LC-ESI-MS/MS Identification of Biologically Active Phenolics in Different Extracts of Alchemilla acutiloba Opiz
Abstract
:1. Introduction
2. Results and Discussion
2.1. Phytochemical Analysis
2.2. Qualitative and Quantitative Analysis
2.3. Antioxidant Activities
2.4. Anti-Inflammatory Activity
2.5. Evaluation of Cytotoxicity
3. Materials and Methods
3.1. Plant Material
3.2. Chemicals and Reagents
3.3. Extraction Procedure
3.4. Total Flavonoid, Phenolic and Phenolic Acids Content
3.5. LC-ESI-MS/MS Analysis
3.6. Antioxidant Activity
3.7. Cyclooxygenase-1 (COX-1) and Cyclooxygenase-2 (COX-2) Inhibitory Activity
3.8. Evaluation of Cytotoxicity
3.9. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Sample | Total Flavonoid Content [mg QE/g DE] | Total Phenolic Content [mg GAE/g DE] | Total Phenolic Acids [mg CAE/g DE] |
---|---|---|---|
AH | 113.79 ± 1.09 | 279.82 ± 1.52 | 39.15 ± 0.18 |
AH-B | 57.09 ± 0.31 | 215.61 ± 2.10 | 72.19 ± 0.39 |
AH-O | 189.25 ± 0.95 | 154.29 ± 1.09 | 11.09 ± 0.01 |
AH-E | 1.57 ± 0.02 | 95.72 ± 0.89 | nd |
AR | 19.85 ± 0.19 | 192.54 ± 0.49 | 6.52 ± 0.02 |
AR-B | 4.63 ± 0.10 | 175.19 ± 1.15 | 9.32 ± 0.03 |
AR-O | 23.18 ± 0.50 | 93.15 ± 0.95 | 2.15 ± 0.10 |
AR-E | 1.92 ± 0.01 | 128.37 ± 1.10 | 5.89 ± 0.05 |
Compound | AH | AR | AH-B | AR-B | AH-O | AR-O | AH-E | AR-E |
---|---|---|---|---|---|---|---|---|
gallic acid | 5.89 ± 0.05 | 2.45 ± 0.10 | <LOQ | <LOQ | <LOQ | nd | 0.00 | 0.00 |
protocatechuic acid | 22.13 ± 0.18 | 0.66 ± 0.02 | 32.88 ± 0.18 | 5.71 ± 0.01 | 19.42 ± 0.12 | 0.80 ± 0.00 | 0.87 ± 0.01 | 2.41 ± 0.03 |
caffeic acid | 23.50 ± 0.18 | 0.00 | 78.88 ± 0.18 | 9.71 ± 0.10 | 9.82 ± 0.02 | <LOQ | 0.28 ± 0.00 | 1.26 ± 0.01 |
syringic acid | 4.98 ± 0.11 | 0.00 | 13.01 ± 0.02 | 1.81 ± 0.13 | 4.40 ± 1.25 | 0.00 | 0.00 | <LOQ |
4-hydroxybenzoic acid | 12.81 ± 0.09 | <LOQ | 70.36 ± 0.18 | 13.04 ± 0.05 | 5.58 ± 0.28 | < LOQ | 1.43 ± 0.02 | 2.16 ± 0.04 |
vanilic acid | 12.56 ± 0.70 | 0.00 | 76.75 ± 0.35 | 9.32 ± 0.05 | <LOQ | 0.00 | 0.00 | 0.00 |
gentisic acid | 0.00 | <LOQ | <LOQ | <LOQ | 3.10 ± 0.05 | 0.24 ± 0.01 | 0.00 | 0.00 |
sinapic acid | 0.00 | 0.00 | <LOQ | <LOQ | 0.00 | 0.00 | 0.00 | 0.00 |
p-coumaric acid | 20.88 ± 0.18 | <LOQ | 187.63 ± 0.18 | 24.79 ± 0.20 | 4.26 ± 0.01 | <LOQ | 1.84 ± 0.01 | 4.47 ± 0.05 |
ferulic acid | 0.00 | 0.00 | 0.00 | 15.93 ± 0.10 | 0.00 | 0.00 | <LOQ | <LOQ |
rosmarinic acid | 7.56 ± 0.52 | 0.14 ± 0.00 | 0.84 ± 0.01 | 0.02 ± 0.00 | 5.87 ± 0.02 | 0.31 ± 0.01 | 0.04 ± 0.00 | 0.05 ± 0.00 |
salicylic acid | 11.59 ± 0.05 | <LOQ | 55.50 ± 0.35 | 6.01 ± 0.02 | 7.52 ± 0.12 | <LOQ | 0.17 ± 0.01 | 0.63 ± 0.00 |
Total of Phenolic Acids | 121.90 | 3.25 | 515.85 | 86.34 | 59.97 | 1.35 | 4.63 | 10.98 |
astragalin | 23.75 ± 0.18 | <LOQ | 7.48 ± 0.11 | 0.33 ± 0.04 | 22.42 ± 0.35 | <LOQ | 0.03 ± 0.00 | 2.25 ± 0.02 |
quercitrin | 294.38 ± 4.12 | 1.19 ± 0.02 | 395.00 ± 3.54 | 57.36 ± 0.10 | 335.83 ± 1.18 | 4.02 ± 0.19 | 10.24 ± 0.12 | 18.58 ± 0.02 |
isoquercitrin | 139.38 ± 0.88 | 1.21 ± 0.03 | 22.59 ± 0.16 | 0.66 ± 0.02 | 147.75 ± 1.06 | 5.86 ± 0.03 | 1.67 ± 0.02 | 4.85 ± 0.02 |
isorhamnetin-3-glucoside | 4.36 ± 0.10 | <LOQ | <LOQ | <LOQ | 2.31±0.01 | <LOQ | <LOQ | <LOQ |
kaempferol-3-rutinoside | 191.88 ± 0.88 | 37.74 ± 0.07 | 7.41 ± 0.16 | 0.63 ± 0.10 | 244.17 ± 3.54 | 53.91 ± 0.04 | 8.35 ± 0.00 | 12.82 ± 0.16 |
rutin | 235.00 ± 3.54 | 83.16 ± 0.00 | 5.05 ± 0.60 | < LOQ | 332.50 ± 1.18 | 97.94 ± 0.42 | 15.99 ± 0.08 | 24.97 ± 0.14 |
narcissoside | 145.00 ± 3.54 | 47.71 ± 0.04 | 5.21 ± 0.12 | 0.58 ± 0.01 | 206.67 ± 4.71 | 61.76 ± 0.00 | 8.05 ± 0.02 | 13.45 ± 0.07 |
hyperoside | 0.00 | 0.00 | 2.47 ± 0.08 | 0.66 ± 0.02 | 0.00 | 0.00 | 0.00 | 1.52 ± 0.01 |
tiliroside | 0.00 | 0.00 | <LOQ | <LOQ | 0.00 | 0.00 | 0.00 | 0.00 |
naringenin-7-glucoside | <LOQ | 0.00 | 0.00 | 0.00 | <LOQ | 0.00 | 0.00 | 0.00 |
Total of Flavonoid Glycosides | 1033.75 | 171.01 | 445.21 | 60.22 | 1291.65 | 223.49 | 44.33 | 78.44 |
luteolin | 0.05 ± 0.00 | 0.00 | 0.51 ± 0.00 | 0.02 ± 0.00 | <LOQ | 0.00 | <LOQ | <LOQ |
kaempferol | 0.00 | 0.00 | 1.23 ± 0.00 | <LOQ | 0.00 | 0.00 | 0.00 | 0.04 ± 0.00 |
3-O-methylquercetin | 0.00 | 0.00 | <LOQ | 0.00 | 0.00 | <LOQ | 0.00 | <LOQ |
naringenin | <LOQ | 0.00 | <LOQ | 0.00 | <LOQ | 0.00 | 0.00 | 0.00 |
eriodictyol | <LOQ | 0.00 | 0.17 ± 0.00 | <LOQ | <LOQ | 0.00 | 0.00 | 0.00 |
quercetin | 0.15 ± 0.00 | <LOQ | 22.38 ± 0.39 | 1.04 ± 0.01 | 0.14 ± 0.00 | <LOQ | 0.00 | 0.00 |
isorhamnetin | <LOQ | 0.00 | 1.89 ± 0.00 | 0.04 ± 0.00 | <LOQ | 0.00 | 0.00 | 0.00 |
rhamnetin | 0.00 | 0.00 | <LOQ | 0.00 | 0.00 | 0.00 | 0.00 | 0.00 |
Total of Flavonoid Aglycones | 0.20 | 0.00 | 26.18 | 1.10 | 0.14 | 0.00 | 0.00 | 0.04 |
AH | AR | AH-B | AR-B | AH-O | AR-O | AH-E | AR-E | AA | Trolox | Na2EDTA*2H2O | |
---|---|---|---|---|---|---|---|---|---|---|---|
DPPH | 18.69 ± 0.04 | 29.87 ± 0.15 | 8.96 ± 0.10 | 12.08 ± 0.18 | 8.83 ± 0.37 | 15.37 ± 0.19 | 41.46 ± 0.32 | 51.42 ± 0.18 | 4.90 ± 0.09 | - | - |
ABTS | 6.17 ± 0.24 | 14.29 ± 0.06 | 1.42 ± 0.18 | 8.78 ± 0.01 | 6.54 ± 0.03 | 10.39 ± 0.15 | 16.28 ± 0.15 | 24.82 ± 0.20 | - | 3.07 ± 0.01 | - |
CHEL | 21.60 ± 0.39 | 25.76 ± 0.03 | 11.43 ± 0.18 | 12.33 ± 0.33 | 18.89 ± 0.94 | 19.30 ± 0.22 | 25.51 ± 0.89 | 44.12 ± 0.24 | - | - | 9.45 ± 0.03 |
COX-1 Inibition [%] ±SD | COX-2 Inhibition [%] ±SD | |||
---|---|---|---|---|
Extract | 50 µg/mL | 100 µg/mL | 50 µg/mL | 100 µg/mL |
AH | 50.17 ± 0.85 | 71.32 ± 2.73 | 43.65 ± 1.36 | 78.52 ± 2.18 |
AR | 31.27 ± 0.76 | 47.80 ± 1.12 | 48.59 ± 2.19 | 54.25 ± 1.23 |
AH-B | 76.82 ± 0.69 | 83.14 ± 1.08 | 79.75 ± 1.29 | 95.10 ± 1.81 |
AR-B | 63.17 ± 1.28 | 78.29 ± 2.25 | 72.64 ± 1.93 | 90.93 ± 2.65 |
AH-O | 52.65 ± 2.19 | 74.50 ± 1.29 | 60.23 ± 0.67 | 80.12 ± 1.73 |
AR-O | 45.73 ± 1.45 | 64.49 ± 2.74 | 58.08 ± 1.79 | 79.15 ± 3.10 |
AH-E | nd | 10.43 ± 0.19 | nd | 28.09 ± 0.13 |
AR-E | nd | 28.71 ± 0.35 | nd | 41.96 ± 1.30 |
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Dos Santos Szewczyk, K.; Pietrzak, W.; Klimek, K.; Grzywa-Celińska, A.; Celiński, R.; Gogacz, M. LC-ESI-MS/MS Identification of Biologically Active Phenolics in Different Extracts of Alchemilla acutiloba Opiz. Molecules 2022, 27, 621. https://doi.org/10.3390/molecules27030621
Dos Santos Szewczyk K, Pietrzak W, Klimek K, Grzywa-Celińska A, Celiński R, Gogacz M. LC-ESI-MS/MS Identification of Biologically Active Phenolics in Different Extracts of Alchemilla acutiloba Opiz. Molecules. 2022; 27(3):621. https://doi.org/10.3390/molecules27030621
Chicago/Turabian StyleDos Santos Szewczyk, Katarzyna, Wioleta Pietrzak, Katarzyna Klimek, Anna Grzywa-Celińska, Rafał Celiński, and Marek Gogacz. 2022. "LC-ESI-MS/MS Identification of Biologically Active Phenolics in Different Extracts of Alchemilla acutiloba Opiz" Molecules 27, no. 3: 621. https://doi.org/10.3390/molecules27030621