Discrimination of Ginseng Origins and Identification of Ginsenoside Markers Based on HPLC-MS Combined with Multivariate Statistical Analysis†

doi:10.7503/cjcu20180452

Abstract

Abstract:

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 included 15 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 contri-buted 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.

Key words: High performance liquid chromatography-mass spectrometry(HPLC-MS), Multivariate statistical analysis, Discrimination of ginseng origin, Ginsenoside

CLC Number:

O657

TrendMD:

ZHAO Huanxi,WANG Qiuying,SUN Xiuli,LI Xue,MIAO Rui,WU Dongxue,LIU Shuying,XIU Yang. Discrimination of Ginseng Origins and Identification of Ginsenoside Markers Based on HPLC-MS Combined with Multivariate Statistical Analysis^†[J]. Chem. J. Chinese Universities, 2019, 40(2): 246.

Figures/Tables 9

References 32

[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

Related Articles 15

[1]	ZHU Ling,WANG Yuchen,ZHAO Jiangyuan,WEN Mengliang,LI Minggang,HAN Xiulin. Transformation of Ginsenoside Rb₃ and C-Mx by Recombinant β-Xylosidase ^† [J]. Chem. J. Chinese Universities, 2020, 41(5): 1010.
[2]	WANG Tianqi,YU Qiongwei,FENG Yuqi. Analysis of Imidazole Propionic Acid in Serum of Patients with Type 2 Diabetes Based on NiO@SiO₂ Solid-phase Extraction Coupled with Liquid Chromatography-Mass Spectrometry ^† [J]. Chem. J. Chinese Universities, 2020, 41(2): 262.
[3]	XIAO Yongkun,LIU Chunying,YU Hongshan,YI Tea-Hoo,XU Longquan,SONG Jianguo,IM Wan-Teak,SUN Changkai,JIN Fengxie. Dynamic Biotransformation of Protopanaxadiol-ginsenosides and Preparation of Minor Ginsenosides C-K or $F 2 †$ [J]. Chem. J. Chinese Universities, 2019, 40(6): 1184.
[4]	ZHAO Lefeng, JIAO Chuanxin, LI Hui, JIAO Lili, MA Yue, WU Wei, LIU Shuying. Chemical Transformation of Protopanaxadiol Type Ginsenoside Rb₁, Rb₂ and Rc Analyzed by RRLC-Q-TOF-MS^† [J]. Chem. J. Chinese Universities, 2018, 39(4): 667.
[5]	QIAO Mengdan, LIU Shang, ZHANG Yan, LI Jing, ZHENG Fei, DAI Yulin, YUE Hao. Biotransformation of Ginsenosides in Fermented Ginseng Using UPLC-Q-Orbitrap MS/MS^† [J]. Chem. J. Chinese Universities, 2018, 39(2): 219.
[6]	LIU Xinru, LIU Chunying, XU Longquan, SONG Jianguo, YU Hongshan. Construction of Ginsenoside-β-glucosidase Gene Vector and Biotransformation in Pichia Pastoris^† [J]. Chem. J. Chinese Universities, 2018, 39(11): 2451.
[7]	LIU Ying, CHEN Yanxin, WU Qian, LI Peng, LI Xuwen, SHI Xiaolei, JIN Yongri. Preparation of 20(S/R)-Ginsenoside Rg₃ and Their Effects on Regulation of Th1/Th2 Imbalance^† [J]. Chem. J. Chinese Universities, 2018, 39(11): 2419.
[8]	MIAO Rui,WU Dongxue,WANG Qiuying,ZHAO Huanxi,LI Xue,XIU Yang,LIU Shuying. Rapid Separation of Ginsenosides Based on Multi-walled Carbon Nanotubes^† [J]. Chem. J. Chinese Universities, 2018, 39(10): 2178.
[9]	LI Ruigang,ZHU Na,ZHAO Huanxi,WANG Nan,SUN Hongmei,YUE Hao,LI Jing. Effects of Ginseng Polysaccharides on the Metabolism of Ginsenoside Re in vivo and Transformation of Ginsenoside Re in vitro^† [J]. Chem. J. Chinese Universities, 2018, 39(10): 2192.
[10]	LI Nan, ZHAO Huanxi, LI Jing, WANG Nan, YU Bohao, YUE Hao, YU Shanshan. Transformation of Total Notoginsenosides by Recombinant Endocellulase Fpendo5A^† [J]. Chem. J. Chinese Universities, 2017, 38(12): 2185.
[11]	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^† [J]. Chem. J. Chinese Universities, 2017, 38(10): 1730.
[12]	LI Lanjie, LI Xuwen, DING Jian, LIU Ying, WU Qian, WANG Xiaozhong, LI Min, JIN Yongri. Ionic Liquid Surfactant-mediated Ultrasonic-assisted Extraction Coupled with HPLC for the Determination of Five Rare Ginsenosides in Panax notoginseng (Burk.) F. H. Chen^† [J]. Chem. J. Chinese Universities, 2016, 37(3): 454.
[13]	XU Chunchun, YU Bohao, WANG Honglei, LI Jing, LIU Shuying, YU Shanshan. Transformation of Minor Ginsenoside Rd and CK by Recombinant Thermostable β-Glucosidase^† [J]. Chem. J. Chinese Universities, 2016, 37(2): 281.
[14]	WANG Yibo, XIAO Dan, LI Xiaoyu, DAI Yulin, YUE Hao, LIU Shuying. Metabolism of Ginsengside Rb1 in Rat Liver Microsomal in vitro by HPLC-MS and Cocktail Probe Substrats Method^† [J]. Chem. J. Chinese Universities, 2015, 36(10): 1894.
[15]	YAO Hua, JIN Yongri, YANG Jie, LI Lanjie, SUN Ting, SHI Xiaolei, LI Xuwen. Conversion Rule of Rare Ginsenosides Produced from Major Ginsenosides by Confined Microiwave Promoted Degradation Method^† [J]. Chem. J. Chinese Universities, 2014, 35(11): 2317.

No.	t_R/min	Ginsenoside	Molecular formula	[M-H]^- ion, m/z	Monoisotopic mass	Ref.
1	1.14	Re1	C₄₈H₈₂O₁₉	961.53	962.54	[23]
2	2.63	Re4	C₄₇H₈₀O₁₈	931.41	932.53	[30]
3	2.84	20-Glc-Rf	C₄₈H₈₂O₁₉	961.52	962.54	[23]
4	2.97	NG-R1 isomer	C₄₇H₈₀O₁₈	931.55	932.53	[23]
5	3.34	NG-R1	C₄₇H₈₀O₁₈	931.53	932.53	-
6	4.24	Re	C₄₈H₈₂O₁₈	945.52	946.55	-
7	4.30	Rg1	C₄₂H₇₂O₁₄	845.44	800.49	-
8	5.41	Ac-Rg1 isomer^a	C₄₄H₇₄O₁₅	841.46	842.50	[23]
9	6.00	m-Re	C₅₁H₈₄O₂₁	1031.56	1032.55	[30]
10	6.06	Ac-Re	C₅₀H₈₄O₁₉	987.54	988.56	[23]
11	6.31	Ac-Rg1	C₄₄H₇₄O₁₅	841.51	542.50	[23]
12	10.57	20(S)-Rf	C₄₂H₇₂O₁₄	799.44	800.49	-
13	10.73	Ra3	C₅₉H₁₀₀O₂₇	1239.64	1240.64	[30]
14	11.13	m-Ra3^b	C₆₂H₁₀₂O₃₀	1325.61	1326.65	[30]
15	11.47	20(S)-NG-R2	C₄₁H₇₀O₁₃	769.44	770.48	-
16	11.62	NG-R4	C₅₉H₁₀₀O₂₇	1239.67	1240.64	[30]
17	11.78	Ra2	C₅₈H₉₈O₂₆	1209.66	1210.63	[30]
18	11.81	m-NG-R4	C₆₂H₁₀₂O₃₀	1325.62	1236.65	[30]
19	11.93	NG-Fa	C₅₉H₁₀₀O₂₇	1239.66	1240.64	[30]
20	12.06	Rb1	C₅₄H₉₂O₂₃	1107.55	1108.60	-
No.	t_R/min	Ginsenoside	Molecular formula	[M-H]^- ion, m/z	Monoisotopic mass	Ref.
21	12.21	m-Ra2	C₆₁H₁₀₀O₂₉	1295.62	1296.63	[30]
22	12.33	20(S)-Rg2	C₄₂H₇₂O₁₃	783.47	784.50	-
23	12.40	m-NG-Fa	C₆₂H₁₀₂O₃₀	1325.59	1326.65	[30]
24	12.52	m-Rb1	C₅₇H₉₄O₂₆	1193.56	1194.60	[30]
25	12.71	Ra1	C₅₈H₉₈O₂₆	1209.56	1210.63	[30]
26	12.80	20(S)-Rh1	C₃₆H₆₂O₉	637.47	638.44	-
27	12.83	Rc	C₅₃H₉₀O₂₂	1077.61	1078.59	-
28	13.11	Ro	C₄₈H₇₆O₁₉	955.49	956.50	-
29	13.26	m-Ra1	C₆₁H₁₀₀O₂₉	1295.66	1296.63	[30]
30	13.42	m-Rc	C₅₆H₉₂O₂₅	1163.50	1164.59	[24]
31	13.66	Rb2	C₅₃H₉₀O₂₂	1077.57	1078.59	-
32	13.94	Rb3	C₅₃H₉₀O₂₂	1077.54	1078.59	-
33	14.19	Ra1 isomer	C₅₈H₉₈O₂₆	1209.58	1210.63	[23]
34	14.34	m-Rb2	C₅₆H₉₂O₂₅	1163.53	1164.59	[30]
35	14.65	m-Rb3	C₅₆H₉₂O₂₅	1163.78	1164.59	[30]
36	14.96	Quinquenoside R1	C₅₆H₉₄O₂₄	1149.62	1150.61	[30]
37	16.20	Rs2	C₅₅H₉₂O₂₃	1119.54	1120.60	[23]
38	16.42	Rd	C₄₈H₈₂O₁₈	945.48	946.55	-
39	16.63	Rs1	C₅₅H₉₂O₂₃	1119.72	1120.60	[23]
40	17.06	Chikusetsusaponin Ⅳa	C₄₂H₆₆O₁₄	793.41	794.44	[23]
41	17.56	m-Rd	C₅₁H₈₄O₂₁	1031.51	1032.55	[30]

No.	t_R/min	Ginsenoside	Molecular formula	[M-H]^- ion, m/z	Monoisotopic mass	Ref.
1	1.14	Re1	C₄₈H₈₂O₁₉	961.53	962.54	[23]
2	2.63	Re4	C₄₇H₈₀O₁₈	931.41	932.53	[30]
3	2.84	20-Glc-Rf	C₄₈H₈₂O₁₉	961.52	962.54	[23]
4	2.97	NG-R1 isomer	C₄₇H₈₀O₁₈	931.55	932.53	[23]
5	3.34	NG-R1	C₄₇H₈₀O₁₈	931.53	932.53	-
6	4.24	Re	C₄₈H₈₂O₁₈	945.52	946.55	-
7	4.30	Rg1	C₄₂H₇₂O₁₄	845.44	800.49	-
8	5.41	Ac-Rg1 isomer^a	C₄₄H₇₄O₁₅	841.46	842.50	[23]
9	6.00	m-Re	C₅₁H₈₄O₂₁	1031.56	1032.55	[30]
10	6.06	Ac-Re	C₅₀H₈₄O₁₉	987.54	988.56	[23]
11	6.31	Ac-Rg1	C₄₄H₇₄O₁₅	841.51	542.50	[23]
12	10.57	20(S)-Rf	C₄₂H₇₂O₁₄	799.44	800.49	-
13	10.73	Ra3	C₅₉H₁₀₀O₂₇	1239.64	1240.64	[30]
14	11.13	m-Ra3^b	C₆₂H₁₀₂O₃₀	1325.61	1326.65	[30]
15	11.47	20(S)-NG-R2	C₄₁H₇₀O₁₃	769.44	770.48	-
16	11.62	NG-R4	C₅₉H₁₀₀O₂₇	1239.67	1240.64	[30]
17	11.78	Ra2	C₅₈H₉₈O₂₆	1209.66	1210.63	[30]
18	11.81	m-NG-R4	C₆₂H₁₀₂O₃₀	1325.62	1236.65	[30]
19	11.93	NG-Fa	C₅₉H₁₀₀O₂₇	1239.66	1240.64	[30]
20	12.06	Rb1	C₅₄H₉₂O₂₃	1107.55	1108.60	-
No.	t_R/min	Ginsenoside	Molecular formula	[M-H]^- ion, m/z	Monoisotopic mass	Ref.
21	12.21	m-Ra2	C₆₁H₁₀₀O₂₉	1295.62	1296.63	[30]
22	12.33	20(S)-Rg2	C₄₂H₇₂O₁₃	783.47	784.50	-
23	12.40	m-NG-Fa	C₆₂H₁₀₂O₃₀	1325.59	1326.65	[30]
24	12.52	m-Rb1	C₅₇H₉₄O₂₆	1193.56	1194.60	[30]
25	12.71	Ra1	C₅₈H₉₈O₂₆	1209.56	1210.63	[30]
26	12.80	20(S)-Rh1	C₃₆H₆₂O₉	637.47	638.44	-
27	12.83	Rc	C₅₃H₉₀O₂₂	1077.61	1078.59	-
28	13.11	Ro	C₄₈H₇₆O₁₉	955.49	956.50	-
29	13.26	m-Ra1	C₆₁H₁₀₀O₂₉	1295.66	1296.63	[30]
30	13.42	m-Rc	C₅₆H₉₂O₂₅	1163.50	1164.59	[24]
31	13.66	Rb2	C₅₃H₉₀O₂₂	1077.57	1078.59	-
32	13.94	Rb3	C₅₃H₉₀O₂₂	1077.54	1078.59	-
33	14.19	Ra1 isomer	C₅₈H₉₈O₂₆	1209.58	1210.63	[23]
34	14.34	m-Rb2	C₅₆H₉₂O₂₅	1163.53	1164.59	[30]
35	14.65	m-Rb3	C₅₆H₉₂O₂₅	1163.78	1164.59	[30]
36	14.96	Quinquenoside R1	C₅₆H₉₄O₂₄	1149.62	1150.61	[30]
37	16.20	Rs2	C₅₅H₉₂O₂₃	1119.54	1120.60	[23]
38	16.42	Rd	C₄₈H₈₂O₁₈	945.48	946.55	-
39	16.63	Rs1	C₅₅H₉₂O₂₃	1119.72	1120.60	[23]
40	17.06	Chikusetsusaponin Ⅳa	C₄₂H₆₆O₁₄	793.41	794.44	[23]
41	17.56	m-Rd	C₅₁H₈₄O₂₁	1031.51	1032.55	[30]

No.	Variable	m/z@t_R	VIP	Molecular ion	Identity
1	2	1107.55@12.06	6.42	[M-H]^-	Rb1
2	3	955.47@13.11	6.03	[M-H]^-	Ro
3	7	1077.61@12.96	5.58	[M-H]^-	Rc
4	4	991.46@4.29	4.70	[M+HCOO]^-	Re
5	1	845.46@4.36	4.62	[M-H]^-	Rg1
6	9	991.49@16.43	4.45	[M+HCOO]^-	Rd
7	6	1123.50@12.56	4.10	[M+HCOO]^-	Rc
8	14	1193.53@12.52	3.87	[M-H]^-	m-Rb1
9	16	1163.49@13.43	3.73	[M-H]^-	m-Rc
10	15	945.52@16.41	3.56	[M-H]^-	Rd
11	12	1209.58@12.71	3.50	[M-H]^-	Ra1
12	5	799.49@10.57	3.27	[M-H]^-	20(S)-Rf
13	20	835.43@4.35	3.02	[M+Cl]^-	Rg1
14	34	1143.57@12.08	2.74	[M+Cl]^-	Rb1
15	18	1239.63@11.96	2.65	[M-H]^-	NG-Fa
16	32	1113.64@12.85	2.58	[M+Cl]^-	Rc
17	11	769.44@11.50	2.48	[M-H]^-	20(S)-NG-R2
18	8	1153.51@12.09	2.38	[M+HCOO]^-	Rb1
19	19	1255.63@11.76	2.34	[M+HCOO]^-	Ra2
20	39	981.41@16.66	2.25	[M+Cl]^-	Rd
21	36	793.41@17.00	2.20	[M-H]^-	Chikusetsusaponin Ⅳa
22	26	1285.55@11.98	2.19	[M+HCOO]^-	NG-Fa
23	10	945.50@4.28	2.14	[M-H]^-	Re
24	31	981.43@4.25	2.09	[M+Cl]^-	Re
25	37	1031.49@17.61	2.05	[M-H]^-	m-Rd
26	52	835.41@10.57	1.81	[M+Cl]^-	20(S)-Rf
27	54	1008.49@2.69	1.72	-	Unknown

No.	Variable	m/z@t_R	VIP	Molecular ion	Identity
1	2	1107.55@12.06	6.42	[M-H]^-	Rb1
2	3	955.47@13.11	6.03	[M-H]^-	Ro
3	7	1077.61@12.96	5.58	[M-H]^-	Rc
4	4	991.46@4.29	4.70	[M+HCOO]^-	Re
5	1	845.46@4.36	4.62	[M-H]^-	Rg1
6	9	991.49@16.43	4.45	[M+HCOO]^-	Rd
7	6	1123.50@12.56	4.10	[M+HCOO]^-	Rc
8	14	1193.53@12.52	3.87	[M-H]^-	m-Rb1
9	16	1163.49@13.43	3.73	[M-H]^-	m-Rc
10	15	945.52@16.41	3.56	[M-H]^-	Rd
11	12	1209.58@12.71	3.50	[M-H]^-	Ra1
12	5	799.49@10.57	3.27	[M-H]^-	20(S)-Rf
13	20	835.43@4.35	3.02	[M+Cl]^-	Rg1
14	34	1143.57@12.08	2.74	[M+Cl]^-	Rb1
15	18	1239.63@11.96	2.65	[M-H]^-	NG-Fa
16	32	1113.64@12.85	2.58	[M+Cl]^-	Rc
17	11	769.44@11.50	2.48	[M-H]^-	20(S)-NG-R2
18	8	1153.51@12.09	2.38	[M+HCOO]^-	Rb1
19	19	1255.63@11.76	2.34	[M+HCOO]^-	Ra2
20	39	981.41@16.66	2.25	[M+Cl]^-	Rd
21	36	793.41@17.00	2.20	[M-H]^-	Chikusetsusaponin Ⅳa
22	26	1285.55@11.98	2.19	[M+HCOO]^-	NG-Fa
23	10	945.50@4.28	2.14	[M-H]^-	Re
24	31	981.43@4.25	2.09	[M+Cl]^-	Re
25	37	1031.49@17.61	2.05	[M-H]^-	m-Rd
26	52	835.41@10.57	1.81	[M+Cl]^-	20(S)-Rf
27	54	1008.49@2.69	1.72	-	Unknown

Origin		Identity	Origin		Identity
XK	HL	Rb1, Rc, Rd, Rg1, Ro	HL	WQ	Rg1, Rb1, m-Rc, m-Rb1
	SH	Rd, Rc, Ro, Rg1		CB	Rg1, Rb1, Rc, Re, m-Rc, m-Rb1
	WQ	Rb1, Rc, Rf, Ra1, Re, m-Rb1	SH	WQ	Rb1, Rc, Rg1, Ro, Ra1, Re
	CB	Rc, Rb1, Rd, Rg1, m-Rc, m-Rb1, Ro		CB	Rb1, Rc, Rd, Rg1, Ra1, Rc, m-Rc
HL	SH	Rg1, Rb1, Rc, Re, m-Rb1	WQ	CB	Re, Rd, Rg1, m-Rc, Ro