Analysis of Chemical Variations between Crude and Salt-Processed Anemarrhenae rhizoma Using Ultra-High-Performance Liquid Chromatography–Mass Spectrometry Methods
Abstract
:1. Introduction
2. Results and Discussion
2.1. Multivariate Data Analyses and Identification of Characteristic Chemical Compositions Differentiation of CAR and SAR
2.2. Determination of the Seven Main Components in CAR and SAR
2.3. Compounds Changed upon Processing
3. Materials and Methods
3.1. Materials and Reagents
3.2. Preparation of Standard Solutions
3.3. Sample Preparation
3.4. UHPLC–QTOF-MS Analysis
3.4.1. Instrument and Chromatographic Conditions
3.4.2. Data Analysis
3.5. Quantification of Seven Compounds by UHPLC-MS
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Samples of the compound timosaponin N is available from the authors. |
Peak | tR | Formula | Negative Ion Mode of ESI-MS (m/z) | Positive Ion Mode of ESI-MS (m/z) | Identification | t-Value | p-Value | Changing Direction c | ||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Precursor Ion | Selective Ion | MS2 Fragmentation | Precursor Ion | Selective Ion | MS2 Fragmentation | |||||||
1 | 9.81 | C45H76O20 | 981.4892 | [M + HCOO]− | 935, 773, 611 | 919.4911 | [M − H2O + H]+ | 757, 595, 433, 415, 289, 271, 253 | Timosaponin N a | 2.77 | 0.01271 | ↓ * |
2 | 9.97 | C45H76O20 | 981.4925 | [M + HCOO]− | 935, 773, 611 | 919.4916 | [M − H2O + H]+ | 757, 595, 433, 415, 289, 271, 253 | Macrostemonoside J [12] | 2.06 | 0.03379 | ↓ * |
3 | 10.78 | C45H76O20 | 981.4914 | [M + HCOO]− | 935, 773, 611 | 919.4918 | [M − H2O + H]+ | 757, 595, 433, 415, 289, 271, 253 | Timosaponin E1 a | 3.25 | 0.00447 | ↓ ** |
4 | 11.08 | C51H86O24 | 1127.5433 | [M + HCOO]− | 1081, 919, 757, 595 | 1065.5482 | [M − H2O + H]+ | 903, 741, 579, 417, 399, 273, 255 | Petunioside N [13] | 2.21 | 0.03999 | ↓ * |
5 | 11.89 | C51H86O23 | 1111.5528 | [M + HCOO]− | 1065, 919, 757, 595 | 1049.5289 | [M − H2O + H]+ | 903, 741, 579, 417, 399, 273, 255 | Curilioside H [14] | 2.92 | 0.00921 | ↓ ** |
6 | 12.03 | C45H76O19 | 965.5029 | [M + HCOO]− | 919, 757, 595 | 903.4982 | [M − H2O + H]+ | 741, 579, 417, 399, 273, 255 | Timosaponin BII a | 4.10 | 0.00067 | ↓ ** |
7 | 12.14 | C45H76O19 | 965.4945 | [M + HCOO]− | 919, 757, 595 | 903.4931 | [M − H2O + H]+ | 741, 579, 417, 399, 273, 255 | 25R-timosaponin BII [12,15] | 4.17 | 0.00058 | ↓ ** |
8 | 12.30 | C45H76O19 | 965.4943 | [M + HCOO]− | 919, 757, 595 | 903.4940 | [M − H2O + H]+ | 741, 579, 417, 399, 273, 255 | 25S-officinalisinin-I [12,15] | 2.48 | 0.03575 | ↓ * |
9 | 12.60 | C56H92O28 | 1257.5733 | [M + HCOO]− | 1211, 1079, 1049, 917, 755, 593 | 1195.4256 | [M − H2O + H]+ | 1033, 901, 739, 577, 433, 415, 271, 253 | Timosaponin H1 [12,16] | 2.73 | 0.01360 | ↓ * |
10 | 12.75 | C56H94O28 | 1259.5853 | [M + HCOO]− | 1213, 1081, 919, 757, 595 | 1197.4558 | [M − H2O + H]+ | 1065, 903, 741, 579, 435, 417, 273, 255 | Timosaponin I1 [12,16] | 2,17 | 0.04392 | ↓ * |
11 | 13.30 | C45H74O19 | 963.4798 | [M + HCOO]− | 917, 755, 593 | 919.4893 | [M + H]+ | 757, 595, 433, 415, 289, 271, 253 | Timosaponin D [12,17] | −5.82 | 1.6189 × 10−5 | ↑ ** |
12 | 13.46 | C45H74O19 | 963.4806 | [M + HCOO]− | 917, 755, 593 | 919.4901 | [M + H]+ | 757, 595, 433, 415, 289, 271, 253 | 25R-timosaponin D [12] | 3.53 | 0.00241 | ↑ ** |
13 | 13.64 | C45H74O19 | 963.4793 | [M + HCOO]− | 917, 755, 593 | 919.4889 | [M + H]+ | 757, 595, 433, 415, 289, 271, 253 | Timosaponin D isomer [12] | −2.02 | 0.03806 | ↑ * |
14 | 13.83 | C51H84O23 | 1109.5370 | [M + HCOO]− | 1063, 901, 739 | 1065.5008 | [M + H]+ | 903, 741, 579, 417, 273, 255 | (25R)-26-O-β-d-glucopyranosyl-5α-furostane-20(22)-en-3β, 26-diol-3-O-β-d-glucopyranosyl-(1→2)-β-d-glucopyranosyl-(1→4)-β-d-galactopyranoside [12,14] | −2.82 | 0.01144 | ↑ * |
15 | 14.82 | C45H74O18 | 947.4865 | [M+HCOO]− | 901, 739, 721, 577 | 903.4984 | [M + H]+ | 741, 579, 417, 273, 255 | Timosaponin C [12,14] | −5.24 | 5.4915 × 10−5 | ↑ ** |
16 | 14.95 | C45H74O18 | 947.4851 | [M + HCOO]− | 901, 739, 577 | 903.4941 | [M + H]+ | 741, 579, 417, 273, 255 | Timosaponin BIII a | −9.15 | 3.4202 × 10−8 | ↑ ** |
17 | 15.14 | C45H74O18 | 947.4854 | [M + HCOO]− | 901, 739, 577 | 903.4958 | [M + H]+ | 741, 579, 417, 273, 255 | 25R-timosaponin BIII [12,14] | −11.53 | 9.5454 ×10−10 | ↑ ** |
18 | 15.33 | C51H84O23 | 1109.5577 | [M + HCOO]− | 1063, 901, 739 | 1065.5078 | [M + H]+ | 903, 741, 579, 417, 273, 255 | Timosaponin B IV [12,14] | −4.44 | 0.00032 | ↑ ** |
19 | 16.85 | C39H66O14 | 803.4412 | [M + HCOO]− | 757, 595, 433 | 741.4397 | [M − H2O + H]+ | 579, 417, 399, 289, 271, 253 | Anemarrhenasaponin I a | 3.23 | 0.00464 | ↓ ** |
20 | 17.03 | C39H66O14 | 803.4415 | [M + HCOO]− | 757, 595, 433 | 741.4398 | [M − H2O + H]+ | 579, 417, 399, 289, 271, 253 | Anemarrhenasaponin II [18] | 2.12 | 0.04854 | ↓ * |
21 | 20.32 | C39H64O13 | 785.4321 | [M + HCOO]− | 739, 577 | 741.4308 | [M + H]+ | 579, 417, 399, 289, 271, 253 | 20(22)-en-5β-furost-3β,15α-diol-3-O-β-d-glucopyranosyl-(1→2)-β-d-galactopyranoside b | −4.71 | 0.00017 | ↑ ** |
22 | 20.77 | C39H64O13 | 785.4323 | [M + HCOO]− | 739, 577 | 741.4303 | [M + H]+ | 579, 417, 399, 289, 271, 253 | 20(22)-en-5β-furost-3β,15β-diol-3-O-β-d-glucopyranosyl-(1→2)-β-d-galactopyranoside b | −2.12 | 0.04033 | ↑ * |
23 | 23.92 | C18H32O3 | 295.2280 | [M − H]− | 277, 195, 171 | Hydroxy-octadecadienoic acid [19] | 4.33 | 0.00040 | ↓ ** | |||
24 | 24.15 | C18H30O3 | 293.2124 | [M − H]− | Hydroxy-octadecatrienoic acid [19] | 2.39 | 0.02800 | ↓ * |
Compounds | Linear Regression | r2 | Linear Range (μg/mL) | LOD (ng/mL) | LOQ (ng/mL) |
---|---|---|---|---|---|
Timosaponin N | y = 68,743x + 27,585 | 0.9997 | 2.0–16.0 | 2.8 | 9.8 |
Timosaponin E1 | y = 100,376x + 13,353 | 0.9997 | 0.1–6.0 | 3.6 | 11.3 |
Timosaponin BII | y = 13,964x + 8444 | 0.9999 | 20.0–120.0 | 4.8 | 19.0 |
Timosaponin BIII | y = 53,259x + 21,310 | 0.9998 | 3.0–60.0 | 2.7 | 8.6 |
Anemarrhenasaponin I | y = 71,064x + 29,916 | 0.9999 | 2.0–16.0 | 3.0 | 10.0 |
Timosaponin AII | y = 58,520x + 17,831 | 0.9998 | 2.0–16.0 | 2.0 | 7.4 |
Timosaponin AIII | y = 38,170x + 32,791 | 0.9998 | 3.0–60.0 | 1.0 | 3.5 |
No. | Timosaponin N | Timosaponin E1 | Timosaponin BII | Timosaponin BIII | Anemarrhenasaponin I | Timosaponin AII | Timosaponin AIII | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
CAR | SAR | CAR | SAR | CAR | SAR | CAR | SAR | CAR | SAR | CAR | SAR | CAR | SAR | |
1 | 11.03 ± 0.28 | 5.20 ± 0.13 ** | 12.57 ± 0.03 | 7.45 ± 0.02 ** | 81.58 ± 1.38 | 72.44 ± 0.61 ** | 1.44 ± 0.18 | 11.18 ± 0.57 ** | 9.16 ± 0.23 | 7.92 ± 0.10 ** | 9.09 ± 0.23 | 9.00 ± 0.23 | 37.82 ± 0.96 | 38.33 ± 0.97 |
2 | 13.44 ± 0.34 | 6.82 ± 0.17 ** | 15.78 ± 0.04 | 10.79 ± 0.03 * | 83.13 ± 1.41 | 72.80 ± 0.62 ** | 2.02 ± 0.26 | 10.53 ± 0.53 ** | 7.54 ± 0.19 | 6.07 ± 0.08 * | 5.63 ± 0.14 | 5.41 ± 0.14 | 26.77 ± 0.68 | 25.79 ± 0.65 |
3 | 12.59 ± 0.32 | 5.26 ± 0.13 ** | 19.70 ± 0.05 | 6.64 ± 0.02 ** | 91.21 ± 1.54 | 63.05 ± 0.53 ** | 1.73 ± 0.22 | 13.04 ± 0.66 ** | 7.15 ± 0.18 | 3.95 ± 0.05 ** | 2.91 ± 0.07 | 4.06 ± 0.10 | 19.94 ± 0.38 | 21.88 ± 0.56 |
4 | 14.47 ± 0.37 | 5.00 ± 0.13 ** | 15.98 ± 0.04 | 8.89 ± 0.02 ** | 84.31 ± 1.43 | 71.33 ± 0.60 ** | 0.63 ± 0.18 | 12.01 ± 0.61 ** | 6.54 ± 0.17 | 5.86 ± 0.07 * | 2.26 ± 0.06 | 2.34 ± 0.06 | 13.49 ± 0.34 | 13.46 ± 0.34 |
5 | 11.53 ± 0.29 | 5.65 ± 0.14 ** | 6.53 ± 0.02 | 4.07 ± 0.01 * | 82.76 ± 1.40 | 72.01 ± 0.61 ** | 6.39 ± 0.15 | 11.58 ± 0.59 ** | 8.24 ± 0.21 | 7.30 ± 0.09 * | 4.23 ± 0.11 | 4.41 ± 0.11 | 35.56 ± 0.90 | 36.99 ± 0.94 |
6 | 9.21 ± 0.23 | 4.11 ± 0.10 ** | 19.37 ± 0.05 | 6.07 ± 0.02 ** | 91.01 ± 1.54 | 60.15 ± 0.51 ** | 1.30 ± 0.17 | 11.92 ± 0.61 ** | 9.78 ± 0.25 | 5.98 ± 0.08 ** | 6.15 ± 0.16 | 7.04 ± 0.18 | 32.63 ± 0.65 | 32.05 ± 0.81 |
7 | 12.58 ± 0.35 | 5.55 ± 0.14 ** | 8.47 ± 0.02 | 4.65 ± 0.01 ** | 71.68 ± 1.21 | 55.24 ± 0.47 ** | 3.32 ± 0.42 | 5.94 ± 0.81 ** | 7.12 ± 0.18 | 5.51 ± 0.07 * | 8.16 ± 0.21 | 9.82 ± 0.25 | 36.65 ± 0.78 | 39.88 ± 1.01 |
8 | 6.89 ± 0.17 | 2.60 ± 0.07 ** | 1.26 ± 0.00 | 0.52 ± 0.01 * | 85.47 ± 1.45 | 68.21 ± 0.58 ** | 1.22 ± 0.15 | 14.20 ± 0.72 ** | 11.69 ± 0.30 | 8.34 ± 0.11 ** | - | - | 6.32 ± 0.12 | 7.22 ± 0.11 |
9 | 10.13 ± 0.26 | 4.87 ± 0.12 ** | 13.84 ± 0.04 | 10.15 ± 0.03 * | 98.30 ± 1.66 | 92.92 ± 0.79 * | 1.06 ± 0.13 | 8.07 ± 0.41 ** | 6.31 ± 0.16 | 4.94 ± 0.08 * | 2.93 ± 0.07 | 3.51 ± 0.09 | 17.71 ± 0.40 | 18.11 ± 0.46 |
10 | 9.06 ± 0.23 | 4.10 ± 0.10 ** | 4.91 ± 0.01 | 2.56 ± 0.01 ** | 73.79 ± 1.25 | 55.96 ± 0.47 ** | 5.99 ± 0.15 | 9.44 ± 0.48 ** | 4.61 ± 0.12 | 3.21 ± 0.04 * | 4.52 ± 0.11 | 4.68 ± 0.12 | 41.02 ± 1.04 | 41.82 ± 1.06 |
No. | Place of Collection | Collection Time | Growing Condition |
---|---|---|---|
1 | Changzhi, Shanxi | 2014.09 | Cultivated |
2 | Anguo, Hebei | 2014.10 | Cultivated |
3 | Bozhou, Anhui | 2014.08 | Cultivated |
4 | Huludao, Liaoning | 2014.11 | wild |
5 | Chifeng, Neimenggu | 2014.12 | wild |
6 | Changzhi, Shanxi | 2015.03 | Cultivated |
7 | Anguo, Hebei | 2015.04 | Cultivated |
8 | Datong, Shanxi | 2015.03 | Cultivated |
9 | Bozhou, Anhui | 2015.04 | Cultivated |
10 | Chifeng, Neimenggu | 2015.05 | wild |
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Ji, D.; Su, X.; Huang, Z.; Su, L.; Li, L.; Lu, T. Analysis of Chemical Variations between Crude and Salt-Processed Anemarrhenae rhizoma Using Ultra-High-Performance Liquid Chromatography–Mass Spectrometry Methods. Molecules 2018, 23, 23. https://doi.org/10.3390/molecules23010023
Ji D, Su X, Huang Z, Su L, Li L, Lu T. Analysis of Chemical Variations between Crude and Salt-Processed Anemarrhenae rhizoma Using Ultra-High-Performance Liquid Chromatography–Mass Spectrometry Methods. Molecules. 2018; 23(1):23. https://doi.org/10.3390/molecules23010023
Chicago/Turabian StyleJi, De, Xiaonan Su, Ziyan Huang, Lialin Su, Lin Li, and Tulin Lu. 2018. "Analysis of Chemical Variations between Crude and Salt-Processed Anemarrhenae rhizoma Using Ultra-High-Performance Liquid Chromatography–Mass Spectrometry Methods" Molecules 23, no. 1: 23. https://doi.org/10.3390/molecules23010023