Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan
Abstract
:1. Introduction
2. Results
2.1. Phytochemical Analysis
2.2. Antibacterial and Antifungal Activity
2.3. Antiviral Activity
2.4. Anticancer Activity
2.4.1. Cytotoxic Evaluation
2.4.2. Assessment of Cell Morphology
2.4.3. Cell Cycle Analysis
2.4.4. Apoptosis Detection
2.4.5. The Level of Cellular Thiols
3. Discussion
4. Materials and Methods
4.1. Plant Material Collection and Preparation
4.2. Extraction Procedure
4.3. Analysis of the CA Extract by RP-HPLC/PDA
4.4. Analysis of the CA Extract by HPLC/ESI-QTOF-MS
4.5. Determination of Antibacterial and Antifungal Activity
4.6. Determination of Antiviral Activity
4.6.1. Cytotoxicity Assessment
4.6.2. Antiviral Assay
4.6.3. End-Point Dilution Assay for HSV-1 Infectious Titre
4.6.4. Real-Time PCR for HSV-1 Viral Load
4.7. Anticancer Activity
4.7.1. Cell Culturing and Treatment
4.7.2. Assessment of Cell Morphology and Cell Cycle Analysis
4.7.3. Detection of Apoptosis
4.7.4. The Research of Oxidative Stress by the Level of Cellular Thiols
4.7.5. Statistical Analysis
5. Conclusions
6. Patents
Author Contributions
Funding
Conflicts of Interest
Sample Availability
References
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No | Compound | Retention Time (min) | Formula | Molecular Ion [M − H]− | Fragmentation Ions |
---|---|---|---|---|---|
1. | Gluconic acid | 1.917 | C6H12O7 | 195.0512 | 177.0421; 129.0195; 99.0094; 75.0095 |
2. | Malic acid | 2.041 | C4H6O5 | 133.0145 | 115.0043; 71.0148 |
3. | 2-Deoxy-2,3-dehydro-n-acetyl- neuraminic acid | 2.273 | C11H17NO8 | 290.0891 | 200.0561; 170.0454; 128.0352 |
4. | Citric acid | 2.459 | C₆H₈O₇ | 191.0200 | 129.0191; 111.0091; 87.0095 |
5. | DH-Crocusatin F | 4.994 | C10H16O4 | 199.0973 | 155.1072; 137.0969; 125.0977; 111.0821 |
6. | Crocusatin F | 5.864 | C10H14O4 | 197.0840 | 153.0939; 137.0985; 125.0987; 111.0831 |
7. | Kaempferol 7-O-tetrahexoside | 12.109 | C39H50O26 | 933.2560 | 771.1928; 609.1475; 446.0885; 284.0339 |
8. | Kaempferol 3-O-acyltetrahexoside | 12.799 | C43H54O26 | 973.2846 | 771.2038; 609.1481; 446.0870; 284.0340 |
9. | 2,4,4-Trimethyl-3-formyl-6-hydroxy-2,5-cyclohexadien-1-one | 13.029 | C10H12O3 | 179.0722 | 135.0801; 120.0578; 109.0305 |
10. | Kaempferol 7-O-dihexosidee | 16.973 | C27H30O16 | 609.1454 | 447.0840; 285.0351 |
11. | Kaempferol 7-O-trihexoside | 20.847 | C33H40O21 | 771.1996 | 447.0847; 285.0373 |
12. | Quercetin 3-O-dihexoside | 20.997 | C27H30O17 | 625.1438 | 300.0292; 271.0247 |
13. | Kaempferol 3,7-rutinoside, dihexoside | 21.331 | C39H50O25 | 917.2568 | 771.1673; 755.1738; 609.1419; 593.1419; 285.0337; 284.0281 |
14. | Kaempferol 3-O-dihexoside | 22.335 | C27H30O16 | 609.1470 | 284.0335; 255.0297 |
15. | Rutoside | 23.172 | C27H30O16 | 609.1486 | 300.0269; 284.0319; 271.0245 |
16. | Kaempferol 7-O-rutinoside | 24.677 | C27H30O15 | 593.1540 | 285.0372 |
17. | Astragalin | 25.514 | C21H20O11 | 447.0944 | 284.0322; 255.0299; 227.0343; 151.0036 |
18. | Nicotiflorin | 26.267 | C27H30O15 | 593.1479 | 284.0311; 255.0292 |
19. | Carboxyvanilic acid | 27.609 | C9H8O6 | 211.0240 | 167.0353; 123.0454; 108.0227 |
20. | Kaempferol | 34.420 | C15H10O6 | 285.0425 | 229.0476; 185.0605; 135.0077; 109.0288 |
21. | Endocrocin | 35.119 | C16H10O7 | 313.0379 | 241.0506; 225.0565; 213.0555; 201.0576; 197.0604 |
22. | Acacetin | 42.691 | C16H12O5 | 283.0614 | 268.0382; 240.0433; 212.0482 |
Compound | Retention Time (min) | Content (μg/mg Dry Extract) | ||
---|---|---|---|---|
Kaempferol 3-O-tetrahexoside | 2.11 | 14.14 | 0.09 a | 0.6 b |
Kaempferol 3-O-acyltetrahexoside | 2.47 | 80.04 | 0.51 a | 0.6 b |
Kaempferol 7-O-trihexoside | 9.10 | 9.32 | 0.22 a | 2.4 b |
Quercetin 3-O-dihexoside | 9.40 | 6.89 | 0.07 a | 1.0 b |
Kaempferol 3,7–rutinoside, dihexoside | 9.54 | 10.51 | 0.09 a | 0.9 b |
Kaempferol 3-O-dihexoside | 12.77 | 150.76 | 0.16 a | 0.1 b |
Rutoside | 13.50 | 5.12 | 0.04 a | 0.7 b |
Kaempferol 7-O-rutinoside | 13.97 | 4.71 | 0.10 a | 2.2 b |
Astragalin | 14.47 | 26.87 | 0.25 a | 0.9 b |
Nicotiflorin | 15.83 | 30.83 | 0.26 a | 0.9 b |
TOTAL (μg/mg dry extract) | 339.19 |
Bacteria | MIC 1 (mg/mL) | MBC 2 (mg/mL) | MBC/MIC Ratio |
Escherichia coli ATCC 25922 | 20 | >20 | Nd 4 |
Staphylococcus aureus ATCC 25923 | 20 | 20 | 1 |
Staphylococcus aureus ATCC BAA-1707 | 20 | 20 | 1 |
Staphylococcus epidermidis ATCC 12228 | 20 | >20 | Nd |
Bacillus cereus ATCC 10876 | 20 | 20 | 1 |
Cutibacterium acnes ATCC 11827 | 20 | 20 | 1 |
Fungi (Yeasts) | MIC (mg/mL) | MFC 3 (mg/mL) | MFC/MIC Ratio |
Candida albicans ATCC 10231 | 10 | 10 | 1 |
Candida glabrata ATCC 90030 | 10 | 20 | 2 |
Concentration (mg/mL) | Reduction in Infectious Titer (Δlog) 1 | Reduction in Viral Load (Δlog’) 2 | ||
---|---|---|---|---|
CVB3 | HSV-1 | CVB3 | HSV-1 | |
2 | 1.14 ± 0.38 | 1.96 ± 0.28 | Nd 3 | 0.85 ± 0.10 |
1 | 0.55 ± 0.31 | 0.90 ± 0.06 | nd | 0.24 ± 0.07 |
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Allambergenova, Z.; Kasela, M.; Adamczuk, G.; Humeniuk, E.; Iwan, M.; Świątek, Ł.; Boguszewska, A.; Rajtar, B.; Józefczyk, A.; Baj, T.; et al. Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan. Molecules 2022, 27, 3468. https://doi.org/10.3390/molecules27113468
Allambergenova Z, Kasela M, Adamczuk G, Humeniuk E, Iwan M, Świątek Ł, Boguszewska A, Rajtar B, Józefczyk A, Baj T, et al. Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan. Molecules. 2022; 27(11):3468. https://doi.org/10.3390/molecules27113468
Chicago/Turabian StyleAllambergenova, Zoya, Martyna Kasela, Grzegorz Adamczuk, Ewelina Humeniuk, Magdalena Iwan, Łukasz Świątek, Anastazja Boguszewska, Barbara Rajtar, Aleksandra Józefczyk, Tomasz Baj, and et al. 2022. "Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan" Molecules 27, no. 11: 3468. https://doi.org/10.3390/molecules27113468