Bioactivity Profiling and Phytochemical Analysis of Carissa carandas Extracts: Antioxidant, Anti-Inflammatory, and Anti-Urinary Tract Infection Properties
Abstract
:1. Introduction
2. Material and Methods
2.1. Collection of Berries
2.2. Solvents and Reagents
2.3. Animals
2.4. Phytochemicals Characterization
2.5. Antioxidant Activity
2.6. Antimicrobial Activity
2.7. Anti-Inflammatory Activity
2.7.1. In Vitro Assessment
2.7.2. In Vivo Assessment
2.8. Bioassay-Guided Fractionation
2.8.1. Liquid-Liquid Partitioning
2.8.2. Method Optimization for Fractionation
2.8.3. RP-HPLC Sub-Fractionation
2.9. LC-ESI-MS/MS Analysis
2.10. Quantification of Tentatively Identified Compounds Using External Standards
2.11. Statistical Analysis
3. Results
3.1. Phytochemical Characterization and Antioxidant Activity
3.2. Anti-UTI Potential of Sequential Crude Extracts
3.3. Anti-Inflammatory Potential of Sequential Crude Extracts
3.4. Bioassay-Guided Fractionation
3.5. LC-ESI-MS/MS Analysis and HPLC Quantification
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Phytochemicals/Antioxidant Activity | DCM | MeOH | 50% MeOH | Quercetin |
---|---|---|---|---|
TPC (mg GAE/g) | 180 ± 0.134 a | 261 ± 0.813 c | 211 ± 0.163 b | -- |
TFC (mg QE/g) | 0.32 ± 0.731 b | 1.22 ± 0.421 c | 0.21 ± 0.512 a | -- |
TAC (mg/kg) | 0.91 ± 0.612 a | 112 ± 2.12 c | 76.0 ± 0.582 b | -- |
DPPH (% inhibition) | 21.0 ± 0.522 a | 73.0 ± 0.403 c | 59.0 ± 0.122 b | 88.0± 0.112 d |
FRAP (mmol/g) | 10.2 ± 0.214 a | 51.5 ± 1.51 c | 43.3 ± 1.11 b | 59.2± 0.121 d |
Microbe Type | Type of Extracts | Standard Drugs | |||
---|---|---|---|---|---|
DCM | MeOH | 50% MeOH | Ciprofloxacin | Nystatin | |
ZOI at 50 µg/mL (mm) | ZOI at 5 μg/mL (mm) | 100 Units/mL (mm) | |||
Escherichia coli (Gram-negative) | NA | 21.0 ± 0.231 c | 11.0 ± 0.126 a | 15.0 ± 0.251 b | -- |
Klebsiella pneumoniae (Gram-negative) | NA | 26.0 ± 0.112 c | 9.0 ± 0.117 a | 19.0 ± 0.512 b | -- |
Candida albicans | NA | 12.0 ± 0.543 b | 4.0 ± 0.237 a | -- | 21.0 ± 0.548 c |
Candida glabrata | NA | 10.0 ± 1.38 a | NA | -- | 16.0 ± 0.547 b |
In Vitro Assays | Type of Extract | Standard | |||
---|---|---|---|---|---|
DCM | MeOH | 50% MeOH | Diclofenac Sodium (In Vitro) | Indomethacin (In Vivo) | |
% Inhibition at 400 µg/mL | % Inhibition at 400 µg/mL | % Inhibition at 100 mg/kg b.w. | |||
Egg albumin denaturation | 11.0 ± 0.01 ns | 76.0 ± 0.131 *** | 51.0 ± 0.932 ** | 91.0 ± 0.126 **** | -- |
Serum albumin denaturation | 14.0 ± 0.02 ns | 78.0 ± 0.546 *** | 54.0 ± 0.351 ** | 93.0 ± 0.124 **** | -- |
Heat-induced hemolysis | 8.0 ± 0.631 ns | 55.0 ± 0.581 ** | 43.0 ± 0.951 ** | 86.0 ± 0.232 **** | -- |
In vivo study | % inhibition at 400 mg/kg b.w. | ||||
Carrageenan-induced paw edema | 21.0 * | 74.0 **** | 39.0 ** | -- | 79.0 **** |
Formaldehyde-induced paw edema | 14.0 ns | 71.0 **** | 32.0 ** | -- | 73.0 **** |
Assay/Activity | LLP Fraction of Methanol Extract | RP-HPLC Sub-Fractions of LLP Fraction B | Standards | ||||
---|---|---|---|---|---|---|---|
B (50 mg/mL) | CCF4 (50 mg/mL) | CCF7 (50 mg/mL) | Qr (125 µg/mL) | Dfs (400 µg/mL) | Cip (5 μg/mL) | Nys (100 units/mL) | |
DPPH (% inhibition) | 76.1 ± 0.211 c | 65.5 ± 0.821 b | 41.4 ± 1.10 a | 88.4 ± 0.541 d | -- | -- | -- |
FRAP (mmol/g) | 59.2 ± 1.10 c | 43.2 ± 0.551 b | 34.2 ± 1.52 a | 59.4 ± 0.223 c | -- | -- | -- |
Anti-inflammtory activity (% inhibition at 400 µg/mL) | |||||||
Egg albumin denaturation | 82.4 ± 0.541 *** | 69.4 ± 0.359 ** | 57.2 ± 0.312 ** | -- | 91.2 ± 0.124 **** | -- | -- |
Serum albumin denaturation | 85.1 ± 1.81 *** | 52.1 ± 0.431 ** | 48.4 ± 0.223 ** | -- | 93.3 ± 0.115 **** | -- | -- |
Heat-induced hemolysis | 66.2 ± 1.52 ** | 29.4 ± 0.611 * | 37.1 ± 0.212 * | -- | 86.1 ± 0.241 **** | -- | -- |
Antibacterial activity (ZOI mm) 50 µg/mL | |||||||
Escherichia coli | 18.4 ± 1.12 d | 8.1 ± 0.148 b | 6.1 ± 0.259 a | -- | -- | 15.4 ± 0.15 mm c | -- |
Klebsiella pneumoniae | 22.2 ± 1.221 d | 7.4 ± 0.124 b | 4.2 ± 0.521 a | -- | -- | 19.1 ± 0.545 mm c | -- |
Candida albicans | 11.4 ± 1.51 a | NA | NA | -- | -- | -- | 21.4 ± 0.541 mm b |
Candida glabrata | 9.1 ± 0.841 a | NA | NA | -- | -- | -- | 16.1 ± 0.543 mm b |
Tenetaive Compound | Detection Mode | Molecular Weight m/z | Fragments | Reference |
---|---|---|---|---|
Quinic acid | Negative | 191 | 191, 173, 127.1, 85.3 | [36] |
Chlorogenic acid | Negative | 353 | 191, 173.1 | [37] |
Ellagic acid | Negative | 301 | 301, 285, 283.1, 256.9, 229, 178.9 | [38] |
Cyanidin-3-galactoside | Negative | 449 | 286.9 | [39] |
Peonidin-3-arabinoside | Positive | 433 | 283 | [39] |
Delphinidin-3-galactoside | Positive | 465 | 303 | [39] |
Delphinidin-3-rutinoside | Positive | 611 | 302.9 | [39] |
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Saeed, W.; Ismail, T.; Qamar, M.; Esatbeyoglu, T. Bioactivity Profiling and Phytochemical Analysis of Carissa carandas Extracts: Antioxidant, Anti-Inflammatory, and Anti-Urinary Tract Infection Properties. Antioxidants 2024, 13, 1037. https://doi.org/10.3390/antiox13091037
Saeed W, Ismail T, Qamar M, Esatbeyoglu T. Bioactivity Profiling and Phytochemical Analysis of Carissa carandas Extracts: Antioxidant, Anti-Inflammatory, and Anti-Urinary Tract Infection Properties. Antioxidants. 2024; 13(9):1037. https://doi.org/10.3390/antiox13091037
Chicago/Turabian StyleSaeed, Wisha, Tariq Ismail, Muhammad Qamar, and Tuba Esatbeyoglu. 2024. "Bioactivity Profiling and Phytochemical Analysis of Carissa carandas Extracts: Antioxidant, Anti-Inflammatory, and Anti-Urinary Tract Infection Properties" Antioxidants 13, no. 9: 1037. https://doi.org/10.3390/antiox13091037