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Chemical composition and antibacterial activity of Azadirachta indica seed oil from Chad

Year 2025, Volume: 12 Issue: 2, 429 - 447

Abstract

This study focuses on extracting oil from the seeds of the Azadirachta indica species A. Juss, known in Chad as mim or neem, utilizing a cold-press extraction method. It aims to characterize the oil's chemical composition through Gas Chromatography-Mass Spectrometry (GC-MS) analysis and evaluate its antibacterial efficacy using the well diffusion and microdilution techniques. The antibacterial potential was assessed against four food-borne pathogens: Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Salmonella sp., and a soil bacterium, Bacillus subtilis. The results from the GC-MS analysis indicated a predominant presence of fatty alcohols, notably Stigmasta-3,5-diene (49.00%), a steroid alcohol, and Tetradecen-11-yn-1-ol (35.37%), a long-chain fatty alcohol. Additionally, lesser quantities of compounds such as (E, E)-7,11,15-Trimethyl-3-methylene-hexadeca-1,6,10,14-tetraene (a diterpene) at 9.68%, Squalene (a triterpene) at 1.77%, and 2,4,4-Trimethyl-3-hydroxymethyl-5a-(3-methyl-but-2-enyl)-cyclohexene (a sesquiterpene) at 4.19% were identified. The analysis showed that neem oil is rich in fatty alcohols and phytosterols, with lower terpenes and phenolic compounds. It showed no significant antibacterial activity against the tested bacteria. This suggests that from cold-press extraction, neem oil may not effectively combat food-borne pathogens and soil bacteria due to its fatty alcohol and phytosterol content, along with the bacteria's resistance. Increasing the concentration of crude oil in the antibacterial test could lead to positive results. The findings indicate a need for further research to isolate stronger antibacterial molecules in neem oil by separating its components, focusing on extraction methods and solvent polarity.

Supporting Institution

Abdelmalek Essaadi University

Thanks

The authors would like to thank the team of Techno-Centre for biochemical and physical analysis in the Faculty of Sciences -Tetouan- for their assistance and help in GC-MS analysis. Our thanks also go to Professor Souhail Badreddine (Laboratory of Water, Environmental Studies and Analysis, Department of Chemistry, Faculty of Sciences).

References

  • ‘Izzah Ibrahim, N., Fairus, S., Zulfarina, M.S., & Mohamed, I.N. (2020). The efficacy of squalene in cardiovascular disease risk‐a systematic review. Nutrients, 12(2), 1–42. https://doi.org/10.3390/nu12020414
  • Abbas, G., Ali, M., Hamaed, A., Al-Sibani, M., Hussain, H., & Al-Harrasi, A. (2020). Azadirachta indica: The medicinal properties of the global problems-solving tree. In Biodiversity and Biomedicine: Our Future. INC, ScienceDirect. https://doi.org/10.1016/B978-0-12-819541-3.00017-7
  • Adedoyin B.J, Okeniyi S.O,G.S., & Salihu L. (2013). Cytotoxicity, antioxidant and antimicrobial activities of essential oil extracted from Euphorbia Heterophylla plant. In Topclass Journal of Herbal Medicine, 2(5),84–89. http://www.topclassglobaljournals.org
  • Alzohairy, M.A. (2016). Therapeutics role of Azadirachta indica (Neem) and their active constituents in diseases prevention and treatment. Evidence-Based Complementary and Alternative Medicine, 2016(Article ID 7382506), 11. https://doi.org/10.1155/2016/7382506
  • Arora, N., Koul, A., & Bansal, M.P. (2011). Chemopreventive activity of Azadirachta indica on two-stage skin carcinogenesis in murine model. Phytotherapy Research, 25(3), 408–416. https://doi.org/10.1002/ptr.3280
  • Arulkumar R, Karthikan S, Gopalasatheeskumar K, & Arulkumaran G. (2019). Neem (Azadirachta Indica): A miraculous medicinal plant from India. International Journal of Universal Pharmacy and Bio Sciences, 8(4), 48–59.
  • Babatunde, D.E., Otusemade, G.O., Efeovbokhan, V.E., Ojewumi, M.E., Bolade, O.P., & Owoeye, T.F. (2019). Chemical composition of steam and solvent crude oil extracts from Azadirachta indica leaves. Chemical Data Collections, 20, 100208. https://doi.org/10.1016/j.cdc.2019.100208
  • Balasubramanian, S., Ganesh, D., & Surya Narayana, V.V.S. (2014). GC-MS analysis of phytocomponents in the methanolic extract of Azadirachta indica (Neem). International Journal of Pharma and Bio Sciences, 5(4), P258–P262.
  • Belmehdi, O., Bouyahya, A., Jekő, J., Cziáky, Z., Zengin, G., Sotkó, G., El baaboua, A., Senhaji, N. S., & Abrini, J. (2021). Synergistic interaction between propolis extract, essential oils, and antibiotics against Staphylococcus epidermidis and methicillin resistant Staphylococcus aureus. International Journal of Secondary Metabolite, 8(3), 195–213. https://doi.org/10.21448/IJSM.947033
  • Blum, F.C., Singh, J., & Merrell, D.S. (2019). In vitro activity of Neem (Azadirachta indica) oil extract against Helicobacter pylori. Journal of Ethnopharmacology, 232, 236–243. https://doi.org/10.1016/j.jep.2018.12.025
  • Bouhdid, S., Abrini, J., Amensour, M., Zhiri, A., Espuny, M.J., & Manresa, A. (2010). Functional and ultrastructural changes in Pseudomonas aeruginosa and Staphylococcus aureus cells induced by Cinnamomum verum essential oil. Journal of Applied Microbiology, 109(4), 1139–1149. https://doi.org/10.1111/j.1365-2672.2010.04740.x
  • Çakaloğlu, B., Özyurt, V.H., & Ötleş, S. (2018). Cold press in oil extraction. A review. Ukrainian Food Journal, 7(4), 640–654. https://doi.org/10.24263/2304-974x-2018-7-4-9
  • Cesa, S., Sisto, F., Zengin, G., Scaccabarozzi, D., Kokolakis, A.K., Scaltrito, M.M., … Basilico, N. (2019). Phytochemical analyses and pharmacological screening of Neem oil. South African Journal of Botany, 120, 331–337. https://doi.org/10.1016/j.sajb.2018.10.019
  • Choupanian, M., Omar, D., Basri, M., & Asib, N. (2017). Preparation and characterization of Neem oil nanoemulsion formulations against Sitophilus oryzae and Tribolium castaneum adults. Journal of Pesticide Science, 42(4), 158–165. https://doi.org/10.1584/jpestics.D17-032
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  • da Silva, R.S., Martins, L.P., Araujo, R. da C., de Sousa, S., & dos Santos, A.F. (2020). The use of Neem oil and chitosan during pre-harvest and in the post-harvest quality of the “Paluma” guava. Revista Ciencia Agronomica, 51(3), 1–10. https://doi.org/10.5935/1806-6690.20200057
  • Das, P., Sharma, N., Puzari, A., Kakati, D.K., & Devi, N. (2021). Synthesis and characterization of Neem (Azadirachta indica) seed oil-based alkyd resins for efficient anticorrosive coating application. Polymer Bulletin, 78(1), 457–479. https://doi.org/10.1007/s00289-020-03120-8
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Chemical composition and antibacterial activity of Azadirachta indica seed oil from Chad

Year 2025, Volume: 12 Issue: 2, 429 - 447

Abstract

This study focuses on extracting oil from the seeds of the Azadirachta indica species A. Juss, known in Chad as mim or neem, utilizing a cold-press extraction method. It aims to characterize the oil's chemical composition through Gas Chromatography-Mass Spectrometry (GC-MS) analysis and evaluate its antibacterial efficacy using the well diffusion and microdilution techniques. The antibacterial potential was assessed against four food-borne pathogens: Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Salmonella sp., and a soil bacterium, Bacillus subtilis. The results from the GC-MS analysis indicated a predominant presence of fatty alcohols, notably Stigmasta-3,5-diene (49.00%), a steroid alcohol, and Tetradecen-11-yn-1-ol (35.37%), a long-chain fatty alcohol. Additionally, lesser quantities of compounds such as (E, E)-7,11,15-Trimethyl-3-methylene-hexadeca-1,6,10,14-tetraene (a diterpene) at 9.68%, Squalene (a triterpene) at 1.77%, and 2,4,4-Trimethyl-3-hydroxymethyl-5a-(3-methyl-but-2-enyl)-cyclohexene (a sesquiterpene) at 4.19% were identified. The analysis showed that neem oil is rich in fatty alcohols and phytosterols, with lower terpenes and phenolic compounds. It showed no significant antibacterial activity against the tested bacteria. This suggests that from cold-press extraction, neem oil may not effectively combat food-borne pathogens and soil bacteria due to its fatty alcohol and phytosterol content, along with the bacteria's resistance. Increasing the concentration of crude oil in the antibacterial test could lead to positive results. The findings indicate a need for further research to isolate stronger antibacterial molecules in neem oil by separating its components, focusing on extraction methods and solvent polarity.

Supporting Institution

Abdelmalek Essaadi University

Thanks

The authors would like to thank the team of Techno-Centre for biochemical and physical analysis in the Faculty of Sciences -Tetouan- for their assistance and help in GC-MS analysis. Our thanks also go to Professor Souhail Badreddine (Laboratory of Water, Environmental Studies and Analysis, Department of Chemistry, Faculty of Sciences).

References

  • ‘Izzah Ibrahim, N., Fairus, S., Zulfarina, M.S., & Mohamed, I.N. (2020). The efficacy of squalene in cardiovascular disease risk‐a systematic review. Nutrients, 12(2), 1–42. https://doi.org/10.3390/nu12020414
  • Abbas, G., Ali, M., Hamaed, A., Al-Sibani, M., Hussain, H., & Al-Harrasi, A. (2020). Azadirachta indica: The medicinal properties of the global problems-solving tree. In Biodiversity and Biomedicine: Our Future. INC, ScienceDirect. https://doi.org/10.1016/B978-0-12-819541-3.00017-7
  • Adedoyin B.J, Okeniyi S.O,G.S., & Salihu L. (2013). Cytotoxicity, antioxidant and antimicrobial activities of essential oil extracted from Euphorbia Heterophylla plant. In Topclass Journal of Herbal Medicine, 2(5),84–89. http://www.topclassglobaljournals.org
  • Alzohairy, M.A. (2016). Therapeutics role of Azadirachta indica (Neem) and their active constituents in diseases prevention and treatment. Evidence-Based Complementary and Alternative Medicine, 2016(Article ID 7382506), 11. https://doi.org/10.1155/2016/7382506
  • Arora, N., Koul, A., & Bansal, M.P. (2011). Chemopreventive activity of Azadirachta indica on two-stage skin carcinogenesis in murine model. Phytotherapy Research, 25(3), 408–416. https://doi.org/10.1002/ptr.3280
  • Arulkumar R, Karthikan S, Gopalasatheeskumar K, & Arulkumaran G. (2019). Neem (Azadirachta Indica): A miraculous medicinal plant from India. International Journal of Universal Pharmacy and Bio Sciences, 8(4), 48–59.
  • Babatunde, D.E., Otusemade, G.O., Efeovbokhan, V.E., Ojewumi, M.E., Bolade, O.P., & Owoeye, T.F. (2019). Chemical composition of steam and solvent crude oil extracts from Azadirachta indica leaves. Chemical Data Collections, 20, 100208. https://doi.org/10.1016/j.cdc.2019.100208
  • Balasubramanian, S., Ganesh, D., & Surya Narayana, V.V.S. (2014). GC-MS analysis of phytocomponents in the methanolic extract of Azadirachta indica (Neem). International Journal of Pharma and Bio Sciences, 5(4), P258–P262.
  • Belmehdi, O., Bouyahya, A., Jekő, J., Cziáky, Z., Zengin, G., Sotkó, G., El baaboua, A., Senhaji, N. S., & Abrini, J. (2021). Synergistic interaction between propolis extract, essential oils, and antibiotics against Staphylococcus epidermidis and methicillin resistant Staphylococcus aureus. International Journal of Secondary Metabolite, 8(3), 195–213. https://doi.org/10.21448/IJSM.947033
  • Blum, F.C., Singh, J., & Merrell, D.S. (2019). In vitro activity of Neem (Azadirachta indica) oil extract against Helicobacter pylori. Journal of Ethnopharmacology, 232, 236–243. https://doi.org/10.1016/j.jep.2018.12.025
  • Bouhdid, S., Abrini, J., Amensour, M., Zhiri, A., Espuny, M.J., & Manresa, A. (2010). Functional and ultrastructural changes in Pseudomonas aeruginosa and Staphylococcus aureus cells induced by Cinnamomum verum essential oil. Journal of Applied Microbiology, 109(4), 1139–1149. https://doi.org/10.1111/j.1365-2672.2010.04740.x
  • Çakaloğlu, B., Özyurt, V.H., & Ötleş, S. (2018). Cold press in oil extraction. A review. Ukrainian Food Journal, 7(4), 640–654. https://doi.org/10.24263/2304-974x-2018-7-4-9
  • Cesa, S., Sisto, F., Zengin, G., Scaccabarozzi, D., Kokolakis, A.K., Scaltrito, M.M., … Basilico, N. (2019). Phytochemical analyses and pharmacological screening of Neem oil. South African Journal of Botany, 120, 331–337. https://doi.org/10.1016/j.sajb.2018.10.019
  • Choupanian, M., Omar, D., Basri, M., & Asib, N. (2017). Preparation and characterization of Neem oil nanoemulsion formulations against Sitophilus oryzae and Tribolium castaneum adults. Journal of Pesticide Science, 42(4), 158–165. https://doi.org/10.1584/jpestics.D17-032
  • CLSI (2012). Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard Eleventh Edition (Vol. 32, Issue 1). CLSI document M02 A11. https://doi.org/M02-A11
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  • Das, P., Sharma, N., Puzari, A., Kakati, D.K., & Devi, N. (2021). Synthesis and characterization of Neem (Azadirachta indica) seed oil-based alkyd resins for efficient anticorrosive coating application. Polymer Bulletin, 78(1), 457–479. https://doi.org/10.1007/s00289-020-03120-8
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There are 58 citations in total.

Details

Primary Language English
Subjects Microbiology (Other)
Journal Section Articles
Authors

Brahim Mahamat Ousman 0000-0002-3112-102X

Omar Belmehdi 0000-0002-7356-1648

Ibtissam Boussaoudi This is me 0009-0003-3285-3403

Boy Brahim Otchom This is me 0009-0005-4975-6700

Saoud Younes This is me 0000-0003-3056-3915

Early Pub Date March 19, 2025
Publication Date
Submission Date March 28, 2024
Acceptance Date December 17, 2024
Published in Issue Year 2025 Volume: 12 Issue: 2

Cite

APA Ousman, B. M., Belmehdi, O., Boussaoudi, I., Otchom, B. B., et al. (2025). Chemical composition and antibacterial activity of Azadirachta indica seed oil from Chad. International Journal of Secondary Metabolite, 12(2), 429-447.
International Journal of Secondary Metabolite

e-ISSN: 2148-6905