Phenolic Profiling and Biological Potential of Ficus curtipes Corner Leaves and Stem Bark: 5-Lipoxygenase Inhibition and Interference with NO Levels in LPS-Stimulated RAW 264.7 Macrophages
Abstract
:1. Introduction
2. Materials and Methods
2.1. General Chemicals and Standards
2.2. Plant Material
2.3. Extraction
2.4. Phenolic Profile Characterization
2.4.1. HPLC-DAD-ESI (Ion Trap)/MSn Qualitative Analysis
2.4.2. UPLC-ESI-QTOF-MS Qualitative Analysis
2.4.3. HPLC-DAD Quantitative Analysis
2.5. 5-LOX Inhibition
2.6. RAW 264.7 Macrophages
2.6.1. Cell Culture
2.6.2. Cell Viability
2.6.3. Determination of NO Levels in Culture Medium
2.6.4. Determination of l-Citrulline Levels in Culture Medium
2.7. Determination of •NO Levels in Non-Cellular System
2.8. Statistical Analysis
3. Results
3.1. Characterization of the Phenolic Profile
3.1.1. HPLC-DAD-ESI (Ion Trap)/MSn Qualitative Analysis
3.1.2. HPLC-DAD Quantitative Analysis
3.2. 5-LOX Inhibition
3.3. Effect on RAW 264.7 Macrophages
3.3.1. Interference with NO Levels
3.3.2. Interference with l-Citrulline Levels
3.4. Nitric Oxide Radical Levels in Non-Cellular System
4. Discussion
4.1. Phenolic Profile Characterization
4.2. 5-LOX Inhibition
4.3. Effect on RAW 264.7 Macrophages and ●NO Scavenging Capacity
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Compounds | Rt (min) | Formula (M) | [M − H]− m/z | MS3[M − H]−, m/z (%) | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|
−60 | −90 | −120 | −162 | Agl2 + 113 | Agl2 + 83 | Agl2 + 71 | Agl2 + 41 | ||||
8 | 10.7 | C27H30O15 | 593.1508 | 533 (2) | 503 (30) | 473 (100) | 383 (45) | 353 (80) | |||
10 | 12.0 | C27H30O15 | 593.1506 | 503 (2) | 473 (30) | 431 (60) | 341 (15) | 311 (100) | |||
11 | 12.3 | C26H28O14 | 563.1396 | 503 (15) | 473 (80) | 443 (100) | 383 (65) | 353 (90) | |||
14 | 13.0 | C26H28O14 | 563.1402 | 503 (7) | 473 (85) | 443 (50) | 383 (75) | 353 (100) | |||
16 | 13.6 | C21H20O10 | 431.0980 | 341 (3) | 311 (100) | ||||||
18 | 14.3 | C21H20O10 | 431.0974 | 341 (30) | 311 (100) |
Compounds | Rt (min) | Formula (M) | [M − H]− m/z | MS3[M − H]− m/z (%) |
---|---|---|---|---|
2 | 7.4 | C15H14O6 | 289.0717 | 245(100), 205(50) |
5 | 8.9 | C30H26O12 | 577.1345 | 425(100), 407(80), 289(25) |
6 | 9.4 | C15H14O6 | 289.0714 | 245(100), 205(35) |
7 | 9.6 | C15H14O6 | 289.0714 | 245(100), 205(30) |
9 | 11.0 | C45H38O18 | 865.1988 | 695(100), 577(90), 425(60), 407(60), 287(30) |
12 | 12.4 | C39H32O15 | 739.1651 | 587(100), 569(25), 435(65), 417(40), 339(30), 289(20) |
13 | 12.6 | C39H32O15 | 739.1678 | 587(100), 569(30), 435(50), 417(10), 339(30), 289(15) |
15 | 13.4 | C24H20O9 | 451.1020 | 341(10) |
17 | 14.3 | C30H26O12 | 577.1347 | 425(100), 407(70), 289(30) |
20 | 15.6 | C24H20O9 | 451.1026 | 341(100) |
21 | 18.5 | C24H20O9 | 451.1024 | 341(10) |
Standard | Regression Equation | Linearity Range (µg/mL) | LOD (µg/mL) | LOQ (µg/mL) | ||
---|---|---|---|---|---|---|
Slope (σ) | Intercept (b) | R2 (n = 3) | ||||
3-O-Caffeoylquinic acid | 44.661 | −17.577 | 0.999 | 2–32 | 0.439 | 1.330 |
Catechin | 16.114 | 22.846 | 0.998 | 141–4.4 | 1.040 | 3.152 |
5-O-Caffeoylquinic acid | 157.450 | 100.020 | 0,999 | 7.5–120 | 2.125 | 6.439 |
Epicatechin | 35.932 | 44.752 | 0.998 | 145–4.5 | 0.691 | 2.094 |
Vicenin-2 | 55.527 | −0.958 | 0.999 | 3–48 | 0.606 | 1.837 |
Epigallocatechin | 3.291 | −5.585 | 0.999 | 250–3.9 | 1.125 | 3.408 |
Vitexin | 80.631 | 73.566 | 0.997 | 3–48 | 0.256 | 0.777 |
Isovitexin | 98.771 | 27.029 | 0.998 | 1–16 | 0.013 | 0.040 |
Compound | SB | LV | |
---|---|---|---|
1 | 3-O-Caffeoylquinic acid | 33.79 ± 2.45 | Nd |
2 | Catechin | 10.17 ± 2.64 | Nd |
3 | Chlorogenic acid isomer | 33.78 ± 2.29 | Nd |
4 | 5-O-Caffeoylquinic acid | 201.23 ± 6.88 | Nd |
5 | Procyanidin type B | 52.17 ± 1.10 | Nd |
6 | Catechin/Epicatechin derivative | 129.38 ± 19.16 | Nd |
7 | Epicatechin | 377.51 ± 21.29 | Nd |
8 | Vicenin-2 | 218.23 ± 11.41 | Nd |
9 | Procyanidin type C | 27.44 ± 0.82 | Nd |
10 | Apigenin-7-O-Hex-6/8-C-Hex | 96.99 ± 6.59 | 152.17 ± 2.83 |
11 | Apigenin-6-C-Pt-8-C-Hex | 204.19 ± 9.89 | 381.20 ± 10.58 |
12 | Cinchonain type II | 280.94 ± 56.52 | Nd |
13 | Cinchonain type II | 727.65 ± 67.62 | Nd |
14 | Apigenin-6-C-Hex-8-C-Pent | 8.46 ± 1.13 | 78.27 ± 16.40 |
15 | Cinchonain type I | 293.45 ± 113.90 | Nd |
16 | Vitexin | 72.73 ± 4.29 | Nd |
17 | Procyanidin type B | 8.81 ± 1.69 | Nd |
18 | Isovitexin | 18.08 ± 2.96 | Nd |
19 | Aviculin | 1024.17 ± 81.73 | Nd |
20 | Cinchonain type I | 77.30 ± 11.35 | Nd |
21 | Cinchonain type I | 1478.00 ± 18.67 | Nd |
Total | 5374.15 ± 436.61 | 611.5 ± 29.81 |
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Andrade, C.; Ferreres, F.; Gomes, N.G.M.; Duangsrisai, S.; Srisombat, N.; Vajrodaya, S.; Pereira, D.M.; Gil-Izquierdo, A.; Andrade, P.B.; Valentão, P. Phenolic Profiling and Biological Potential of Ficus curtipes Corner Leaves and Stem Bark: 5-Lipoxygenase Inhibition and Interference with NO Levels in LPS-Stimulated RAW 264.7 Macrophages. Biomolecules 2019, 9, 400. https://doi.org/10.3390/biom9090400
Andrade C, Ferreres F, Gomes NGM, Duangsrisai S, Srisombat N, Vajrodaya S, Pereira DM, Gil-Izquierdo A, Andrade PB, Valentão P. Phenolic Profiling and Biological Potential of Ficus curtipes Corner Leaves and Stem Bark: 5-Lipoxygenase Inhibition and Interference with NO Levels in LPS-Stimulated RAW 264.7 Macrophages. Biomolecules. 2019; 9(9):400. https://doi.org/10.3390/biom9090400
Chicago/Turabian StyleAndrade, Catarina, Federico Ferreres, Nelson G. M. Gomes, Sutsawat Duangsrisai, Nattawut Srisombat, Srunya Vajrodaya, David M. Pereira, Angel Gil-Izquierdo, Paula B. Andrade, and Patrícia Valentão. 2019. "Phenolic Profiling and Biological Potential of Ficus curtipes Corner Leaves and Stem Bark: 5-Lipoxygenase Inhibition and Interference with NO Levels in LPS-Stimulated RAW 264.7 Macrophages" Biomolecules 9, no. 9: 400. https://doi.org/10.3390/biom9090400