35~37
[31]Lin L, Wu C, Yan Z, et al. Chromatographia, 1998,47: 689~694
[32]LIU Ying, CHEN Yuan-yin, ZENG Zhao-niu, et al. Chinese Journal of
Chromatography, 1998,16(5): 393~395
[33]Zeng Z R, Liu M. Chromatographia, 1998,48:817~821
[34]WU Jian-qiang, ZHU Dao-qian, WANG Qing-hai, et al. Chinese Journal of
Analytical Chemistry, 1999,27(2): 174~177
[35]JIANG Yan-jie, ZHAO Chun-xia, WANG Qing-hai, et al. Chinese Journal of
Analytical Chemistry, 1999,27(9): 1 022~1 025
[36]YUAN Li-ming FU Ruo-nong, DAI Rong-ji, et al. Chemical Journal of Chinese
University, 1998,19(2): 207~209
[37]Zhang H, Yuan X, Fu R, et al. J Chromatogr A, 1998,809:65~73
[38]LIANG Li, WANG Jia-liang. Chinese Journal of Analytical Chemistry,
1999,27(3): 271~275
[39]ZHOU Dong-mei, ZOU Han-fa, YANG Li, et al. Chinese Journal of Analytical
Chemistry, 1999,27(12): 1 370~1 375
[40]SHANG Zhen-hua, ZHOU Dong-mei, GUO Wei, et al. Journal of Instrumental
Analysis, 1998,17(1): 1~4
[41]SHANG Zhen-hua,ZHOU Dong-mei, GUO Wei, et al. Journal of Instrumental
Analysis, 1998,17(2):8~12
56
参考文献
[42]WANG Jin-fang, MENG, Zi-hui, ZHOU Liang-mo, et al. Chinese Journal of
Analytical Chemistry, 1999,27(7): 745~748
[43]ZHANG Jin-hui, JIANG Zhong-hua, CHEN Hui-peng, et al. Chiese Journal of
Chromatograph, 1999,17(3): 253~256
[44]LIU Tong, GENG Xin-du. Chiese Journal of Chromatograph, 1998,16(1): 30~34
[45]朱良漪，分析仪器手册，北京：化学工业出版社，1997，841
[46]Bulingame A L,Boyd R K,Gaskell S J.Mass spectrometry [J].Anal Chem,
1994,66(12):634~683
[47]于良涛，三七叶甙中人参皂甙单体Rb3分离纯化的研究：[硕士学位论文]，天
津；天津大学，2002
[48]李超生，潘书洋等，高效液相色谱法测定红参中人参皂甙 Rg1 含量，特定研
究，1998，1：13~15
[49]王旭，周富荣，HPLC 法测定西洋参中人参皂甙 Rb1 的含量，中国中药杂志，
1997，24（4）：227~228
[50]秦桂莲，徐飞等，RP-HPLC 测定洋参醇皂甙注射液中人参皂甙 Rb3 的含量，
中成药，2000，22（5）：338~339
[51]李毅，三七叶甙类新药生产工艺研究：[硕士学位论文]，天津；天津大学，
2004
[52]徐智秀，肖红斌等，高效液相色谱－质谱－质谱法分析人参皂甙，色谱，2000，
18（6）：521~524
[53]Nowakowski R. Computer simulation of non-line preparative chromatography
processed[J]. Chromatographia,1989,28(5/6):293~302
[54]J. C. Gidding, Dynamis of Chromatography, M. Dekker, New Youk,NY.1965
[55]B. C. Lin, Z. Ma, S. Golshan－Shirazi, G. Guiochon, Optimizing mobile phase
composition its flow rate and column temperature in HPLC using taboo search. J.
Chromatography., 1990,185:500~511
[56]Helfferich F, Klein G. Multicomponent chromatography-theory of interference.
Marcel Dekker, New York, 1970,35~45
[57]Ruthven DM. Principles of adsorption and adsorption process. New York:
Wiley,1980, 84~94
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