Phytochemical Analysis by HPLC–HRESI-MS and Anti-Inflammatory Activity of Tabernaemontana catharinensis
Abstract
:1. Introduction
2. Results
2.1. Thin-Layer Chromatography (TLC) Profile of the Hydroethanolic Extract and Organic Fractions of T. catharinensis Leaves
2.2. High-Performance Liquid Chromatography–High-Resolution Electrospray Ionization-Mass Spectrometry (HPLC–HRESI-MS) Analysis of the Hydroethanolic Extract of T. catharinensis Leaves
2.3. Cytotoxicity Assay
2.4. Anti-Inflammatory Effect of the Pretreatment with the Hydroethanolic Extract and the Fractions of T. catharinensis Leaves in the Carrageenan-Induced Paw Edema Model
2.5. Anti-Inflammatory Effect of the Post-Treatment with the Hydroethanolic Extract and the Fractions of T. catharinensis Leaves in the Carrageenan-Induced Paw Edema Model
2.6. Evaluation of the Anti-Inflammatory Effect of the Hydroethanolic Extract and the Fractions of T. catharinensis Leaves in the Zymosan-Induced Air-Pouch Model
2.7. Evaluation of the Effect of the Hydroethanolic Extract and the Fractions of T. catharinensis Leaves on the Levels of Myeloperoxidase and Cytokines
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Extract Preparation
4.3. Thin-Layer Chromatography (TLC) Profile of the Leaves Extract
4.4. High-Performance Liquid Chromatography–High-Resolution Electrospray Ionization-Mass Spectrometry (HPLC–HRESI-MS) Profile of the Leaves Extract
4.5. MTT Assay
4.6. Animals
4.7. Carrageenan-Induced Paw Edema Model
4.8. Zymosan-Induced Air-Pouch Model
4.9. Differential Cell Count in the Exudate
4.10. Determination of Total Proteins
4.11. Quantitative Determination of Myeloperoxidase Levels
4.12. Determination of Cytokines Concentration
4.13. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflict of Interest
References
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Peak | (TR) Retention Time (min) | (MS) Mass Spectrometry (M/Z) | MS2 | UV Max (nm) | Compounds |
---|---|---|---|---|---|
5 | 13.5 | [M − H]—739.17 | 739.17–575.09; 500.15; 473.04; 393.02; 338.92; 284.92; 256.94 | kaempferol-3-robinobioside-7-rhamnoside | |
6 | 13.8 | [M − H]—609.07 | 609.07–564.41; 488.97; 462.96; 408.87; 344.06; 300.95; 270.97; 216.99 | 211, 264 | quercetin-O-rutinoside |
7 | 14.5 | [M – H]—769.12 | 769.12–623.13; 477.09; 357.00; 314.98 | 211, 351 | 7-methylquercetin-3-galactoside-6″-rhamnoside-3′′′-rhamnoside |
8 | 15.0 | [M – H]—769.12 | 769.12–754.11; 701.12; 638.06; 623.12; 605.10; 579.02; 503.00; 400.05; 356.95; 314.94 | 211, 351 | isorhamnetin-3-galactoside-6″-rhamnoside-3′′′-rhamnoside |
12 | 16.8 | [M – H]—593.06 | 593.06–326.98; 298.96; 284.91 | 211, 253 | kaempferol-O-rutinoside |
13 | 16.8 | [M – H]—623.05 | 623.05–595.09; 477.02; 411.93; 356.95; 327.96; 314.95; 299.89; 270.87 | isorhamnetin-3-hexoside-6″-rhamnoside | |
14 | 19.4 | [M – H]—477.00 | 477.00–357; 326.95; 284.95; 254.93 | 249 | isorhamnetin-3-O-glicoside |
15 | 20.1 | [M − H]—477.00 | 477.00–449.05; 356.95; 315.00 | 210, 253 | isorhamnetin-3-O-glicoside |
16 | 20.5 | [M − H]—477.00 | 477.00–356.96; 314.92 | 211, 251 | isorhamnetin-3-O-glicoside |
12 | 22.3 | [M − H]—563.18 | 563.18–360.71 | isoschaftoside |
Groups | Dose (mg/kg) | Cell Migration (×106/mL) | Inhibition (%) |
---|---|---|---|
Saline | - | 31.00 ± 4.000 | - |
Dexamethasone | 2 | 12.80 ± 1.300 *** | 59 |
Hydroethanolic extract | 50 | 28.40 ± 1.965 | 9 |
Hydroethanolic extract | 100 | 11.25 ± 2.780 *** | 64 |
Hydroethanolic extract | 150 | 7.125 ± 2.461 *** | 77 |
EtOAc fraction | 50 | 7.400 ± 2.040 *** | 76 |
BuOH fraction | 50 | 17.75 ± 2.602 ** | 43 |
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Marques, J.I.; Alves, J.S.F.; Torres-Rêgo, M.; Furtado, A.A.; Siqueira, E.M.d.S.; Galinari, E.; Araújo, D.F.d.S.; Guerra, G.C.B.; Azevedo, E.P.d.; Fernandes-Pedrosa, M.D.F.; et al. Phytochemical Analysis by HPLC–HRESI-MS and Anti-Inflammatory Activity of Tabernaemontana catharinensis. Int. J. Mol. Sci. 2018, 19, 636. https://doi.org/10.3390/ijms19020636
Marques JI, Alves JSF, Torres-Rêgo M, Furtado AA, Siqueira EMdS, Galinari E, Araújo DFdS, Guerra GCB, Azevedo EPd, Fernandes-Pedrosa MDF, et al. Phytochemical Analysis by HPLC–HRESI-MS and Anti-Inflammatory Activity of Tabernaemontana catharinensis. International Journal of Molecular Sciences. 2018; 19(2):636. https://doi.org/10.3390/ijms19020636
Chicago/Turabian StyleMarques, José Ivan, Jovelina Samara Ferreira Alves, Manoela Torres-Rêgo, Allanny Alves Furtado, Emerson Michell da Silva Siqueira, Eder Galinari, Daline Fernandes de Souza Araújo, Gerlane Coelho Bernardo Guerra, Eduardo Pereira de Azevedo, Matheus De Freitas Fernandes-Pedrosa, and et al. 2018. "Phytochemical Analysis by HPLC–HRESI-MS and Anti-Inflammatory Activity of Tabernaemontana catharinensis" International Journal of Molecular Sciences 19, no. 2: 636. https://doi.org/10.3390/ijms19020636