Bioactive Metabolites in Melaleuca leucadendra Leaves Extract: Phytochemical Profilling and GC-MS Characterization for Antimicrobial Relevance
DOI:
https://doi.org/10.22437/chp.v10i1.49078Keywords:
Antibacterial, bioactive compounds, GC-MS analysis, Melaleuca leucadendraAbstract
The global escalation of antimicrobial resistance (AMR) necessitates the exploration of natural sources of antibacterial agents. This study examined the ethanolic leaf extract of Melaleuca leucadendra's phytochemical composition, chemical profile, and antibacterial activity. Alkaloids, flavonoids, saponins, tannins, phenolics, terpenoids, and steroids were all detected by phytochemical screening. The quantitative total flavonoid content (TFC) of the extract was 17.78 mg QE/g, while the total phenolic content (TPC) was 292.43 mg GAE/g extract. Gas Chromatography–Mass Spectrometry (GC–MS) analysis identified oxygenated esters (18.86%), oxygenated aromatics (15.22%), phenolic derivatives (13.44%), and flavonoids (12.86%), methoxylated aromatics and terpenoids (5.34%), phytol, and fatty acid derivatives. These metabolite classes are widely reported to exert antibacterial effects through multiple mechanisms such as membrane disruption, enzyme inactivation, and inhibition of nucleic acid synthesis. Antibacterial testing against Staphylococcus aureus and Escherichia coli using the agar diffusion method showed a concentration-dependent response, with the highest activity at 80% extract (15.67 ± 0.58 mm and 15.33 ± 0.58 mm inhibition zones, respectively). Raman spectroscopy confirmed the interaction between polymer molecules and secondary metabolite compounds in the extract, thereby potentially enhancing antibacterial properties. These findings highlight that the antibacterial activity of M. leucadendra is mediated by the synergistic interplay of phenolics, flavonoids, methoxylated aromatics, terpenoids, and fatty acids. The findings offer compelling proof that M. leucadendra is a viable natural source for the creation of antibacterial compounds that will lessen antibiotic resistance.
References
[1] Yan, B., Huang, F., Ying, J., Zhou, D., Norouzi, S., Zhang, X., Wang, B., Liu, F. Global antibiotic hotspots and risks: A One Health assessment. Environmental Science and Ecotechnology. 2025. 25, 100564, DOI:10.1016/j.ese.2025.100564.
[2] Wang, N.C., and Liu, Y. Going shopping or consulting in medical visits: Caregivers’ roles in pediatric antibiotic prescribing in China. Social Science & Medicine. 2021. 290,114075, DOI:10.1016/j.socscimed.2021.114075.
[3] H. A. Sheerah et al., “Strengthening global health security through antimicrobial resistance control: Insights from Saudi Arabia. Journal of Infection and Public Health. 2025. 18(7); 102788, DOI: 10.1016/j.jiph.2025.102788
[4] Lakhani, A., Jindal, K., Khatri, K. Antimicrobial resistance (AMR) in Orthopaedic surgeries: A Complex issue and global threat. Journal of Orthopaedic Reports. 2025. 4(4), 100466, DOI:10.1016/j.jorep.2024.100466
[5] Rolta, R., Sharma, A., Sourirajan, A., Mallikarjunan, P.K., Dev, K. Combination between antibacterial and antifungal antibiotics with phytocompounds of Artemisia annua L: A strategy to control drug resistance pathogens. Journal of Ethnopharmacology. 2021, 266, 113420, DOI:10.1016/j.jep.2020.113420
[6] Abushaheen, M.A., Muzaheed, Fatani, A.J., Alosaimi, M., Mansy, W., George, M., Acharya, S., Rathod, S., Divakar, D.D., Jhugroo, C., Vellappally, S., Khan, A.A., Shaik, J., Jhugroo, P. Antimicrobial resistance, mechanisms and its clinical significance. Disease-a-Month. 2020. 66(6); 100971, DOI:10.1016/j.disamonth.2020.100971
[7] Atina., Royani, I., Idah, A., Arsyad, F.S. Antibacterial Properties of Taro : Extraction , Antibacterial Testing Method , Modification and Application. Science & Technology Indonesia. 2025.10(2):374–401, DOI:10.26554/sti.2025.10.2.374-401
[8] Feng, J., Zheng, Y., Ma, W., Ihsan, A., Hao, H., Cheng, G., Wang, X. Multitarget antibacterial drugs: An effective strategy to combat bacterial resistance,” Pharmacology & Therapeutics. 2023. 252, 108550, DOI:10.1016/j.pharmthera.2023.108550
[9] Sudiansyah, M.I., Yusro, F., Mariani, Y. Aktivitas Antibakteri Ekstrak Kulit Batang Gelam (Melaleuca leucadendra Linn.) terhadap Salmonella enterica serovar Typhimurium. Jurnal Serambi Engineering. 2023. 8(3): 6161–6167. DOI: 10.32672/jse.v8i3.6104
[10] Jothinathan, D.B., Suseem, S.R. Fabrication, characterization, and physicochemical properties of curcumin incorporated new P. vulgari spolysaccharides bio-composite film ; potent drug carrier , antibacterial , and antioxidant properties. International Journal of Biological Macromolecules. 2025, 330, 148165, DOI:10.1016/j.ijbiomac.2025.148165
[11] Tennalli, G.B., Yallappa, S., Hungund, B.S., Joseph, S., Divate, M.N., Upadhyaya, K. Biogenic synthesis of copper oxide nanoparticles from Ricinus communis L . leaf extract and their cytotoxic and pro-apoptotic effects on tumor. Result in Chemistry. 2025. 18, 102896, 2025. DOI: 10.1016/j.rechem.2025.102896
[12] Zeng, H., Wang, X., Tang, J., Liu, P., Zhang, S., Chu, H., Chen, B., Ma, M. Proteomic and metabolomic analyses reveal the antibacterial mechanism of Cannabidiol against gram-positive bacteria. Journal of Proteomics. 2025. 315,105411. DOI: 10.1016/j.jprot.2025.105411
[13] Dan, W., Gao, J., Zhang,, J., Cao, Y., Liu, J., Sun, Y., Wang, J., Dai, J. Antibacterial activity and mechanism of action of canthin-6-one against Staphylococcus aureus and its application on beef preservation. Food Control. 2023. 147, 109604. DOI:10.1016/j.foodcont.2023.109604
[14] Zamzami, A.D.R.A., Isnaini., Yasmina, A. Uji Toksisitas Ekstrak Metanol Kulit Kayu dan Daun Galam dengan Metode BSLT. Homeostatis. 2021. 4(1):43–48, 2021. DOI: 10.20527/ht.v4i1.3321
[15] Wardhani, R.R.A.A.K., Akhyar, A., Prasiska, E. Skrining Fitokimia, Aktivitas Antioksidan, dan Kadar Total Fenol-Flavonoid Ekstrak Daun dan Buah Tanaman Galam Rawa Gambut (Melaleuca cajuputi ROXB). QUANTUM: Jurnal Inovasi Pendidikan Sains. 2018. 9(2); 133–143, 2018. DOI: 10.20527/quantum.v9i2.5571
[16] Vila Nova, B.G., Silva, L.D.S., Andrade, M.D.S., de Santana, A.V.S., da Silva, L.C.T., Sá, G.C., Zafred, I.F., Moreira, P.H.A., Monteiro, C.A., da Silva, L.C.N., Abreu, A.G. The essential oil of Melaleuca alternifolia incorporated into hydrogel induces antimicrobial and anti-inflammatory effects on infected wounds by Staphylococcus aureus. Biomedicine & Pharmacotherapy. 2024. 173,116389, DOI: 10.1016/j.biopha.2024.116389
[17] Bugarcic, A., Bowles, E.J., Summer, K., Agnew, T., Barkla, B., Lauche, R. Australian Tea Tree (Melaleuca alternifolia) oil: an updated review of antimicrobial and other medicinal properties. Phytomedicine Plus. 2025. 5(3); 100846, DOI:10.1016/j.phyplu.2025.100846
[18] Naccari, C., Cicero, N., Orlandella, B.M., Naccari, V., Palma, E. Antimicrobial activity of essential oils (Citrus bergamia Risso & Poiteau, Melaleuca alternifolia and Chenopodium botrys) on pathogen strains isolated in milk samples from mastitic sheep. Natural Product Research. 2025 39(10);2963–2969, 2025, DOI:10.1080/14786419.2023.2300041
[19] Idjan, M.K.N.A., Fatmawati, Almafie, M.R., Dani, R., Subandrate, Fudholi, A., Sriyanti, I. 2025. Physicochemical properties and antioxidant activity of PVP/CA membranes nanofiber loaded Arcangelisia flava L. Merr. stem extract using electrospinning technique. Next Materials. 2025. 8,100839, DOI: 10.1016/j.nxmate.2025.100839
[20] Miksusanti, M., Apriani, E.F., Ahmadi, A., Shiyan, S., Wijaya, D.P., Risanli, V.A. Formulation and Characterization of Sambiloto Extract Orally Dissolving Films: a Dual Mechanism for Probiotic Growth and Antibacterial Action,” International Journal of Applied Pharmaceutics. 2025. 17(2):198–203, DOI:10.22159/ijap.2025v17i2.52902
[21] Miksusanti, M., Apriani, E.F., Aprida, N. Formulation and Optimization Peel-Off Gel Mask with Polyvinyl Alcohol and Whey Protein-Based using Factorial Design from Ethanolic Extract of Mangosteen Peel (Garcinia Mangostana) as Antioxidant,” Research Journal of Pharmacy and Technology. 2023. 16(2): 870–878, DOI: 10.52711/0974-360X.2023.00148
[22] Pfoze, B.K., Singh, R.R., Pekosii, B.K., Singh, T.P., Singh, O.M. Phytochemical screening and assessment of the in-vitro wound healing activity of Bidens pilosa leaves,” Pharmacological Research - Natural Products. 2025. 8.100331, DOI:10.1016/j.prenap.2025.100331
[23] Sharma, A., and Kaur, A. Catharanthus roseus leaf-based green synthesis of silver oxide nanoparticles: Characterization, phytochemicals screening and antimicrobial activity. The Microbe. 2025. 8,100469, DOI:10.1016/j.microb.2025.100469
[24] Motlhalamme, T., Kaningini, A.G., Maropeng, R., Thema, F.T., Mohale, K.C., Maaza, M. Tomato lycopene, total flavonoids, vitamin C content and mineral composition as affected by biosynthesized calcium carbonate nanoparticles foliar application. Journal of Food Composition and Analysis. 2025.139, 107173, DOI:10.1016/j.jfca.2024.107173
[25] Ferdous, U.T., Nurdin, A., Ismail, S., Shaari, K., Yusof, Z.N.B. A comparative study on antioxidant properties, total phenolics, total flavonoid contents, and cytotoxic properties of marine green microalgae and diatoms. Journal of Genetic Engineering and Biotechnology. 2025. 23(1);100456, DOI:10.1016/j.jgeb.2024.100456
[26] Dzigbor, A., Neglo, D., Dzah, C.S., and Sraha, R. Total phenolic content, phytochemical screening, antioxidant and antimicrobial activities of Borassus flabellifer and Borassus aethiopum fruits,” Food Chemistry Advances. 2025. 7.100937, DOI: 10.1016/j.focha.2025.100937
[27] Eleazu, C.O. Characterization of the natural products in cocoyam (Colocasia esculenta) using GC–MS. Pharmaceutical Biology. 2016. 54(12): 2880–2885. DOI:10.1080/13880209.2016.1190383
[28] Tchaikovskaya, O., Bocharnikova, E., Bezlepkina, N., Solomonov, V., Makarova, A., Spirina, A., Chaikovsky, S. The total phenolic content in an aqueous solution of chloramphenicol under exposure to UV and e-beam irradiation. Chemosphere. 2025. 386, 144602, DOI: 10.1016/j.chemosphere.2025.144602
[29] Endris, Y. A., Abdu, K. Y., and Abate, S. G., Investigation of bioactive phytochemical compounds of the Ethiopian medicinal plant using GC-MS and FTIR. 2024. Heliyon. 10(15)e34687. DOI:10.1016/j.heliyon.2024.e34687
[30] Alallam, B., Abdulameed, H.T., and Lim, V. Unbiased Metabolomic and Chemometric profiles of three Sargassum polycystum extracts using GCMS and LCMS/MS: content analysis, correlation analysis and molecular docking. Food Chemistry. 2025. 470, 142666.
DOI:10.1016/j.foodchem.2024.142666
[31] Elsadek, M.F., Al-Numair, K.S. Profiling of phytochemical constituents of terminalia chebula fruit extract by different solvent effects and synchronized analysis of FTIR and GCMS. Journal of King Saud University-Science. 2024. 36(9),103414, DOI: 10.1016/j.jksus.2024.103414
[32] Abate, S.G., Endris, Y.A., Abdu, K.Y., Mekonnen, K.D. Integrated analysis of Zehneria scabra leaf extract: phytochemical screening, GC–MS, FT-IR spectroscopy, and antibacterial activity evaluation. Natural Product Research. 2025. 1–4, DOI:10.1080/14786419.2025.2491117
[33] Palladini, G., Garbarino, C., Luppi, A., Russo, S., Filippi, A., Arrigoni, N., Massella, E., Ricchi, M. Comparison between broth microdilution and agar disk diffusion methods for antimicrobial susceptibility testing of bovine mastitis pathogens. Journal of Microbiological Methods. 2023. 212, 106796. DOI: 10.1016/j.mimet.2023.106796
[34] Carvalho, G.M., Silva, B.A., Xavier, R.G.C., Zanon, I.P., Vilela, E.G., Nicolino, R.R., Tavares, G.C., Silva, R.O.S. Evaluation of disk diffusion method for testing the rifampicin, erythromycin, and tetracycline susceptibility of Clostridioides (prev. clostridium) difficile. 2023. Anaerobe. 80. 102720,.DOI:10.1016/j.anaerobe.2023.102720
[35] Elfita, E., Mardiyanto, M., Fitrya, F., Larasati, J.E., Julinar, J., Widjajanti, H., Muharni, M. Antibacterial activity of Cordyline fruticosa leaf extracts and its endophytic fungi extracts. Biodiversitas,. 2019. 20(12):3804–3812. DOI:10.13057/biodiv/d201245
[36] Duan, M., Sun, J., Huang, Y., Jiang, H., Hu, Y., Pang, J., Wu, C. Electrospun gelatin / chitosan nanofi bers containing curcumin for multifunctional food packaging. Food Science and Human Wellness. 2023.12(2): 614–621, DOI:10.1016/j.fshw.2022.07.064
[37] Mochtar, C.F., Aisyiyah, N.M., Husna, Q.A., Hamzah, H., Bakhtiar, M.I., Devi, R.S., Varizza, F.P., Faradillah, A., Hafidzah, E., Suriati, S., Damis, N.L. Antipyretic And Antiinflammatory Activities Of Bopot Leaf Extract From Kutai Kartanegara District,” Indonesian Journal of Pharmaceutical Science and Technology. 2023. 1(1):80–89, DOI: 10.24198/ijpst.v0i0.46188
[38] Padalia, R.C., Verma, R.S., Chauhan, A., and Chanotiya, C.S. The essential oil composition of Melaleuca leucadendra L. grown in India: A novel source of (E)-nerolidol. Industrial Crops and Products. 2015. 69: 224–227, DOI: 10.1016/j.indcrop.2015.02.019
[39] S. Siddique, Z. Parveen, Firdaus-e-Bareen, and S. Mazhar, “Chemical composition, antibacterial and antioxidant activities of essential oils from leaves of three Melaleuca species of Pakistani flora. Arabian Journal of Chemistry. 2020. 13(1): 67–74, DOI:10.1016/j.arabjc.2017.01.018
[40] Herlina, T., Rizaldi Akili, A.W., Nishinarizki, V., Hardianto, A., and Latip, J.B. Review on antibacterial flavonoids from genus Erythrina: Structure-activity relationship and mode of action. Heliyon. 2025. 11(1),e41395, DOI:10.1016/j.heliyon.2024.e41395
[41] Liu J, Xia F, Ouyang H, Wang W, Li T, Shi Y, Yan X, Yan X, He S. Nardosinane-related antimicrobial terpenoids from Lemnalia sp. soft coral. Phytochemistry. 2022. 196, 113088, DOI:10.1016/j.phytochem.2022.113088
[42] Siddiqui, T., Khan, M.U., Sharma, V., and Gupta, K. Terpenoids in essential oils: Chemistry, classification, and potential impact on human health and industry. Phytomedicine Plus. 2024. 4(2)100549, DOI: 10.1016/j.phyplu.2024.100549
[43] Zhang, D., Liu, Y., Li, X., Xiao, J., Sun, J., Guo, L. Inactivation of Escherichia coli on broccoli sprouts via plasma activated water and its effects on quality attributes,” Lwt. 2022. 154, 112761, DOI: 10.1016/j.lwt.2021.112761
[44] Martins, N., Costa-Oliveira, S., Melo, L.D.R., Ferreira, I.C.F.R., Azeredo, J., Henriques, M., Silva, S. Susceptibility testing of Candida albicans and Candida glabrata to Glycyrrhiza glabra L. Industrial Crops and Products. 2017. 108: 480–484, DOI: 10.1016/j.indcrop.2017.07.008
[45] Sundaram R. L. and Vasanthi, H. R. Spermacoce hispida Linn: A critical review on pharmacognosy, phytochemistry, and pharmacology based on traditional claims,” Phytomedicine Plus. 2022. 2(1): 100143, DOI: 10.1016/j.phyplu.2021.100143
[46] López-González D, Graña E, Teijeira M, Verdeguer M, Reigosa MJ, Sánchez-Moreiras AM, Araniti F. Similarities on the mode of action of the terpenoids citral and farnesene in Arabidopsis seedlings involve interactions with DNA binding proteins. Plant Physiology and Biochemistry. 196: 507–519, 2023, DOI: 10.1016/j.plaphy.2023.02.004
[47] Schäfer, N., Pokajewicz, K., Balwierz, R., Byrski, A., Biernat, P., Lipok, J. Chemical composition and antibacterial activity of chaulmoogra oil from Hydnocarpus wightiana: A detailed analysis of volatile and fatty acid fractions. Phytochemistry Letters. 2024. 65: 102–112, DOI: 10.1016/j.phytol.2024.12.008
[48] Arellano, H., Swebocki, T., Le Coeur, C., Prevost, S., Abdallah, M., Nardello-Rataj, V., Fameau, A.L. Influence of critical micelle concentration of choline-based long chain fatty acid soaps on their antibacterial activity against Methicillin resistant Staphylococcus aureus. Journal of Colloid and Interface Science. 2025. 677(A), 314–323. DOI: 10.1016/j.jcis.2024.07.218
[49] Liu, D., Pan, S., Sun, J. Natural Phenolic Acids as Promising Antimicrobial Candidates in Food Industry: A Review. International Journal of Food Microbiology. 2025. 111413. DOI: 10.1016/j.ijfoodmicro.2025.111413
[50] Yahia, Y., Zaghdoud, C., Tlahig, S., Bouzidi, A., Boufahja, F., Elfalleh, W. Phenolic Profilling and Antioxidant-antimicrobial Activity of Spontaneous Plants Naturally Grown in Tunisia,” Food Bioscience. 2025. 107470, DOI: 10.1016/j.fbio.2025.107470.
[51] Z. Yu et al., “Insights into relationship between microbial community-induced polycyclic aromatic hydrocarbons degradation and soluble organic matter transformation during vermiculite-amended sewage sludge composting. Journal of Cleaner Production. 2024. 472(9):143491. DOI: 10.1016/j.jclepro.2024.143491
[52] Khan U.M, Sameen A., Decker E.A., Shabbir M.A., Hussain, S., Latif, A., Abdi G, Aadil RM. Implementation of plant extracts for cheddar-type cheese production in conjunction with FTIR and Raman spectroscopy comparison,” Food Chemistry. 2024. X, 22, 101256. DOI: 10.1016/j.fochx.2024.101256
[53] Gómez, D.A., Coello, J., Maspoch, S. “The influence of particle size on the intensity and reproducibility of Raman spectra of compacted samples,” Vibrational Spectroscopy. 2019. 100; 48–56, DOI: 10.1016/j.vibspec.2018.10.011.
