In vitro Antifungal Activity of Medicinal Plant Extracts Against Mucorales and Chemical Characterization of Garlic Extract by Gas Chromatography–Mass Spectrometry
Abstract
The medicinal plant exhibits antimicrobial activities and therapeutic properties (antibacterial, antiviral, fungicidal, anti-cardiovascular disease, anticancer, antidiabetic, antihypertensive) over a wide range of microorganisms. This broad- spectrum was designed to evaluate the in vitro antimycotic activity of 5 medicinal plants against four selected fungal strains (Rhizopus sp., Mucor sp., Absidia blakesleeana and Rhizomucor pusillus). In this research, the 20 plant extracts of 5 plant species were prepared using four solvents (petroleum ether, aqueous, ethanol and methanol) of required concentration by following maceration process. The antifungal activity of plants was determined by performing agar well diffusion method. Petroleum ether extract of Allium sativum showed more activity with zone of inhibition against Rhizopus sp. (15mm), Mucor sp. (25mm), Absidia blakesleeana (18mm) and Rhizomucor pusillus (19mm) as compared to other plants. Qualitative Phytochemicals analysis was screened of most potent antifungal plant, Allium sativum. Garlic petroleum ether solvent showed the presence of alkaloids, flavonoids, saponins and phenols. The minimum inhibitory concentration of most promising agent Allium sativum was examined by two-fold macro-dilution agar plate method against four fungal strains (Rhizopus sp. 0.67 mg/ml, Mucor sp. 10 mg/ml, Absidia blakesleeana1.25 mg/ml, and Rhizomucor pusillus 1.25 mg/ml).GC-MS analysis of petroleum ether extracts of Allium sativum (garlic) revealed various sulphur-containing compounds, including diallyl disulfide, diallyl trisulfide, and allyl methyl trisulfide, which are responsible for its antifungal activity. The major bioactive compounds present in the GC-MS analysis carried on petroleum ether extract of Allium sativum were 3-Vinyl-1,2- dithiacyclohex-5-ene (97.37% probability), 3-Vinyl-1,2- dithiacyclohex-5-ene (dithiins) (92.94%), 3-Vinyl-1,2- dithiacyclohex-5-ene (92.46%),1-Hexanone, 5-methyl-1- phenyl (86.8%), 3-Vinyl-1,2- dithiacyclohex-5-ene (86.20%). This work suggests that, following appropriate in vivo research, garlic extracts may be utilized as an antibiotic substitute.
Keywords:
Antimycotic, Gas Chromatography–Mass Spectrometry (GC–MS), Mucormycosis, Medicinal Plants, Phytochemical Screening
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References
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Ashraf, S. A., Khan, M. A., Awadelkareem, A. M., Tajuddin, S., Ahmad, M. F., & Hussain, T. (2019). GC-MS analysis of commercially available Allium sativum and Trigonella foenum-graecum essential oils and their antimicrobial activities. J Pure Appl Microbiol, 13(4), 2545-2552. https://doi.org/10.22207/JPAM.13.4.69
Barbu, I. A., Ciorîță, A., Carpa, R., Moț, A. C., Butiuc-Keul, A., &Pârvu, M. (2023). Phytochemical characterization and antimicrobial activity of several Allium extracts. Molecules, 28(10), 3980. https://doi.org/10.3390/molecules28103980
Batiha, G. E., Beshbishy, A. M., G. Wasef, L. G., Elewa, Y. H. A., Al-Sagan, A. A., El-Hack, M. E. A., .....& Devkota, H. P. (2020). Chemical constituents and pharmacological activities of garlic (Allium sativum L.): A review. Nutrients, 12(3), 872. https://doi.org/10.3390/nu12030872
Benayache, S., Demirtas, I., Altun, M., Gül, F., Sarri, F. B., Benayache, S., Mekkiou, R. (2017). HPLC analysis, anti-oxidant activity of Genista ferox and its anti-proliferative effect in HeLa cell line. Bangladesh Journal of Pharmacology, 12, 260-267. https://doi.org/10.3329/bjp.v12i3.32310
Brunet, K., &Rammaert, B. (2020). Mucormycosis treatment: Recommendations, latest advances, and perspectives. Journal de Mycologie Medicale, 30(3), 101007. https://doi.org/10.1016/j.mycmed.2020.101007
Chen, C., Liu, C. H., Cai, J., Zhang, W., Qi, W. L., Wang, Z., ... & Yang, Y. (2018). Broad-spectrum antimicrobial activity, chemical composition and mechanism of action of garlic (Allium sativum) extracts. Food Control, 86, 117-125. https://doi.org/10.1016/j.foodcont.2017.11.015
Choudhary, S., Noor, M. U., Hussain, M. S., Mishra, M., & Tyagi, S. (2022). Pharmacological properties and phytoconstituents of garlic (Allium sativum L.): A review. Biological Sciences, 2(4), 338-346. https://doi.org/10.55006/biolsciences.2022.2402
Corbu, V. M., Gheorghe, I., Marinaș, I. C., Geană, E. I., Moza, M. I., Csutak, O., &Chifiriuc, M. C. (2021). Demonstration of Allium sativum extract inhibitory effect on biodeteriogenic microbial strain growth, biofilm development, and enzymatic and organic acid production. Molecules, 26(23), 7195. https://doi.org/10.3390/molecules26237195
Elsharawy, N. T., Baghdadi, A. M., Saddiq, A. A., & Naidu, R. (2024). Exploring antifungal activities of Tamarixaphylla and Zygophyllum album extracts against human mucormycosis. International Journal of Biosciences, 24(5), 203–210. http://dx.doi.org/10.12692/ijb/24.5.203-210
Faqer, O. E., Rais, S., Ouadghiri, Z., Faqer, A. E., Benchama, Z., Ouaddari, A. E., ... &Mtairag, E. M. (2024). Physicochemical properties, GC–MS profiling, and antibacterial potential of Allium sativum essential oil: In vitro and in silico approaches. Scientific African, 26, e02484. https://doi.org/10.1016/j.sciaf.2024.e02484
Faway, E., Staerck, C., Danzelle, C., Vroomen, S., Courtain, C., Mignon, B., &Poumay, Y. (2021). Towards a standardized procedure for the production of infective spores to study the pathogenesis of dermatophytosis. Journal of Fungi, 7(12), 1029. https://doi.org/10.3390/jof7121029
Fufa, B. K. (2019). Anti-bacterial and anti-fungal properties of garlic extract (Allium sativum): A review. Microbiology Research Journal International, 28(3), 1-5. https://hal.science/hal-05368481/
Ghajiri, H., Amal, A. G., Saadon, H., & Al-Mankhee, A. A. (2023). Isolating and Identifying Some Types of Bacteria and Fungi from wheat grains and studying the effect of some types of antibiotics and the aqueous and alcoholic extract of the Alium sativum plant on them. University of Thi-Qar Journal of Science, 10(2), 211-215. https://doi.org/10.32792/utq/utjsci/v10i2.1144
Hashem, A. H., Shehabeldine, A. M., Abdelaziz, A. M., Amin, B. H., & Sharaf, M. H. (2022). Antifungal activity of endophytic Aspergillus terreus extract against some fungi causing mucormycosis: ultrastructural study. Applied Biochemistry and Biotechnology, 194(8), 3468-3482. https://doi.org/10.1007/s12010-022-03876-x
Hu, B., Hu, H., Peng, D., Wei, Z., Wang, Q., & Kuang, H. (2024). Phytochemical components analysis and hypolipidemic effect on hyperlipidemia mice of the aerial parts from Allium sativum. Frontiers in Nutrition, 11, 1422857. https://doi.org/10.3389/fnut.2024.1422857
Hussain, M. K., Ahmed, S., Khan, A., Siddiqui, A. J., Khatoon, S., & Jahan, S. (2023). Mucormycosis: A hidden mystery of fungal infection, possible diagnosis, treatment and development of new therapeutic agents. European Journal of Medicinal Chemistry, 246, 115010. https://doi.org/10.1016/j.ejmech.2022.115010
Jangid, R., & Begum, T. (2022). Antimycotic activity of some medicinal plants against Mucor circinelloides. BioMed Research International, 2022(1), 3523920. https://doi.org/10.1155/2022/3523920
Kobacy, W., Gong, X., Elshamandy, N. K., Zahran, H. A., Osman, A. I., & Rashwan, A. K. (2025). ATR-FTIR, DSC, GC–MS of garlic (Allium sativum), parsley (Petroselinum crispum), and thyme (Thymus vulgaris) essential oils and their synergistic effects on antioxidant and antimicrobial properties. Food Bioscience, 71, 107262. https://doi.org/10.1016/j.fbio.2025.107262
Kumar, A., Gupta, P. K., & Srivastava, A. (2020). A review of modern technologies for tackling COVID-19 pandemic. Diabetes & Metabolic Syndrome: Clinical Research & Reviews, 14(4), 569-573. https://doi.org/10.1016/j.dsx.2020.05.008
Kumar, S., Aharwal, R. P., Shukla, H., Rajak, R. C., & Sandhu, S. S. (2014). Endophytic fungi: As a source of antimicrobials bioactive compounds. World Journal of Pharmacy and Pharmaceutical Sciences, 3(2),1179-1197. https://www.wjpps.com/Wjpps_controller/abstract_id/810
Kutawa, A. B., Danladi, M. D., & Haruna, A. (2018). Regular article antifungal activity of garlic (Allium sativum) extract on some selected fungi. Journal of Medicinal Herbs and Ethnomedicine, 4, 12-14. https://doi.org/10.25081/JMHE.2018.V4.3383
Lata, K., Priyanka, Sen, I., Azmi, W. (2023). Selected Medicinal Plants for Treatment of Mucormycosis, in India. In: Máthé, Á., Khan, I.A. (eds) Medicinal and Aromatic Plants of India Vol. 2. Medicinal and Aromatic Plants of the World, vol 9. Cham: Springer International Publishing.https://doi.org/10.1007/978-3-031-44914-7_12
Mane, D., &Wadikar, B. (2025). The study was aimed to determine the antifungal activity of Allium sativum (garlic) on some selected fungi. World Journal of Pharmaceutical Research, 14(9), 1049-1059. https://wjpr.s3.ap-south-1.amazonaws.com/article_issue/61c073a77187dbfb9ef269fe508f9560.pdf
Mohsen, A. H., Mohsen, I. H., Najm, A. B., &Sgheer, S. K. (2024). Study of the Effectiveness of Extracts of Black Pepper Fruits (Piper nigrum) on the Growth of Fungi Isolated from Some Food Products. European Journal of Medical and Health Research, 2(4), 240-247. https://doi.org/10.59324/ejmhr.2024.2(4).31
Nimoshini, G., Nataraj, P., Rameshkumar, A., Krishnan, R., & Savithri, S. (2023). Inhibitory effect of garlic and omam extracts on Mucor circinelloides, a fungus causing mucormycosis: An: in vitro: study. Journal of Oral and Maxillofacial Pathology, 27(1), 71-75. https://doi.org/10.4103/jomfp.jomfp_45_22
Panda, N., Panigrahi, S. P., Gupta, M. K., Kumari, A., Mehta, D., Goswami, T. D., ... & Gangopadhyay, A. (2024). Phytochemical screening and pharmacological study of antidiabetic potential and bioactive compounds present in Allium sativum. Journal of Chemical Health Risks, 14, 2646-2654. https://www.jchr.org/index.php/JCHR/article/view/3098/2123
Pârvu, M., Moţ, C. A., Pârvu, A. E., Mircea, C., Stoeber, L., Roşca-Casian, O., &Ţigu, A. B. (2019). Allium sativum extract chemical composition, antioxidant activity and antifungal effect against Meyerozymaguilliermondii and Rhodotorulamucilaginosa causing onychomycosis. Molecules, 24(21), 3958. https://doi.org/10.3390/molecules24213958
Patel, M. K., Pandey, S., Kumar, M., Haque, M. I., Pal, S., & Yadav, N. S. (2021). Plants metabolome study: Emerging tools and techniques. Plants, 10(11), 2409. https://doi.org/10.3390/plants10112409
Rahmoune, I., Karoune, S., Azzam, C., Saad, S., Foughalia, A., Sarri, M., Chebrouk, F., Abidat, H. &Kechebarr, A. S. M. (2024). In vitro antioxidant, antimicrobial and antidiabetic properties of the organic fraction of distillate of Salviahispanica seeds. Revista Agraria Academica, 7(5), 19-33.https://doi.org/10.32406/v7n5/2024/19-33/agrariacad
Sarkar, T., Bharadwaj, K. K., Salauddin, M., Pati, S., & Chakraborty, R. (2022). Phytochemical characterization, antioxidant, anti-inflammatory, anti-diabetic properties, molecular docking, pharmacokinetic profiling, and network pharmacology analysis of the major phytoconstituents of raw and differently dried Mangifera indica (Himsagar cultivar): An in vitro and in silico investigations. Applied Biochemistry and Biotechnology, 194(2), 950-987. https://doi.org/10.1007/s12010-021-03669-8
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Jain, M., Pundir, R. and Yogi, R. (2026) “In vitro Antifungal Activity of Medicinal Plant Extracts Against Mucorales and Chemical Characterization of Garlic Extract by Gas Chromatography–Mass Spectrometry”, International Journal of Advancement in Life Sciences Research, 9(2), pp. 207-219. doi: https://doi.org/10.31632/ijalsr.2026.v09i02.016.
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