Polyphenols Investigation and In Vitro Antioxidant Capacity of Romanian Wild-Grown Geranium spp. (Geraniaceae)
Abstract
1. Introduction
2. Results
2.1. ABTS and DPPH IC50
2.2. FRAP Assay
2.3. TPC and TFC Assay
2.4. HPTLC Fingerprinting and Effect-Directed DPPH Assay
2.5. Phenolic Acids Profile (UHPLC/UV–MS Analysis)
2.6. Relationships Between Phenolic Profiles and Antioxidant Capacity
3. Discussion
3.1. Correlation Between TPC, TFC, and Antioxidant Capacity
3.2. Study Limitations
4. Materials and Methods
4.1. Plant Material
4.2. Chemicals and Reagents
4.3. Extraction Procedure
4.4. Standards Preparation
4.5. Antioxidant Capacity Assays
4.5.1. DPPH Antioxidant Assay
4.5.2. ABTS Antioxidant Assay
4.5.3. FRAP Antioxidant Assay
4.6. Total Polyphenols and Flavonoids
4.6.1. TPC Assay
4.6.2. TFC Assay
4.7. HPTLC Fingerprinting for Antioxidant Capacity
4.8. UHPLC Analysis of Phenolic Acids
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ABTS | 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) |
AlCl3 | Aluminum chloride |
ANOVA | Analysis of variance |
DPPH | 2,2-Diphenyl-1-picrylhydrazyl |
FeCl3 | Ferric chloride |
FeSO4·7H2O | Ferrous sulfate heptahydrate |
FRAP | Ferric-reducing antioxidant power |
G1 | Geranium dissectum |
G2 | Geranium lucidum |
G3 | Geranium pussilum |
G4 | Geranium robertianum |
GAE | Gallic acid equivalents |
HCl | Hydrochloric acid |
HPTLC | High-performance thin-layer chromatography |
IC50 | Half-maximal inhibitory concentration |
m/z | Mass-to-charge ratio |
MS | Mass spectrometry |
NP–PEG | Natural products–polyethylene glycol |
PDA | Photodiode array |
QE | Quercetin equivalents |
Rf | Retention factor |
RT | Room temperature |
SD | Standard deviation |
TFC | Total flavonoid content |
TPC | Total phenolic content |
TPTZ | 2,4,6-Tris(2-pyridyl)-1,3,5-triazine |
tR | Retention time |
UHPLC | Ultra-high-performance liquid chromatography |
UV | Ultraviolet |
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Sample | ABTS IC50 (mg/mL) | DPPH IC50 (mg/mL) | FRAP (mM Fe2+) | TPC (mg GAE/g) | TFC (mg QE/g) |
---|---|---|---|---|---|
G1 | 1.283 ± 0.048 ns | 0.664 ± 0.029 ns | 27.727 ± 0.762 **** | 68.170 ± 1.520 **** | 5.462 ± 0.200 ns |
G2 | 0.640 ± 0.023 ns | 0.344 ± 0.014 ns | 38.049 ± 1.484 **** | 82.930 ± 2.670 **** | 4.060 ± 0.090 ns |
G3 | 0.404 ± 0.014 ns | 0.245 ± 0.015 ns | 52.483 ± 1.818 **** | 120.360 ± 2.730 **** | 6.716 ± 0.234 ns |
G4 | 0.321 ± 0.011 ns | 0.170 ± 0.006 ns | 60.492 ± 1.934 **** | 140.140 ± 3.480 **** | 5.967 ± 0.162 ns |
Sample | Caffeic Acid (μg/g) | Chlorogenic Acid (μg/g) | p-Coumaric Acid (μg/g) | Ferulic Acid (μg/g) | Gallic Acid (μg/g) | Protocatechuic Acid (μg/g) | Syringic Acid (μg/g) | Vanillic Acid (μg/g) |
---|---|---|---|---|---|---|---|---|
G1 | 650.928 ± 23.223 **** | 87.821 ± 1.956 ns | 280.226 ± 5.703 ** | 112.034 ± 2.915 ** | 0.000 ± 0.000 **** | 198.007 ± 6.636 **** | 459.751 ± 16.764 **** | 270.588 ± 9.994 **** |
G2 | 518.406 ± 16.950 **** | 62.373 ± 2.463 ns | 129.846 ± 4.912 **** | 71.654 ± 1.787 *** | 91.316 ± 2.243 **** | 401.116 ± 8.381 **** | 162.736 ± 3.340 **** | 183.357 ± 5.725 **** |
G3 | 256.377 ± 6.405 **** | 283.606 ± 8.806 **** | 244.255 ± 6.277 ** | 155.413 ± 5.988 **** | 331.612 ± 8.327 **** | 709.509 ± 19.062 **** | 60.062 ± 1.454 **** | 130.633 ± 3.801 **** |
G4 | 411.969 ± 14.485 **** | 87.959 ± 2.377 ns | 971.760 ± 25.246 **** | 84.219 ± 3.186 *** | 425.105 ± 11.872 **** | 1236.165 ± 40.646 **** | 95.272 ± 3.084 ** | 77.700 ± 2.101 **** |
Sample | Species/Vegetal Product | Date/Collection Site (Southwest Romania Flora; Geographic Coordinates) | Voucher Specimen |
---|---|---|---|
G1 | G. dissectum/herba | 20 April 2024/Stroeşti Commune, Vâlcea County (45°4′39.65″ N, 23°54′34.16″ E) | GER-DIS-2024-0420-2 |
G2 | G. lucidum/herba | 16 April 2024/Băile Herculane City, Caraş Severin County (44°53′54.69″ N, 22°25′46.91″ E) | GER-LUC-2024-0416-1 |
G3 | G. pussilum/herba | 28 April 2024/Cârcea Commune, Dolj County (44°16′29.84″ N, 23°52′32.89″ E) | GER-PUS-2024-0428-1 |
G4 | G. robertianum/herba | 16 April 2024/Băile Herculane City, Caraş Severin County (44°54′26.97″ N, 22°25′55.47″ E) | GER-ROB-2024-0416-2 |
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Bejenaru, C.; Segneanu, A.-E.; Biţă, A.; Bejenaru, L.E.; Hovaneţ, M.-V.; Ciocîlteu, M.V.; Tîrnă, A.C.; Blendea, A.; Mogoşanu, G.D. Polyphenols Investigation and In Vitro Antioxidant Capacity of Romanian Wild-Grown Geranium spp. (Geraniaceae). Plants 2025, 14, 3190. https://doi.org/10.3390/plants14203190
Bejenaru C, Segneanu A-E, Biţă A, Bejenaru LE, Hovaneţ M-V, Ciocîlteu MV, Tîrnă AC, Blendea A, Mogoşanu GD. Polyphenols Investigation and In Vitro Antioxidant Capacity of Romanian Wild-Grown Geranium spp. (Geraniaceae). Plants. 2025; 14(20):3190. https://doi.org/10.3390/plants14203190
Chicago/Turabian StyleBejenaru, Cornelia, Adina-Elena Segneanu, Andrei Biţă, Ludovic Everard Bejenaru, Marilena-Viorica Hovaneţ, Maria Viorica Ciocîlteu, Adriana Cosmina Tîrnă, Antonia Blendea, and George Dan Mogoşanu. 2025. "Polyphenols Investigation and In Vitro Antioxidant Capacity of Romanian Wild-Grown Geranium spp. (Geraniaceae)" Plants 14, no. 20: 3190. https://doi.org/10.3390/plants14203190
APA StyleBejenaru, C., Segneanu, A.-E., Biţă, A., Bejenaru, L. E., Hovaneţ, M.-V., Ciocîlteu, M. V., Tîrnă, A. C., Blendea, A., & Mogoşanu, G. D. (2025). Polyphenols Investigation and In Vitro Antioxidant Capacity of Romanian Wild-Grown Geranium spp. (Geraniaceae). Plants, 14(20), 3190. https://doi.org/10.3390/plants14203190