Effect of Different Processing Methods on the Chemical Constituents of Scrophulariae Radix as Revealed by 2D NMR-Based Metabolomics
Abstract
:1. Introduction
2. Results and Discussion
2.1. Samples Collection and Processing
2.2. Optimization of Extraction Conditions
2.2.1. Selecting the Optimum Extraction Solvent
2.2.2. Optimization of the Solid/Liquid Ratio of the Extraction
2.2.3. Optimum Sample Amount Used per NMR Sample
2.3. Comparison of SR Being Steamed Processing for Different Time
2.3.1. Multivariate Statistical Analysis
2.3.2. Change of Chemical Constituents of SR Steamed for Different Durations
Feature | δH | δC | Assignment | Feature | δH | δC | Assignment | Feature | δH | δC | Assignment |
---|---|---|---|---|---|---|---|---|---|---|---|
F115 | 2.72 | 46.66 | aucubin | F121 | 3.02 | 49.38 | aucubin | F124 | 5.05 | 99.01 | aucubin |
F128 | 5.81 | 131.75 | aucubin | F127 | 5.12 | 107.85 | aucubin | F135 | 6.32 | 142.93 | aucubin |
F179 | 4.33 | 62.72 | aucubin | F196 | 4.35 | 62.72 | aucubin | F283 | 3.84 | 79.19 | harpagoside |
F299 | 5.01 | 108.14 | harpagoside | F346 | 2.95 | 56.61 | harpagoside | F372 | 6.16 | 96.43 | harpagoside |
F408 | 6.47 | 145.71 | harpagoside | F146 | 1.52 | 24.36 | harpagoside | F558 | 2.29 | 47.69 | harpagoside |
F581 | 2.04 | 47.68 | harpagoside | F773 | 6.55 | 121.43 | harpagoside, cinnamic acid | F225 | 7.63 | 131.07 | harpagoside, cinnamic acid |
F679 | 7.69 | 148.31 | harpagoside, cinnamic acid | F101 | 4.72 | 101.44 | aucubin, harpagoside | F118 | 1.26 | 26.82 | harpagide |
F242 | 5.03 | 109.70 | harpagide | F216 | 3.78 | 79.47 | harpagide | F219 | 4.67 | 101.09 | harpagide |
F279 | 6.37 | 144.22 | harpagide | F254 | 2.57 | 60.43 | harpagide | F273 | 5.73 | 95.26 | harpagide |
F375 | 1.95 | 48.66 | harpagide | F294 | 1.85 | 48.66 | harpagide | F607 | 1.83 | 48.66 | harpagide |
F495 | 1.97 | 48.66 | harpagide | F106 | 3.30 | 76.01 | aucubin, harpagoside, harpagide | F268 | 3.31 | 75.64 | aucubin, harpagoside, harpagide |
F244 | 3.35 | 72.89 | aucubin, harpagoside, harpagide, glucose | F220 | 3.77 | 61.50 | 6-O-methyl-catalpol | F218 | 2.35 | 38.86 | 6-O-methyl-catalpol |
F229 | 6.40 | 143.71 | 6-O-methyl-catalpol | F207 | 5.08 | 105.85 | 6-O-methyl-catalpol | F520 | 4.18 | 63.00 | 6-O-methyl-catalpol |
F138 | 4.82 | 101.35 | 6-O-methyl-catalpol | F201 | 5.05 | 97.23 | 6-O-methyl-catalpol | F123 | 4.20 | 62.79 | 6-O-methyl-catalpol |
F184 | 3.35 | 76.10 | 6-O-methyl-catalpol | F194 | 2.60 | 44.63 | 6-O-methyl-catalpol | F178 | 4.23 | 62.72 | 6-O-methyl-catalpol |
F111 | 3.48 | 78.90 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol, glucuronic acid | F107 | 3.38 | 79.20 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol, glucose | F108 | 3.37 | 79.39 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol |
F42 | 3.45 | 79.22 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol, glucuronic acid | F300 | 3.41 | 79.25 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol | F228 | 3.41 | 79.45 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol, glucose |
F649 | 1.53 | 27.47 | isoleucine | F367 | 3.39 | 78.91 | aucubin, harpagoside, harpagide, 6-O-methyl-catalpol, glucose | F460 | 3.61 | 62.61 | isoleucine |
F238 | 0.96 | 14.06 | isoleucine | F611 | 1.98 | 39.00 | isoleucine | F224 | 1.03 | 17.53 | isoleucine |
F159 | 7.18 | 133.57 | tyrosine | F330 | 2.65 | 32.01 | methionine | F1423 | 3.00 | 38.63 | tyrosine |
F659 | 3.20 | 38.64 | tyrosine | F147 | 6.85 | 118.65 | tyrosine | F248 | 3.88 | 59.18 | tyrosine |
F339 | 2.05 | 28.38 | glutamine | F357 | 3.01 | 38.65 | tyrosine | F389 | 3.22 | 38.65 | tyrosine |
F186 | 3.53 | 63.50 | threonine, valine | F261 | 2.09 | 28.39 | glutamine | F145 | 2.07 | 28.41 | glutamine |
F596 | 1.76 | 43.04 | leucine | F95 | 1.35 | 22.64 | threonine | F235 | 0.99 | 25.00 | leucine |
F297 | 1.73 | 27.11 | leucine, arginine | F230 | 0.97 | 23.83 | leucine | F637 | 1.66 | 43.03 | leucine |
F198 | 1.94 | 30.71 | arginine | F350 | 1.67 | 27.09 | arginine | F470 | 3.21 | 43.69 | arginine |
F40 | 3.83 | 75.61 | sucrose | F213 | 1.90 | 30.67 | arginine | F622 | 3.82 | 62.76 | sucrose |
F31 | 3.80 | 63.37 | sucrose | F53 | 3.44 | 72.46 | sucrose | F28 | 3.83 | 65.34 | sucrose |
F19 | 3.76 | 75.90 | sucrose | F55 | 3.78 | 63.39 | sucrose | F18 | 3.54 | 74.19 | sucrose |
F5 | 3.81 | 65.32 | sucrose, stachyose, raffinose | F2 | 3.66 | 64.87 | sucrose, stachyose | F12 | 3.78 | 65.34 | sucrose, stachyose, raffinose |
F11 | 5.43 | 95.10 | sucrose, stachyose, raffinose | F6 | 4.18 | 79.84 | sucrose, stachyose, raffinose | F9 | 3.85 | 84.66 | sucrose, stachyose, raffinose |
F23 | 4.95 | 101.65 | stachyose, raffinose | F65 | 5.41 | 95.23 | sucrose, stachyose, raffinose | F20 | 4.12 | 71.75 | stachyose |
F25 | 3.67 | 69.01 | stachyose, raffinose | F22 | 4.02 | 69.01 | stachyose, raffinose | F21 | 4.06 | 74.43 | stachyose, raffinose |
F46 | 3.23 | 77.36 | glucose, glucuronic acid | F56 | 3.71 | 76.20 | sucrose, glucose, glucuronic acid | F51 | 5.21 | 95.42 | glucuronic acid |
F258 | 3.25 | 77.38 | glucose | F116 | 3.81 | 74.47 | glucose | F54 | 3.48 | 74.80 | glucose |
F137 | 3.92 | 63.97 | glucose | F103 | 4.58 | 99.25 | β-glucose | F102 | 3.21 | 77.44 | glucose |
F97 | 4.01 | 66.26 | fructose | F155 | 3.46 | 74.92 | glucose | F57 | 4.03 | 66.26 | fructose |
F328 | 3.70 | 67.08 | fructose | F44 | 3.81 | 70.65 | fructose | F180 | 3.98 | 79.17 | fructose |
F41 | 3.71 | 67.08 | fructose | F74 | 3.70 | 66.25 | fructose | F154 | 4.07 | 85.43 | fructose |
F61 | 3.55 | 65.84 | fructose | F35 | 3.53 | 67.08 | fructose | F321 | 3.51 | 67.08 | fructose |
F48 | 3.68 | 66.24 | fructose | F68 | 4.08 | 78.69 | fructose | F163 | 3.79 | 70.65 | fructose |
F71 | 3.53 | 65.82 | fructose | F692 | 3.02 | 41.93 | 4-aminobutyric acid | F199 | 2.32 | 37.41 | 4-aminobutyric acid |
F175 | 3.01 | 42.37 | 4-aminobutyric acid | F192 | 1.91 | 26.53 | 4-aminobutyric acid | F624 | 2.29 | 32.19 | valine |
F237 | 1.06 | 20.98 | valine | F232 | 1.02 | 19.63 | valine | F223 | 3.92 | 79.83 | |
F260 | 3.24 | 56.82 | F386 | 1.94 | 25.75 | F182 | 3.21 | 56.76 | |||
F546 | 4.05 | 58.63 | F758 | 2.67 | 39.47 | F92 | 4.14 | 61.30 | |||
F34 | 4.62 | 99.40 | F946 | 2.83 | 39.45 | F975 | 3.12 | 44.34 | |||
F791 | 3.81 | 60.65 | F52 | 3.73 | 68.65 | F503 | 3.51 | 70.54 | |||
F473 | 3.86 | 55.36 | F740 | 6.78 | 118.64 | F864 | 2.81 | 39.47 | |||
F292 | 2.50 | 28.39 | F156 | 2.48 | 28.41 | F309 | 2.41 | 36.86 | |||
F872 | 6.67 | 123.63 | F176 | 2.40 | 32.70 | F853 | 6.79 | 119.26 | |||
F90 | 2.38 | 32.74 | F181 | 4.02 | 84.29 | F204 | 2.36 | 32.69 | |||
F285 | 2.46 | 28.39 | F278 | 5.04 | 110.79 | F642 | 3.59 | 49.68 | |||
F708 | 2.11 | 32.99 | F288 | 3.59 | 72.65 | F49 | 3.98 | 68.66 | |||
F36 | 3.85 | 72.91 | F315 | 5.11 | 110.38 | F120 | 3.50 | 74.41 | |||
F487 | 1.30 | 19.68 | F322 | 4.00 | 86.92 | F500 | 3.67 | 56.35 | |||
F502 | 4.18 | 85.47 | F354 | 4.24 | 82.63 | F545 | 2.82 | 37.70 | |||
F505 | 3.82 | 55.44 | F480 | 5.06 | 110.70 | F772 | 3.40 | 58.90 | |||
F636 | 2.04 | 24.55 | F557 | 4.25 | 84.47 | F1400 | 5.43 | 75.92 | |||
F405 | 3.25 | 77.58 | F608 | 3.96 | 80.14 | F1430 | 4.97 | 72.81 | |||
F699 | 2.21 | 32.98 | F616 | 4.10 | 84.97 | F1483 | 5.43 | 74.44 | |||
F447 | 2.75 | 39.54 | F645 | 4.09 | 84.02 | F117 | 3.51 | 60.15 | |||
F43 | 3.59 | 77.32 | F646 | 3.91 | 76.55 | F188 | 3.76 | 74.20 | |||
F723 | 3.42 | 57.74 | F777 | 1.79 | 29.00 | F195 | 3.75 | 89.84 | |||
F66 | 3.80 | 84.11 | F921 | 2.15 | 23.05 | F739 | 6.56 | 138.61 | |||
F72 | 4.00 | 73.00 | F745 | 1.91 | 33.03 | F287 | 4.18 | 77.06 | |||
F89 | 4.08 | 77.78 | F1209 | 5.90 | 128.65 | F457 | 4.06 | 84.65 | |||
F349 | 2.14 | 16.84 | F13 | 4.06 | 77.06 | F29 | 3.83 | 71.78 | |||
F32 | 4.16 | 71.74 | F125 | 4.48 | 84.00 | F206 | 4.06 | 79.80 | |||
F174 | 3.87 | 73.28 | F422 | 3.56 | 63.35 | F750 | 5.43 | 85.29 | |||
F193 | 3.61 | 66.25 | F845 | 4.29 | 84.69 | F684 | 3.36 | 49.67 | |||
F256 | 4.10 | 84.45 | F932 | 4.16 | 85.44 |
2.4. Comparison of SR Processed by SW, HD, and S04
2.4.1. Multivariate Statistical Analysis
2.4.2. Identification of Differential Metabolites of SW, HD, and S04
Feature | δH | δC | Assignment | Feature | δH | δC | Assignment | Feature | δH | δC | Assignment |
---|---|---|---|---|---|---|---|---|---|---|---|
F115 | 2.72 | 46.66 | aucubin | F121 | 3.02 | 49.38 | aucubin | F124 | 5.05 | 99.01 | aucubin |
F135 | 6.32 | 142.93 | aucubin | F128 | 5.81 | 131.75 | aucubin | F179 | 4.33 | 62.72 | aucubin |
F127 | 5.12 | 107.85 | aucubin | F685 | 2.27 | 47.68 | harpagoside | F299 | 5.01 | 108.14 | harpagoside |
F408 | 6.47 | 145.71 | harpagoside | F372 | 6.16 | 96.43 | harpagoside | F346 | 2.95 | 56.61 | harpagoside |
F146 | 1.52 | 24.36 | harpagoside | F625 | 2.06 | 47.68 | harpagoside | F283 | 3.84 | 79.19 | harpagoside |
F712 | 7.66 | 148.31 | harpagoside, cinnamic acid | F225 | 7.63 | 131.07 | harpagoside, cinnamic acid | F773 | 6.55 | 121.43 | harpagoside, cinnamic acid |
F671 | 6.53 | 121.43 | harpagoside, cinnamic acid | F101 | 4.72 | 101.44 | aucubin, harpagoside | F244 | 3.35 | 72.89 | aucubin, harpagoside, glucose |
F138 | 4.82 | 101.35 | 6-O-methyl-catalpol | F207 | 5.08 | 105.85 | 6-O-methyl-catalpol | F194 | 2.60 | 44.63 | 6-O-methyl-catalpol |
F123 | 4.20 | 62.79 | 6-O-methyl-catalpol | F184 | 3.35 | 76.10 | 6-O-methyl-catalpol | F229 | 6.40 | 143.71 | 6-O-methyl-catalpol |
F201 | 5.05 | 97.23 | 6-O-methyl-catalpol | F218 | 2.35 | 38.86 | 6-O-methyl-catalpol | F220 | 3.77 | 61.50 | 6-O-methyl-catalpol |
F42 | 3.45 | 79.22 | aucubin, harpagoside, 6-O-methyl-catalpol, glucose | F333 | 3.39 | 79.72 | aucubin, harpagoside, 6-O-methyl-catalpol, glucose | F111 | 3.48 | 78.90 | aucubin, harpagoside, 6-O-methyl-catalpol |
F108 | 3.37 | 79.39 | aucubin, harpagoside, 6-O-methyl-catalpol | F367 | 3.39 | 78.91 | aucubin, harpagoside, 6-O-methyl-catalpol, glucose | F130 | 3.46 | 79.14 | aucubin, harpagoside, 6-O-methyl-catalpol |
F107 | 3.38 | 79.20 | aucubin, harpagoside, 6-O-methyl-catalpol, glucose | F211 | 3.44 | 79.30 | aucubin, harpagoside, 6-O-methyl-catalpol | F494 | 3.80 | 56.89 | methionine |
F699 | 2.21 | 32.98 | methionine | F349 | 2.14 | 16.84 | methionine | F330 | 2.65 | 32.01 | methionine |
F708 | 2.11 | 32.99 | methionine | F659 | 3.20 | 38.64 | tyrosine | F1423 | 3.00 | 38.63 | tyrosine |
F248 | 3.88 | 59.18 | tyrosine | F103 | 4.58 | 99.25 | β-glucose | F150 | 5.19 | 95.24 | α-glucose |
F102 | 3.21 | 77.44 | glucose | F50 | 3.73 | 57.21 | F1209 | 5.90 | 128.65 | ||
F88 | 3.98 | 72.38 | F667 | 5.14 | 98.33 | F125 | 4.48 | 84.00 | |||
F77 | 3.91 | 74.04 | F1055 | 1.58 | 30.33 | F340 | 1.24 | 17.22 | |||
F117 | 3.51 | 60.15 | F195 | 3.75 | 89.84 | F514 | 1.94 | 24.72 | |||
F622 | 3.82 | 62.76 | F47 | 1.35 | 23.18 | F610 | 2.36 | 37.79 | |||
F723 | 3.42 | 57.74 | F606 | 4.01 | 82.57 | F631 | 4.45 | 105.80 | |||
F988 | 1.93 | 29.66 | F689 | 7.38 | 132.36 | F741 | 3.13 | 51.58 | |||
F717 | 1.95 | 29.64 | F727 | 1.99 | 29.63 | F760 | 3.35 | 76.83 | |||
F761 | 3.26 | 55.60 | F772 | 3.40 | 58.90 | F989 | 4.81 | 101.63 | |||
F777 | 1.79 | 29.00 | F794 | 3.35 | 55.66 | F800 | 4.02 | 66.89 | |||
F182 | 3.21 | 56.76 | F503 | 3.51 | 70.54 | F546 | 4.05 | 58.63 |
3. Materials and Methods
3.1. Chemicals and Reagents
3.2. Samples Collection and Processing
3.2.1. Fresh SR Raw Material Collection
3.2.2. SR Raw Material Samples Processing
3.3. Optimization of Extraction Conditions
3.4. NMR Sample Preparation and Data Acquisition
3.5. NMR Spectrum Processing and Data Preprocessing
3.6. Chemometric Analysis
3.7. Metabolite Identification and Verification
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
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Duan, X.; Zhang, M.; Du, H.; Gu, X.; Bai, C.; Zhang, L.; Chen, K.; Hu, K.; Li, Y. Effect of Different Processing Methods on the Chemical Constituents of Scrophulariae Radix as Revealed by 2D NMR-Based Metabolomics. Molecules 2022, 27, 4687. https://doi.org/10.3390/molecules27154687
Duan X, Zhang M, Du H, Gu X, Bai C, Zhang L, Chen K, Hu K, Li Y. Effect of Different Processing Methods on the Chemical Constituents of Scrophulariae Radix as Revealed by 2D NMR-Based Metabolomics. Molecules. 2022; 27(15):4687. https://doi.org/10.3390/molecules27154687
Chicago/Turabian StyleDuan, **aohui, Mina Zhang, Huan Du, **u Gu, Caihong Bai, Liuqiang Zhang, Kaixian Chen, Kaifeng Hu, and Yiming Li. 2022. "Effect of Different Processing Methods on the Chemical Constituents of Scrophulariae Radix as Revealed by 2D NMR-Based Metabolomics" Molecules 27, no. 15: 4687. https://doi.org/10.3390/molecules27154687