基于HPLC-HRMS/MSn/QqQ技术的人参皂苷Rb1化学转化产物的结构与途径分析

doi:10.7503/cjcu20170216

高等学校化学学报 ›› 2017, Vol. 38 ›› Issue (10): 1730.doi: 10.7503/cjcu20170216

基于HPLC-HRMS/MSⁿ/QqQ技术的人参皂苷Rb₁化学转化产物的结构与途径分析

李雪¹, 赵幻希¹, 苗瑞¹, 李文影¹, 修洋¹(), 刘淑莹^1,²()

1. 长春中医药大学, 吉林省人参科学研究院, 长春 130117
2. 中国科学院长春应用化学研究所, 长春 130022

收稿日期:2017-04-11 出版日期:2017-10-10 发布日期:2017-09-20
作者简介:联系人简介: 刘淑莹, 女, 博士, 教授, 博士生导师, 主要从事有机质谱方面的研究. E-mail:syliu@ciac.ac.cn;修洋, 男, 博士, 助理研究员, 主要从事无机功能材料与中药化学方面的研究. E-mail:ys830805@sina.com
基金资助:
国家自然科学基金(批准号: 21475012)和吉林省科技发展计划项目(批准号: 20160520123JH, 20160309002YY)资助

Structure and Pathway Research on Chemical Transformation of Ginsenoside Rb₁ via HPLC-HRMS/MSⁿ/QqQ Technique^†

LI Xue¹, ZHAO Huanxi¹, MIAO Rui¹, LI Wenying¹, XIU Yang^1,^*(), LIU Shuying^1,^2,^*()

1. Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun 130117, China
2. Changchun Institute of Applied Chemistry, Chinese Academy of Science, Changchun 130022, China

Received:2017-04-11 Online:2017-10-10 Published:2017-09-20
Contact: XIU Yang,LIU Shuying E-mail:ys830805@sina.com;syliu@ciac.ac.cn
Supported by:
† Supported by the National Natural Science Foundation of China(No.21475012) and the Science and Technology Development Plan Project of Jilin Province, China(Nos.20160520123JH, 20160309002YY)

摘要/Abstract

摘要：

利用液相色谱-质谱联用技术分析了Keggin型12-磷钨酸化学转化人参皂苷Rb₁产物的结构与转化途径. 基于高效液相色谱对转化产物的快速分离, 利用Q Exactive高分辨质谱的FullMS-AIF模式快速鉴定了产物结构, 并利用多级串联质谱进行结构验证. 进一步结合人参皂苷异构体在反向C₁₈色谱柱上的相对保留时间, 快速分析鉴定出Rb₁的10种转化产物为20(S)-Rg₃, 20(R)-Rg₃, 20(S)-25-OH-Rg₃, 20(R)-25-OH-Rg₃, 25-OH-Rk₁, 25-OH-Rg₅, Rg₅, Rk₁, (20S,25)-环氧-Rg₃和(20R,25)-环氧-Rg₃. 根据转化产物的结构初步推断了人参皂苷的转化途径: 在12-磷钨酸产生的酸性环境中, Rb₁主要通过C₂₀位去糖基化、差向异构化和烯烃链的水合、消除及环合反应转化为稀有皂苷. 采用三重四极杆质谱的选择反应监测模式准确定量分析了Rb₁的转化效率和稀有皂苷20(S)-Rg₃, 20(R)-Rg₃, Rk₁和Rg₅的产率. 定量分析结果显示, 与生物转化相比, 12-磷钨酸对Rb₁有更高的转化效率, 反应40 min后转化率达到100%. 本文结果表明, HPLC-HRMS/MSⁿ/QqQ技术是人参皂苷等天然产物结构解析与定量分析的有效方法.

关键词: 人参皂苷Rb1, HPLC-HRMS/MSn/QqQ, 杂多酸, 化学转化

Abstract:

Keggin-type dodeca tungstophosphoric acid(HPW) was used in the chemical transformation of ginsenoside Rb₁. The aim of this research is to analyze the structure of the transformed products and transformation pathways based on high-performance liquid chromatography coupled with three kinds of mass spectrometry(HPLC-MS), the potential of which in qualitative and quantitative analysis of natural product was also developed. Q Exactive, a kind of high-resolution MS(HRMS), was used for the rapid structural identification of transformed products. Multistage tandem MS(MSⁿ) was further applied to verifying the identified structure. Ten transformed products, 20(S)-Rg₃, 20(R)-Rg₃, 20(S)-25-OH-Rg₃, 20(R)-25-OH-Rg₃, 25-OH-Rk₁, 25-OH-Rg₅, Rg₅, Rk₁, (20S,25)-epoxy-Rg₃ and(20R,25)-epoxy-Rg₃ were rapidly identified in all. In HPW-generated acidic solution, ginsenoside Rb₁ was chemically transformed through deglycosylation, epimerization at C₂₀ as well as hydration, elimination and cyclization reactions on the aglycone. Triple quadrupole(QqQ) MS was further used in its SRM mode for the accurately quantitative analysis of the conversion of Rb₁ and the yield of 20(S)-Rg₃, 20(R)-Rg₃, Rk₁ and Rg₅. Rb₁ was completely transformed to rare ginsenosides within 40 min. Based on the quantitative analysis results, HPW exhibited a much higher activity towards the transformation of Rb₁ in comparison with biotransformation, and it is a kind of efficient reagent for the chemical transformation of ginsenoside Rb₁. What’s more, HPLC-HRMS/MSⁿ/QqQ technique was proved to be effective for the qualitative and quantitative analysis of the natural products such as ginsenosides.

Key words: Ginsenoside Rb1, HPLC-HRMS/MSn/QqQ, Heteroploy acid, Chemical transformation

中图分类号:

O657.6

TrendMD:

李雪, 赵幻希, 苗瑞, 李文影, 修洋, 刘淑莹. 基于HPLC-HRMS/MSⁿ/QqQ技术的人参皂苷Rb₁化学转化产物的结构与途径分析. 高等学校化学学报, 2017, 38(10): 1730.

LI Xue, ZHAO Huanxi, MIAO Rui, LI Wenying, XIU Yang, LIU Shuying. Structure and Pathway Research on Chemical Transformation of Ginsenoside Rb₁ via HPLC-HRMS/MSⁿ/QqQ Technique^†. Chem. J. Chinese Universities, 2017, 38(10): 1730.

图/表 9

Fig.1 Total ion chromatogram of chemical transformation for ginsenoside Rb1

Fig.2 MS/MS spectrum of the [M-H]- ion at m/z 1107 of ginsenoside Rb1

Fig.3 MS/MS spectrum of the [M-H]- ion at m/z 801 of ginsenoside 20(S)-25-OH-Rg3

Fig.4 MS/MS spectrum of the [M-H]- ion at m/z 783 of ginsenoside 25-OH-Rk1

Fig.5 MS/MS spectrum of the [M-H]- ion at m/z 783 of ginsenoside 20(S)-Rg3

Fig.6 MS/MS spectrum of the [M-H]- ion at m/z 783 of (20S,25)-epoxy-Rg3

Fig.7 MS/MS spectrum of the [M-H]- ion at m/z 765 of ginsenoside Rk1

Scheme 1 Chemical transformation pathways of ginsenoside Rb1 by 12-H3PW12O40

Fig.8 Conversion of ginsenosides Rb1, 20(S)-Rg3, 20(R)-Rg3, Rk1 and Rg5

参考文献 37

[1]	Ganesan P., Ko H. M., Kim I. S., Choi D. K., RSC Adv., 2015, 5, 98634—98642
[2]	Wong A. S. T., Che C. M., Leung K. W., Nat. Prod. Rep., 2015, 32, 256—272
[3]	Li K. K., Gong X. J., RSC Adv., 2015, 5, 47353—47366
[4]	Kim D. H., J. Ginseng Res., 2012, 36, 1—15
[5]	Baek S. H., Bae O. N., Park J. H., J. Ginseng Res., 2012, 36, 119—134
[6]	Christensen L. P., Adv. Food Nutr. Res., 2009, 55, 1—99
[7]	Yuan Y., Hu Y. B., Hu C. X., Leng J. Y., Chen H. L., Zhao X. S., Gao J. Zhou Y. F., J. Mol. Catal. B: Enzym., 2015, 120, 60—67
[8]	Wang J. R., Yau L. F., Zhang R., Xia Y., Ma J., Ho H. M., Hu P., Hu M., Liu L. Jiang Z. H., J. Agric. Food Chem., 2014, 62, 2558—2573
[9]	Yu S. S., Zhou X., Li F., Xu C., Zheng F., Li J., Zhao H., Dai Y., Liu S., Feng Y., J. Sci. Rep., 2017, 7, 138
[10]	Wu W., Qin Q. J., Guo Y. Y., Sun J. H., Liu S. Y., J. Agric. Food Chem., 2012, 60, 10007—10014
[11]	Xiu Y, Zhao H. X., Gao Y, Liu W. L., Liu S. Y., New J. Chem., 2016, 40, 9073—9080
[12]	Zhang X., Song F., Cui M., Liu Z., Liu S., J. Planta Med., 2007, 73, 1225—1229
[13]	Cheng L. Q., Jin Y. Z., Liang D. C., Chem. J. Chinese Universities,2011, 32(1), 67—73
	(成乐琴, 金瑜真, 梁德春. 高等学校化学学报, 2011, 32(1), 67—73)
[14]	Zhou Q. L., Zhu D. N., Yang Y. F., Xu W., Yang X. W., J. Pharmaceut. Biomed., 2017, 137, 1—12
[15]	Qiu S., Yang W. Z., Yao C. L., Shi X. J., Li J. Y., Lou Y., Duan Y. N., Wu W. Y., Guo D. A., J. Nat. Prod., 2017, 80, 899—908
[16]	Cui Y. C., Pan C. S., Yan L., Li L., Hu B. H., Chang X., Liu Y. Y., Fan J. Y., Sun K., Li Q., Han J. Y., Sci. Rep., 2017, 7, 44579
[17]	Gao Y., J. Korean Soc. Appl. Bi., 2017, 25, 202—212
[18]	Wang Y. B., Xiao D., Li X. Y., Dai Y. L., Yue H., Liu S. Y., Chem. J. Chinese Universities,2015, 36(10), 1894—1899
	(王一博, 肖丹, 李晓宇, 戴雨霖, 越皓, 刘淑莹. 高等学校化学学报, 2015, 36(10), 1894—1899)
[19]	Lee S. J., Lee W. J., Chang S. E., Lee G. Y., J. Ginseng. Res., 2015, 39, 238—242
[20]	Teng S., Wang Y., Li P., Liu J. H., Wei A. H., Wang H. T., Meng X. K., Pan D., Zhang X. M., Mol. Med. Rep., 2017, 15, 2029—2038
[21]	Yayeh T., Yun K., Jang S., Oh S., J. Ginseng Res., 2016, 40, 445—452
[22]	Fu Y., Yin Z. H., Yin Y. C., J. Appl. Microbiol., 2017, 122, 1579—1585
[23]	Chang K. H., Mi N. J., Kim K. T., Paik H. D., J. Ginseng Res., 2014, 38, 47—51
[24]	Han Y., Sun B., Jiang B., Hu X., Spranger M. I., Zhang Y., Zhao Y., J. Appl. Microbiol., 2010, 109, 792—798
[25]	Wang D., Liao P. Y., Zhu H. T., Chen K. K., Xu M., Zhang Y. J., Yang C. R., Food Chem., 2012, 132, 1808—1813
[26]	Vo H. T., Cho J. Y., Choi Y. E., Choi Y. S., Jeong Y. H., J. Ginseng Res., 2015, 39, 304—313
[27]	Wang H., Wang J. Y., Wu X. P., Shen H. Y., Luo Y., Dai H. F., Mei W. L., Chem. Res. Chinese Universities,2015, 31(11), 38—43
[28]	Li L. J., Li X. W., Ding J., Liu Y., Wu Q., Wang X. Z., Li M., Jin Y. R., Chem. J. Chinese Universities,2016, 37(3), 454—459
	(李兰杰, 李绪文, 丁健, 刘迎, 吴谦, 王晓中, 李敏, 金永日. 高等学校化学学报, 2016, 37(3), 454—459)
[29]	Li S., Lai S., Song J., Qiao C., Liu X., Zhou Y., Cai H., Cai B., Xu H., J. Pharm. Biomed. Anal., 2010, 53, 946—957
[30]	Domon B., Costello C. E., Glycoconjugate J., 1988, 5, 397—409
[31]	Chattaraj P. K., Sarkar U., Roy D. R., Chem. Rev., 2006, 106, 2065—2091
[32]	Liu S., Rong C., Lu T., J. Phys. Chem. A,2014, 118, 3698—3704
[33]	Wang J., Yau L. F., Zhang R., Xia Y., Ma J., Ho H. M., Hu P., Hu M., Liu L., Jiang Z., J. Agric. Food Chem., 2014, 62, 2558—2573
[34]	Yang H., Lee Y. D., Kang K. B., Kim J. Y., Kim S. O., Yoo Y. H., Sung S. H., J. Pharm. Biomed. Anal., 2015, 109, 91—104
[35]	Kwon S. W., Han S. B., Park I. H., Kim J. M., Park M. K., Park J. H., J. Chromatogr. A,2001, 921, 335—339
[36]	Rosado-Reyes C. M., Tsang W., Alecu I. M., Merchant S. S., Green W. H., J. Chromatogr. A,2013, 117, 6724—6736
[37]	Toteva M. M., Richard J. P., J. Am. Chem. Soc., 1996, 118, 11434—11445

基于HPLC-HRMS/MSⁿ/QqQ技术的人参皂苷Rb₁化学转化产物的结构与途径分析

Structure and Pathway Research on Chemical Transformation of Ginsenoside Rb₁ via HPLC-HRMS/MSⁿ/QqQ Technique^†

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