Background
Methods
Plant materials
No. | Species | Source (geographical location) | Collection time | Deposition numbers |
---|---|---|---|---|
BC1 |
Bupleurum chinense DC. | Wei Village, Fen Cheng town, Xiang fen County, Shanxi Province, cultivated for three years | 2013–8-7 | SX-0807 |
BC2 |
Bupleurum chinense DC. | Long Xing Village, Kao Lao town, Wan Rong County, Shanxi Province, cultivated for three years | 2013–8-8 | SX-0808 |
BC3 |
Bupleurum chinense DC. | Xiang Quan County, Chen Cang District, Bao Ji City, Shaanxi Province, wild | 2013–9-4 | SX-0917 |
BC4 |
Bupleurum chinense DC. | Xin Min County, Chen Cang District, Bao Ji City, Shaanxi Province, cultivated for three years | 2013–9-4 | SX-0918 |
BY1 |
Bupleurum yinchowense Shan et Y. Li | An Yi Town, Long Xi County, Gansu Province, cultivated for two years | 2013–9-2 | GS-0903 |
BY2 |
Bupleurum yinchowense Shan et Y. Li | Lu Ming Village, Wu Zhu Town, Wei Yuan County, Gansu Province, cultivated for two years | 2013–9-2 | GS-0905 |
BY3 |
Bupleurum yinchowense Shan et Y. Li | Da Hong Yan Village, San Fen Town, Zhang County, Gansu Province, cultivated for two years | 2013–9-3 | GS-0908 |
BF |
Bupleurum falcatum L. | Han Yuan County, Sichuan Province, cultivated for one years | 2013–9-5 | SC-0919 |
Chemicals and reagents
Preparation of sample solution
UHPLC-QTOF-MS analysis
Data analysis
Results and discussion
Chemical profiling
Peak No. | Identification | tR (min) | Molecular formula | Molecular ions (m/z) (mass accuracy (ppm)) | Fragments (m/z) | BC | BY | BF | |||
---|---|---|---|---|---|---|---|---|---|---|---|
Root | Aerial | Root | Aerial | Root | Aerial | ||||||
1 | Kaempferol-3-O-glucopyranoside-7-O-rhamnopyranoside | 6.90 | C27H30O15
| 593.1537 [M-H]− (4.2) | 431.1086 [M-H-(Glc-H2O)]−, 285.0458 [M-H-(Glc-H2O)-(Rha-H2O)]−
| + | + | ||||
2 | Rutin | 7.03 | C27H30O16
| 609.1439 [M-H]− (−3.61) | 1219.3206 [2 M–H]−, 300.0334 [M-H-(Glc-H2O)-(Rha-H2O)]−
| + | + | + | + | + | + |
3 | Isoquercitrin | 7.30 | C21H20O12
| 463.0871 [M-H]− (−2.4) | 300.2900 [M-H-(Glc-H2O)]−
| + | + | + | + | + | |
4 | Kaempferitrin | 7.67 | C27H30O14
| 577.1589 [M-H]− (4.5) | 431.1079 [M-H-(Rha-H2O)]−, 285.0477 [M-H-2(Rha-H2O)]−
| + | + | + | |||
5 | Quercetin 3′-glucoside-7-acetat | 7.84 | C23H22O13
| 505.0992 [M-H]− (−0.8) | 300.0311 [M-2H-Acetyl-(Glc-H2O)]−
| + | + | ||||
6 | Quercetin 3,7-diglucoside | 9.05 | C27H30O17
| 625.1433 [M-H]− (3.7) | 463.1016 [M-H-(Glc-H2O)]−
| + | + | ||||
7 | Quercetin 4′-glucoside | 9.89 | C21H20O12
| 463.0884 [M-H]− (0.4) | 300.283 [M-H-(Glc-H2O)]- | + | |||||
8 | Unknown | 10.01 | C42H66O15
| 809.4367 [M-H]− (4.7) 855.4347 [M + HCOO]− (−4.3) | 647.3920 [M-H-(Glc-H2O)]−
| + | + | ||||
9 | Rotundifolioside I/Rotundioside O | 10.86 | C47H76O16
| 895.5017 [M-H]− (−4.9) | / | + | |||||
10 | Rotundioside N | 11.23 | C48H78O18
| 941.5142 [M-H]− (2.87) 987.5146 [M + HCOO]− (−2.43) | 633.4307 [M-H-(Glc-H2O)-(Fuc-H2O)]−
| + | |||||
11 | Hydroxy-SSa | 12.27 | C42H70O14
| 797.4646 [M-H]− (−5.9) 843.4705 [M + HCOO]− (−5.1) | 635.4320 [M-H-(Glc-H2O)]−
| + | |||||
12 | Hydroxy-SSd | 12.67 | C42H70O14
| 797.4656 [M-H]− (−4.6)) 843.4711 [M + HCOO]− (−4.4) | 635.4365 [M-H-(Glc-H2O)]−
| + | + | ||||
13 | Rotundioside W | 13.12 | C48H78O18
| 941.5195 [M-H]− (8.5) 987.5156 [M + HCOO]− (−1.4) | 796.4688 [M-H- (Rha-H2O)]−
| + | |||||
14 | Acetyl-hydroxy-SSa | 13.22 | C44H72O15
| 839.4793 [M-H]− (−0.6) 885.4824 [M + HCOO]− (−3.28)) | 797.4801 [M-H-Acetyl]−; 635.4276 [M-H-Acetyl-(Glc-H2O)]−
| + | |||||
15 | Acetyl-hydroxy-SSd | 13.29 | C44H72O15
| 839.4777 [M-H]− (−2.5) 885.4822 [M + HCOO]− (−3.5) | 797.4856 [M-H-Acetyl]−; 635.4268 [M-H-Acetyl-(Glc-H2O)]−
| + | |||||
16 | Malonyl-acetyl-hydroxy-SSa/Malonyl-acetyl-hydroxy-SSd | 13.37 | C45H72O17
| 883.4663 [M-H]− (−3.9) | 797.4801 [M-H-Malonyl-Acetyl]−; 635.4276 [M-H-Malonyl-Acetyl- (Glc-H2O)]−
| + | + | ||||
17 | Malonyl-SSn | 13.61 | C51H80O21
| 1027.5194 [M-H]− (6.21) | 985.5383 [M-H-Acetyl]−, 941.5287 [M-H-Acetyl-CO2]−
| + | |||||
18 | Unknown | 14.06 | C51H90O26
| 1117.5640 [M-H]− (−0.7) | 955.5135 [M-H-(Glc-H2O)]−
| + | |||||
19 | Unknown | 14.15 | C51H90O26
| 1117.5654 [M-H]− (−0.5) | 955.5135 [M-H-(Glc-H2O)]−
| + | |||||
20 | SSc | 14.73 | C48H78O17
| 925.5168 [M-H]− (0.2) 971.5208 [M + HCOO]− (1.3) | 779.4734 [M-H-(Rha-H2O)]−
| + | + | + | + | ||
21 | SSf | 14.87 | C48H80O17
| 927.5313 [M-H]− (1.1) 973.5376 [M + HCOO]− (0.2) | 781.4895 [M-H-(Rha-H2O)]−
| + | + | + | + | ||
22 | Malonyl-Ssc | 15.23 | C51H80O20
| 1011.5162 [M-H]− (−0.8) | 967.5447 [M-H-CO2]−, 779.4838 [M-H-Malonyl-(Rha-H2O)]−
| + | + | + | + | ||
23 | Malonyl-Ssf | 15.37 | C51H82O20
| 1013.5322 [M-H]− (−0.5) | 969.5611 [M-H-CO2]−, 927.5527 [M-H- Malonyl]−, 781.4795 [M-H-Malonyl-(Rha-H2O)]−, 765.4884 [M-H-Malonyl-(Glc-H2O)]−
| + | + | ||||
24 | Malonyl-acetyl-rotundifolioside B | 15.59 | C52H82O21
| 1041.5205 [M-H]− (−6.8) | 997.5215 [M-H-CO2]−, 835.4661 [M-H-CO2-(Glc-H2O)]−
| + | |||||
25 | SSb3 / SSb4 | 16.34 | C43H72O14
| 811.4847 [M-H]− (−0.3) 857.4985 [M + HCOO]− (0.9) | 649.4344 [M-H-(Glc-H2O)]−
| + | |||||
26 | SSn | 17.31 | C48H78O18
| 941.5196 [M-H]− (0.8) 987.5127 [M + HCOO]− (0.4) | 779.4722 [M-H-(Glc-H2O)]−
| + | |||||
27 | Malonyl-SSb3 / SSb4 | 17.38 | C46H74O17
| 897.4823 [M-H]− (3.3) | 853.5090 [M-H-CO2]−, 811.5024 [M-H- Malonyl]−, 649.4453 [M-H-Malonyl-(Glc-H2O)]−
| + | |||||
28 | SSa | 17.83 | C42H68O13
| 779.4558 [M-H]− (−0.4) 825.4629 [M + HCOO]− (−0.2) | 617.4186 [M-H-(Glc-H2O)]−
| + | + | + | + | ||
29 | Acetyl-SSa | 18.65 | C44H70O14
| 821.4668 [M-H]− (−3.0) 867.4713 [M + HCOO]− (−4.0) | 779.4742 [M-H-Acetyl]−, 617.4236 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | |||
30 | Acetyl-SSa | 18.77 | C44H70O14
| 821.4660 [M-H]− (−4.0) 867.4718 [M + HCOO]− (−3.4) | 779.4746 [M-H-Acetyl]−, 617.4209 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | |||
31 | Acetyl-SSa | 18.93 | C44H70O14
| 821.4649 [M-H]− (−5.3) 867.4714 [M + HCOO]− (−3.9) | 779.4752 [M-H-Acetyl]−, 617.4177 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | |||
32 | Malonyl-SSa | 19.01 | C45H70O16
| 865.4576 [M-H]− (−1.7) | 821.4851 [M-H-CO2]−, 779.4747 [M-H- Malonyl]−, 617.4182 [M-H- Malonyl-(Glc-H2O)]−
| + | + | + | + | ||
33 | Acetyl-SSa | 19.55 | C44H70O14
| 821.4657 [M-H]− (−4.4) 867.4713 [M + HCOO]− (−4.0) | 779.4746 [M-H-Acetyl]−, 617.4188 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | |||
34 | Dimalonyl-SSa | 19.68 | C52H72O16
| 951.4765 [M-H]− (1.8) | 907.4863 [M-H-CO2]−, 779.4687 [M-H-2 Malonyl]−, 617.4188 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
35 | Dimalonyl-SSa | 20.08 | C52H72O16
| 951.4771 [M-H]− (2.4) | 907.4873 [M-H-CO2]−, 779.4757 [M-H-2 Malonyl]−, 617.4244 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
36 | Dimalonyl-SSa | 20.22 | C52H72O16
| 907.4874 [M-H-CO2]− (2.8) | 779.4717 [M-H-2 Malonyl]−, 617.4192 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
37 | Dimalonyl-SSa | 20.43 | C52H72O16
| 907.4880 [M-H-CO2]− (3.4) | 779.4764 [M-H-2 Malonyl]−, 617.4199 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
38 | Dimalonyl-SSa | 20.64 | C52H72O16
| 907.4865 [M-H-CO2]− (1.8) | 779.4799 [M-H-2 Malonyl]−, 617.4156 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
39 | Dimalonyl-acetyl-SSa | 21.31 | C54H74O17
| 993.4864 [M-H]− (1.1) | 949.4979 [M-H-CO2]−, 864.5019 [M-2H-Malonyl-Acetyl]−, 821.4803 [M-2Malonyl]−, 761.4609 [M-H-Malonyl-(Fuc-H2O)]−
| + | + | ||||
40 | SSs | 21.44 | C59H74O10
| 941.5269 [M-H]− (6.4) 987.5245 [M + HCOO]− (−1.9) | 780.4802 [M-(Glc-H2O)]−
617.4202 [M-H-2(Glc-H2O)]−
| + | |||||
41 | Dimalonyl-SSa / SSd | 21.53 | C52H72O16
| 907.4865 [M-H-CO2]− (1.8) | 779.4799 [M-H-2 Malonyl]−, 617.4156 [M-H-2 Malonyl-(Glc-H2O)]−
| + | |||||
42 | Malonyl-SSe | 21.68 | C45H70O15
| 849.4689 [M-H]− (5.5) | 805.4903 [M-H-CO2]−, 763.4800 [M-H-Malonyl]−, 643.4376 [M-H-CO-(Glc-H2O)]−
601.4253 [M-H-Malonyl-(Glc-H2O)]−
| + | + | + | |||
43 | SSd | 22.62 | C42H68O13
| 779.4547 [M-H]− (−5.1) 825.4606 [M + HCOO]− (−4.4) | 617.4186 [M-H-(Glc-H2O)]−
| + | + | + | + | ||
44 | Acetyl-SSd | 23.60 | C44H70O14
| 821.4653 [M-H]− (−4.9) 867.4756 [M + HCOO]− (0.9) | 779.4739 [M-H-Acetyl]−, 617.4196 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | |||
45 | Malonyl-SSd | 23.85 | C45H70O16
| 865.4571 [M-H]− (−2.3) | 821.4859 [M-H-CO2]−, 779.4716 [M-H- Malonyl]−, 617.4199 [M-H- Malonyl-(Glc-H2O)]−
| + | + | + | + | ||
46 | Acetyl-SSd | 24.22 | C44H70O14
| 821.4648 [M-H]− (−5.5) 867.4722 [M + HCOO]− (−3.0) | 779.4745 [M-H-Acetyl]−, 617.4238 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | + | ||
47 | Acetyl-SSd | 24.49 | C44H70O14
| 821.4671 [M-H]− (−2.7) 867.4717 [M + HCOO]− (−3.6) | 779.4723 [M-H-Acetyl]−, 617.4218 [M-H-Acetyl-(Glc-H2O)]−
| + | + | + | |||
48 | Dimalonyl-SSd | 24.62 | C52H72O16
| 951.4771 [M-H]− (2.4) | 907.4859 [M-H-CO2]−, 821.4865[M-H- Malonyl-CO2]−, 779.4799 [M-H-2 Malonyl]−, 617.4156 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
49 | Dimalonyl-SSd | 25.14 | C52H72O16
| 907.4863 [M-H-CO2]− (1.5) | 821.4924[M-H- Malonyl-CO2]−, 779.4829 [M-H-2 Malonyl]−, 617.4177 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | |||
50 | Dimalonyl-SSd | 25.60 | C52H72O16
| 907.4873 [M-H-CO2]− (2.6) | 821.4856 [M-H- Malonyl-CO2]−, 779.4761 [M-H-2 Malonyl]−, 617.4191 [M-H-2 Malonyl-(Glc-H2O)]−
| + | + | + | + | ||
51 | Dimalonyl-acetyl-SSd | 26.40 | C54H74O17
| 993.4828 [M-H]− (−2.5) | 949.4978 [M-H-CO2]−, 821.4838 [M-2Malonyl]−
| + | + | ||||
52 | Dimalonyl-acetyl-SSd | 26.52 | C54H74O17
| 993.4896 [M-H]− (4.0) | 949.4976 [M-H-CO2]−, 864.5027 [M-2H-Malonyl-Acetyl]−, 821.4860 [M-2Malonyl]−, 761.4615 [M-H-Malonyl-(Fuc-H2O)]−, | + | + | + | |||
53 | Malonyl-diacetyl-SSd | 27.81 | C53H73O15
| 949.4971 [M-H]− (1.7) | 863.5013 [M-H-Malonyl]−, 779.4689 [M-H-Malonyl-2Acetyl]−
| + | + | ||||
54 | Isorhamnetin-3-O-rutinoside | 8.20 | C28H32O16
| 623.1651 [M-H]− (5.3) | / | + | + | + | + | + | |
55 | Quercetin-3-O-rhamnoside | 8.32 | C21H20O11
| 447.0936 [M-H]− (0.7) | 300.0349 [M-H-(Rha-H2O)]−
| + | + | ||||
56 | Quercetin 3-(6″-acetylglucoside) | 8.54 | C23H22O13
| 505.0993 [M-H]− (1.0) | 300.0330 [M-2H-Acetyl-(Glc-H2O)]−
| + |
Principal component analysis (PCA)
Potential differentiating markers
Peak No. | Compounds | R.T. (mins) |
p-value | Log FC |
---|---|---|---|---|
3 | Isoquercitrin | 7.30 | 2.09 × 10−6
| −15.798277 |
23 | Malonyl-Ssf | 15.37 | 7.41 × 10−6
| 15.704748 |
28 | Saikosaponin a | 17.83 | 6.74 × 10−6
| 17.776505 |
32 | Malonyl-Ssa | 19.01 | 6.61 × 10−6
| 16.194246 |
43 | Saikosaponin d | 22.62 | 7.21 × 10−6
| 17.776505 |
44 | Acetyl-Ssd | 23.60 | 7.90 × 10−6
| 15.572599 |
45 | Malonyl-Ssd | 23.85 | 1.33 × 10−16
| 20.250618 |
46 | Acetyl-Ssd | 24.22 | 6.35 × 10−6
| 17.99062 |
52 | Dimalonyl-acetyl-Ssd | 26.52 | 6.61 × 10−6
| 16.017168 |