Azadirachta indica leaf extract: Phytochemical composition and antioxidant, antibacterial, and antifungal activities

Year 2025, Volume: 12 Issue: 3, 615 - 629, 04.09.2025

Abadi Abderrezzak Wassima Lakhdari , Salah Neghmouche Nacer , Abderrahmene Dehliz Hafida Khelafi Ridha Ben Salem Moussaoui Younes

https://doi.org/10.21448/ijsm.1511401

Abstract

The chemical components and natural traits of Azadirachta indica A. Juss. leaf extract from arid environments has been studied. LC-MS analysis identified quercetin, oleanolic acid, salicin, and catechin among the many bioactive compounds in the sample. It was observed that at a concentration of 25.01 ± 0.71 µg/mL, DPPH exhibited antioxidant activity with an IC50 value that is higher than that of β-carotene bleaching (22.29 ± 0.66 µg/mL), ABTS assay (45.09 ± 1.23 µg/mL), respectively. Bacillussubtilis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa showed susceptibility to the extract, with MICs ranging from VHC to 30 mg/mL while Alternaria sp., Fusarium solani, and Thielaviopsis paradox represent the fungal strains tested for their sensitivity to this extract. Therefore, Azadirachta indica leaf extract from dry regions exhibits significant effects, potentially explaining its traditional use in medicine and agriculture.

Keywords

Azadirachtaindica , Phytochemical composition , Antioxidant , Antibacterial , Antifungal

Thanks

The authors gratefully acknowledge the Al-Majd Laboratory of El Oued for their valuable scientific support in the completion of this work. Ridha Ben Salem and Younes Moussaoui greatly acknowledge the financial support of the Ministry of Higher Education and Scientific Research of Tunisia.

Azadirachta indica leaf extract: phytochemical composition and antioxidant, antibacterial, and antifungal activities

Abstract

The chemical components and natural traits of Azadirachta indica A. Juss. leaf extract from arid environments has been studied. LC-MS analysis identified quercetin, oleanolic acid, salicin, and catechin among the many bioactive compounds in the sample. It was observed that at a concentration of 25.01 ± 0.71 µg/mL, DPPH exhibited antioxidant activity with an IC50 value that is higher than that of β-carotene bleaching (22.29 ± 0.66 µg/mL), ABTS assay (45.09 ± 1.23 µg/mL), respectively. Bacillussubtilis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa showed susceptibility to the extract, with MICs ranging from VHC to 30 mg/mL while Alternaria sp., Fusarium solani, and Thielaviopsis paradox represent the fungal strains tested for their sensitivity to this extract. Therefore, Azadirachta indica leaf extract from dry regions exhibits significant effects, potentially explaining its traditional use in medicine and agriculture.

Keywords

Azadirachtaindica , Phytochemical composition , Antioxidant , Antibacterial , Antifungal

Thanks

The authors gratefully acknowledge the Al-Majd Laboratory of El Oued for their valuable scientific support in the completion of this work. Ridha Ben Salem and Younes Moussaoui greatly acknowledge the financial support of the Ministry of Higher Education and Scientific Research of Tunisia.

References

• Abid, I., Laghfiri, M., Bouamri, R., Aleya, L., Bourioug, M. (2021). Integrated pest management (IPM) for Ectomyelois ceratoniae on date palm. Current Opinion in Environmental Science & Health, 19, 100219. https://doi.org/10.1016/j.coesh.2020.10.007
• Ahmad, S. (2023). phytochemicals as alternative anthelmintics against poultry parasites: A review. Agrobiological Records, 12, 34–45. https://doi.org/10.47278/journal.abr/2023.015
• Ahmed, M., Marrez, D.A., Abdelmoeen, N.M., Mahmoud, E.A., Ali, M.A., Decsi, K., Tóth, Z. (2023). Studying the Antioxidant and the Antimicrobial Activities of Leaf Successive Extracts Compared to the Green-Chemically Synthesized Silver Nanoparticles and the Crude Aqueous Extract from Azadirachta indica. Processes, 11(6), 1644. https://doi.org/10.3390/pr11061644
• Alsharif, W., Saad, M.M., & Hirt, H. (2020). Desert microbes for boosting sustainable agriculture in extreme environments. Frontiers in Microbiology, 11. https://doi.org/10.3389/fmicb.2020.01666
• Álvarez-Caballero, J.M., & Coy-Barrera, E. (2019). Chemical and Antifungal Variability of Several Accessions of Azadirachta indica A. Juss. from Six Locations Across the Colombian Caribbean Coast: Identification of Antifungal Azadirone Limonoids. Plants, 8(12), 555. https://doi.org/10.3390/plants8120555
• Ariom, T.O., Dimon, E., Nambeye, E., Diouf, N.S., Adelusi, O.O., & Boudalia, S. (2022). Climate-Smart Agriculture in African Countries: A review of strategies and impacts on smallholder farmers. Sustainability, 14(18), 11370. https://doi.org/10.3390/su141811370
• Avasiloaiei, D.I., Calara, M., Brezeanu, P.M., Murariu, O.C., & Brezeanu, C. (2023). On the Future Perspectives of Some Medicinal Plants within Lamiaceae Botanic Family Regarding Their Comprehensive Properties and Resistance against Biotic and Abiotic Stresses. Genes, 14(5), 955. https://doi.org/10.3390/genes14050955
• Batista, F.L.A., Lima, L.M.G., Abrante, I.A., de Araújo, J.I.F., Batista, F.L.A., Abrante, I.A., …, Magalhães, F.E.A. (2018). Antinociceptive activity of ethanolic extract of Azadirachta indica A. Juss (Neem, Meliaceae) fruit through opioid, glutamatergic and acid-sensitive ion pathways in adult zebrafish (Danio rerio). Biomedicine & Pharmacotherapy, 108, 408–416. https://doi.org/10.1016/j.biopha.2018.08.160
• Benkova, M., Soukup, O., & Marek, J. (2020). Antimicrobial susceptibility testing: currently used methods and devices and the near future in clinical practice. Journal of Applied Microbiology, 129(4), 806–822. https://doi.org/10.1111/jam.14704
• Bolaji, N.O., Abolade, N.Y.A., Aduwa, N.S., Isiaka, N.A.B., Durodola, N.O., Adeoye, N.A., …, Siame, N.T. (2024). Potential health and environmental benefits of the identified phytochemicals screening of (Azadirachta indica) neem leaves in Bauchi Metropolis, Bauchi State, Nigeria. GSC Biological and Pharmaceutical Sciences, 26(3), 068–083. https://doi.org/10.30574/gscbps.2024.26.3.0037
• Boudjaber, K., Miri, Y.B., Benabdallah, A., Bennia, N., Hamadi, C., Soumati, B., Djenane, D., & Simal-Gandara, J. (2023). Evaluation of Antifungal and anti-aflatoxin B1 efficacy of some crude extracts of Chamaerops humilis L. against Aspergillus flavus isolated from peanuts (Arachis hypogea L.). Food Control, 152, 109858. https://doi.org/10.1016/j.foodcont.2023.109858
• Boukeloua, A., Leila, S., Bendif, H., Mustafa, A.M., Boga, M., Serralheiro, M.L., …, Garzoli, S. (2024). A comprehensive study on chemical and biological profiles of Algerian Azadirachta indica (A. Juss). Phytochemistry Letters, 61, 89 96. https://doi.org/10.1016/j.phytol.2024.03.017
• Chaudhry, S., Sidhu, & G.P.S. (2021). Climate change regulated abiotic stress mechanisms in plants: a comprehensive review. Plant Cell Reports, 41(1), 1 31. https://doi.org/10.1007/s00299-021-02759-5
• Darkwah, W., Puplampu, J., Biney, E., & Nkoom, M. (2020). High-performance liquid chromatography analysis and antioxidant activities of extract of Azadirachta indica (Neem) leaves. Pharmacognosy Research, 12(1), 29. https://doi.org/10.4103/pr.pr_14_19
• El Aanachi, S., Gali, L., Nacer, S.N., Bensouici, C., Dari, K., & Aassila, H. (2020). Phenolic contents and in vitro investigation of the antioxidant, enzyme inhibitory, photoprotective, and antimicrobial effects of the organic extracts of Pelargonium graveolens growing in Morocco. Biocatalysis and Agricultural Biotechnology, 29, 101819. https://doi.org/10.1016/j.bcab.2020.101819
• El-Chaghaby, G.A., Mohammed, F.S., Rashad, S., Uysal, I., Koçer, O., Lekesiz, Ö., …, Sevindik, M. (2024). Genus Hypericum: General Properties, Chemical Contents and Biological Activities. Egyptian Journal of Botany, 64(1), 1 26. https://dx.doi.org/10.21608/ejbo.2023.217116.2378
• Gupta, S.C., Prasad, S., Tyagi, A.K., Kunnumakkara, A.B., & Aggarwal, B.B. (2017). Neem (Azadirachta indica): An indian traditional panacea with modern molecular basis. Phytomedicine, 34, 14–20. https://doi.org/10.1016/j.phymed.2017.07.001
• Hasnat, A., Moheman, A., Usmani, M.A., Ansari, A., Bhawani, S.A., Tariq, A., & Alotaibi, K.M. (2023). Solvent extraction of natural products. In Elsevier eBooks (pp. 91–110). https://doi.org/10.1016/b978-0-12-823349-8.00003-4
• Hassan, M.U., Chattha, M.U., Khan, I., Chattha, M.B., Barbanti, L., Aamer, M., & Aslam, M.T. (2020). Heat stress in cultivated plants: nature, impact, mechanisms, and mitigation strategies a review. Plant Biosystems, 155(2), 211 234. https://doi.org/10.1080/11263504.2020.1727987
• Humphries, R., Bobenchik, A.M., Hindler, J.A., & Schuetz, A.N. (2021). Overview of changes to the Clinical and Laboratory Standards Institute Performance Standards for Antimicrobial Susceptibility Testing, M100, 31st Edition. Journal of Clinical Microbiology, 59(12). https://doi.org/10.1128/jcm.00213-21
• Hussain, M.I., Farooq, M., Syed, Q.A., Ishaq, A., Al-Ghamdi, A.A., & Hatamleh, A.A. (2021). Botany, nutritional value, phytochemical composition and biological activities of quinoa. Plants, 10(11), 2258. https://doi.org/10.3390/plants10112258
• Islas, J.F., Acosta, E., G-Buentello, Z., Delgado-Gallegos, J.L., Moreno-Treviño, M.G., Escalante, B., & Moreno-Cuevas, J.E. (2020). An overview of Neem (Azadirachta indica) and its potential impact on health. Journal of Functional Foods, 74, 104171. https://doi.org/10.1016/j.jff.2020.104171
• Khattabi, L., Chettoum, A., Hemida, H., Boussebaa, W., Atanassova, M., & Messaoudi, M. (2023). Pirimicarb Induction of Behavioral Disorders and of Neurological and Reproductive Toxicities in Male Rats: Euphoric and Preventive Effects of Ephedra alata Monjauzeana. Pharmaceuticals, 16(3), 402. https://doi.org/10.3390/ph16030402
• Kumar, S., Singh, N., Devi, L.S., Kumar, S., Kamle, M., Kumar, P., & Mukherjee, A. (2022). Neem oil and its nanoemulsion in sustainable food preservation and packaging: Current status and future prospects. Journal of Agriculture and Food Research, 7, 100254. https://doi.org/10.1016/j.jafr.2021.100254
• Kumar, S., Yadav, A., Yadav, M., & Yadav, J.P. (2017). Effect of climate change on phytochemical diversity, total phenolic content and in vitro antioxidant activity of Aloe vera (L.) Burm.f. BMC Research Notes, 10(1). https://doi.org/10.1186/s13104-017-2385-3
• Manach, C., Scalbert, A., Morand, C., Rémésy, C., & Jiménez, L. (2004). Polyphenols: food sources and bioavailability. The American Journal of Clinical Nutrition, 79(5), 727–747. https://doi.org/10.1093/ajcn/79.5.727
• Maury, G.L., Rodríguez, D.M., Hendrix, S., Arranz, J.C.E., Boix, Y.F., Pacheco, A.O., …, Cuypers, A. (2020). Antioxidants in plants: A valorization potential emphasizing the need for the conservation of plant biodiversity in Cuba. Antioxidants, 9(11), 1048. https://doi.org/10.3390/antiox9111048
• Mohammed, F.S., Sevindik, M., Uysal, İ., Çesko, C., & Koraqi, H. (2024). Chemical composition, biological activities, uses, nutritional and mineral contents of Cumin (Cuminum cyminum). Measurement: Food, 100157. https://doi.org/10.1016/j.meafoo.2024.100157
• Monyela, S., Kayoka, P.N., Ngezimana, W., & Nemadodzi, L.E. (2024). Evaluating the metabolomic profile and Anti-Pathogenic properties of cannabis species. Metabolites, 14(5), 253. https://doi.org/10.3390/metabo14050253
• Nacer, S.N., Zobeidi, A., Bensouici, C., Amor, M.L.B., Haouat, A., Louafi, F., …, Elboughdiri, N. (2023). In vitro antioxidant and antibacterial activities of ethanolic extracts from the leaves and stems of Oudneya Africana R. growing in the El Oued (Algeria). Biomass Conversion and Biorefinery 14, 29911–29922. https://doi.org/10.1007/s13399-023-04856-9
• Ngoumen, D.J.N., Mandob, D.E., Ella, F.A., Ambamba, B.D.A., Nanhah, J.V.K., Fonkoua, M., & Ngondi, J.L. (2022). Flavonoid-enrich extract of Autranella congolensis (Sapotaceae) protects against aluminium chloride-mediated Alzheimer’s disease-like oxidative stress in rat through the antioxidant properties. Metabolic Brain Disease, 38(3), 1025–1034. https://doi.org/10.1007/s11011-022-01142-x
• Oke, E., Adeyi, O., Okolo, B., Adeyi, J., Ayanyemi, J., Osoh, K., & Adegoke, T. (2020). Phenolic compound extraction from Nigerian Azadirachta Indica leaves: Response surface and neuro-fuzzy modelling performance evaluation with Cuckoo Search multi-objective optimization. Results in Engineering, 8, 100160. https://doi.org/10.1016/j.rineng.2020.100160
• Olugbami, J.O., Gbadegesin, M.A., & Odunola, O.A. (2015). In vitro free radical scavenging and antioxidant properties of ethanol extract of Terminalia glaucescens. Pharmacognosy Research, 7(1), 49. https://doi.org/10.4103/0974-8490.147200
• Patil, S.S., Deshannavar, U.B., Gadekar-Shinde, S.N., Gadagi, A.H., & Kadapure, S.A. (2023). Optimization studies on batch extraction of phenolic compounds from Azadirachta indica using genetic algorithm and machine learning techniques. Heliyon, 9(11), e21991. https://doi.org/10.1016/j.heliyon.2023.e21991
• Pierce, V.M., & Mathers, A.J. (2021). Setting Antimicrobial Susceptibility Testing Breakpoints: A primer for pediatric infectious diseases specialists on the Clinical and Laboratory Standards Institute approach. Journal of the Pediatric Infectious Diseases Society, 11(2), 73–80. https://doi.org/10.1093/jpids/piab106
• Prabuseenivasan, S., Jayakumar, M., & Ignacimuthu, S. (2006). In vitro antibacterial activity of some plant essential oils. BMC Complementary and Alternative Medicine, 6(1). https://doi.org/10.1186/1472-6882-6-39
• Sarkar, S., Singh, R.P., & Bhattacharya, G. (2021). Exploring the role of Azadirachta indica (neem) and its active compounds in the regulation of biological pathways: an update on molecular approach. 3 Biotech, 11(4), 178. https://doi.org/10.1007/s13205-021-02745-4
• Seleiman, M.F., Al-Suhaibani, N., Ali, N., Akmal, M., Alotaibi, M., Refay, Y., …, Battaglia, M.L. (2021). Drought stress impacts on plants and different approaches to alleviate its adverse effects. Plants, 10(2), 259. https://doi.org/10.3390/plants10020259
• Shikov, A.N., Mikhailovskaya, I.Y., Narkevich, I.A., Flisyuk, E.V., & Pozharitskaya, O.N. (2022). Methods of extraction of medicinal plants. In Elsevier eBooks (pp. 771–796). https://doi.org/10.1016/b978-0-323-85542-6.00029-9
• Singh, V., Roy, M., Garg, N., Kumar, A., Arora, S., & Malik, D.S. (2021). An Insight into the dermatological applications of neem: A review on traditional and modern aspect. Recent Advances in Anti infective Drug Discovery, 16(2), 94 121. https://doi.org/10.2174/2772434416666210604105251
• Sirichaiwetchakoon, K., Lowe, G.M., & Eumkeb, G. (2020). The Free Radical Scavenging and Anti-Isolated Human LDL Oxidation Activities of Pluchea indica (L.) Less. Tea Compared to Green Tea (Camellia sinensis). BioMed Research International, 2020, 1–12. https://doi.org/10.1155/2020/4183643
• Sood, M., Kapoor, D., Kumar, V., Kalia, N., Bhardwaj, R., Sidhu, G.P.S., & Sharma, A. (2021). Mechanisms of plant defense under pathogen stress: A review. Current Protein and Peptide Science/Current Protein & Peptide Science, 22(5), 376 395. https://doi.org/10.2174/1389203722666210125122827
• Telli, A., Chedad, A., & Sadine, S.E. (2022). Ethnobotanical study of medicinal plants used for scorpion sting envenoming treatments in Algerian Septentrional Sahara. International Journal of Environmental Studies, 81(3), 1071 1093. https://doi.org/10.1080/00207233.2022.2055345
• Uysal, İ., Mohammed, F.S., & Sevindik, M. (2024). Genus Capparis: Chemical, nutritional composition and biological activity. Studies in Natural Products Chemistry, 81, 367–386. https://doi.org/10.1016/B978-0-443-15628-1.00009-X
• Vats, S. (2015). Effect of Initial Temperature Treatment on Phytochemicals and Antioxidant Activity of Azadirachta indica A. Juss. Applied Biochemistry and Biotechnology, 178(3), 504–512. https://doi.org/10.1007/s12010-015-1890-x
• Zahedi, S.M., Karimi, M., & Venditti, A. (2019). Plants adapted to arid areas: specialized metabolites. Natural Product Research, 35(19), 3314 3331. https://doi.org/10.1080/14786419.2019.1689500