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. 2019 Mar 18;24(6):1053. doi: 10.3390/molecules24061053

Non-Targeted Metabolomic Analysis of Methanolic Extracts of Wild-Simulated and Field-Grown American Ginseng

Hongqiang Lin 1, Hailin Zhu 1, Jing Tan 1, Han Wang 1, Qinghai Dong 1, Fulin Wu 1, Yunhe Liu 1, Pingya Li 1,*, Jinping Liu 1,*
PMCID: PMC6470646  PMID: 30889792

Abstract

Aiming at revealing the structural diversity of secondary metabolites and the different patterns in wild-simulated American ginseng (WsAG) and field-grown American ginseng (FgAG), a comprehensive and unique phytochemical profile study was carried out. In the screening analysis, a total of 121 shared compounds were characterized in FgAG and WsAG, respectively. The results showed that both of these two kinds of American ginseng were rich in natural components, and were similar in terms of the kinds of compound they contained. Furthermore, in non-targeted metabolomic analysis, when taking the contents of the constituents into account, it was found that there indeed existed quite a difference between FgAG and WsAG, and 22 robust known biomarkers enabling the differentiation were discovered. For WsAG, there were 12 potential biomarkers including two ocotillol-type saponins, two steroids, six damarane-type saponins, one oleanane-type saponins and one other compound. On the other hand, for FgAG, there were 10 potential biomarkers including two organic acids, six damarane-type saponins, one oleanane-type saponin, and one ursane. In a word, this study illustrated the similarities and differences between FgAG and WsAG, and provides a basis for explaining the effect of different growth environments on secondary metabolites.

Keywords: wild-simulated American ginseng, field-grown American ginseng, screening analysis, metabolomic analysis

1. Introduction

American ginseng (Panax quinquefolius L.) is grouped into four categories: wild, wild-simulated, woods-grown, and field-grown [1,2]. The herb growing in its native habitat is called wild American ginseng. Wild-simulated American ginseng (WsAG) refers to a method of growing ginseng in a hardwood forest environment under natural conditions without any other human intervention [2,3,4]. As such, WsAG roots are indeed indistinguishable from the wild roots due to the similar characteristics. When the seeds are planted in hardwood forests, and are grown in prepared rows or beds, or with removed ground vegetation or fertilizer and pesticides being available [5,6,7], it is called as wood-grown ginseng. This variety of American ginseng requires 6 to 9 years to mature before harvesting [8]. Different from wild-simulated category, the quality of wood-grown ginseng is between that of the wild and field-grown categories. That means, wood-grown American ginseng cannot be considered a substitute of the wild one. Field-grown American ginseng (FgAG), also called cultivated American ginseng, is intensely cultivated under artificial shade structures using fertilizers and pesticides [4,9]. Generally speaking, FgAG is harvested after 3–4 years, while WsAG is collected at least after 10–20 years or longer [10].

Modern pharmacological studies have shown that American ginseng has immunomodulatory [11], anti-tumor [12], anti-fatigue [13,14], anti-diabetic [15,16,17], anti-oxidant effects [18] and the functions of improving impaired memory and learning functions [14,19], etc. Furthermore, it is the traditional belief that roots from the wild are more medicinally efficacious, more potent and more valuable than those from cultivated sources, and wild roots thus command much higher prices on the Chinese medicine market [20,21,22]. But, since the late 18th century, natural wild American ginseng resources suffered a sharp decline due to predatory exploitation in North America under the influence of economic interests, and are nearing extinction now [23]. Meanwhile WsAG, with high quality and four to ten times the retail value of field-grown roots, is similar to the wild one [4]. Actually, planting wild-simulated ginseng is encouraged with the aim of reducing harvest pressure on wild populations [24]. Wild and wild-simulated roots could share the same export and trade regulations due to the similar morphology phenotype and market value [8,25]. Recently, because the so-called wild-simulated American ginseng could capitalize on the premium paid for wild-appearing roots, and the species appeared well suited to the practice of forest farming, American ginseng has been recommended as an agroforestry crop candidate [26]. With the continuous expansion of the folk and clinical applications of American ginseng, it is necessary to conduct an in-depth study on the chemical constituents of American ginseng aiming to clarify the material basis of efficacy. So far, there are a few comparative analysis on FgAG and wild American ginseng [27,28]. These results showed that ginsenosides are different between them [26], especially, the ratios of Rg1/Rd, Rg1/Rb1 and (Rg1 + Re)/Rd are characteristic markers for differentiating these two groups [28]. However, a comparative study on the chemical composition between FgAG and WsAG does not exist.

Untargeted metabolomics, being able to profile diverse classes of metabolites, has been successfully applied to compare and identify the overall small-molecule components of different groups of samples [29]. Ultra-high performance liquid chromatography (UPLC) combined with quadrupole time-of-flight tandem mass spectrometry (QTOF-MS) and multivariate statistical analyses are often applied to profile the different groups. For multivariate statistical analyses, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) are the most common statistical methods. Meanwhile, UPLC-QTOF-MS combined with the automated data processing software UNIFI was often applied recently in the characterization of chemical components of herbal medicines [30,31,32,33,34,35]. When the coeluting constituents possessed different m/z values, HR-MS can provide a specific and accurate mass. While, UNIFI, a highly comprehensive, high throughput, efficient and simple platform, offers a method for integrating data acquisition, data mining, library searching and reporting generation.

Aiming to find out the similarities and differences between FgAG and WsAG, and to provide a reference for quality control and material basis of efficacy, the screening and the comparative analysis of chemical constituents in FgAG and WsAG is conducted for the first time in this paper. The shared constituents would be evaluated with the UPLC-QTOF-MS method combined with UNIFI. The characteristic components were to be found using the untargeted metabolomics method. The results will also be helpful in explaining the different pharmacological activities and controlling the quality of FgAG and WsAG.

2. Results

2.1. Identification of Components from FgAG and WsAG Based on the UNIFI Platform

As a result of our screening analysis, a total of 121 compounds were identified or tentatively characterized in both positive and negative mode from FgAG and WsAG (Table 1), the base peak intensity (BPI) chromatograms are shown in Figure 1, and their chemical structures are shown in Figure 2.

Table 1.

Compounds identified from FgAG and WsAG by UPLC-QTOF-MSE.

No. tR (min) Formula Calculated Mass (Da) TheoreticalMass (Da) Mass Error (ppm) MSE Fragmentation Identification Sources Ref.
1 0.57 C7H12O6 192.0636 192.0634 1.0 191.0563[M − H]−, 173.0454[M − H-H2O]−, 127.0407[M − H-H2O-HCOOH]−, 109.0452[M − H-2H2O-HCOOH]−, 91.0352[M − H-3H2O-HCOOH]− Quinic acid WsAG, FgAG s
2 0.64 C12H22O11 342.1162 342.1165 0.8 341.1092[M − H]−, 179.0562[M − H-Glu]− α-Maltose WsAG, FgAG s
3 0.77 C10H13N5O4 267.0959 267.0968 −3.3 268.1031[M+H]+, 237.0874[M + H-CH2OH]+, 226.0898[M + H-CN2H2]+, 136.0612[M + H-Rib]+, 130.0495[M + H-CH2OH-C4N4H3]+ Adenosine WsAG, FgAG s
4 0.93 C12H22O11 342.1168 342.1162 1.7 341.1095[M − H]−, 287.1097[M − H-3H2O]−, 179.0563[M − H-Glu]− Sucrose WsAG, FgAG s
5 0.95 C9H11NO2 165.0777 165.0782 −3.0 166.0850[M + H]+, 150.0589[M + H-NH2]+, 132.0486[M + H-H2O-NH2]+, 120.0807[M + H-HCOOH]+, 91.0559[M + H-CH-NH2-HCOOH]+ L-Phenylalanine WsAG, FgAG s
6 * 1.02 C14H18O10 346.0903 346.0900 1.0 345.0830[M − H]−, 327.0598[M − H-H2O]−, 309.0728[M − H-2H2O]−, 165.0195[M − H-Glu]−, 150.0115[M − H-Glu-CH3]− Methyl gallate 3-O-β-d-glucoside WsAG > FgAG
VIP: 14.18
p < 0.001		 s
7 1.51 C11H12N2O2 204.0899 204.0899 −0.1 203.0826[M − H]−, 141.0660[M − H-HCOOH-NH2]−, 129.0506[M − H-C3H6O2]− L-Tryptophane WsAG, FgAG s
8 4.71 C17H20O9 368.1106 368.1107 −0.4 367.1033[M − H]−, 191.0754[M − H-C10H9O3]−, 193.0466[M − H-GluA]−, 177.0758[M − H-GluA-CH3]−, 127.0350[M − H-GluA-H2O-OCH3]− 3-O-trans-Feruloylquinic Acid WsAG, FgAG [36]
9 4.92 C48H82O19 962.5440 962.5450 −1.0 1007.5432[M + HCOO]−, 763.2898[M − H-2H2O-Glu]−, 815.4784[M − H-Rha]−, 781.3155[M − H-Glu]−, 635.2307[M − H-Glu-Rha-H2O]−, 437.1863[M − H-2Glu-Rha-3H2O]− Majoroside F6 WsAG, FgAG [37]
10 5.18 C36H58O8 618.4130 618.4132 −0.3 619.4203[M + H]+, 439.3712[M + H-Glu]+, 422.3451[M + H-Glu-OH]+, 383.2823[M + H-Glu-C4H8]+, 297.2336[M + H-Glu-C9H16O]+ Oleanolic acid -28-O-β-d-glucopyranoside WsAG, FgAG s
11 5.32 C48H82O20 978.5399 978.5397 −0.3 1023.5379[M + HCOO]−, 997.5331[M − H]−,815.4972[M − H-Glu]−, 797.4718[M − H-Glu-H2O]−, 653.3389[M − H-2Glu]−, 491.2724[M − H-3Glu]− Yesanchinoside B WsAG, FgAG [38]
12 * 5.54 C47H80O19 948.5305 948.5294 1.2 993.5270[M + HCOO]−, 815.4921[M − H-Ara]−, 653.3392[M − H-2Glu-Ara]−, 473.3030[M − H-2Glu-Ara]−, 455.3922[M − H-2Glu-Ara-H2O]−, 391.2582[M − H-2Glu-Ara-C6H12O]− Yesanchinoside C WsAG > FgAG
VIP: 6.18
p < 0.001		 [38]
13 # 5.78 C48H82O19 962.5443 962.5450 −1.0 1007.5436[M + HCOO]−, 961.5388[M − H]−, 799.4784[M − H-Glu]−, 637.2307[M − H-2Glu]−, 475.5863[M − H-3Glu]− Notoginsenoside N WsAG < FgAG
VIP: 11.83
p < 0.001		 [39]
14 # 5.94 C42H74O15 818.5031 818.5028 0.4 863.5006[M + HCOO]−, 667.4323[M − H-Rha]−, 533.2329[M − H-C6H13O2-C11H19O]−, 506.3845[M − H-Glu-Rha]−, 477.2169[M − H-C20H36O4]− Quinquenoside L9 WsAG < FgAG
VIP: 7.20
p = 0.0002		 s
15 5.94 C42H72O14 800.4915 800.4922 −0.9 801.4988[M + H]+, 621.4983[M + H-Glu-H2O]+, 459.3659[M + H-2Glu]+, 423.3450[M + H-2Glu-3H2O]+ Majoroside F2 WsAG, FgAG [40]
16 6.25 C26H34O11 522.2097 522.2101 −0.7 567.2079[M + HCOO]−, 521.2204[M − H]−, 458.2935[M − H-H2O-C2H5O]−, 341.1396[M − H-Glu-H2O]−, 178.0559[M − H-C20H23O5]− Urolignoside WsAG, FgAG [41]
17 6.48 C54H92O23 1108.6034 1108.6029 0.4 1153.6016[M + HCOO]−, 961.5452[M − H-Rha]−, 799.4902[M − H-Glu-Rha]−, 637.2950[M − H-2Glu-Rha]−, 475.2681[M − H-3Glu-Rha]− Yesanchinoside E WsAG, FgAG [42]
18 6.77 C47H80O18 932.5348 932.5345 0.3 977.5318[M + HCOO]−, 931.5271[M − H]−, 799.4697[M − H-Ara]−, 769.4734[M − H-Glu]−, 637.3146[M − H-Glu-Ara]− Quinquenoside F6 WsAG, FgAG [37]
19 6.88 C48H82O19 962.5450 962.5450 −0.1 1007.5432[M + HCOO]−, 859.4881[M − H-C5H10O2]−, 799.4204[M − H-Glu]−, 696.4328[M − H-Glu-C5H9O]−, 637.3158[M − H-2Glu]−, 601.2316[M − H-2Glu-2H2O]− Quinquenoside L2 WsAG, FgAG [43]
20 7.01 C42H72O14 800.4914 800.4922 −1.0 845.4896[M + HCOO]−, 799.4836[M − H]−, 653.4319[M − H-Rha]−, 491.2475[M − H-Glu-Rha]− (24S)-Pseudoginsenoside F11 WsAG, FgAG s
21 * 7.05 C47H80O18 932.5349 932.5345 0.4 977.5346[M + HCOO]−, 840.4930[M − H-2H2O-C4H7]−, 799.4859[M − H-Xyl]−, 769.4735[M − H-Glu]−, 637.4321[M − H-Glu-Xyl]− Notoginsenoside R1 WsAG > FgAG
VIP: 24.59
p < 0.001		 s
22 * 7.10 C28H48O 400.3723 400.3705 4.3 423.3620[M+Na]+, 382.2862[M + H-CH3]+, 339.2934[M + H-H2O-C3H7]+, 255. 2948[M + H-H2O-C9H19]+ Methylcholesta-7-en-3β-ol WsAG > FgAG
VIP: 4.69
p < 0.001		 [44]
23 7.12 C48H82O19 962.5435 962.5450 −1.5 1007.54171[M + HCOO]−, 961.5329[M − H]−, 799.3722[M − H-Glu]−, 637.4321[M − H-2Glu]−, 475.3722[M − H-3Glu]−, 391.4833[M − H-3Glu-C6H12]− Notoginsenoside R6 WsAG, FgAG [42]
24 7.25 C47H80O18 932.5335 932.5345 −1.0 977.5317[M + HCOO]−, 931.5282[M − H]−, 799.4701[M − H-Xyl]−, 673.3294[M − H-Xyl-Glu]−, 475.2148[M − H-Xyl-2Glu]− Notoginsenoside ST5 WsAG, FgAG [45]
25 7.29 C54H90O24 1122.5818 1122.5822 0.4 1167.5812[M + HCOO]−, 1121.5747[M − H]−, 959.5120[M − H-Glu]−, 797.4669[M − H-2Glu]−, 473.4334[M − H-4Glu]− Quinquenoside IV WsAG, FgAG [42]
26 7.40 C42H72O14 800.4918 800.4922 −0.5 845.4900[M + HCOO]−, 784.4683[M − H-CH3]−, 637.4340[M − H-Glu]−, 471.3787[M − H-2Glu]− Ginsenoside Rg1 WsAG, FgAG s
27 7.47 C48H82O18 946.5491 946.5501 −1.0 991.5473[M + HCOO]−, 945.5413[M − H]−, 783.5142[M − H-Glu]−, 637.4125[M − H-Glu-Rha]−, 475.5147[M − H-2Glu-Rha]− Ginsenoside Re WsAG, FgAG s
28 7.47 C28H48O 400.3715 400.3705 2.4 423.3617[M+Na]+, 401.3540[M + H]+, 383.2861[M + H-H2O]+, 325.2982[M + H-H2O-CH3-C3H7]+, 284.1420[M + H-H2O-C7H15]+, 175.1221[M + H-H2O-C15H28]+ Campesterol WsAG, FgAG a
29 7.69 C45H74O17 886.4926 886.4925 −0.1 885.4853[M − H]−, 799.4748[M − H-Mal]−, 637.4303[M − H-Mal-Glu]−, 475.3751[M − H-Mal-2Glu]− Malonyl-ginsenoside Rg1 WsAG, FgAG [46]
30 7.85 C42H72O14 800.4929 800.4922 0.8 845.4911[M + HCOO]−, 799.4837[M − H]−, 653.3687[M − H-Rha]−, 491.2354[M − H-Rha-Glu]− Quinquenoside L11 WsAG, FgAG s
31 # 7.84 C51H84O21 1032.5505 1032.5532 2.6 1031.5414[M − H]−, 945.5212[M − H-Mal]−, 783.4173[M − H-Mal-Glu]−, 637.4385[M − H-Mal-Rha-Glu-Ac]−, 475.3932[M − H-Mal-2Glu-Rha]− Malonyl-ginsenoside Re WsAG < FgAG
VIP: 5.65
p = 0.0060		 [39]
32 7.94 C47H80O19 948.5282 948.5294 −1.2 947.5209[M − H]−, 815.4786[M − H-Xyl]−, 653.2758[M − H-Glu-Xyl]−, 491.1787[M − H-2Glu-Xyl]− Vinaginsenoside R6 WsAG, FgAG [39]
33 8.15 C47H80O17 916.5409 916.5396 1.4 961.5377[M + HCOO]−, 915.5306[M − H]−, 783.4819[M − H-Ara]−, 753.4732[M − H-Glu]−, 621.4290[M − H-Ara-Glu]−, 459.4687[M − H-2Glu-Ara]− Quinquenoside L14 WsAG, FgAG [47]
34 8.23 C44H74O15 842.4996 842.5028 −3.6 887.4978[M + HCOO]−, 841.4939[M − H]−, 799.4833[M − H-Ac]−, 695.4459[M − H-Glu]−, 653.4321[M − H-Ac-Rha]−, 684.3932[M − H-CH3-C8H14O2]−, 491.4219[M − H-Rha-Glu-Ac]− Vinaginsenoside R1 WsAG, FgAG [48]
35 * 8.26 C54H94O24 1126.6162 1126.6135 2.4 1171.6110[M + HCOO]−, 1125.6094[M − H]−, 975.5349 [M − H-Xyl-H2O]−, 963.5502[M − H-Glu]−, 801.3547[M − H-2Glu]−, 831.3214[M − H-Glu-Xyl]−, 507.3214[M − H-3Glu-Xyl]− Quinquenoside F3 WsAG > FgAG
VIP: 3.35
p < 0.001		 s
36 8.57 C44H74O15 842.5028 842.5012 −1.7 887.4995[M + HCOO]−, 841.4939[M − H]−, 637.4321[M − H-Glu-Ac]−, 475.3030[M − H-2Glu-Ac]−, 391.4158[M − H-2Glu-Ac-C6H11]− Acetyl-Ginsenoside Rg1 WsAG, FgAG [39]
37 * 8.59 C41H70O13 770.4808 770.4816 −1.0 815.4798[M + HCOO]−, 769.4722[M − H]−, 637.4321[M − H-Ara]−, 475.2678[M − H-Ara-Glu]−, 391.1748[M − H-Ara-Glu-C6H11]− Notoginsenoside R2 WsAG > FgAG
VIP: 4.83
p < 0.001		 [42]
38 8.63 C48H82O19 962.5423 962.5450 −2.7 1007.5405[M + HCOO]−, 961.5371[M − H]−, 815.4317[M − H-Rha]−, 799.4622[M − H-Glu]−, 653.4385[M − H-Glu-Rha]−, 617.4316[M − H-Glu-Rha-H2O]−, 491.2912[M − H-2Glu-Rha]− Majoroside F5 WsAG, FgAG [37]
39 8.64 C30H48O2 440.3646 440.3654 −1.9 441.3719[M + H]+, 423.3606[M + H-H2O]+, 339.2908[M + H-HCOOH-C4H8]+, 248.2948[M + H-C14H24]+, 203.1849[M + H-HCOOH-C14H24]+ Deoxyoleanolic acid WsAG, FgAG [46]
40 8.78 C48H80O18 944.5338 944.5345 −0.6 989.5320[M + HCOO]−, 943.5250[M − H]−, 781.4541[M − H-Glu]−, 619.4143[M − H-2Glu]−, 457.5876[M − H-3Glu]− Quinquenoside L1 WsAG, FgAG [49]
41 8.83 C53H88O23 1092.5718 1092.5716 0.2 1137.5696[M + HCOO]−, 1091.5641[M − H]−, 959.5571[M − H-Xyl]−, 929.4601[M − H-Glu]−, 797.4852[M − H-Glu-Xyl]− Yesanchinoside G WsAG, FgAG [50]
42 * 8.89 C47H78O17 914.5225 914.5239 −1.5 959.5225[M + HCOO]−, 913.5147[M − H]−, 733.2547[M − H-H2O-Glu]−, 619.4527[M − H-Glu-Xyl]−, 457.3254[M − H-2Glu-Xyl]− Quinquenoside L8 WsAG > FgAG
VIP: 3.97
p = 0.0004		 s
43 8.97 C48H82O19 962.5438 962.5450 −1.2 1007.5420[M + HCOO]−, 946.5212[M − H-CH3]−, 781.4533[M − H-Glu-H2O]−, 637.4321[M − H-2Glu]−, 475.3932[M − H-3Glu]− Majoroside F1 WsAG, FgAG [40]
44 * 9.14 C42H70O13 782.4325 782.4816 1.2 781.4747[M − H]−, 619.4181[M − H-Glu]−, 457.4798[M − H-2Glu]−, 376.4797[M − H-2Glu-C6H9]− Quinquenoside F1 WsAG > FgAG
VIP: 4.10
p = 0.0004		 [51]
45 9.27 C15H10O6 286.0480 286.0477 0.8 285.0407[M − H]−, 227.0521[M − H-C2H2O2]−, 151.0037[M − H-C8H6O2]−, 106.0148[M − H-C9H7O4]−, 112.0351[M − H-C9H5O5]− Kaempferol WsAG, FgAG s
46 # 9.32 C53H86O24 1118.5514 1118.5509 0.5 1117.5436[M − H]−, 1040.5481[M − H-CH3OH]−, 955.3219[M − H-Glu]−, 793.2905[M − H-2Glu]−, 453.1095[M − H-3Glu-GluA]− Ginsenoside ROA WsAG < FgAG
VIP: 12.60
p < 0.001		 [52]
47 * 9.54 C41H70O14 786.4775 786.4766 1.1 831.4775[M + HCOO]−, 767.4297[M − H-H2O]−, 653.4318[M − H-Xyl]−, 491.2015[M − H-Glu-Xyl]− Majonoside R2 WsAG > FgAG
VIP: 25.80
p < 0.001		 [39]
48 9.61 C48H80O19 960.5285 960.5294 −0.9 1005.5267[M + HCOO]−, 941.5316[M − H-H2O]−, 797.4287[M − H-Glu]−, 635.3221[M − H-2Glu]−, 473.2684[M − H-3Glu]− Notoginsenoside G WsAG, FgAG [42]
49 9.68 C42H72O14 800.4922 800.4922 0.0 845.4904[M + HCOO]−, 799.4844[M − H]−, 783.2451[M − H-Rha]−, 621.3547[M − H-Glu-Rha]− Pseudo-ginsenoside F11 WsAG, FgAG s
50 9.73 C36H62O10 654.4340 654.4343 −0.4 699.4322[M + HCOO]−, 653.4262[M − H]−, 635.4312[M − H-H2O]−, 491.3254[M − H-Glu]− Pseudo-ginsenoside RT5 WsAG, FgAG s
51 9.78 C36H62O10 654.4337 654.4343 −0.9 655.4410[M + H]+, 599.4418[M − H-3H2O]+, 493.3437[M − H-Glu]+, 457.2651[M − H-Glu-2H2O]+ Pseudo-ginsenoside RT4 WsAG, FgAG [39]
52 9.82 C59H100O27 1240.6458 1240.6452 0.5 1239.6380[M − H]−, 1107.6376[M − H-Xyl]−, 954.6930[M − H-Xyl-Glu]−, 783.4833[M − H-Xyl-2Glu]−, 621.4431[M − H-Xyl-3Glu]−, 459.4943[M − H-Xyl-4Glu]− Ginsenoside Ra3 WsAG, FgAG [53]
53 9.96 C41H70O13 770.4810 770.4816 −0.8 815.4804[M + HCOO]−, 751.4804[M − H-H2O]−, 637.4904[M − H-Ara]−, 475.3804[M − H-Glu-Ara]− Ginsenoside F5 WsAG, FgAG [39]
54 9.98 C58H98O26 1210.6350 1210.6346 0.3 1209.6272[M − H]−, 1077.5814[M − H-Xyl]−, 945.4706[M − H-Xyl-Ara]−, 783.4803[M − H-Xyl-Ara-Glu]−, 459.4799[M − H-Xyl-Ara-3Glu]− Ginsenoside Ra2 WsAG, FgAG [53]
55 10.02 C41H66O11 734.4590 734.4605 −2.1 735.4663[M + H]+, 589.3646[M + H-Rha]+, 457.3705[M + H-Rha-Ara]+, 441.5712[M + H-Rha-Ara-HCOOH]+ Eleutheroside K WsAG, FgAG [54]
56 10.04 C48H80O18 944.5320 944.5345 −2.6 989.5302[M + HCOO]−, 943.5263[M − H]−, 781.4839[M − H-Glu]−, 619.4206[M − H-2Glu]−, 457.5701[M − H-3Glu]− Quinquenoside L6 WsAG, FgAG -
57 10.07 C30H48O2 440.3638 440.3654 −3.6 441.3717[M + H]+, 394.3508[M + H-H2O-CHO]+, 328.3504[M + H-CHO-C6H12]+, 219.1792[M + H-C15H26O]+, 205.1619[M + H-H2O-C15H22O]+ 3β-Hydroxyolean-12-en-28-al WsAG, FgAG a
58 10.14 C59H100O27 1240.6452 1240.6462 0.8 1285.6444[M + HCOO]−, 1107.5976[M − H-Xyl]−, 945.4900[M − H-Xyl-Glu]−, 783.4835[M − H-Xyl-2Glu]−, 459.4929[M − H-Xyl-4Glu]− Notoginsenoside Fa WsAG, FgAG [53]
59 10.24 C54H92O23 1108.6039 1108.6029 0.8 1153.6021[M + HCOO]−, 1107.5961[M − H]−, 943.5414[M − H-Glu]−, 763.4784[M − H-2Glu]−, 615.4417[M − H-3Glu]− Ginsenoside Rb1 WsAG, FgAG s
60 10.24 C30H48O 424.3692 424.3705 −3.1 425.3765[M + H]+, 409.3102[M + H-H2O]+, 371.3759[M + H-CH3-C3H5]+, 189.1614[M + H-C16H26O]+, 205.1775[M + H-C15H26O]+ Olean-18-en-3-one WsAG, FgAG [55]
61 10.31 C57H94O26 1194.6054 1194.6033 1.7 1193.5981[M − H]−, 1077.5402[M − H-mal]−, 945.5097[M − H-mal-Glu]−, 783.4906[M − H-mal-2Glu]−, 621.4906[M − H-mal-3Glu]− Malonyl-ginsenoside Rb1 WsAG, FgAG [53]
62 10.33 C42H72O13 784.4975 784.4973 0.3 829.4957[M + HCOO]−, 768.4744[M − H-CH3]−, 635.4330[M − H-Rha]−, 471.3782[M − H-Glu-Rha]− 20(R)-Ginsenoside Rg2 WsAG, FgAG s
63 10.35 C36H62O9 638.4391 638.4394 −0.4 683.4373[M + HCOO]−, 637.4313[M − H]−, 475.2658[M − H-Glu]−, 457.2235[M − H-Glu-H2O]− 20(S)-Ginsenoside Rh1 WsAG, FgAG s
64 10.36 C41H70O13 770.4817 770.4816 0.0 815.4799[M + HCOO]−, 678.4450[M − H-2H2O-C4H7]−, 637.4321[M − H-Ara]−, 590.2706[M − H-C4H7-C9H16]−, 475.2622[M − H-Glu-Ara]− Ginsenoside F3 WsAG, FgAG [39]
65 10.39 C53H90O22 1078.5931 1078.5924 0.6 1123.5913[M + HCOO]−, 943.5423[M − H-Araf]−, 854.4890[M − H-H2O-Araf-C4H7]−, 763.4850[M − H-Glu-Araf]− Ginsenoside Rc WsAG, FgAG s
66 10.42 C36H60O8 620.4276 620.4288 −1.9 621.4349[M + H]+, 603.4238[M + H-H2O]+, 441.3714[M + H-Glu]+, 423.3612[M + H-Glu-H2O]+, 350.2971[M + H-Glu-2H2O-C4H7]+, 341.1160[M + H-Glu-C6H12O]+ Ginsenoside Rh4 WsAG, FgAG [56]
67 10.46 C58H98O26 1210.6353 1210.6346 0.6 1209.6275[M − H]−, 1077.5914[M − H-Xyl]−, 945.4807[M − H-Xyl-Ara]−, 783.4687[M − H-Xyl-Ara-Glu]−, 459.4329[M − H-Xyl-Ara-3Glu]− Ginsenoside Ra1 WsAG, FgAG [53]
68 10.58 C56H92O25 1164.5932 1164.5928 0.3 1163.5859[M − H]−, 1119.5976[M − H-CO2]−, 1077.6021[M − H-Mal]−, 1031.5694[M − H-Araf]−, 945.4900[M − H-Araf-Mal]−, 783.4835[M − H-Glu-Araf-Mal]− Malonyl-ginsenoside Rc WsAG, FgAG [53]
69 10.62 C48H76O19 956.4976 956.4981 −0.5 955.4903[M − H]−, 783.4214[M − H-GluA]−, 631.4157[M − H-2Glu]−, 459.4174[M − H-2Glu-GluA]− Ginsenoside Ro WsAG, FgAG s
70 # 10.69 C30H46O2 438.3486 438.3498 −2.7 439.3563[M + H]+, 424.3600[M + H-CH3]+, 411.1114[M + H-CO]+, 233.1676[M + H-C15H24]+, 205.1928[M + H-C15H22O2]+, 190.1778[M + H-C16H23O2]+ 3,11-dioxo-β-amyrene WsAG < FgAG
VIP: 6.49
p < 0.001		 [57]
71 10.70 C53H84O23 1088.5402 1088.5403 −0.1 1087.5326[M − H]−, 955.5235[M − H-Ara]−, 925.4610[M − H-Glu]−, 793.2350[M − H-Ara-Glu]−, 455.4611[M − H-Ara-2Glu-GluA]− Stipuleanoside R2 WsAG, FgAG [39]
72 10.78 C53H90O22 1078.5924 1078.5924 0.0 1123.5906[M + HCOO]−, 913.5403[M − H-Glu]−, 779.4886[M − H-Glu-Ara]−, 615.4431[M − H-2Glu-Ara]− Ginsenoside Rb2 WsAG, FgAG s
73 10.79 C53H90O22 1078.5924 1078.5924 0.0 1123.5320[M + HCOO]−, 913.4581[M − H-Glu]−, 779.3696[M − H-Glu-Xyl]−, 615.4912[M − H-2Glu-Xyl]−, 451.3672[M − H-3Glu-Xyl]− Ginsenoside Rb3 WsAG, FgAG s
74 10.89 C55H92O23 1120.6009 1120.6029 −1.8 1165.5991[M + HCOO]−, 1077.3151[M − H-Xyl]−, 945.5076[M − H-Ara-Xyl]−, 783.3942[M − H-Ara-Xyl-Glu]−, 621.4742[M − H-Ara-Xyl-2Glu]− Notoginsenoside Fc WsAG, FgAG [53]
75 10.94 C56H92O25 1164.5937 1164.5928 0.8 1163.5864[M − H]−, 1077.5570[M − H-Mal]−, 945.5302[M − H-Ara-Mal]−, 783.4540[M − H-Glu-Ara-Mal]−, 621.4570[M − H-2Glu-Ara-Mal]− Malonyl-ginsenoside Rb2 WsAG, FgAG [53]
76 11.01 C56H94O24 1150.6138 1150.6135 0.3 1195.6120[M + HCOO]−, 1149.6060[M − H]−, 1107.4997[M − H-Ac]−, 987.4976[M − H-Glu]−, 945.6047[M − H-Glu-Ac]−, 783.4864[M − H-2Glu-Ac]− Quinquenoside R1 WsAG, FgAG [53]
77 11.02 C47H74O18 926.4864 926.4875 −1.2 925.4791[M − H]−, 793.4272[M − H-Ara]−, 612.3784[M − H-GluA-Ara]−, 540.3784[M − H-Glu-C14H21O]−, 455.2841[M − H-Glu-Ara-GluA]− Chikusetsu saponin IV WsAG, FgAG [39]
78 11.14 C56H92O25 1164.4967 1164.4958 0.8 1163.4925[M − H]−, 1077.5760[M − H-Mal]−, 945.5503[M − H-Mal-Xyl]−, 783.4735[M − H-Mal-Xyl-Glu]−, 621.4269[M − H-Mal-Xyl-2Glu]− Malonyl-ginsenoside Rb3 WsAG, FgAG [53]
79 11.16 C36H62O9 638.4395 638.4394 0.2 683.4366[M + HCOO]−, 637.4317[M − H]−, 475.2574[M − H-Glu]−, 457.2147[M − H-Glu-H2O]− 20(R)-ginsenoside Rh1 WsAG, FgAG s
80 11.18 C43H72O15 828.4864 828.4871 −0.8 873.4846[M + HCOO]−, 784.4798[M − H-COCH3]−, 695.2912[M − H-Xyl]−, 491.4938[M − H-Xyl-Glu-Ac]−, 455.2535[M − H-Xyl-Glu-Ac-2H2O]− Vinaginsenoside R2 WsAG, FgAG [39]
81 # 11.20 C48H82O17 930.5546 930.5552 −0.7 929.5474[M − H]−, 767.4642[M − H-Glu]−, 605.4365[M − H-2Glu]−, 443.1196[M − H-3Glu]− Vinaginsenosides R3 WsAG < FgAG
VIP: 7.60
p < 0.001		 [58,59]
82 11.34 C42H66O14 794.4447 794.4453 −0.8 793.4368[M − H]−, 631.3279[M − H-Glu]−, 613.4222[M − H-Glu-H2O]−, 569.2927[M − H-Glu-HCOOH]−, 455.1562[M − H-Glu-GluA]− Chikusetsu saponin II WsAG, FgAG [39]
83 11.36 C48H82O18 946.5508 946.5501 0.7 991.5490[M + HCOO]−, 945.5430[M − H]−, 783.5147[M − H-Glu]−, 459.3241[M − H-3Glu]− Ginsenoside Rd WsAG, FgAG s
84 11.39 C55H92O23 1120.6029 1120.6049 1.7 1119.5941[M − H]−, 1077.5699[M − H-Ac]−, 943.4874[M − H-Ac-Ara]−, 779.1224[M − H-Ac-Ara-Glu]−, 451.2649[M − H-Ac-Ara-3Glu]− Ginsenoside Rs1 WsAG, FgAG s
85 11.52 C51H84O21 1032.5503 1032.5505 −0.2 1031.5425[M − H]−, 987.5520[M − H-CO2]−, 945.5192[M − H-mal]−, 783.3540[M − H-mal-Glu]−, 621.2570[M − H-mal-2Glu]−, 459.3458[M − H-mal-3Glu]− Malonyl-ginsenoside Rd WsAG, FgAG [53]
86 11.70 C55H92O23 1120.6039 1120.6029 0.8 1165.6021[M + HCOO]−, 1119.5961[M − H]−, 987.3684[M − H-Ara]−, 914.4587[M − H-Glu-Ac]−, 458.5471[M − H-3Glu-Ara-Ac]− Ginsenoside Rs2 WsAG, FgAG s
87 11.88 C48H82O18 946.5501 946.5500 −0.1 991.5482[M + HCOO]−, 783.4871[M − H-Glu]−, 603.4416[M − H-2Glu]− Gypenoside XVII WsAG, FgAG s
88 12.20 C19H36O5 344.2565 344.2563 0.6 343.2486[M − H]−, 329.0232[M − H-CH3]−, 311.2112[M − H-H2O-CH3]−, 294.1609[M − H-H2O-OCH3]−, 255.1494[M − H-OCH3-C4H9]−, 242.1610[M − H-OCH3-C5H11]−, 228.2523[M − H-OCH3-C6H12]− Methyl-9,10,11-trihydroxy-12- octadecenoate WsAG, FgAG [60]
89 12.27 C47H80O17 916.5398 916.5396 0.2 961.5380[M + HCOO]−, 866.4901[M − H-H2O-CH2OH]−, 783.4749[M − H-Ara]−, 753.4664[M − H-Glu]−, 621.4616[M − H-Glu-Ara]−, 459.4008[M − H-2Glu-Ara]− Notoginsenoside Fe WsAG, FgAG [39]
90 12.38 C50H84O19 988.5594 988.5607 −1.3 1033.5576[M + HCOO]−, 987.5539[M − H]−, 945.5428[M − H-COCH3]−, 809.4326[M − H-2H2O-C8H14O2]−, 797.4813[M − H-Glu-C2H4]− Quinquenoside III WsAG, FgAG [61]
91 # 12.48 C47H80O17 916.5385 916.5396 −1.1 961.5385[M + HCOO]−, 900.5146[M − H-CH3]−, 783.49.6[M − H-Ara]−, 630.4290[M − H-Glu-2H2O-C5H9]−, 621.3500[M − H-Glu-Ara]−, 459.2328[M − H-2Glu-Ara]− Chikusetsu saponin III WsAG < FgAG
VIP: 12.78
p < 0.001		 [39]
92 12.57 C30H46O2 766.4855 766.4867 −1.6 767.4928[M + H]+, 749.3674[M + H-H2O]+, 621.2398[M + H-Rha]+, 459.2280[M + H-Glu-Rha]+, 207.1780[M + H-Glu-Rha-C16H26O]+ (20E)-Ginsenoside F4 WsAG, FgAG [62]
93 12.68 C47H80O17 916.5399 916.5396 0.3 961.5381[M + HCOO]−, 814.4616[M − H-H2O-C6H11]−, 783.4923[M − H-Xyl]−, 621.4390[M − H-Glu-Xyl]− Gypenoside IX WsAG, FgAG [53]
94 13.14 C42H70O14 798.4748 798.4765 −2.1 797.4676[M − H]−, 651.4246[M − H-Rha]−, 489.4158[M − H-Glu-Rha]− Ginsenoside Rg8 WsAG, FgAG [63]
95 13.18 C52H86O19 1014.5756 1014.5763 −0.6 1059.5738[M + HCOO]−, 1013.5685[M − H]−, 945.5933[M − H-C4H5O]−, 851.4510[M − H-Glu]−, 833.4903[M − H-Glu-H2O]−, 620.2875[M − H-2Glu-C4H5O]−, 458.2663[M − H-3Glu-C4H5O]− Quinquenoside I WsAG, FgAG [61]
96 13.31 C48H82O17 930.5548 930.5552 −0.4 929.5470[M − H]−, 783.4642[M − H-Rha]−, 767.4695[M − H-Glu]−, 621.4365[M − H-Glu-Rha]−, 459.3956[M − H-2Glu-Rha]− Gypenoside X WsAG, FgAG -
97 13.43 C42H70O12 766.4861 766.4867 −0.8 765.4783[M + HCOO]−, 610.2361[M − H-CH2OH-C8H13-CH3]−, 603.2375[M − H-Glu]−, 441.1811[M − H-2Glu]−, 340.2323[M − H-C30H49O]− Ginsenoside Rk1 WsAG, FgAG [39]
98 13.50 C47H74O18 926.4862 926.4875 −1.4 925.4789[M − H]−, 793.4879[M − H-Ara]−, 731.4457 [M − H-Ara-HCOOH]−, 727.4338[M − H-Glu-H2O]−, 659.4254[M − H-Glu-C3H4-HCOOH]−, 569.4945 [M − H-Ara-Glu-HCOOH]−, 455. 4979 [M − H-Ara- Glu-GluA]− Chikusetsu saponin Ib WsAG, FgAG [39]
99 # 13.52 C42H72O13 784.4974 784.4973 −0.1 783.4896[M − H]−, 737.4755[M − H-CH2OH-CH3]−, 660.4330[M − H-3H2O-C5H9]−, 621.4361[M − H-Glu]−, 459.3782[M − H-2Glu]− Ginsenoside F2 WsAG < FgAG
VIP: 17.01
p < 0.001		 [39]
100 13.57 C18H30O4 310.2141 310.2144 −1.0 309.2068[M − H]−, 291.1960[M − H-H2O]−, 185.1181[M − H-COOH-C6H9]−, 171.1024[M − H-CH2COOH-C6H9]− 13S-hydroperoxy-9Z,11E,15Z- octadecatrienoic acid WsAG, FgAG [64]
101 13.62 C36H60O7 604.4334 604.4339 −0.8 605.4407[M + H]+, 586.4285[M + H-H2O]+, 443.3860[M + H-Glu]+, 405.3657[M + H-Glu-H2O]+, 333.0939[M + H-2H2O-C16H26O]+, 296.1006[M + H-Glu-H2O-C8H13]+ Isoginsenoside Rh3 WsAG, FgAG [65]
102 * 13.93 C42H66O14 794.4449 794.4453 −0.5 793.4387[M − H]−, 613.3751[M − H-Glu]−, 569.3830[M − H-Glu-H2O-CO2]− Chikusetsusaponin Iva WsAG > FgAG
VIP:16.17
p < 0.001		 [53]
103 14.51 C42H72O13 784.4967 784.4973 −0.7 829.4951[M + HCOO]−, 783.4892[M − H]−, 621.4442[M − H-Glu]−, 459.3684[M − H-2Glu]− 20(R)-Ginsenoside Rg3 WsAG, FgAG s
104 14.64 C17H30O2 266.2246 266.2246 −0.1 311.2228[M + HCOO]−, 168.1023[M − H-C7H13]−, 154.1074[M − H-C8H15]−, 137.2250[M − H-C7H13-OCH3]−, 115.0352[M − H-C4H7-C6H9O]−, 96.0352[M − H-C11H21O]− 5-Hexenoic acid, 10-undecenyl ester WsAG, FgAG a
105 # 14.74 C18H28O2 276.2081 276.2089 −2.9 277.2156[M + H]+, 150.1312[M + H-C7H11O2]+, 137.0951[M + H-H2O-C9H14]+, 110.1017[M + H-C10H15O2]+ Palmitoleic acid WsAG < FgAG
VIP: 8.57
p < 0.001		 s
106 14.75 C42H72O13 784.4940 784.4973 −4.1 829.4951[M + HCOO]−, 783.4865[M − H]−, 621.4942[M − H-Glu]−, 459.4578[M − H-2Glu]−, 441.5214[M − H-2Glu-H2O]− 20(S)-Ginsenoside Rg3 WsAG, FgAG s
107 14.90 C41H70O12 754.4874 754.4867 0.8 799.4856[M + HCOO]−, 621.3141[M − H-Xyl]−, 459.3887[M − H-Glu-Xyl]−, 351.2556[M − H-Xyl-H2O-C16H28O2]−, 275.1442[M − H-Glu-Xyl-2C6H11]− Gypenoside XIII WsAG, FgAG [66]
108 14.98 C41H64O13 764.4345 764.4347 −0.3 763.4272[M − H]−, 613.3766[M − H-Xyl]−, 569.3856[M − H-Xyl-HCOOH]− Pseudo-ginsenoside Rp1 WsAG, FgAG [39]
109 15.05 C17H30O2 266.2244 266.2246 −0.7 311.2226[M + HCOO]−, 222.1128[M − H-C3H7]−, 139.0826[M − H-C9H19]−, 127.1127[M − H-C8H11O2]− (2E,4E)-Hydroprene WsAG, FgAG a
110 15.75 C43H68O14 808.4610 808.4609 0.1 807.4532[M − H]−, 609.3820[M − H-Glu-H2O]−, 455.3519[M − H-Glu-Glu acid methyl ester]−, 319.1792[M − H-Glu acid methyl ester-C21H32]− Chikusetsusaponin IVa methyl ester WsAG, FgAG [39]
111 15.98 C18H30O3 294.2194 294.2195 −0.3 293.2122[M − H]−, 275.2013[M − H-H2O]−, 171.1024[M − H-C9H15]−, 121.1020[M − H-C9H15O3]− (E,E)-9-Oxooctadeca-10,12-dienoic acid WsAG, FgAG [34]
112 * 16.90 C36H62O8 622.4431 622.4445 −2.1 667.4442[M + HCOO]−, 621.4360[M − H]−, 459.2656[M − H-Glu]−, 441.4772[M − H-Glu-H2O]− Ginsenoside Rh2 WsAG > FgAG
VIP: 4.68
p = 0.0032		 s
113 17.34 C18H32O3 296.2347 296.2351 1.4 295.2274[M − H]−, 278.2172[M − H-H2O]−, 233.2273[M − H-HCOOH-O]−, 184.1182[M − H-C8H15]−, 171.1023[M − H-C9H16]−, 148.1125[M − H-C8H15O2]−, 125.1174[M − H-H2O-C10H17O]− 9-Hydroxyoctadeca-10,12-dienoic acid WsAG, FgAG [67]
114 17.37 C42H70O12 766.4860 766.4867 −0.9 811.4933[M + HCOO]−, 747.4834[M − H-H2O]−, 603.4833[M − H-Glu]−, 585.4309[M − H-Glu]−, 459.0768[M − H-Glu-Rha]−, 421.4457[M − H-Glu-Rha]− Ginsenoside Rg5 WsAG, FgAG [53]
115 # 17.45 C18H30O2 278.2245 278.2246 −0.1 279.2321[M + H]+, 218.1936[M + H-HCOOH-CH3]+, 184.1479[M + H-C7H11]+ α-Linolenic Acid WsAG < FgAG
VIP: 5.24
p < 0.001		 s
116 18.24 C32H50O4 498.3722 498.3709 2.4 521.3614[M+Na]+, 484.3365[M + H-CH3]+, 439.3322[M + H-C2H3O2]+, 303.3080[M-C12H20O2]+, 263.2783[M-C15H24O2]+, 248.2610[M + H-C16H26O2]+, 203.0991[M + H-C2H3O2-C15H22O2]+ 3-O-Acetyloleanolic acid WsAG, FgAG a
117 * 18.50 C19H24O2 284.1773 284.1776 −1.1 285.1843[M + H]+, 259.2243[M + H-C2H2]+, 243.1701[M + H-C2H2O]+, 159.1308[M + H-C8H13O]+, 122.1168[M + H-C11H11O]+ Androsta-1,4-diene-3,17-dione WsAG > FgAG
VIP: 6.16
p < 0.001		 a
118 19.97 C16H28O3 268.2039 268.2038 0.4 291.1950[M+Na]+, 223.1536[M + H-HCOOH]+, 123.0441[M + H-H2O-C7H13O2]+, 95.0141[M + H-C4H9O-C5H9O2] 13-Hydroxy-9,11-hexadecanedioic acid WsAG, FgAG [34]
119 20.15 C17H24O2 260.1766 260.1776 −3.8 261.1839[M + H]+, 243.1708[M + H-H2O]+, 221.1479[M + H-CH3-C2H3]+, 159.0791[M + H-H2O-C6H13]+ Panaxydol WsAG, FgAG [68]
120 20.49 C17H26O3 280.3138 280.3130 3.3 303.3030[M+Na]+, 252.2401[M + H-C2H5]+, 149.1310[M + H-C10H21]+,140.1322[M + H-C10H21]+, 97.1025[M + H-C13H27]+ 1-Eicosene WsAG, FgAG a
121 22.88 C19H38O4 330.2766 330.2770 −1.2 353.2658[M+Na]+, 313.2725[M + H-H2O]+, 280.2603[M + H-2H2O-CH3]+, 239.2352[M + H-C3H7O3]+, 99.0871[M + H-C4H7O4-C8H17]+ Monopalmitin WsAG, FgAG [30]
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* Characteristic component in WsAG. # Characteristic component in FgAG. s Identified with a standard, a Compared with spectral data obtained from Wiley Subscription Services, Inc. (New York, NY, USA).

Figure 1.

Figure 1

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The representative base peak intensity (BPI) chromatograms of FgAG and WsAG in negative and positive modes.

Figure 2.

Figure 2

Figure 2

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Chemical structures of compounds identified in FgAG and WsAG.

In FgAG and WsAG, these compounds were all shared constituents, including 47 protopanaxdiol-type saponins, 23 protopanaxtriol-type saponins, 15 oleanane- type saponins, 10 ocotillol-type saponins, one ursane, one flavonoid, one lignin, 12 organic acids and organic acid esters, three steroids, and eight other compounds.

2.2. Biomarker Discovery for FgAG and WsAG

The QC injections were clustered tightly in PCA indicating a satisfactory stability of the system. The PCA 2D plots of the samples from FgAG and WsAG groups were classified into two clusters according to their common spectral characteristics (Figure 3), with the FgAG samples of different years clustered into one group, while the WsAG samples were clustered into another group. The FgAG and WsAG samples were clearly separated, indicating that these two herb species could be easily differentiated.

Figure 3.

Figure 3

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The PCA of FgAG and WsAG in positive mode and negative mode.

After OPLS-DA plots (Figure 4A and Figure 5A) in both negative and positive modes were generated, the maximum separation between MsAG and FgAG groups was available. In the sufficient permutation test, the lines of grouping samples were significantly located underneath the random sampling lines (Figure 4B and Figure 5B), which indicated a fine validity for the following characteristic metabolites biomarkers identification [42]. S-plots were then created to explore the potential chemical markers that contributed to the differences. Based on p values (p < 0.05) and VIP values (VIP > 3) [30,61] from univariate statistical analysis, 22 robust known biomarkers enabling the differentiation between FgAG and WsAG, were marked and listed (Figure 4C and Figure 5C and Table 2). Additionally, a heatmap was generated from these biomarkers in order to systematically evaluate the biomarkers (Figure 6), which visually showed the intensities of potential biomarkers between two species.

Figure 4.

Figure 4

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The OPLS-DA (A); permutation tests (B) and S-plot (C) in negative mode.

Figure 5.

Figure 5

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The OPLS-DA (A); permutation tests (B) and S-plot (C) in positive mode.

Table 2.

Details of FgAG and WsAG samples.

Species and the Morphological Features Source Growth Year Collection Time Batch No.
FgAGs
Main roots 9~15 cm (length) × 1.5~3.0 cm (diameter); 2~3 branch roots with diameters of 2~3.5 cm; fibrous roots with diameters of 0.1~0.2 cm; 3~4 stem scars in rhizomes; no adventitious roots. 		Ji’an City, Jilin Province, China 3, 4 2017.09–2017.10 FgAG1, 11
Fusong County, Jilin Province, China 3, 4 2017.09–2017.10 FgAG2, 12
Tonghua City, Jilin Province, China 3, 4 2017.09–2017.10 FgAG3,13
Jingyu Country, Jilin Province, China 3, 4 2017.09–2017.10 FgAG4, 14
Antu Country, Jilin Province, China 3, 4 2017.09–2017.10 FgAG5, 15
Hunchun City, Jilin Province, China 3, 4 2017.09–2017.10 FgAG6, 16
Helong City, Jilin Province, China 3, 4 2017.09–2017.10 FgAG7, 17
Huadian City, Jilin Province, China 3, 4 2017.09–2017.10 FgAG8, 18
Huairou District, Beijing Province, China 3, 4 2017.10–2017.11 FgAG9, 19
Wendeng Area, Shandong Province, China 3, 4 2017.10–2017.11 FgAG10, 20
WsAGs
Main roots 5.0~6.0 cm (length) × 1.5~2.0 cm (diameter); 2~3 branch roots with diameters of 0.5~0.9 cm; fibrous roots with diameters of 0.1~0.2 cm; 15~25 stem scars in rhizomes; adventitious roots with diameters of 0.5~0.8 cm 		Lawton Coumtry, Michigan State, American >15 2017.09–2017.11 WsAG1, 3, 6
Schoharie County, Catskill region, American >15 2017.09–2017.10 WsAG2, 4, 8
Monongalia County, West Virginia, American >15 2017.10–2017.11 WsAG5, 7
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Figure 6.

Figure 6

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The heatmap visualizing the intensities of potential biomarkers.

3. Discussion

In the screening analysis, 121 compounds were characterized in FgAG and WsAG, respectively. The results showed that both of these kinds of American ginseng were rich in natural components. These 121 compounds were all shared constituents in FgAG and WsAG, which means that they were similar in terms of the kinds of compound they contained. It has been reported that there are high ginsenoside contents in American ginseng. In this study, ginsenosides were also the main chemical components. Besides the most common dammarane-type and oleanane-type saponins, the ocotillol-type saponins are also occupying a notable proportion. The ocotillol-type is the characteristic type of saponin enabling American ginseng to be differentiated from Asian ginseng. So far, the studies on the mechanism of biosynthesis were focused on dammarane-type and oleanane-type ginsenosides. For example, dammaranediol was obtained by DS (dammarenediol synthase), and then modified by CYP450 to obtain dammarane-type saponins. Another example, oleanane-type ginsenosides were obtained by modifing β-amyrin with CYP450 and UGT (UDP-glycosyltransferase). Actually, there were little literature about the mechanism of ocotillol-type ginsenoside biosynthesis. The phytochemicals in WsAG and FgAG might provide a material basis for mechanistic studies. In short, this comprehensive and unique phytochemical profile study revealed the structural diversity of secondary metabolites and the similar patterns in FgAG and WsAG.

Furthermore, in non-targeted metabolomic analysis, when taking the contents of the constituents into account, it was found that there indeed existed quite a few differences between FgAG and WsAG, and 22 robust known biomarkers enabling the differentiation were discovered. This study illustrated the differences between FgAG and WsAG, and provided a basis for explaining the effect of different growth environments on secondary metabolites. For WsAG, there are 12 potential biomarkers, including two ocotillol-type saponins (12, 47), two steroids (22, 117), six damarane-type saponins (21, 35, 37, 42, 44, 112), one oleanane-type saponin (102) and one other compound (6). The contents of these 12 components in WsAG were much greater than in FgAG. On the other hand, for FgAG, there are 10 potential biomarkers including two organic acids (105, 115), six damarane-type saponins (13, 14, 31, 46, 81, 91, 99), one oleanane-type saponin (46), and one ursane (70), which contents in FgAG were much greater than in WsAG. It has been reported that wild American ginseng has better biological activity than the FgAG. As is known, biological activity is caused by the high contents of phytochemicals. Correlation studies between potential markers and biological activities could be performed in the future.

Even so, there are still several unresolved issues. For example, as shown in BPI chromatograms, though 121 compounds were identified, there are still some unidentified components. Further research should be carried on based on the formula of these unknown compounds.

4. Materials and Methods

4.1. Materials and Reagents

Twenty eight batches of commercially available FgAGs and WsAGs root products were collected or purchased from different cultivation areas in China and American, including 20 batches of FgAGs and eight batches of WsAGs. A detailed sample list is provided in Table 2.

For FgAGs, six roots of each sample were selected for analysis, while for WsAGs, 2–3 roots of each sample were analyzed. All the herbs were authenticated by the authors and the corresponding voucher specimens have been deposited in the Research Center of Natural Drug, School of Pharmaceutical Sciences, Jilin University, China.

A total of 25 saponins were isolated in our laboratory and identified by spectroscopic data. Among of these saponins, ginsenoside Ro [69], 15 ginsenosides [70,71] (Rb1, Rb2, Rb3, Rc, Rd, Re, 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rh2, Rg1, 20(R)-Rg2, 20(S)-Rh1, 20(R)-Rh1, pseudo-ginsenoside F11, pseudo-ginsenoside RT5) and another six saponins [71] (quinquenoside L8, L9, L11, F3, 24(S)-pseudo- ginsenoside F11, gypenoside XVII) were isolated and identified by our group.

Oleanolic acid-28-O-β-d-glucopyranoside, ginsenoside Rs1, -Rs2 and methyl gallate-3-O-β-d-glucoside were also isolated in our laboratory and identified by NMR spectroscopy. Adenosine, α-maltose, l-tryptophan, notoginsenoside R1, kaempferol, l-phenylalanine, sucrose, palmitoleic acid, quinic acid and α-linolenic acid were purchased from Beijing Zhongke Quality Inspection Biotechnology Co., Ltd. (Beijing, China).

Acetonitrile and methanol suitable for UPLC-MS were purchased from Fisher Chemical Company (Geel, Belgium). Formic acid was purchased from Sigma-Aldrich Company (St. Louis, MO, USA). Deionized water was purified using a Millipore water purification system (Millipore, Billerica, MA, USA). All other chemicals were of analytical grade.

4.2. Sample Preparation and Extraction

All samples were respectively air-dried, grinded and sieved (Chinese National Standard Sieve No. 3, R40/3 series) to get a homogeneous powder. Then each fine powder was accurately weighed (0.2 g) and soaked with 10 mL of methanol overnight. On the second day, each powder was extracted in an ultrasonic bath (power of 250 W, frequency of 40 kHz) for half an hour. After cooling to room temperature, the lost weight was replenished with methanol. After filtering through a syringe filter (0.22 μm), the extracts were injected directly into the UPLC system. In addition, to ensure the stability and suitability consistency of MS analysis, a quality control (QC) sample was prepared by pooling the same volume (50 μL) from every sample and five QC injections were performed randomly through the whole worklist. The volumes injected for samples and QC were all 5 μL for each run.

4.3. UPLC-QTOF-MS

UPLC-QTOF-MSE analysis was performed on a Waters Xevo G2-XS QTOF mass spectrometer (Waters Co., Milford, MA, USA) equipped with a UPLC system through an electrospray ionization (ESI) interface. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm) from Waters Corporation (Milford, MA, USA). The mobile phases were composed of eluent A (0.1% formic acid in water, v/v) and eluent B (0.1% formic acid in acetonitrile, v/v) with flow rate of 0.4 mL/min. The elution conditions applied were: 0→2 min, 10% B; 2→26 min, 10~100% B; 26→29 min, 100% B; 29→29.1 min, 100~10% B; 29.1→32 min, 10% B. The temperature of the autosampler and the UPLC column manager were set at 15 °C and 30 °C respectively. Mixtures of 90/10 and 10/90 water/acetonitrile were used as the weak wash solvent and the strong wash solvent respectively. The mass spectrum was acquired from 100 to 1500 Da in MSE mode. The positive mode conditions were: capillary voltage, 2.6 kV; cone voltage, 40 V; source temperature, 150 °C; desolvation temperature, 400 °C; cone gas flow, 50 L/h; desolvation gas flow, 800 L/h. Negative mode conditions were identical to the positive mode conditions except for capillary voltage (2.2 kV). In MSE mode, data acquisition was performed via the mass spectrometer by rapidly switching from a low-collision energy (CE) scan to a high-CE scan during a single LC run. The low energy function was set to 6 V, while ramp collision energy of high energy function was set to 20~40 V. Leucine enkephalin (m/z 556.2771 in positive mode and 554.2615 in negative mode) was used as external reference of Lock Spray™ infused at a constant flow of 10 μL/min. During acquisition, data were collected in continumn mode for the screening analysis, and in centroid mode for the metabolomics analysis.

4.4. Chemical Information Database for the Components of FgAG and WsAG

Besides the in-house Traditional Medicine Library in the UNIFI software, a systematic investigation of chemical components was conducted [34]. A self-built database of compounds that were isolated from FgAG and WsAG was established by searching online databases or internet search engines such as PubMed, Full-Text Database (CNKI), ChemSpider, Web of Science and Medline. Chemical information including the component name, molecular formula and structure of the components from the herbs were obtained from the database [56].

4.5. The Screening Analysis by the UNIFI Platform

To quickly identify the chemical compounds, the MS raw data, compressed with Waters Compression and Archival Tool v1.10, was automatically screened and identified by using the streamlined workflow of UNIFI 1.7.0 software (Waters, Manchester, UK) [30]. The parameters were as follows: the minimum peak area of 200 was set for 2D peak detection; the peak intensity of low energy over 1000 counts and the peak intensity of high energy over 200 counts were selected for 3D peak detection. Mass error up to ±5 ppm for identified compounds, retention time in the range of ±0.1 min was allowed to match the reference substance. The matching compounds would be generated predicted fragments from structure. The negative adducts containing +COOH and -H and positive adducts containing +H and +Na were selected in the analysis. Leucine-enkephalin was selected as the reference compound, and [M − H]− 554.2620 was used for negative ion and [M + H]+ 556.2766 was used for positive ion [72].

4.6. The Metabolomics Analysis

The raw data were processed by MarkerLynx XS V4.1 software (Waters, Milford, CT, USA) for alignment, deconvolution, data reduction, etc. [73]. A MarkerLynx processing method was firstly created, and the main parameters were as follows: retention time range 0–26 min, mass range 100–1500 Da, mass tolerance 0.10, minimum intensity 5%, mass window 0.10, retention time window 0.20, marker intensity threshold 2000 counts and noise elimination level 6. After processing the data, the results could be showed in the Extended Statistics (XS) Viewer. m/z-RT pairs with corresponding intensities for all the detected peaks from each data file were listed. The same value of RT and m/z in different batched of samples were regarded as the same component. Then, multivariate statistical analysis was performed. Firstly, principal component analysis (PCA) was used to show the pattern recognition and maximum variation aiming to obtain the overview and classification. Secondly, orthogonal projections to latent structures discriminant analysis (OPLS-DA) in both positive and negative modes was performed in order to get the maximum separation between MsAG and FgAG group and to explore the potential chemical markers that contributed to the differences. Then, S-plots was created to provide visualization of the OPLS-DA predictive component loading to facilitate model interpretation. Meawhile, variable importance for the projection (VIP) was helpful to screen the different components, and the metabolites with VIP value (>3.0) were considered as potential markers [29]. In addition, the permutation test was performed to provide reference distributions of the R2/Q2-values that could indicate the statistical significance [30,31,32,33]. Simca 15.0 software (Umetrics, Malmö, Sweden) was used to show the analysis results [56,74].

5. Conclusions

In a comprehensive and unique phytochemical profile study, a total of 121 chemical compounds with different structural types were identified from WsAG and FgAG. The structural patterns included protopanaxdiol-type saponins, protopanaxtriol-type saponins, ocotillol-type saponins, oleanane-type saponins and other glyosides, organic acid and organic acid esters, steroids, etc. The results showed that WsAG and FgAG were rich in natural components. Furthermore, these 121 compounds were all shared constituents in them, meaning that they were similar in the kinds of compounds they contain. In metabolomic analysis, it was found that there indeed existed several differences in the contents of the constituents between FgAG and WsAG, and 22 robust known biomarkers enabling the differentiation were discovered. In a word, the results will fill the data gap in the study on the chemical constituents of WsAG and provide a reference for quantitative determinations in its quality control.

Author Contributions

The individual contributions of authors are specified as following: Data curation, Investigation, Writing-original draft, H.L.; Methodology, Software, H.Z.; Formal analysis, Writing-original draft, J.T.; Formal analysis, Writing editing, H.W.; Conceptualization, Methodology, Q.D.; Investigation, F.W.; Data curation, Y.L.; Funding acquisition, P.L.; Supervision, J.L.

Funding

This research is supported by the Bethune Plan Research Project of Jilin University [Grant No. 2018B22] and the Graduate Innovation Fund of Jilin University [Grant No. 101832018C08, and the Ph.D. Interdisciplinary Research Project Funding of Jilin University [Grant No. 10183201847].

Conflicts of Interest

The authors declare that they have no conflicts of interest concerning this article.

Footnotes

Sample Availability: Samples of the compounds are not available from the authors.

References

  • 1.Ko S.K., Cho O.S., Bae H.M., Sohn U.D., Im B.O., Cho S.H., Chung S.H., Lee B.Y. Change of Ginsenoside Composition of Various American Ginseng Roots. J. Korean Soc. Appl. Biol. Chem. 2009;52:198–201. doi: 10.3839/jksabc.2009.036. [DOI] [Google Scholar]
  • 2.Lee D.P. Production Procedures and Economics of the American Ginseng. J. Ginseng Res. 2006;30:172–180. [Google Scholar]
  • 3.Chen Y.J., Zhao Z.Z., Chen H.B., Brand E., Yi T., Qin M.J., Liang Z.T. Determination of ginsenosides in Asian and American ginsengs by liquid chromatography-quadrupole/time-of-flight MS: Assessing variations based on morphological characteristics. J. Ginseng Res. 2017;41:10–22. doi: 10.1016/j.jgr.2015.12.004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Nadeau I., Simard R.R., Olivier A. The impact of lime and organic fertilization on the growth of wild-simulated American ginseng. Can. J. Plant Sci. 2003;83:603–609. doi: 10.4141/P02-044. [DOI] [Google Scholar]
  • 5.Lim W., Mudge K.W., Vermeylen F. Effects of population, age, and cultivation methods on ginsenoside content of wild American ginseng (Panax quinquefolium) J. Agric. Food Chem. 2005;53:8498–8505. doi: 10.1021/jf051070y. [DOI] [PubMed] [Google Scholar]
  • 6.Charest P., Dorais M., Gauthier L., Khanizadeh S. The influence of soil preparation, seedling rates and organic mulch on the production of woods-cultivated ginseng. Acta Hortic. 2000;523:87–96. doi: 10.17660/ActaHortic.2000.523.11. [DOI] [Google Scholar]
  • 7.US Fish and Wildlife Service American Ginseng. [(accessed on 7 January 2019)]; Available online: https://www.fws.gov/international/permits/by-species/american-ginseng.html.
  • 8.US Fish and Wildlife Service American Ginseng Production in Woodlots. [(accessed on 6 January 2019)]; Available online: http://digitalcommons.unl.edu/agroforestnotes/
  • 9.Schlag E.M., Mcintosh M.S. RAPD-based assessment of genetic relationships among and within American ginseng (Panax quinquefolius L.) populations and their implications for a future conservation strategy. Genet. Resour. Crop Evol. 2012;59:1553–1568. doi: 10.1007/s10722-011-9784-4. [DOI] [Google Scholar]
  • 10.Burkhart E.P. American ginseng (Panax quinquefolius L.) floristic associations in Pennsylvania: Guidance for identifying calcium-rich forest farming sites. Agrofor. Syst. 2013;87:1157–1172. doi: 10.1007/s10457-013-9627-8. [DOI] [Google Scholar]
  • 11.Zhu W., Han B., Sun Y., Wang Z.Y., Yang X.H. Immunoregulatory effects of a glucogalactan from the root of Panax quinquefolium L. Carbohydr. Polym. 2012;87:2725–2729. doi: 10.1016/j.carbpol.2011.11.066. [DOI] [Google Scholar]
  • 12.Duda R.B., Zhong Y., Navas V., Li M.Z., Toy B.R., Alavarez J.G. American ginseng and breast cancer therapeutic agents synergistically inhibit MCF-7 breast cancer cell growth. J. Surg. Oncol. 2015;72:230–239. doi: 10.1002/(SICI)1096-9098(199912)72:4&#x0003c;230::AID-JSO9&#x0003e;3.0.CO;2-2. [DOI] [PubMed] [Google Scholar]
  • 13.Barton D.L., Soori G.S., Bauer B.A., Sloan J.A., Johnson P.A., Figueras C., Duane S., Mattar B., Liu H., Atherton P.J., et al. Pilot study of Panax quinquefolius (American ginseng) to improve cancer-related fatigue: A randomized, double-blind, dose-finding evaluation: NCCTG trial N03CA. Support. Care Cancer. 2010;18:179–187. doi: 10.1007/s00520-009-0642-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Assinewe V.A., Baum B.R., Gagnon D., Arnason J.T. Phytochemistry of wild populations of Panax quinquefolius L. (North American ginseng) J. Agric. Food Chem. 2003;51:4549–4553. doi: 10.1021/jf030042h. [DOI] [PubMed] [Google Scholar]
  • 15.Vuksan V., Sievenpiper J.L., Wong J., Xu Z., Beljan-Zdravkovic U., Arnason J.T., Assinewe V., Stavro M.P., Jenkins A.L., Leiter L.A., et al. American ginseng (Panax quinquefolius L.) attenuates postprandial glycemia in a time-dependent but not dose-dependent manner in healthy individuals. Am. J. Clin. Nutr. 2001;73:753–758. doi: 10.1093/ajcn/73.4.753. [DOI] [PubMed] [Google Scholar]
  • 16.Oshima Y., Sato K., Hikino H. Isolation and hypoglycemic activity of quinquefolans A, B, and C, glycans of Panax quinquefolium roots. J. Nat. Prod. 1987;50:188–190. doi: 10.1021/np50050a010. [DOI] [PubMed] [Google Scholar]
  • 17.Vuksan V., Sievenpiper J.L., Koo V.Y., Francis T., Beljan-Zdravkovic U., Xu Z., Vidgen E. American ginseng (Panax quinquefolius L) reduces postprandial glycemia in nondiabetic subjects and subjects with type 2 diabetes mellitus. Arch. Intern. Med. 2000;160:1009–1013. doi: 10.1001/archinte.160.7.1009. [DOI] [PubMed] [Google Scholar]
  • 18.Kitts D.D., Wijewickreme A.N., Hu C. Antioxidant properties of a North American ginseng extract. Mol. Cell. Biochem. 2000;203:1–10. doi: 10.1023/A:1007078414639. [DOI] [PubMed] [Google Scholar]
  • 19.Li Z., Guo Y.Y., Wu C.F., Li X., Wang J.H. Protective Effects of Pseudoginsenoside-F11 on Scopolamine-induced Memory Impairment in Mice and Rats. J. Pharm. Spharmacol. 2010;51:435–440. doi: 10.1211/0022357991772484. [DOI] [PubMed] [Google Scholar]
  • 20.Robbins C.S. Comparative Analysis of Management Regimes and Medicinal Plant Trade Monitoring Mechanisms for American Ginseng and Goldenseal. Conserv. Biol. 2000;14:1422–1434. doi: 10.1046/j.1523-1739.2000.99100.x. [DOI] [Google Scholar]
  • 21.Grubbs H.J., Case M.A. Allozyme variation in American ginseng (Panax quinquefolius L.): Variation, breeding system, and implications for current conservation practice. Conserv. Genet. 2004;5:13–523. doi: 10.1023/B:COGE.0000014064.44592.bc. [DOI] [Google Scholar]
  • 22.Crusesanders J.M., Hamrick J.L. Genetic diversity in harvested and protected populations of wild American ginseng, Panax quinquefolius L. (Araliaceae) Am. J. Bot. 2004;91:540–548. doi: 10.3732/ajb.91.4.540. [DOI] [PubMed] [Google Scholar]
  • 23.Voort M.E.V.D., Mcgraw J.B. Effects of harvester behavior on population growth rate affects sustainability of ginseng trade. Biol. Conserv. 2006;130:505–516. doi: 10.1016/j.biocon.2006.01.010. [DOI] [Google Scholar]
  • 24.Beyfuss B. New York State Wild Simulated American Ginseng has been Selling for well over $1000 per Pound. [(accessed on 18 January 2019)]; Available online: http://www.ginsenggeek.org/new-york-state-wild-simulated-american-ginseng-Has-been-selling-for-well-over-1000-per-pound/
  • 25.Anderson R.C., Anderson M.R., Houseman G. Wild American Ginseng. Native Plants J. 2002;3:93–105. [Google Scholar]
  • 26.Predy G.N., Goel V., Lovlin R. Efficacy of an extract of North American ginseng containing poly-furanosyl-pyranosyl-saccharides for preventing upper respiratory tract infections: A randomized controlled trial. Cmaj. 2005;173:1043–1048. doi: 10.1503/cmaj.1041470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Zhao H., Xu J., Ghebrezadik H., Hylands P.J. Metabolomic quality control of commercial Asian ginseng, and cultivated and wild American ginseng using 1H NMR and multi-step PCA. J. Pharm. Biomed. Anal. 2015;114:113–120. doi: 10.1016/j.jpba.2015.05.010. [DOI] [PubMed] [Google Scholar]
  • 28.Wang J.R., Leung C.Y., Ho H.M., Chai S., Yau L.F., Zhao Z.Z., Jiang Z.H. Quantitative Comparison of Ginsenosides and Polyacetylenes in Wild and Cultivated American Ginseng. Chem. Biodivers. 2010;7:975–983. doi: 10.1002/cbdv.200900264. [DOI] [PubMed] [Google Scholar]
  • 29.Wang C.Z., Zhang N.Q., Wang Z.Z., Qi Z., Zhu H.L., Zheng B.Z., Li P.Y., Liu J.P. Nontargeted Metabolomic Analysis of Four Different Parts of Platycodon grandiflorum Grown in Northeast China. Molecules. 2017;22:1280. doi: 10.3390/molecules22081280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Wang C.Z., Zhang N.Q., Wang Z.Z., Qi Z., Zheng B.Z., Li P.Y., Liu J.P. Rapid characterization of chemical constituents of Platycodon grandiflorum and its adulterant Adenophora stricta by UPLC-QTOF-MS/MS. J. Mass Spectrom. 2017;52:643–656. doi: 10.1002/jms.3967. [DOI] [PubMed] [Google Scholar]
  • 31.Zhang F.X., Li M., Qiao L.R., Yao Z.H., Li C., Shen X.Y., Wang Y., Yu K., Yao X.S., Dai Y. Rapid characterization of Ziziphi Spinosae Semen by UPLC/Q-tof MS with novel informatics platform and its application in evaluation of two seeds from Ziziphus species. J. Pharm. Biomed. Anal. 2016;122:59–80. doi: 10.1016/j.jpba.2016.01.047. [DOI] [PubMed] [Google Scholar]
  • 32.Deng L., Shi A.M., Liu H.Z., Meruva N., Liu L., Hu H., Yang Y., Huang C., Li P., Wang Q. Identification of chemical ingredients of peanut stems and leaves extracts using UPLC-QTOF-MS coupled with novel informatics UNIFI platform. J. Mass Spectrom. 2016;51:1157–1167. doi: 10.1002/jms.3887. [DOI] [PubMed] [Google Scholar]
  • 33.Tang J.F., Li W.X., Tan X.J., Li P., Xiao X.H., Wang J.B., Zhu M.J., Li X.L., Meng F. A novel and improved UHPLC-QTOF/MS method for the rapid analysis of the chemical constituents of Danhong Injection. Anal. Methods. 2016;8:2904–2914. doi: 10.1039/C5AY03173G. [DOI] [Google Scholar]
  • 34.Wang Y.R., Wang C.Z., Lin H.Q., Liu Y.H., Li Y.M., Zhao Y., Li P.Y., Liu J.P. Discovery of the Potential Biomarkers for Discrimination between Hedyotis diffusa and Hedyotis corymbosa by UPLC-QTOF/MS Metabolome Analysis. Molecules. 2018;23:1525. doi: 10.3390/molecules23071525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Tan J., Wang C.Z., Zhu H.L., Zhou B.S., Xiong L.X., Wang F., Li P.Y., Liu J.P. Comprehensive Metabolomics Analysis of Xueshuan Xinmaining Tablet in Blood Stasis Model Rats Using UPLC-Q/TOF-MS. Molecules. 2018;23:1650. doi: 10.3390/molecules23071650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Yang X., Yang L., Xiong A., Li D., Wang Z. Authentication of senecio scandens and s. vulgaris based on the comprehensive secondary metabolic patterns gained by uplc–dad/esi-ms. J. Pharm. Biomed. Anal. 2011;56:165–172. doi: 10.1016/j.jpba.2011.05.004. [DOI] [PubMed] [Google Scholar]
  • 37.Wang D.Q., Feng B.S., Wang X.B., Yang C.R., Zhou J. Further study on dammarane saponins of leaves of panax japonicus var.major collected in qinling m ountains china. Acta Pharm. Sin. 1989;24:633–637. [PubMed] [Google Scholar]
  • 38.Zou K., Zhu S., Tohda C., Cai S., Komatsu K. Dammarane-type triterpene saponins from Panax japonicus. J. Nat. Prod. 2002;65:346–351. doi: 10.1021/np010354j. [DOI] [PubMed] [Google Scholar]
  • 39.Du Z., Li J., Zhang X., Pei J., Huang L. An integrated LC-MS-based strategy for the quality assessment and discrimination of three panax species. Molecules. 2018;23:2988. doi: 10.3390/molecules23112988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Feng B.S., Wang X.B., Wang D.Q., Yang C.R., Zhou J. Dammarane saponins of Panax japonicus var. major collected in Qinling mountain, China. Acta Bot. Yunnanica. 1987;28:633–636. [PubMed] [Google Scholar]
  • 41.Meng H.Y., Wang X.W., Zhai C.M., Jiang H., Yang C.J., Song Y., Wang Z.B. Isolation and Identification of Lignans from the Fruits of Acanthopanax Sessiliflorus. Inf. Tradit. Chin. Med. 2016;30:1–4. [Google Scholar]
  • 42.Wang L.L., Han L.F., Yu H.S., Sang M.M., Liu E.W., Zhang Y., Fang S.M., Wang T., Gao X.M. Analysis of the Constituents in “Zhu She Yong Xue Shuan Tong” by Ultra High Performance Liquid Chromatography with Quadrupole Time-of-Flight Mass Spectrometry Combined with Preparative High Performance Liquid Chromatography. Molecules. 2015;20:20518–20537. doi: 10.3390/molecules201119712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Wang J.H. Ph.D. Thesis. Shenyang Pharmaceutical University; Shenyang, China: 1999. Studies on Chemical Constituents and Biological Activities of Stems and Leaves of American Ginseng. [Google Scholar]
  • 44.Ye D.Y. Master’s Thesis. Fujian Agriculture And Forestry University; Fujian, China: 2015. Extraction, Separation and Purification of Sterols from Abalone Gland. [Google Scholar]
  • 45.Liao P.Y., Wang D., Zhang Y.J., Yang C.R. Dammarane-Type Glycosides from Steamed Notoginseng. J. Agric. Food Chem. 2008;56:1751–1756. doi: 10.1021/jf073000s. [DOI] [PubMed] [Google Scholar]
  • 46.Zang Y.W. Studies on the chemical constituents of Schizonepeta mulifida (L.) Briq. China J. Chin. Mater. Med. 1989;14:44–45. [PubMed] [Google Scholar]
  • 47.Li G.Y., Zeng Y.M., Meng H., Li X., Wang J.H. A new triterpenoid saponin from the leaves and stems of Panax quinquefolium L. Chin. Chem. Lett. 2009;20:1207–1210. doi: 10.1016/j.cclet.2009.05.017. [DOI] [Google Scholar]
  • 48.Ha L.T., Pawlicki-Jullian N., Pillon-Lequart M., Boitel-Conti M., Duong H.X., Gontier E. Hairy root cultures ofpanax vietnamensis, a promising approach for the production of ocotillol-type ginsenosides. Plant Cell Tissue Org. Cult. 2016;126:93–103. doi: 10.1007/s11240-016-0980-y. [DOI] [Google Scholar]
  • 49.Wang J.H., Lia W., Sha Y., Tezuka Y., Kadota S., Li X. Triterpenoid Saponins from Leaves and Stems of Panax Quinquefolium L. J. Asian Nat. Prod. Res. 2001;3:123–130. doi: 10.1080/10286020108041379. [DOI] [PubMed] [Google Scholar]
  • 50.Zou K., Zhu S., Meselhy M.R., Tohda C., Cai S., Komatsu K. Dammarane-Type Saponins from Panax japonicus and Their Neurite Outgrowth Activity in SK-N-SH Cells. J. Nat. Prod. 2002;65:1288–1292. doi: 10.1021/np0201117. [DOI] [PubMed] [Google Scholar]
  • 51.Li P.Y. Ph.D. Thesis. Shenyang Pharmaceutical University; Shenyang, China: 1999. Study on Chemical Constituents and Biological Activities of American Ginseng. [Google Scholar]
  • 52.Liu J.Y., Xiao S.Y., Shang W.F., Xu L.Z., Yang S.L. A new minor triterpene saponin from kaixin-san prescription. J. Asian Nat. Prod. Res. 2005;7:643–648. doi: 10.1080/1028602032000169550. [DOI] [PubMed] [Google Scholar]
  • 53.Yuan J.B., Chen Y., Liang J., Wang C.Z., Liu X.F., Yan Z.H., Tang Y., Li J.K. Component analysis and target cell-based neuroactivity screening of Panax ginseng, by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. J. Chromatogr. B. 2016;1038:1–11. doi: 10.1016/j.jchromb.2016.10.014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Lu J.C., Xu B.B., Zhang X.Y., Sun Q.S. Study on chemical constituents of rhizome of Anemone raddeana. Acta Pharm. Sin. 2002;37:709–715. [PubMed] [Google Scholar]
  • 55.Assimopoulou A.N., Papageorgiou V.P. GC-MS analysis of penta-and tetra-cyclic triterpenes from resins of Pistacia species. Part I. Pistacia lentiscus var. Chia. Biomed. Chromatogr. 2005;19:285–311. doi: 10.1002/bmc.454. [DOI] [PubMed] [Google Scholar]
  • 56.Zhu H.L., Lin H.Q., Tan J., Wang H., Wu F.L., Dong Q.H., Liu Y.H., Li P.Y., Liu J.P. UPLC-QTOF/MS-Based Nontargeted Metabolomic Analysis of Mountain- and Garden-Cultivated Ginseng of Different Ages in Northeast China. Molecules. 2019;24:33. doi: 10.3390/molecules24010033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 57.Liang G.Y., Zhou Y., Cao P.X., Xu B.X. Studies on chemical constituents of sabia schumanniana. Chin. Pharm. J. 2005;39:900–901. [Google Scholar]
  • 58.Zhu T.T., Li F., Chen B., Deng Y., Wang M.K., Li L.H. Studies on the saponins from the leaves of Studies on the saponins from the leaves of Panax ginseng. Chin. J. Appl. Environ. Biol. 2016;22:70–74. [Google Scholar]
  • 59.Duc N.M., Nguyen M.D., Minh N.N.T., Kasai R., Ohtani K., Kasai R. Saponins from Vietnamese Ginseng, Panax vietnamensis Haet Grushv. Collected in Central Vietnam. II. Chem. Pharm. Bull. 1994;42:115–122. doi: 10.1248/cpb.42.634. [DOI] [PubMed] [Google Scholar]
  • 60.Zhao P.J., Gan F.Y., Zhu N., Shen Y.M. Studies on the Tissue Culture of Cynanchum otophyllum and Calli Chemical Constituents. Chin. Bull. Bot. 2003;20:565–571. [Google Scholar]
  • 61.Yoshikawa M., Murakami T., Yashiro K., Yamahara J., Matsuda H., Saijoh R., Tanaka O. Bioactive Saponins and Glycosides. XI. Structures of New Dammarane-Type Triterpene Oligoglycosides, Quinquenosides I, II, III IV, and V, from American Ginseng, the Roots of Panax quinquefolium L. Chem. Pharm. Bull. 1998;46:647–654. doi: 10.1248/cpb.46.647. [DOI] [PubMed] [Google Scholar]
  • 62.Li S.L., Lai S.F., Song J.Z., Qiao C.F., Liu X., Zhou Y., Cai H., Cai B.C., Xu H.X. Decocting-induced chemical transformations and global quality of Du-Shen-Tang, the decoction of ginseng evaluated by UPLC-Q-TOF-MS/MS based chemical profiling approach. J. Pharm. Biomed. Anal. 2010;53:946–957. doi: 10.1016/j.jpba.2010.07.001. [DOI] [PubMed] [Google Scholar]
  • 63.Dou D., Li W., Guo N., Fu R., Pei Y., Koike K., Nikaido T. Ginsenoside Rg8, a New Dammarane-Type Triterpenoid Saponin from Roots of Panax quinquefolium. Chem. Pharm. Bull. 2006;54:751–753. doi: 10.1248/cpb.54.751. [DOI] [PubMed] [Google Scholar]
  • 64.Ritter A., Goulitquer S., Salaün J.P., Tonon T., Correa J.A., Potin P. Stress Induces Biosynthesis of Octadecanoid and Eicosanoid Oxygenated Derivatives in the Brown Algal Kelp Laminaria digitata. New Phytol. 2008;180:809–821. doi: 10.1111/j.1469-8137.2008.02626.x. [DOI] [PubMed] [Google Scholar]
  • 65.Wang J.Y., Li X.G., Zheng Y.N., Yang X.W. Isoginsenoside-rh3, a new triterpenoid saponin from the fruits of panax ginseng CA Mey. J. Asian Nat. Prod. Res. 2004;6:289–293. doi: 10.1080/10286020310001595980. [DOI] [PubMed] [Google Scholar]
  • 66.He K., Liu Y., Yang Y., Peng L., Ling Y. A Dammarane Glycoside Derived from Ginsenoside Rb3. Chem. Pharm. Bull. 2005;53:177–179. doi: 10.1248/cpb.53.177. [DOI] [PubMed] [Google Scholar]
  • 67.Sun M., Salomon R.G. Oxidative Fragmentation of Hydroxy Octadecadienoates Generates Biologically Active γ-Hydroxyalkenals. J. Am. Chem. Soc. 2004;126:5699–5708. doi: 10.1021/ja038756w. [DOI] [PubMed] [Google Scholar]
  • 68.Xu G.H., Choo S.J., Ryoo I.J., Kim Y.H., Paek K.Y., Yoo I.D. Polyacetylenes from the Tissue Cultured Adventitious Roots of Panax ginseng C.A. Meyer. Nat. Prod. Sci. 2008;14:177–181. [Google Scholar]
  • 69.Qiu N.N., Li P.Y. Master’s Thesis. Jilin University; Jilin, China: 2013. Studies on the Chemical Constituents, Fingerprint and Bioactivities of Purple Red Ginseng. [Google Scholar]
  • 70.Liu J.P., Li P.Y. Ph.D. Thesis. Shenyang Pharmaceutical University; Shenyang, China: 2005. Studies on Isolation, Structure Modification and Pharmacological Activities of Saponins from the Leaves and Stems of Panax quiquefolium L. Cultivated in China. [Google Scholar]
  • 71.Li P., Liu J.P., Lu D. Standard NMR Spectrum of Ginsenosides. Chemical Industry Press; Beijing, China: 2012. [Google Scholar]
  • 72.Lee J.W., Ji S.H., Choi B.R., Choi D.J., Lee Y.G., Kim H.G., Kim G.S., Kim K., Lee Y.H., Baek N.I., et al. UPLC-QTOF/MS-Based Metabolomics Applied for the Quality Evaluation of Four Processed Panax ginseng Products. Molecules. 2018;23:2062. doi: 10.3390/molecules23082062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 73.Zhao Y.Y., Cheng X.L., Wei F., Xiao X.Y., Sun W.J., Zhang Y.M., Lin R.C. Serum metabonomics study of adenine-induced chronic renal failure in rats by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Biomarkers. 2012;17:48–55. doi: 10.3109/1354750X.2011.637180. [DOI] [PubMed] [Google Scholar]
  • 74.Pang Z.Q., Wang G.Q., Ran N., Lin H.Q., Wang Z.Y., Guan X.W., Yuan Y.Z., Fang K.Y., Liu J.P., Wang F. Inhibitory Effect of Methotrexate on Rheumatoid Arthritis Inflammation and Comprehensive Metabolomics Analysis Using Ultra-Performance Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry (UPLC-Q/TOF-MS) Int. J. Mol. Sci. 2018;19:2894. doi: 10.3390/ijms19102894. [DOI] [PMC free article] [PubMed] [Google Scholar]

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