HPLC-MS结合多元统计分析区分人参产地及筛选皂苷类标志物
|
摘要
利用高效液相色谱-质谱联用(HPLC-MS)技术结合多元统计分析方法,区分中国人参主产区5个不同产地的45个人参样本,筛选出差异性皂苷类标志物.根据人参总皂苷在反相C18色谱柱中的洗脱顺序,结合串联质谱分析和标准品比对,在提取的人参总皂苷中鉴定出15种原人参三醇型、24种原人参二醇型和2种齐墩果酸型共41种皂苷.对人参总皂苷的HPLC-MS全扫描数据进行了多元统计分析.正交偏最小二乘-判别分析(OPLS-DA)结果表明,所建立的分析模型具有良好的数据描述能力和预测能力.所有人参样本能够根据产地被区分,并筛选得到同时区分5个产地的差异性皂苷类组分18种;能够区分任意2个产地人参样本的差异性组分主要为在人参中含量较高的人参皂苷Rb1,Rg1,Re,Rc,Rd,Ro和m-Rb1等.分层聚类分析(HCA)结果显示,黑龙江和吉林两省的样本能够独自聚类,但是绥化市的样本更接近于吉林省.初步推断原因为绥化市地理位置较接近吉林省,两地人参生长环境相似并可能存在种质资源交换.
High performance liquid chromatography-mass spectrometry combined with multi-variate statistical analysis was employed to discriminate 45 ginseng samples harvested from 5 different main cultivation areas in Northeast China and to identify the differential ginsenoside markers. A total of 41 ginsenosides,which included15 protopanaxatriol type,24 protopanaxadiol type and 2 oleanolic type ginsenosides,were identified based on the elution order of total ginsenosides in the reverse-phase C18 column coupled with the tandem MS analysis and comparison with authentic standard. Multivariate statistical analysis was further used to extract the information of HPLC-MS data sets. Orthogonal partial least squares-discriminate analysis revealed that the established analysis model had high goodness of fit and predictability. All the 45 ginseng samples were discriminated according to their origins. And 18 ginsenosides were identified as the differential markers,which contributed most to the simultaneous discrimination of the 5 ginseng origins. In addition,the differential ginsenoside markers which could distinguish any two origins were mainly those of high content in wild ginseng,such as Rb1,Rg1,Re,Rc,Rd,Ro,and m-Rb1. In the results of hierarchical clustering analysis,the ginseng samples of Heilongjiang and Jilin Province gathered separately except for the samples from Suihua city,which showed similarity to those of Jilin Province. This discrepancy may be attributed to the geographical location.Suihua city is relatively closer to Jilin Province and hence results in the similar growth environment of ginseng and the facility to exchange germplasm resources.
High performance liquid chromatography-mass spectrometry combined with multi-variate statistical analysis was employed to discriminate 45 ginseng samples harvested from 5 different main cultivation areas in Northeast China and to identify the differential ginsenoside markers. A total of 41 ginsenosides,which included15 protopanaxatriol type,24 protopanaxadiol type and 2 oleanolic type ginsenosides,were identified based on the elution order of total ginsenosides in the reverse-phase C18 column coupled with the tandem MS analysis and comparison with authentic standard. Multivariate statistical analysis was further used to extract the information of HPLC-MS data sets. Orthogonal partial least squares-discriminate analysis revealed that the established analysis model had high goodness of fit and predictability. All the 45 ginseng samples were discriminated according to their origins. And 18 ginsenosides were identified as the differential markers,which contributed most to the simultaneous discrimination of the 5 ginseng origins. In addition,the differential ginsenoside markers which could distinguish any two origins were mainly those of high content in wild ginseng,such as Rb1,Rg1,Re,Rc,Rd,Ro,and m-Rb1. In the results of hierarchical clustering analysis,the ginseng samples of Heilongjiang and Jilin Province gathered separately except for the samples from Suihua city,which showed similarity to those of Jilin Province. This discrepancy may be attributed to the geographical location.Suihua city is relatively closer to Jilin Province and hence results in the similar growth environment of ginseng and the facility to exchange germplasm resources.
引文
[1] Yun T. K.,Lancet Oncol.,2001,2(1),49—55
[2] Kim D. H.,J. Ginseng Res.,2012,36(1),1—15
[3] Liu Z. Q.,Chem. Rev.,2012,112(6),3329—3355
[4] Wong A. S. T.,Che C. M.,Leung K. W.,Nat. Prod. Rep.,2015,32(2),256—272
[5] Liu Z.,Wang C. Z.,Zhu X. Y.,Wan J. Y.,Zhang J.,Li W.,Ruan C. C.,Yuan C. S.,Molecules,2017,22(5),734
[6] Pace R.,Martineli E. M.,Sardone N.,Combarieu E. D. E.,Fitoterapia,2015,101,80—91
[7] Xiao D.,Yue H.,Xiu Y.,Sun X. L.,Wang Y. B.,Liu S. Y.,J. Ginseng Res.,2015,39(4),338—344
[8] Cheng C.,Yuan Q.,Zhou H.,Huang L.,Microsc. Res. Tech.,2016,79(2),98—105
[9] Chen Y.,Zhao Z.,Chen H.,Brand E.,Yi T.,Qin M.,Liang Z.,J. Ginseng Res.,2017,41(1),10—22
[10] Kwon S. W.,Han S. B.,Park I. H.,Kim J. M.,Park M. K.,Park J. H.,J. Chromatogr. A,2001,921(2),335—339
[11] Luo Z. Y.,Zhou G.,Zhou S. Q.,Chen X. H.,Luo J. Q.,Hu W. X.,Acta Pharm. Sin.,2000,35(8),626—629
[12] Wen J.,Zimmer E. A.,Mol. Phylogenet. Evol.,1996,6(2),167—177
[13] Kim J. H.,Yi Y. S.,Kim M. Y.,Cho J. Y.,J. Ginseng Res.,2016,41(4),435—443
[14] Li K. K.,Gong X. J.,RSC Adv.,2015,5,47353—47366
[15] Li X.,Zhao H. X.,Miao R.,Li W. Y.,Xiu Y.,Liu S. Y.,Chem. J. Chinese Universities,2017,38(10),1730—1736(李雪,赵幻希,苗瑞,李文影,修洋,刘淑莹.高等学校化学学报,2017,38(10),1730—1736)
[16] Shin B. K.,Kwon S. W.,Park J. H.,J. Ginseng Res.,2015,39(4),287—298
[17] Chen S.,Kong H.,Lu X.,Li Y.,Yin P.,Zeng Z.,Xu G.,Anal. Chem.,2013,85(17),8326—8333
[18] Cheng Y.,Liu Y. M.,Huang F. J.,Chen T. L.,Zheng X. J.,Zhao A. H.,He P. G.,Jia W.,Chem. J. Chinese Universities,2013,34(1),77—83(成玉,刘玉敏,黄凤杰,陈天璐,郑晓皎,赵爱华,何品刚,贾伟.高等学校化学学报,2013,34(1),77—83)
[19] Huang Y.,Gu C. Y.,Wu H. Z.,Xia X. S.,Li X.,Chem. J. Chinese Universities,2017,38(10),1742—1750(黄玉,谷彩云,吴翰钟,夏晓爽,李新.高等学校化学学报,2017,38(10),1742—1750)
[20] Chen Y.,Zhao Z.,Chen H.,Yi T.,Qin M.,Liang Z.,Phytochem. Anal.,2015,26(2),145—160
[21] Sun X.,Chen P.,Cook S. L.,Jackson G. P.,Harnly J. M.,Harrington P. B.,Anal. Chem.,2012,84(8),3628—3634
[22] Cho I. H.,Lee H. J.,Kim Y. S.,J. Agric. Food Chem.,2012,60(31),7616—7622
[23] Wang H. P.,Zhang Y. B.,Yang X. W.,Zhao D. Q.,Wang Y. P.,J. Ginseng Res.,2016,40(4),382—394
[24] Song H. H.,Moon J. Y.,Ryu H. W.,Noh B. S.,Kim J. H.,Lee H. K.,Oh S. R.,J. Ginseng Res.,2014,38(3),187—193
[25] Xu X.,Cheng X.,Lin Q.,Li S.,Jia Z.,Han T.,Lin R.,Wang D.,Wei F.,Li X.,J. Ginseng Res.,2016,40(4),344—350
[26] Shan S. M.,Luo J. G.,Huang F.,Kong L. Y.,J. Pharm. Biomed. Anal.,2014,89,76—82
[27] Xiu Y.,Li X.,Sun X.,Xiao D.,Miao R.,Zhao H.,Liu S.,J. Ginseng Res.,2017,DOI:10.1016/j.jgr.2017.12.001
[28] Xiu Y.,Zhao H.,Gao Y.,Liu W.,Liu S.,New J. Chem.,2016,40(11),9073—9080
[29] Song F.,Liu Z.,Liu S.,Cai Z.,Anal. Chim. Acta,2005,531(1),69—77
[30] Xu X. F.,Xu S. Y.,Zhang Y.,Zhang H.,Liu M. N.,Liu H.,Gao Y.,Xue X.,Xiong H.,Lin R. C.,Li X. R.,Molecules,2017,22(5),717
[31] Worley B.,Powers R.,Curr. Metabolomics,2013,1(1),92—107
[32] Triba M. N.,Moyec L. L.,Amathieu R.,Goossens C.,Bouchemal N.,Nahon P.,Rutledge D. N.,Savarin P.,Mol. Bio. Syst.,2015,11(1),13—19
[2] Kim D. H.,J. Ginseng Res.,2012,36(1),1—15
[3] Liu Z. Q.,Chem. Rev.,2012,112(6),3329—3355
[4] Wong A. S. T.,Che C. M.,Leung K. W.,Nat. Prod. Rep.,2015,32(2),256—272
[5] Liu Z.,Wang C. Z.,Zhu X. Y.,Wan J. Y.,Zhang J.,Li W.,Ruan C. C.,Yuan C. S.,Molecules,2017,22(5),734
[6] Pace R.,Martineli E. M.,Sardone N.,Combarieu E. D. E.,Fitoterapia,2015,101,80—91
[7] Xiao D.,Yue H.,Xiu Y.,Sun X. L.,Wang Y. B.,Liu S. Y.,J. Ginseng Res.,2015,39(4),338—344
[8] Cheng C.,Yuan Q.,Zhou H.,Huang L.,Microsc. Res. Tech.,2016,79(2),98—105
[9] Chen Y.,Zhao Z.,Chen H.,Brand E.,Yi T.,Qin M.,Liang Z.,J. Ginseng Res.,2017,41(1),10—22
[10] Kwon S. W.,Han S. B.,Park I. H.,Kim J. M.,Park M. K.,Park J. H.,J. Chromatogr. A,2001,921(2),335—339
[11] Luo Z. Y.,Zhou G.,Zhou S. Q.,Chen X. H.,Luo J. Q.,Hu W. X.,Acta Pharm. Sin.,2000,35(8),626—629
[12] Wen J.,Zimmer E. A.,Mol. Phylogenet. Evol.,1996,6(2),167—177
[13] Kim J. H.,Yi Y. S.,Kim M. Y.,Cho J. Y.,J. Ginseng Res.,2016,41(4),435—443
[14] Li K. K.,Gong X. J.,RSC Adv.,2015,5,47353—47366
[15] Li X.,Zhao H. X.,Miao R.,Li W. Y.,Xiu Y.,Liu S. Y.,Chem. J. Chinese Universities,2017,38(10),1730—1736(李雪,赵幻希,苗瑞,李文影,修洋,刘淑莹.高等学校化学学报,2017,38(10),1730—1736)
[16] Shin B. K.,Kwon S. W.,Park J. H.,J. Ginseng Res.,2015,39(4),287—298
[17] Chen S.,Kong H.,Lu X.,Li Y.,Yin P.,Zeng Z.,Xu G.,Anal. Chem.,2013,85(17),8326—8333
[18] Cheng Y.,Liu Y. M.,Huang F. J.,Chen T. L.,Zheng X. J.,Zhao A. H.,He P. G.,Jia W.,Chem. J. Chinese Universities,2013,34(1),77—83(成玉,刘玉敏,黄凤杰,陈天璐,郑晓皎,赵爱华,何品刚,贾伟.高等学校化学学报,2013,34(1),77—83)
[19] Huang Y.,Gu C. Y.,Wu H. Z.,Xia X. S.,Li X.,Chem. J. Chinese Universities,2017,38(10),1742—1750(黄玉,谷彩云,吴翰钟,夏晓爽,李新.高等学校化学学报,2017,38(10),1742—1750)
[20] Chen Y.,Zhao Z.,Chen H.,Yi T.,Qin M.,Liang Z.,Phytochem. Anal.,2015,26(2),145—160
[21] Sun X.,Chen P.,Cook S. L.,Jackson G. P.,Harnly J. M.,Harrington P. B.,Anal. Chem.,2012,84(8),3628—3634
[22] Cho I. H.,Lee H. J.,Kim Y. S.,J. Agric. Food Chem.,2012,60(31),7616—7622
[23] Wang H. P.,Zhang Y. B.,Yang X. W.,Zhao D. Q.,Wang Y. P.,J. Ginseng Res.,2016,40(4),382—394
[24] Song H. H.,Moon J. Y.,Ryu H. W.,Noh B. S.,Kim J. H.,Lee H. K.,Oh S. R.,J. Ginseng Res.,2014,38(3),187—193
[25] Xu X.,Cheng X.,Lin Q.,Li S.,Jia Z.,Han T.,Lin R.,Wang D.,Wei F.,Li X.,J. Ginseng Res.,2016,40(4),344—350
[26] Shan S. M.,Luo J. G.,Huang F.,Kong L. Y.,J. Pharm. Biomed. Anal.,2014,89,76—82
[27] Xiu Y.,Li X.,Sun X.,Xiao D.,Miao R.,Zhao H.,Liu S.,J. Ginseng Res.,2017,DOI:10.1016/j.jgr.2017.12.001
[28] Xiu Y.,Zhao H.,Gao Y.,Liu W.,Liu S.,New J. Chem.,2016,40(11),9073—9080
[29] Song F.,Liu Z.,Liu S.,Cai Z.,Anal. Chim. Acta,2005,531(1),69—77
[30] Xu X. F.,Xu S. Y.,Zhang Y.,Zhang H.,Liu M. N.,Liu H.,Gao Y.,Xue X.,Xiong H.,Lin R. C.,Li X. R.,Molecules,2017,22(5),717
[31] Worley B.,Powers R.,Curr. Metabolomics,2013,1(1),92—107
[32] Triba M. N.,Moyec L. L.,Amathieu R.,Goossens C.,Bouchemal N.,Nahon P.,Rutledge D. N.,Savarin P.,Mol. Bio. Syst.,2015,11(1),13—19