不同产地人参根和根茎中人参皂苷的含量分析

目的：研究不同产地人参根和根茎中14个人参皂苷的含量，为制定人参皂苷的质量标准提供科学依据。方法：采用Diamonsil^® ODS C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈和乙腈-水-0.1%磷酸水溶液（5：90：8，v/v/v）为流动相，梯度洗脱，203 nm波长处二极管阵列检测器检测，外标对照品法测定人参根和根茎中人参皂苷（G）Ra₁、Ra₂、Rb₁、Rb₂、Rb₃、Rc、Rd、Re、Rf、Rg₁、Ro，20-O-葡萄糖基人参皂苷Rf（20-GG-Rf），三七人参皂苷（NG）R₁、R₂ 14个化合物。结果：不同产地人参根和根茎中人参皂苷含量的变化较大，26批次五年生人参根和根茎样品中14个人参皂苷的含量（μg·g^-1，人参根和根茎）变化幅度分别为G-Ra₁，0~2 958；G-Ra₂，0~1 320；G-Rb₁，1 195~7 129；G-Rb₂，835~6 993；G-Rb₃，178~1 812；G-Rc，414~5 569；G-Rd，393~5 529；G-Re，603~3 564；G-Rf，301~1 504；G-Rg₁，738~4 710；G-Ro，893~8 569；20-GG-Rf，110~656；NG-R₁，74~778；NG-R₂，0~844。结论：G-Ra₁、Ra₂、Rb₁、Rb₂、Rb₃、Rc和Rd为典型的原人参二醇型人参皂苷，G-Re、Rf、Rg₁，20-GG-Rf、NG-R₁和R₂为典型的原人参三醇型人参皂苷，G-Ro为典型的齐墩果酸型人参皂苷，三大类人参皂苷能代表体现人参药物活性的皂苷成分。其中，又以G-Rb₁、Rb₂、Rb₃、Rc、Rd、Re、Rf、Rg₁、Ro和20-GG-Rf含量更高，作为判定人参根和根茎中人参皂苷质量的指标性成分更科学。

Objective: To determine the contents of fourteen ginsenosides in the roots and rhizomes of Panax ginseng C. A.Meyer from different regions,so as to provide a scientific basis for quality standard establishment of ginsenosides.Methods: The analysis was carried out on a Diamonsil^TM ODS C₁₈ column (250 mm×4.6 mm,5 μm) with a gradient elution with acetonitrile (A) and acetonitrile-water-0.1% phosphoric acid aqueous solution (5:90:8,v/v/v) (B).The detection wavelength was at 203 nm and the detector was diode array detector (DAD).External standard as well as standard reference methods were adopted to determine the contents of fourteen ginsenosides including ginsenosides (G) Ra₁,Ra₂,Rb₁,Rb₂,Rb₃,Rc,Rd,Re,Rf,Rg₁,Ro,20-O-glucopyranosylginsenoside Rf (20-GG-Rf),notoginsenosides (NG) R₁ and R₂.Results: The investigated ginsenoside contents varied greatly among the 26 batches of five-year old ginseng roots and rhizomes samples collected from different localities,and the change ranges of fourteen ginsenoside contents (μg·g^-1 ginseng roots and rhizomes) were 0-2 958 for G-Ra₁,0-1 320 for G-Ra₂,1 195-7 129 for G-Rb₁,835-6 993 for G-Rb₂,178-1 812 for G-Rb₃,414-5 569 for G-Rc,393-5 529 for G-Rd,603-3 564 for G-Re,301-1 504 for G-Rf,738-4 710 for G-Rg₁,893-8 569 for G-Ro,110-656 for 20-GG-Rf,74-778 for NG-R₁,and 0-844 for NG-R₂,respectively.Conclusion: G-Ra₁,Ra₂,Rb₁,Rb₂,Rb₃,Rc,and Rd represent the typical protopanaxadiol-type ginsenoside,whereas G-Re,Rf,Rg₁,and 20-GG-Rf,NG-R₁,as well as NG-R₂ represent typical protopanaxatriol-type ginsenoside,and G-Ro represents typical oleanolic acid-type ginsenoside,which has been claimed to be responsible for the wide pharmacological activities of total saponins in the roots and rhizomes of P. ginseng.Among them,the contents of G-Rb₁,Rb₂,Rb₃,Rc,Rd,Re,Rf,Rg₁,Ro,and 20-GG-Rf were found to be much higher than other ginsenosides.These ten ginsenosides should be selected as markers to evaluate the quality of total saponins in the roots and rhizomes of P. ginseng.

[1] 中国药典2015年版.一部[S].2015:8 ChP 2015.Vol Ⅰ [S].2015:8
[2] 杨秀伟,富力.人参中三萜类化学成分的生物学活性和药理学作用[J].中国现代中药,2016,18(1):36 YANG XW,FU L.Biological and pharmacological activities of triterpenoids from of Panax ginseng[J].Mod Chin Med,2016,18(1):36
[3] 杨秀伟.人参化学成分的药物代谢动力学研究[J].中国现代中药,2016,18(1):16 YANG XW.Pharmacokinetic studies of chemical constituents of ginseng[J].Mod Chin Med,2016,18(1):16
[4] 王洪平,杨鑫宝,杨秀伟,等.吉林人参根和根茎的化学成分研究[J].中国中药杂志,2013,38(17):2807 WANG HP,YANG XB,YANG XW,et al.Chemical constituents from roots and rhizomes of Panax ginseng cultivated in Jilin province[J].China J Chin Mater Med,2013,38(17):2807
[5] WANG HP,YANG XB,YANG XW,et al.Ginsenjilinol,a new protopanaxatriol type saponin with inhibitory activity on LPS-activated NO production in macrophage RAW 264.7 cells from the roots and rhizomes of Panax ginseng[J].J Asian Nat Prod Res,2013,15(5):579
[6] 杨秀伟.人参中三萜类化学成分的研究[J].中国现代中药,2016,18(1):7 YANG XW.Triterpenoids in Panax ginseng[J].Mod Chin Med,2016,18(1):7
[7] WANG HP,ZHANG YB,YANG XW,et al.High-performance liquid chromatography with diode array detector and electrospray ionization ion trap time-of-flight tandem mass spectrometry to evaluate ginseng roots and rhizomes from different regions[J].Molecules,2016,21(5):603
[8] WANG HP,ZHANG YB,YANG XW,et al.Rapid characterization of ginsenosides in the roots and rhizomes of Panax ginseng by UPLC-DAD-QTOF-MS/MS and simultaneous determination of 19 ginsenosides by HPLC-ESI-MS[J].J Ginseng Res,2016,40(4):382
[9] ZHOU QL,YANG XW.Four new ginsenosides from red ginseng with inhibitory activity on melanogenesis in melanoma cells[J].Bioorg Med Chem Lett,2015,25:3112
[10] 周琪乐,徐嵬,杨秀伟.中国红参化学成分研究[J].中国中药杂志,2016,41(2):233 ZHOU QL,XU W,YANG XW.Chemical constituents of Chinese red ginseng[J].China J Chin Mater Med,2016,41(2):233
[11] 杨秀伟.中药物质基础研究是中药继承、发展、创新的关键科学问题[J].中国中药杂志,2015,40(17):3429 YANG XW.Substance basis research on Chinese materia medica is one of key scientific problems of inheriting,development and innovation of Chinese materia medica[J].China J Chin Mater Med,2015,40(17):3429
[12] CORTHOUT J,NAESSENS T,APERS S,et al.Quantitative determination of ginsenosides from Panax ginseng roots and ginseng preparations by thin layer chromatography-densitometry[J].J Pharm Biomed Anal,1999,21(1):187
[13] SHI W,WANG YT,LI J,et al.Investigation of ginsenosides in different parts and ages of Panax ginseng[J].Food Chem,2007,102(3):664
[14] SUN BS,GU LJ,FANG ZM,et al.Simultaneous quantification of 19 ginsenosides in black ginseng developed from Panax ginseng by HPLC-ELSD[J].J Pharm Biomed Anal,2009,50(1):15
[15] LEE DY,CHO JG,LEE MK,et al.Discrimination of Panax ginseng roots cultivated in different areas in Korea using HPLC-ELSD and principal component analysis[J].J Ginseng Res,2011,35(1):31
[16] JI QC,HARKEY MR,HENDERSON GL,et al.Quantitative determination of ginsenosides by high-performance liquid chromatography-tandem mass spectrometry[J].Phytochem Anal,2001,12(5):320
[17] XIE YY,LUO D,CHENG YJ,et al.Steaming induced chemical transformations and holistic quality assessment of red ginseng derived from Panax ginseng by means of HPLC-ESI-MS/MSn based multicomponent quantification fingerprint[J].J Agric Food Chem,2012,60(33):8213
[18] 张高菊,沈勇,孟珍贵,等.HPLC法测定姜状三七5种皂苷含量[J].药物分析杂志,2016,36(3):500 ZHANG GJ,SHEN Y,MENG ZG,et al.Determination of 5 saponins in Panax zingiberensis by HPLC[J].Chin J Pharm Anal,2016,36(3):500
[19] 杨鑫宝,杨秀伟,刘建勋.人参中皂苷类化学成分的研究[J].中国现代中药.2013,15(5):349 YANG XB,YANG XW,LIU JX.Study on ginsenoside in the roots and rhizomes of Panax ginseng[J].Mod Chin Med,2013,15(5):349
[20] GUI FJ,YANG XW,LI LY,et al.Simultaneous enantiomer determination of 20(R)-and 20(S)-ginsenoside-Rg₂ in rat plasma after intravenous administration using HPLC method[J].J Chromatogr B,2007,850(1-2):1
[21] 杨秀伟,桂方晋,田建明,等.人参皂苷-Rg₂在大鼠体内的药代动力学[J].中国药理学通报,2009,25(7):967 YANG XW,GUI FJ,TIAN JM,et al.Pharmacokinetics of ginsenoside-Rg₂ in rats[J].Chin Pharmacol Bull,2009,25(7):967
[22] JIANG Z,WANG YH,ZHANG XY,et al.Protective effect of ginsenoside Ro on anoxic and oxidative[J].Biomol Ther,2012,20(6):544
[23] 么厉,程恵珍,杨智.中药材规范化种植(养殖)技术指南[M].北京:中国农业出版社,2006:293 YAO L,CHENG HZ,YANG Z.Guidelines for Good Agricultural Practice of Chinese Crude Drugs[M].Beijing:China Agriculture Press,2006:293