A New UPLC-MS/MS Method for the Characterization and Discrimination of Polysaccharides from Genus Ephedra Based on Enzymatic Digestions
Abstract
:1. Introduction
2. Experimental
2.1. Materials and Reagents
2.2. Preparation of Polysaccharides from Genus Ephedra
2.3. Specific Enzymatic Digestion of Ephedra Polysaccharides
2.4. Derivatization with PMP Reagent
2.5. UPLC Apparatus and Conditions
2.6. MS Apparatus and Conditions
2.7. Data Processing
3. Results and Discussion
3.1. Derivatization and MS Conditions
3.2. Quantification and Validation
3.2.1. Effects of Columns
3.2.2. Effects of Additives
3.2.3. Effects of Column Temperature and Flow Rate
3.3. Method Validations
3.4. Application to Real Samples
3.5. Discrimination of Ephedra Polysaccharides Based on PCA
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Compounds 1–22 are available from the authors. |
Peaks | Original Mw | PMP Derivatives (m/z) | Q1 (m/z) | Q3 (m/z) | DP (V) | CE (V) |
---|---|---|---|---|---|---|
1 | 180.2 | [M − H]− | 509.1 | 215.1 | −49.2 | −28.1 |
2 | 215.6 | [M − H]− | 508.2 | 214.0 | −65.9 | −29.8 |
3 | 215.6 | [M − H]− | 508.4 | 213.7 | −64.5 | −26.0 |
4 | 1153.0 | [M − H]− | 1481.9 | 1308.0 | −96.4 | −35.1 |
5 | 150.1 | [M − H]− | 478.7 | 215.0 | −58.4 | −29.9 |
6 | 164.2 | [M − H]− | 493.2 | 214.9 | −60.6 | −29.0 |
7 | 990.9 | [M − H]− | 1319.7 | 869.8 | −99.5 | −61.2 |
8 | 215.6 | [M − H]− | 508.2 | 213.8 | −67.9 | −33.2 |
9 | 828.7 | [M − H]− | 1157.7 | 707.6 | −90.3 | −51.7 |
10 | 666.6 | [M − H]− | 995.6 | 214.8 | −80.8 | −52.6 |
11 | 212.2 | [M − H]− | 523.2 | 215.0 | −55.9 | −30.3 |
12 | 194.1 | [M − H]− | 523.1 | 215.0 | −57.5 | −47.4 |
13 | 504.4 | [M − H]− | 833.6 | 215.3 | −98.4 | −46.8 |
14 | 221.2 | [M − H]− | 550.2 | 256.2 | −63.9 | −32.9 |
15 | 360.3 | [M − H]− | 671.5 | 214.8 | −61.2 | −34.1 |
16 | 182.2 | [M − H]− | 511.2 | 214.6 | −63.7 | −28.0 |
17 | 180.2 | [M − H]− | 509.0 | 214.7 | −59.9 | −26.1 |
18 | 221.2 | [M − H]− | 550.3 | 256.0 | −69.4 | −34.1 |
19 | 180.2 | [M − H]− | 509.2 | 215.0 | −65.0 | −29.9 |
20 | 150.1 | [M − H]− | 479.0 | 215.1 | −48.4 | −33.9 |
20 | 150.1 | [M − H]− | 479.1 | 215.0 | −63.1 | −28.1 |
21 | 164.2 | [M − H]− | 493.1 | 215.1 | −54.5 | −30.4 |
22 | 126.1 | [M − H]− | 455.4 | 173.1 | −60.9 | −14.1 |
No. | Calibration Curves | R2 | Linear Range (μM) | LOD (nM) | LOQ (nM) |
---|---|---|---|---|---|
1 | y = 0.2634x + 0.1916 | 0.9938 | 0.02–49.60 | 3.00 | 8.00 |
2 | y = 0.1721x + 0.0360 | 0.9982 | 0.19–49.60 | 24.00 | 58.00 |
3 | y = 0.2135x + 0.2631 | 0.9931 | 0.05–49.60 | 12.00 | 34.00 |
4 | y = 0.0002x + 0.0002 | 0.9995 | 3.10–49.60 | 780.00 | 2240.00 |
5 | y = 0.4846x + 0.2466 | 0.9960 | 0.02–49.60 | 3.00 | 92.00 |
6 | y = 0.1496x + 0.1560 | 0.9952 | 0.19–49.60 | 48.00 | 140.00 |
7 | y = 0.0012 x + 0.0013 | 0.9938 | 0.78–49.60 | 193.00 | 640.00 |
8 | y = 0.1692x + 0.2124 | 0.9920 | 0.05–49.60 | 12.00 | 48.00 |
9 | y = 0.0042x + 0.0009 | 0.9985 | 0.19–49.60 | 97.00 | 260.00 |
10 | y = 0.0325x − 0.0110 | 0.9952 | 0.10–49.60 | 12.00 | 34.00 |
11 | y = 0.0661 x + 0.0919 | 0.9955 | 0.05–49.60 | 24.00 | 48.00 |
12 | y = 0.0223x + 0.0342 | 0.9935 | 0.39–49.60 | 32.00 | 96.00 |
13 | y = 0.0144x − 0.0040 | 0.9967 | 0.10–49.60 | 48.00 | 16.00 |
14 | y = 0.1207x + 0.0956 | 0.9938 | 0.05–49.60 | 12.00 | 34.00 |
15 | y = 0.0317x − 0.0026 | 0.9989 | 0.02–49.60 | 48.00 | 14.00 |
17 | y = 0.3381x + 0.2201 | 0.9973 | 0.02–49.60 | 3.00 | 6.00 |
18 | y = 0.3737x + 0.1974 | 0.9954 | 0.02–49.60 | 12.00 | 36.00 |
19 | y = 0.6961x + 0.4415 | 0.9954 | 0.02–49.60 | 3.00 | 10.00 |
20 | y = 0.6961x + 0.4415 | 0.9954 | 0.02–49.60 | 3.00 | 10.00 |
21 | y = 0.3787x + 0.4894 | 0.9925 | 0.19–49.60 | 24.00 | 72.00 |
22 | y = 0.4953x − 0.2220 | 0.9972 | 0.05–49.60 | 2.00 | 5.00 |
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Xia, Y.-G.; Wang, T.-L.; Sun, L.-M.; Liang, J.; Yang, B.-Y.; Kuang, H.-X. A New UPLC-MS/MS Method for the Characterization and Discrimination of Polysaccharides from Genus Ephedra Based on Enzymatic Digestions. Molecules 2017, 22, 1992. https://doi.org/10.3390/molecules22111992
Xia Y-G, Wang T-L, Sun L-M, Liang J, Yang B-Y, Kuang H-X. A New UPLC-MS/MS Method for the Characterization and Discrimination of Polysaccharides from Genus Ephedra Based on Enzymatic Digestions. Molecules. 2017; 22(11):1992. https://doi.org/10.3390/molecules22111992
Chicago/Turabian StyleXia, Yong-Gang, Tian-Long Wang, Li-Ming Sun, Jun Liang, Bing-You Yang, and Hai-Xue Kuang. 2017. "A New UPLC-MS/MS Method for the Characterization and Discrimination of Polysaccharides from Genus Ephedra Based on Enzymatic Digestions" Molecules 22, no. 11: 1992. https://doi.org/10.3390/molecules22111992