Chemical profile by GC-MS and protective effect of Algerian cloves (Syzygium aromaticum) against Lactobacillus spp. and Streptococcus spp. isolated from dental caries
Year 2024,
Volume: 11 Issue: 4, 833 - 852, 03.11.2024
Ouahiba Benhamada
,
Nabila Benhamada
,
Lilia Boussouf
Essaid Leghouchi
Abstract
The oral cavity is home to a large and diversified microbial flora, which plays an important role in the genesis of multiple diseases, including tooth decay. Indeed, tooth decay is the most common ailment in the world, with almost everyone having experienced it at least once in their lifetime. Examining plants used in traditional medicine is one of the research approaches used to discover novel, potent antibacterial chemicals with a broad spectrum of action, as present antibacterials have significant drawbacks. This study aims to examine the chemical composition of Algerian clove Syzygium aromaticum using GC-MS and to evaluate the antibacterial activity of the methanol extract against bacteria isolated from dental caries caused by Streptococcus spp. and Lactobacillus spp. The results show a strong extraction yield of 29.7%, with high amounts of polyphenols and flavonoids calculated at 178.82 mg GAEQ/g and 24.13 mg QEQ/g. The principal chemical elements of S. aromaticum peel methanol extract were identified as eugenol (61.23%) and eugenol acetate (26.45%) based on mass spectrum data and retention times. The methanol extract has a significant antibacterial effect against tested strains, with MICs ranging from 111.37 to 445.5 mg/mL. Higher concentrations of polyphenols resulted in a significant increase in inhibition zone diameter against S1 (r2 = 0.94, p < 0.001), L3 (r2 = 0.94, p < 0.001), L5 (r2 = 0.93, p < 0.001), and L9 (r2 = 0.96, p < 0.001).
Thanks
The authors would also like to thank the Algerian Ministry of Higher Education and Scientific Research and the General Directorate of Research and Development Technologies (DGRSDT) of Algeria.
References
- Ajobiewe, H.F., Elisha, E., Ibrahim, A.E., Ajobiewe, J.O., Salami, A.O., Umeji, L.C., … Alau, K.K. (2022). Antimicrobial activity of clove plant flower bud extract (Syzygium aromaticum) on Escherichia coli. Scholars Journal of Applied Medical Sciences, 10(5), 673-684. https://doi.org/10.36347/sjams.2022.v10i05.002
- Alghazzaly, A., Elsherbiny, G., Moghannem, S., & Sharaf, M. (2022). Antibacterial, antibiofilm, antioxidants and phytochemical profiling of Syzygium aromaticum extract. Egyptian Journal of Aquatic Biology and Fisheries, 26(5), 207-218.
- Almaamori, A.M. (2023). Oral health and dental caries. World bulletin of public health, 14
- Amadi, E.K., Kareru, P.G., Keriko, J.M., & Kiptoo, J. (2016). Antimicrobial activity of selected medicinal plants’ extracts against streptococcus pneumoniae. International Journal of Botany Studies, 1(1), 20-22.
- Aneja, K.R., & Joshi, R. (2010). Antimicrobial activity of Syzygium aromaticum and its bud oil against dental cares causing microorganisms. Ethnobotanical Leajlets, 14, 960-975.
- Aneja, K.R., Joshi, R., & Sharma, C. (2009). Antimicrobial activity of Dalchini (Cinnamomum zeylanicum bark) extracts on some dental caries pathogens. Journal of Pharmacy Research, 2(9), 1387-1390.
- Arya, S.S., Rookes, J.E., Cahill, D.M., & Lenka, S.K. (2021). Vanillin: a review on the therapeutic prospects of a popular flavouring molecule. ADV TRADIT MED (ADTM), 21(3), 1–17. https://doi.org/10.1007/s13596-020-00531-w
- Bai, J., Li, J., Chen, Z., Bai, X., Yang, Z., Wang, Z., & Yang, Y. (2023). Antibacterial activity and mechanism of clove essential oil against foodborne pathogens. LWT- Food science and technology, 173, 114249. https://doi.org/10.1016/j.lwt.2022.114249
- Basli, A., Chibane, M., Madani, K., & Oukil, N. (2012). Activité antibactérienne des polyphénols extraits d’une plante médicinale de la flore d’Algérie: Origanum glandulosum Desf. Phytothérapie, 10, 2-9. https://doi.org/10.1007/s10298-012-0683-9
- Benhamada, O., Benhamada, N., & Leghouchi, E. (2022). Polyphenols and antibacterial activity of Xanthoria parietina (L.) Th. Fr. methanol extract under lead stress. Journal of Applied Biological Sciences, 16(3), 537-552. https://doi.org/10.5281/zenodo.7114289
- Bhuiyan, M.N.I., Begum, J, Nandi, N.C., & Akter, F. (2010). Constituents of the essential oil from leaves and buds of clove (Syzigium caryophyllatum (L.) Alston). African Journal of Plant Science, 4(11), 451-454.
- Brustugun, J., Tonnesen, H.H., Edge, R., & Navaratnam, S. (2005). Formation and reactivity of free radicals in 5-hydroxymethyl-2-furaldehyde–the effect on isoprenaline photostability. Journal of Photochemistry and Photobiology, 79(2), 109 119. https://doi.org/10.1016/j.jphotobiol.2004.12.005
- Cai, J-N., & Kim, D. (2023). Biofilm ecology associated with dental caries: understanding of microbial interactions in oral communities leads to development of therapeutic strategies targeting cariogenic biofilms. Advances in Applied Microbiology, 122, 27-75. https://doi.org/10.1016/bs.aambs.2023.02.001
- Cheikhyoussef, A., Cheikhyoussef, N., Rahman, A., & Hussein, A.A. (2022). Clove (Syzygium Aromaticum): Chapter 14 - Clove (Syzygium aromaticum) phenolics: Extraction, compositions, and biological activities. Chemistry, Functionality and Applications, 215-233. https://doi.org/10.1016/B978-0-323-85177-0.00036-7
- Chen, Z., Liu, Q., Zhao, Z., Bai, B., Sun, Z., Cai, L., … Xi, G. (2021). Effect of hydroxyl on antioxidant properties of 2,3 dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one to scavenge free radicals. RSC Advances, 11(55), 34456–34461. https://doi.org/10.1039/d1ra06317k
- Cortés-Rojas, D.F., de Souza, C.R.F., & Oliveira, W.P. (2014). Clove (Syzygium aromaticum): a precious spice. Asian Pacific Journal of Tropical Biomedicine, 4(2), 90-96. https://doi.org/10.1016/S2221-1691(14)60215-X
- de Freitas, T.S., da Cunha Xavier, J., Pereira, R.L.S., Rocha, J.E., Muniz, D.F., da Silva, P.T., … Coutinho, H.D.M. (2020). Direct antibacterial and antibiotic resistance modulatory activity of chalcones synthesized from the natural product 2-hydroxy-3,4,6-trimethoxyacetophenone. FEMS Microbiology Letters, 367(15). https://doi.org/10.1093/femsle/fnaa124
- Delarras, C. (2007). Microbiologie pratique pour le laboratoire d’analyse ou de contrôle sanitaire. Ed. Lavoisier. P, 128, 129, 269, 271.
- de Oliveira Carvalho, I., Purgato, G.A., Píccolo, M.S., Pizziolo, V.R., Coelho, R.R., Diaz-Muñoz, G., & Alves Nogueira Diaz, M. (2020). In vitro anticariogenic and antibiofilm activities of toothpastes formulated with essential oils. Archives of Oral Biology, 117, 104834. https://doi.org/10.1016/j.archoralbio.2020.104834
- Devi, B., Ramasubramaniaraja, R., Carism, & Devi, B. (2009). Dental caries and medicinal plants –An overview. Journal of Pharmacy Research, 2, 1669-1675.
- Diastuti, H., Chasani, M., & Suwandri, S. (2019). Antibacterial Activity of Benzyl Benzoate and Crotepoxide from Kaempferia rotunda L. Rhizome. Indonesian Journal of Chemistry, 20(1), 9. https://doi.org/10.22146/ijc.37526
- Dogruoz, N., Zeybek, Z., & Karagoz, A. (2008). Antibacterial Activity of Some Plant Extracts. IUFS Journal of Biology, 67(1), 17-21.
- Duvauchelle, V., Majdi, C., Bénimélis, D., Dunyach-Remy, C., Meffre, P., & Benfodda, Z. (2021). Synthesis, structure elucidation, antibacterial activities, and synergistic effects of novel juglone and naphthazarin derivatives against clinical methicillin-resistant Staphylococcus aureus Strains. Frontier in Chemistry, 9, 773981. https://doi.org/10.3389/fchem.2021.773981
- El-Haci, I.A., Atik-Bekkara, F., Didi, A., Gherib, M., & Didi, M.A. (2012). Teneurs en polyphénols et pouvoir antioxydant d’une plante médicinale endémique du Sahara algérien. Phytothérapie, 10, 280-285. https://doi.org/10.1007/s10298-012-0726-x
- El-Sherbiny, G.M., & Elbestawy, M.K.M. (2022). A review – plant essential oils active against Helicobacter pylori. Journal of Essential Oil Research, 34(3), 203 215. https://doi.org/10.1080/10412905.2022.2025464
- Featherstone, J.D.B. (2000). The science and the practice of caries prevention. The Journal of the American Dental Association, 131(7), 887 899. https://doi.org/10.14219/jada.archive.2000.0307
- Foda, A.M., Kalaba, M.H., El-Sherbiny, G.M., Moghannem, S.A., & El-Fakharany, E.M. (2022). Antibacterial activity of essential oils for combating colistin-resistant bacteria. Expert Review of Anti infective Therapy, 20(10), 1351 1364. https://doi.org/10.1080/14787210.2022.2101997
- Freitas, T.S., da Cunha Xavier, J., Pereira, R.L.S., Rocha, J.E., Muniz, D.F., da Silva, P.T., … Coutinho, H.D.M. (2020). Direct antibacterial and antibiotic resistance modulatory activity of chalcones synthesized from the natural product 2 hydroxy 3,4,6 trimethoxyacetophenone. FEMS-Microbiology Letters, 367(15). https://doi.org/10.1093/femsle/fnaa124
- Frohlich, P.C., Santos, K.A., Ascari, J., dos Santos Refati, J.R., Palú, F., Cardozo-Filho, L., & da Silva, E.A. (2023). Antioxidant compounds and eugenol quantification of clove (Syzygium aromaticum) leaves extracts obtained by pressurized liquid extraction and supercritical fluid extraction. The Journal of Supercritical Fluids, 196, 105865. https://doi.org/10.1016/j.supflu.2023.105865
- Gengatharan, A., & Abd Rahim, M.H. (2023). The application of clove extracts as a potential functional component in active food packaging materials and model food systems: A mini-review. Applied Food Research, 3(1), 100283. https://doi.org/10.1016/j.afres.2023.100283
- Gupta, S., Park, S.E., Mozaffari, S., El-Aarag, B., Parang, K., & Tiwari, R.K. (2023). Design, Synthesis, and Antiproliferative Activity of Benzopyran-4-One-Isoxazole Hybrid Compounds. Molecules, 28(10), 4220. https://doi.org/10.3390/molecules28104220
- Gyrdymova, Y., & Rubtsova, S. (2021). Caryophyllene and caryophyllene oxide: a variety of chemical transformations and biological activities. Chemical Papers. 76(2). https://doi.org/10.1007/s11696-021-01865-8
- Hemeda, N.A., Hegazy, G.E., Abdelgalil, S., Soliman, N.A., Abdel-Meguid, D.I., & El-Assar, S.A. (2022). Maximization of red pigment production from Streptomyces sp. LS1 structure elucidation and application as antimicrobial/antifouling against human pathogens and marine microbes. Journal of Genetic Engineering and Biotechnology, 20, 168. https://doi.org/10.1186/s43141-022-00452-y
- Huang, L., Zhu, X., Zhou, S., Cheng, Z., Shi, K., Zhang, C., & Shao, H. (2021). Phthalic Acid Esters: Natural Sources and Biological Activities. Toxins (Basel), 13(7), 495. https://doi.org/10.3390/toxins13070495
- Hugar, S., M Patel, P., Nagmoti, J., Uppin, C., Mistry, L., & Dhariwal, N. (2017). An in vitro comparative evaluation of efficacy of disinfecting ability of garlic oil, neem oil, clove oil, and tulsi oil with autoclaving on ndodontic K Files tested against Enterococcus faecalis. International Journal of Clinical Pediatric Dentistry, 10(3), 283 288. https://doi.org/10.5005/jp-journals-10005-1451
- Jadhav, J.J., Jadeja, G.C., & Desai, M.A. (2022). Clove (Syzygium aromaticum): Chapter 28 - Effect of extraction techniques on the yield, composition, and quality of clove (Syzygium aromaticum) essential oil. Chemistry, Functionality and Applications, 485-500. https://doi.org/10.1016/B978-0-323-85177-0.00027-6
- Joseph, B., & Sujatha, S. (2011). Bioactive Compounds and its autochthonous microbial activities of extract and clove oil (Syzygium aromaticum L.) on some food borne pathogens. Asian Journal of Biological Sciences, 4(1), 35-43. https://doi.org/10.3923/ajbs.2011.35.43
- Kablan, B.J., Adiko, M., & Abrogoua, D.P. (2008). Évaluation in vitro de l'activité antimicrobienne de Kalanchoe crenata et de Manotes longiflora utilisées dans les ophtalmies en Côte d'ivoire. Phytothérapie, 6(5), 282-288. https://doi.org/10.1007/s10298-008-0332-0
- Kabra, P., Loomba, K., Kabra, S.K., Majumdar, D.S.P., & Kuma, N. (2012). Medicinal plants in the treatment of dental caries. Asian Journal of Oral Health & Allied Sciences, 2(1), 12-16.
- Kaur, D., & Chandrul, K. (2017). Syzygium aromaticum L. (Clove): A vital herbal drug used in periodontal disease. Indian Journal of Pharmaceutical and Biological Research, 5(2), 45-51. https://doi.org/10.30750/ijpbr.5.2.9
- Khan, M. K., Hassan, S., Imran, M., & Ahmad, M.H. (2022). Clove (Syzygium aromaticum): Chapter 22 - Extraction of bioactive compounds from clove (Syzygium aromaticum). Chemistry. Functionality and Applications, 405-417. https://doi.org/10.1016/B978-0-323-85177-0.00010-0
- Kiralan, M., & Ketenoglu, O. (2022). Clove (Syzygium aromaticum): Chapter 36 - Food applications of clove (Syzygium aromaticum) extracts. Chemistry, Functionality and Applications, 607-617. https://doi.org/10.1016/B978-0-323-85177-0.00030-6
- Kuete, V., Dongfack, M.D., Mbaveng, A.T., Lallemand, M-C., Van-Dufat, H.T., Wansi, J-D., … Wandji, J. (2010). Antimicrobial activity of the methanolic extract and compounds from the stem bark of Drypetes tessmanniana. Chinese Journal of Integrative Medicine, 16(4), 337-343. https://doi.org/10.1007/s11655-010-0527-8
- Li, Y.H., & Lu, X.Y. (2005). Investigation on the origin of 5-HMF in Shengmaiyin decoction by RP-HPLC method. Journal of Zhejiang University Science B, 6, 1015–1021. https://doi.org/10.1631/jzus.2005.B1015
- Mallya, P.S., & Mallya, S. (2020). Microbiology and clinical implications of dental caries – a review. Journal of Evolution of Medical and Dental Sciences, 9(48), 3670-3675. https://doi.org/10.14260/jemds/2020/805
- Mazumder, K., Nabila, A., Aktar, A., & Farahnaky, A. (2020). Bioactive Variability and In Vitro and In Vivo Antioxidant Activity of Unprocessed and Processed Flour of Nine Cultivars of Australian lupin Species: A Comprehensive Substantiation. Antioxidants (Basel), 9(4), 282. https://doi.org/10.3390/antiox9040282
- Meda, A., Lamien, C.E., Romito, M., Millogo, J., & Nacoulma, O.G. (2005). Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan Honey, as well as their radical scavenging activity. Food Chemistry, 91(3), 571 577. https://doi.org/10.1016/j.foodchem.2004.10.006
- Mohammadi Ziarani, G., Kheilkordi, Z., & Mohajer, F. (2020). Recent advances in the application of acetophenone in heterocyclic compounds synthesis. Journal of the Iranian Chemical Society, 17, 247–282. https://doi.org/10.1007/s13738-019-01774-4
- Moroh, J-L.A. (2008). Étude de l’activité antibactérienne de l’extrait acétatique (EAC) de Morinda morindoides (Baker) milne-redheat (rubiaceae) sur la croissance in-vitro des souches d’Escherichia coli. Bulletin de la Societe Royale des Sciences de Liege, 77. 44 - 61.
- Mostafa, A.A., Al-Askar, A.A., Almaary, K.S., Dawoud, T.M., Sholkamy, E.N., & Bakri, M.M. (2018). Antimicrobial activity of some plant extracts against bacterial strains causing food poisoning diseases. Saudi Journal of Biological Sciences, 25(2), 361 366. https://doi.org/10.1016/j.sjbs.2017.02.004
- Munson, M.A., Banerjee, A., Watson, T.F., & Wade, W.G. (2004). Molecular analysis of the microflora associated with dental caries. Journal of clinical microbiology, 42(7), 3023-3029. https://doi.org/10.1128/JCM.42.7.3023-3029.2004
- Nakilcioglu, T.E., & Otles, S. (2021). Influence of extraction solvents on thepolyphenol contents, compositions, and antioxidant capacities of fig (Ficus carica L.) seeds. Annals of the Brazilian Academy of Sciences Printed, 93(1), 1678-2690. https://doi.org/10.1590/0001-3765202120190526
- Nkere, C.K., & Iroegbu, C.U. (2005). Antibacterial screening of the root, seed and stem bark extracts of Picralima nitida. African Journal of Biotechnology, 4(6), 522-526.
- Oliveira, L.C.C., Rodrigues, F.A.A., dos Santos Barbosa, C.R., dos Santos, J.F.S., Macêdo, N.S., de Sousa Silveira, Z., Coutinho, H.D.M., & da Cunha, F.A.B. (2022). Antibacterial Activity of the Pyrogallol against Staphylococcus aureus Evaluated by Optical Image. Biologics, 2, 139-150. https://doi.org/10.3390/biologics2020011
- Oshomoh, E.O., ldu, M., & Udinyiwe, O.C. (2015). Phytochemical screening and antimicrobial sensitivity of clove flower (Syzygium aromaticum, L. Merrill and Perry) bud on dental pathogens. International Journal of Pharmacy and Pharmaceutical Research, 3(2), 1-12.
- Owen, P.L., & Johns, T. (1999). Xanthine oxidase inhibitory activity of northeastern North American plant remedies used forgout. Journal of Ethnopharmacology, 64,149-160. https://doi.org/10.1016/s0378-8741(98)00119-6
- Penot, E. (2016). Analyse technico-économique de la filière amont de production d’essence de girofle à Fénérive- Est, Madagascar: de la feuille à l’alambic.
- Prajapati, R.A., & Raol, B.V. (2014). Characterization of microbial flora isolated from human teeth carious lesions. Life Sciences Leajlets, 47, 01-08.
- Qasim, M., Aziz, I., Rasheed, M., Gul, B., & Khan, M. (2016). Effect of extraction solvents on polyphenols and antioxidant activity of medicinal halophytes. Pakistan Journal of Botany, 48(2), 621-627.
- Ratri, P.J., Ayurini, M., Khumaini, K., & Rohbiya, A. (2020). Clove oil extraction by steam distillation and utilization of clove buds waste as potential candidate for Eco-Friendly Packaging. Jurnal Bahan Alam Terbarukan, 9(1), 47 54. https://doi.org/10.15294/jbat.v9i1.24935
- Reddy, D.S., & Rogawski, M.A. (2012). “Neurosteroids - Endogenous Regulators of Seizure Susceptibility and Role in the Treatment of Epilepsy”. In Noebels JL, Avoli M, Rogawski MA, et al. (eds.). Jasper’s Basic Mechanisms of the Epilepsies [Internet]. 4th Edition. Bethesda (MD): National Center for Biotechnology Information (US). National Center for Biotechnology Information (US). PMID 22787590
- Riaz̽, A., Umar, M., & Farah, N. (2023). Prevalence, causes and management of dental caries. Journal of Xi’an Shiyou University, Natural Science Edition, 19(5), 1033-1053.
- Rodrigues, L.B., Martins, A.O.B.P.B., Cesário, F.R.A.S., Castro, F.F., de Albuquerque, T.R., Fernandes, M.N.M., … de Menezes, I.R.A. (2016). Anti-inflammatory and antiedematogenic activity of the Ocimum basilicum essential oil and its main compound estragole: In vivo mouse models. Chemico-Biological Interactions, 257, 14-25. https://doi.org/10.1016/j.cbi.2016.07.026
- Ryu, C.K., Kim, D.H., Kim, H.J., & Chung, S.Y. (1993). The antimicrobial activities of some 1,4 naphthalenediones (IV). Archives of Pharmacal Research, 16, 327 330. https://doi.org/10.1007/BF02977525
- Sabriu – Haxhijaha, A., Jordanovska, S., & Popovska, O. (2021). Analysis of clove (Syzygium aromaticum L.) oil obtained with hydrodistillation and ultrasound-assisted extraction. UDC582.776.2-113.551:542.61. https://doi.org/10.20544/HORIZONS.B.09.2.P07
- Saikumari, D., Shiva Rani, S.K., & Saxena, N. (2016). Antibacterial Activity of Syzigium aromaticum L. (Clove). International Journal of Current Microbiology and Applied Sciences, 5 (11), 484-489. http://dx.doi.org/10.20546/ijcmas.2016.511.056
- Saini, M., Alam, S., Prajapati, R., Kr, L., & Thakur (2019). Analysis and estimation of eugenol content in microemulsion formulation containing clove oil (Syzigium aromaticum). Journal of Applicable Chemistry, 8(1), 195-202.
- Shafira, K., Azad, A.K., Zubair, L., & Helal Uddin, A.B.M. (2020). Extraction and quantification of eugenol from clove buds using HPLC. Current Chromatography, 07(1) https://doi.org/10.2174/2213240607999200818161356
- Sharaf, M.H., El-Sherbiny, G.M., Moghannem, S.A., Abdelmonem, M., Islam, A., Elsehemy, I.A., … Kalaba, M.H. (2021). New combination approaches to combat methicillin-resistant Staphylococcus aureus (MRSA). Scientific Reports, 11(1), 1 16. https://doi.org/10.1038/s41598-021-82550-4
- Shehadi, M., Awada, F., Oleik, R., Chokr, A., Hamze, K., Hamdan, H.A., & Kobaissi, A. (2014). Comparative analysis of the anti-bacterial activity of four plant extracts. International Journal of Current Research and Academic Review, 2(6), 83-94.
- Shrivastava, K., Sahu, S., Mishra, S.K., & De, K. (2014). In vitro antimicrobial activity and phytochemical screening of Syzygium aromaticum. Asian Journal of Research in Phannaceutical Science, 4(1), 12-15.
- Slinkard, K., & Singleton, V.L. (1977). Total phenol analysis: automation and comparison with manual methods, American Journal of Enology and Viticulture, 28(1),49-55.
- Souad, A., Chalghem, I., Kassah-Laouar, A., Laroui, S., & Souad, K. (2010). Activité antioxydante et antimicrobienne d'extraits de Cuminum cyminum L. Lebanese Science Journal, 11, 69-81.
- Srikacha, N., & Ratananikom, K. (2020). Antibacterial activity of plant extracts in different solvents against pathogenic bacteria: An in vitro experiment. Journal of Acute Disease, 9(5), 223-226. https://doi.org/10.4103/2221-6189.291288
- Sultana, B., Anwar, F., Mushtaq, M., Aslam, M., & Ijaz, S. (2014). In vitro antimutagenic, antioxidant activities and total phenolics of clove (Syzygium aromaticum L.) seed extracts. Pakistan Journal of Pharmaceutical Sciences, 27(4), 893-899.
- Tinh, T.H. Nuidate, T., Vuddhakul, V., & Rodkhum, C. (2016). Antibacterial Activity of Pyrogallol, a Polyphenol Compound against Vibrio parahaemolyticus Isolated from The Central Region of Thailand. Procedia Chemistry, 18, 162 168. https://doi.org/10.1016/j.proche.2016.01.025
- Ulanowska, M., & Olas, B. (2021). Biological Properties and Prospects for the Application of Eugenol - A Review. International Journal of Molecular Sciences, 22(7), 3671. https://doi.org/0.3390/ijms22073671
- Vijisaral Elezabeth, D., & Arumugam, S. (2014). GC-MS analysis of bioactive constituents of Indigofera suffruticosa leaves. Journal of Chemical and Pharmaceutical Research, 6(8), 294-300.
- Wichtl, M., & Anton, R. (1999). Plantes thérapeutiques. 2nd Ed. Lavoisier, Paris, 106-108.
- Xu, Q., Li, Y.H., & Lu, X.Y. (2007). Investigation on influencing factors of 5-HMF content in Schisandra. Journal of Zhejiang University Science B., 8(6), 439 345. https://doi.org/10.1631/jzus.2007.B0439
- Yakubu Bello, R., Umar, S., Wakil, S., & Bello, A. (2022). Antibacterial activity of clove extracts (Syzygium aromaticum) against Bacillus cereus. International Journal of Advanced Academic Research, 8(8), 33-46. https://doi.org/10.46654
- Zolkowska, D., Dhir, A., Krishnan, K., Covey, D.F., & Rogawski, M.A. (2014). Anticonvulsant potencies of the enantiomers of the neurosteroids androsterone and etiocholanolone exceed those of the natural forms. Psychopharmacology, 231 (17), 3325 3332. https://doi.org/10.1007/s00213-014-3546-x
Chemical profile by GC-MS and protective effect of Algerian cloves (Syzygium aromaticum) against Lactobacillus spp. and Streptococcus spp. isolated from dental caries
Year 2024,
Volume: 11 Issue: 4, 833 - 852, 03.11.2024
Ouahiba Benhamada
,
Nabila Benhamada
,
Lilia Boussouf
Essaid Leghouchi
Abstract
The oral cavity is home to a large and diversified microbial flora, which plays an important role in the genesis of multiple diseases, including tooth decay. Indeed, tooth decay is the most common ailment in the world, with almost everyone having experienced it at least once in their lifetime. Examining plants used in traditional medicine is one of the research approaches used to discover novel, potent antibacterial chemicals with a broad spectrum of action, as present antibacterials have significant drawbacks. This study aims to examine the chemical composition of Algerian clove Syzygium aromaticum using GC-MS and to evaluate the antibacterial activity of the methanol extract against bacteria isolated from dental caries caused by Streptococcus spp. and Lactobacillus spp. The results show a strong extraction yield of 29.7%, with high amounts of polyphenols and flavonoids calculated at 178.82 mg GAEQ/g and 24.13 mg QEQ/g. The principal chemical elements of S. aromaticum peel methanol extract were identified as eugenol (61.23%) and eugenol acetate (26.45%) based on mass spectrum data and retention times. The methanol extract has a significant antibacterial effect against tested strains, with MICs ranging from 111.37 to 445.5 mg/mL. Higher concentrations of polyphenols resulted in a significant increase in inhibition zone diameter against S1 (r2 = 0.94, p < 0.001), L3 (r2 = 0.94, p < 0.001), L5 (r2 = 0.93, p < 0.001), and L9 (r2 = 0.96, p < 0.001).
Thanks
The authors would also like to thank the Algerian Ministry of Higher Education and Scientific Research and the General Directorate of Research and Development Technologies (DGRSDT) of Algeria.
References
- Ajobiewe, H.F., Elisha, E., Ibrahim, A.E., Ajobiewe, J.O., Salami, A.O., Umeji, L.C., … Alau, K.K. (2022). Antimicrobial activity of clove plant flower bud extract (Syzygium aromaticum) on Escherichia coli. Scholars Journal of Applied Medical Sciences, 10(5), 673-684. https://doi.org/10.36347/sjams.2022.v10i05.002
- Alghazzaly, A., Elsherbiny, G., Moghannem, S., & Sharaf, M. (2022). Antibacterial, antibiofilm, antioxidants and phytochemical profiling of Syzygium aromaticum extract. Egyptian Journal of Aquatic Biology and Fisheries, 26(5), 207-218.
- Almaamori, A.M. (2023). Oral health and dental caries. World bulletin of public health, 14
- Amadi, E.K., Kareru, P.G., Keriko, J.M., & Kiptoo, J. (2016). Antimicrobial activity of selected medicinal plants’ extracts against streptococcus pneumoniae. International Journal of Botany Studies, 1(1), 20-22.
- Aneja, K.R., & Joshi, R. (2010). Antimicrobial activity of Syzygium aromaticum and its bud oil against dental cares causing microorganisms. Ethnobotanical Leajlets, 14, 960-975.
- Aneja, K.R., Joshi, R., & Sharma, C. (2009). Antimicrobial activity of Dalchini (Cinnamomum zeylanicum bark) extracts on some dental caries pathogens. Journal of Pharmacy Research, 2(9), 1387-1390.
- Arya, S.S., Rookes, J.E., Cahill, D.M., & Lenka, S.K. (2021). Vanillin: a review on the therapeutic prospects of a popular flavouring molecule. ADV TRADIT MED (ADTM), 21(3), 1–17. https://doi.org/10.1007/s13596-020-00531-w
- Bai, J., Li, J., Chen, Z., Bai, X., Yang, Z., Wang, Z., & Yang, Y. (2023). Antibacterial activity and mechanism of clove essential oil against foodborne pathogens. LWT- Food science and technology, 173, 114249. https://doi.org/10.1016/j.lwt.2022.114249
- Basli, A., Chibane, M., Madani, K., & Oukil, N. (2012). Activité antibactérienne des polyphénols extraits d’une plante médicinale de la flore d’Algérie: Origanum glandulosum Desf. Phytothérapie, 10, 2-9. https://doi.org/10.1007/s10298-012-0683-9
- Benhamada, O., Benhamada, N., & Leghouchi, E. (2022). Polyphenols and antibacterial activity of Xanthoria parietina (L.) Th. Fr. methanol extract under lead stress. Journal of Applied Biological Sciences, 16(3), 537-552. https://doi.org/10.5281/zenodo.7114289
- Bhuiyan, M.N.I., Begum, J, Nandi, N.C., & Akter, F. (2010). Constituents of the essential oil from leaves and buds of clove (Syzigium caryophyllatum (L.) Alston). African Journal of Plant Science, 4(11), 451-454.
- Brustugun, J., Tonnesen, H.H., Edge, R., & Navaratnam, S. (2005). Formation and reactivity of free radicals in 5-hydroxymethyl-2-furaldehyde–the effect on isoprenaline photostability. Journal of Photochemistry and Photobiology, 79(2), 109 119. https://doi.org/10.1016/j.jphotobiol.2004.12.005
- Cai, J-N., & Kim, D. (2023). Biofilm ecology associated with dental caries: understanding of microbial interactions in oral communities leads to development of therapeutic strategies targeting cariogenic biofilms. Advances in Applied Microbiology, 122, 27-75. https://doi.org/10.1016/bs.aambs.2023.02.001
- Cheikhyoussef, A., Cheikhyoussef, N., Rahman, A., & Hussein, A.A. (2022). Clove (Syzygium Aromaticum): Chapter 14 - Clove (Syzygium aromaticum) phenolics: Extraction, compositions, and biological activities. Chemistry, Functionality and Applications, 215-233. https://doi.org/10.1016/B978-0-323-85177-0.00036-7
- Chen, Z., Liu, Q., Zhao, Z., Bai, B., Sun, Z., Cai, L., … Xi, G. (2021). Effect of hydroxyl on antioxidant properties of 2,3 dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one to scavenge free radicals. RSC Advances, 11(55), 34456–34461. https://doi.org/10.1039/d1ra06317k
- Cortés-Rojas, D.F., de Souza, C.R.F., & Oliveira, W.P. (2014). Clove (Syzygium aromaticum): a precious spice. Asian Pacific Journal of Tropical Biomedicine, 4(2), 90-96. https://doi.org/10.1016/S2221-1691(14)60215-X
- de Freitas, T.S., da Cunha Xavier, J., Pereira, R.L.S., Rocha, J.E., Muniz, D.F., da Silva, P.T., … Coutinho, H.D.M. (2020). Direct antibacterial and antibiotic resistance modulatory activity of chalcones synthesized from the natural product 2-hydroxy-3,4,6-trimethoxyacetophenone. FEMS Microbiology Letters, 367(15). https://doi.org/10.1093/femsle/fnaa124
- Delarras, C. (2007). Microbiologie pratique pour le laboratoire d’analyse ou de contrôle sanitaire. Ed. Lavoisier. P, 128, 129, 269, 271.
- de Oliveira Carvalho, I., Purgato, G.A., Píccolo, M.S., Pizziolo, V.R., Coelho, R.R., Diaz-Muñoz, G., & Alves Nogueira Diaz, M. (2020). In vitro anticariogenic and antibiofilm activities of toothpastes formulated with essential oils. Archives of Oral Biology, 117, 104834. https://doi.org/10.1016/j.archoralbio.2020.104834
- Devi, B., Ramasubramaniaraja, R., Carism, & Devi, B. (2009). Dental caries and medicinal plants –An overview. Journal of Pharmacy Research, 2, 1669-1675.
- Diastuti, H., Chasani, M., & Suwandri, S. (2019). Antibacterial Activity of Benzyl Benzoate and Crotepoxide from Kaempferia rotunda L. Rhizome. Indonesian Journal of Chemistry, 20(1), 9. https://doi.org/10.22146/ijc.37526
- Dogruoz, N., Zeybek, Z., & Karagoz, A. (2008). Antibacterial Activity of Some Plant Extracts. IUFS Journal of Biology, 67(1), 17-21.
- Duvauchelle, V., Majdi, C., Bénimélis, D., Dunyach-Remy, C., Meffre, P., & Benfodda, Z. (2021). Synthesis, structure elucidation, antibacterial activities, and synergistic effects of novel juglone and naphthazarin derivatives against clinical methicillin-resistant Staphylococcus aureus Strains. Frontier in Chemistry, 9, 773981. https://doi.org/10.3389/fchem.2021.773981
- El-Haci, I.A., Atik-Bekkara, F., Didi, A., Gherib, M., & Didi, M.A. (2012). Teneurs en polyphénols et pouvoir antioxydant d’une plante médicinale endémique du Sahara algérien. Phytothérapie, 10, 280-285. https://doi.org/10.1007/s10298-012-0726-x
- El-Sherbiny, G.M., & Elbestawy, M.K.M. (2022). A review – plant essential oils active against Helicobacter pylori. Journal of Essential Oil Research, 34(3), 203 215. https://doi.org/10.1080/10412905.2022.2025464
- Featherstone, J.D.B. (2000). The science and the practice of caries prevention. The Journal of the American Dental Association, 131(7), 887 899. https://doi.org/10.14219/jada.archive.2000.0307
- Foda, A.M., Kalaba, M.H., El-Sherbiny, G.M., Moghannem, S.A., & El-Fakharany, E.M. (2022). Antibacterial activity of essential oils for combating colistin-resistant bacteria. Expert Review of Anti infective Therapy, 20(10), 1351 1364. https://doi.org/10.1080/14787210.2022.2101997
- Freitas, T.S., da Cunha Xavier, J., Pereira, R.L.S., Rocha, J.E., Muniz, D.F., da Silva, P.T., … Coutinho, H.D.M. (2020). Direct antibacterial and antibiotic resistance modulatory activity of chalcones synthesized from the natural product 2 hydroxy 3,4,6 trimethoxyacetophenone. FEMS-Microbiology Letters, 367(15). https://doi.org/10.1093/femsle/fnaa124
- Frohlich, P.C., Santos, K.A., Ascari, J., dos Santos Refati, J.R., Palú, F., Cardozo-Filho, L., & da Silva, E.A. (2023). Antioxidant compounds and eugenol quantification of clove (Syzygium aromaticum) leaves extracts obtained by pressurized liquid extraction and supercritical fluid extraction. The Journal of Supercritical Fluids, 196, 105865. https://doi.org/10.1016/j.supflu.2023.105865
- Gengatharan, A., & Abd Rahim, M.H. (2023). The application of clove extracts as a potential functional component in active food packaging materials and model food systems: A mini-review. Applied Food Research, 3(1), 100283. https://doi.org/10.1016/j.afres.2023.100283
- Gupta, S., Park, S.E., Mozaffari, S., El-Aarag, B., Parang, K., & Tiwari, R.K. (2023). Design, Synthesis, and Antiproliferative Activity of Benzopyran-4-One-Isoxazole Hybrid Compounds. Molecules, 28(10), 4220. https://doi.org/10.3390/molecules28104220
- Gyrdymova, Y., & Rubtsova, S. (2021). Caryophyllene and caryophyllene oxide: a variety of chemical transformations and biological activities. Chemical Papers. 76(2). https://doi.org/10.1007/s11696-021-01865-8
- Hemeda, N.A., Hegazy, G.E., Abdelgalil, S., Soliman, N.A., Abdel-Meguid, D.I., & El-Assar, S.A. (2022). Maximization of red pigment production from Streptomyces sp. LS1 structure elucidation and application as antimicrobial/antifouling against human pathogens and marine microbes. Journal of Genetic Engineering and Biotechnology, 20, 168. https://doi.org/10.1186/s43141-022-00452-y
- Huang, L., Zhu, X., Zhou, S., Cheng, Z., Shi, K., Zhang, C., & Shao, H. (2021). Phthalic Acid Esters: Natural Sources and Biological Activities. Toxins (Basel), 13(7), 495. https://doi.org/10.3390/toxins13070495
- Hugar, S., M Patel, P., Nagmoti, J., Uppin, C., Mistry, L., & Dhariwal, N. (2017). An in vitro comparative evaluation of efficacy of disinfecting ability of garlic oil, neem oil, clove oil, and tulsi oil with autoclaving on ndodontic K Files tested against Enterococcus faecalis. International Journal of Clinical Pediatric Dentistry, 10(3), 283 288. https://doi.org/10.5005/jp-journals-10005-1451
- Jadhav, J.J., Jadeja, G.C., & Desai, M.A. (2022). Clove (Syzygium aromaticum): Chapter 28 - Effect of extraction techniques on the yield, composition, and quality of clove (Syzygium aromaticum) essential oil. Chemistry, Functionality and Applications, 485-500. https://doi.org/10.1016/B978-0-323-85177-0.00027-6
- Joseph, B., & Sujatha, S. (2011). Bioactive Compounds and its autochthonous microbial activities of extract and clove oil (Syzygium aromaticum L.) on some food borne pathogens. Asian Journal of Biological Sciences, 4(1), 35-43. https://doi.org/10.3923/ajbs.2011.35.43
- Kablan, B.J., Adiko, M., & Abrogoua, D.P. (2008). Évaluation in vitro de l'activité antimicrobienne de Kalanchoe crenata et de Manotes longiflora utilisées dans les ophtalmies en Côte d'ivoire. Phytothérapie, 6(5), 282-288. https://doi.org/10.1007/s10298-008-0332-0
- Kabra, P., Loomba, K., Kabra, S.K., Majumdar, D.S.P., & Kuma, N. (2012). Medicinal plants in the treatment of dental caries. Asian Journal of Oral Health & Allied Sciences, 2(1), 12-16.
- Kaur, D., & Chandrul, K. (2017). Syzygium aromaticum L. (Clove): A vital herbal drug used in periodontal disease. Indian Journal of Pharmaceutical and Biological Research, 5(2), 45-51. https://doi.org/10.30750/ijpbr.5.2.9
- Khan, M. K., Hassan, S., Imran, M., & Ahmad, M.H. (2022). Clove (Syzygium aromaticum): Chapter 22 - Extraction of bioactive compounds from clove (Syzygium aromaticum). Chemistry. Functionality and Applications, 405-417. https://doi.org/10.1016/B978-0-323-85177-0.00010-0
- Kiralan, M., & Ketenoglu, O. (2022). Clove (Syzygium aromaticum): Chapter 36 - Food applications of clove (Syzygium aromaticum) extracts. Chemistry, Functionality and Applications, 607-617. https://doi.org/10.1016/B978-0-323-85177-0.00030-6
- Kuete, V., Dongfack, M.D., Mbaveng, A.T., Lallemand, M-C., Van-Dufat, H.T., Wansi, J-D., … Wandji, J. (2010). Antimicrobial activity of the methanolic extract and compounds from the stem bark of Drypetes tessmanniana. Chinese Journal of Integrative Medicine, 16(4), 337-343. https://doi.org/10.1007/s11655-010-0527-8
- Li, Y.H., & Lu, X.Y. (2005). Investigation on the origin of 5-HMF in Shengmaiyin decoction by RP-HPLC method. Journal of Zhejiang University Science B, 6, 1015–1021. https://doi.org/10.1631/jzus.2005.B1015
- Mallya, P.S., & Mallya, S. (2020). Microbiology and clinical implications of dental caries – a review. Journal of Evolution of Medical and Dental Sciences, 9(48), 3670-3675. https://doi.org/10.14260/jemds/2020/805
- Mazumder, K., Nabila, A., Aktar, A., & Farahnaky, A. (2020). Bioactive Variability and In Vitro and In Vivo Antioxidant Activity of Unprocessed and Processed Flour of Nine Cultivars of Australian lupin Species: A Comprehensive Substantiation. Antioxidants (Basel), 9(4), 282. https://doi.org/10.3390/antiox9040282
- Meda, A., Lamien, C.E., Romito, M., Millogo, J., & Nacoulma, O.G. (2005). Determination of the total phenolic, flavonoid and proline contents in Burkina Fasan Honey, as well as their radical scavenging activity. Food Chemistry, 91(3), 571 577. https://doi.org/10.1016/j.foodchem.2004.10.006
- Mohammadi Ziarani, G., Kheilkordi, Z., & Mohajer, F. (2020). Recent advances in the application of acetophenone in heterocyclic compounds synthesis. Journal of the Iranian Chemical Society, 17, 247–282. https://doi.org/10.1007/s13738-019-01774-4
- Moroh, J-L.A. (2008). Étude de l’activité antibactérienne de l’extrait acétatique (EAC) de Morinda morindoides (Baker) milne-redheat (rubiaceae) sur la croissance in-vitro des souches d’Escherichia coli. Bulletin de la Societe Royale des Sciences de Liege, 77. 44 - 61.
- Mostafa, A.A., Al-Askar, A.A., Almaary, K.S., Dawoud, T.M., Sholkamy, E.N., & Bakri, M.M. (2018). Antimicrobial activity of some plant extracts against bacterial strains causing food poisoning diseases. Saudi Journal of Biological Sciences, 25(2), 361 366. https://doi.org/10.1016/j.sjbs.2017.02.004
- Munson, M.A., Banerjee, A., Watson, T.F., & Wade, W.G. (2004). Molecular analysis of the microflora associated with dental caries. Journal of clinical microbiology, 42(7), 3023-3029. https://doi.org/10.1128/JCM.42.7.3023-3029.2004
- Nakilcioglu, T.E., & Otles, S. (2021). Influence of extraction solvents on thepolyphenol contents, compositions, and antioxidant capacities of fig (Ficus carica L.) seeds. Annals of the Brazilian Academy of Sciences Printed, 93(1), 1678-2690. https://doi.org/10.1590/0001-3765202120190526
- Nkere, C.K., & Iroegbu, C.U. (2005). Antibacterial screening of the root, seed and stem bark extracts of Picralima nitida. African Journal of Biotechnology, 4(6), 522-526.
- Oliveira, L.C.C., Rodrigues, F.A.A., dos Santos Barbosa, C.R., dos Santos, J.F.S., Macêdo, N.S., de Sousa Silveira, Z., Coutinho, H.D.M., & da Cunha, F.A.B. (2022). Antibacterial Activity of the Pyrogallol against Staphylococcus aureus Evaluated by Optical Image. Biologics, 2, 139-150. https://doi.org/10.3390/biologics2020011
- Oshomoh, E.O., ldu, M., & Udinyiwe, O.C. (2015). Phytochemical screening and antimicrobial sensitivity of clove flower (Syzygium aromaticum, L. Merrill and Perry) bud on dental pathogens. International Journal of Pharmacy and Pharmaceutical Research, 3(2), 1-12.
- Owen, P.L., & Johns, T. (1999). Xanthine oxidase inhibitory activity of northeastern North American plant remedies used forgout. Journal of Ethnopharmacology, 64,149-160. https://doi.org/10.1016/s0378-8741(98)00119-6
- Penot, E. (2016). Analyse technico-économique de la filière amont de production d’essence de girofle à Fénérive- Est, Madagascar: de la feuille à l’alambic.
- Prajapati, R.A., & Raol, B.V. (2014). Characterization of microbial flora isolated from human teeth carious lesions. Life Sciences Leajlets, 47, 01-08.
- Qasim, M., Aziz, I., Rasheed, M., Gul, B., & Khan, M. (2016). Effect of extraction solvents on polyphenols and antioxidant activity of medicinal halophytes. Pakistan Journal of Botany, 48(2), 621-627.
- Ratri, P.J., Ayurini, M., Khumaini, K., & Rohbiya, A. (2020). Clove oil extraction by steam distillation and utilization of clove buds waste as potential candidate for Eco-Friendly Packaging. Jurnal Bahan Alam Terbarukan, 9(1), 47 54. https://doi.org/10.15294/jbat.v9i1.24935
- Reddy, D.S., & Rogawski, M.A. (2012). “Neurosteroids - Endogenous Regulators of Seizure Susceptibility and Role in the Treatment of Epilepsy”. In Noebels JL, Avoli M, Rogawski MA, et al. (eds.). Jasper’s Basic Mechanisms of the Epilepsies [Internet]. 4th Edition. Bethesda (MD): National Center for Biotechnology Information (US). National Center for Biotechnology Information (US). PMID 22787590
- Riaz̽, A., Umar, M., & Farah, N. (2023). Prevalence, causes and management of dental caries. Journal of Xi’an Shiyou University, Natural Science Edition, 19(5), 1033-1053.
- Rodrigues, L.B., Martins, A.O.B.P.B., Cesário, F.R.A.S., Castro, F.F., de Albuquerque, T.R., Fernandes, M.N.M., … de Menezes, I.R.A. (2016). Anti-inflammatory and antiedematogenic activity of the Ocimum basilicum essential oil and its main compound estragole: In vivo mouse models. Chemico-Biological Interactions, 257, 14-25. https://doi.org/10.1016/j.cbi.2016.07.026
- Ryu, C.K., Kim, D.H., Kim, H.J., & Chung, S.Y. (1993). The antimicrobial activities of some 1,4 naphthalenediones (IV). Archives of Pharmacal Research, 16, 327 330. https://doi.org/10.1007/BF02977525
- Sabriu – Haxhijaha, A., Jordanovska, S., & Popovska, O. (2021). Analysis of clove (Syzygium aromaticum L.) oil obtained with hydrodistillation and ultrasound-assisted extraction. UDC582.776.2-113.551:542.61. https://doi.org/10.20544/HORIZONS.B.09.2.P07
- Saikumari, D., Shiva Rani, S.K., & Saxena, N. (2016). Antibacterial Activity of Syzigium aromaticum L. (Clove). International Journal of Current Microbiology and Applied Sciences, 5 (11), 484-489. http://dx.doi.org/10.20546/ijcmas.2016.511.056
- Saini, M., Alam, S., Prajapati, R., Kr, L., & Thakur (2019). Analysis and estimation of eugenol content in microemulsion formulation containing clove oil (Syzigium aromaticum). Journal of Applicable Chemistry, 8(1), 195-202.
- Shafira, K., Azad, A.K., Zubair, L., & Helal Uddin, A.B.M. (2020). Extraction and quantification of eugenol from clove buds using HPLC. Current Chromatography, 07(1) https://doi.org/10.2174/2213240607999200818161356
- Sharaf, M.H., El-Sherbiny, G.M., Moghannem, S.A., Abdelmonem, M., Islam, A., Elsehemy, I.A., … Kalaba, M.H. (2021). New combination approaches to combat methicillin-resistant Staphylococcus aureus (MRSA). Scientific Reports, 11(1), 1 16. https://doi.org/10.1038/s41598-021-82550-4
- Shehadi, M., Awada, F., Oleik, R., Chokr, A., Hamze, K., Hamdan, H.A., & Kobaissi, A. (2014). Comparative analysis of the anti-bacterial activity of four plant extracts. International Journal of Current Research and Academic Review, 2(6), 83-94.
- Shrivastava, K., Sahu, S., Mishra, S.K., & De, K. (2014). In vitro antimicrobial activity and phytochemical screening of Syzygium aromaticum. Asian Journal of Research in Phannaceutical Science, 4(1), 12-15.
- Slinkard, K., & Singleton, V.L. (1977). Total phenol analysis: automation and comparison with manual methods, American Journal of Enology and Viticulture, 28(1),49-55.
- Souad, A., Chalghem, I., Kassah-Laouar, A., Laroui, S., & Souad, K. (2010). Activité antioxydante et antimicrobienne d'extraits de Cuminum cyminum L. Lebanese Science Journal, 11, 69-81.
- Srikacha, N., & Ratananikom, K. (2020). Antibacterial activity of plant extracts in different solvents against pathogenic bacteria: An in vitro experiment. Journal of Acute Disease, 9(5), 223-226. https://doi.org/10.4103/2221-6189.291288
- Sultana, B., Anwar, F., Mushtaq, M., Aslam, M., & Ijaz, S. (2014). In vitro antimutagenic, antioxidant activities and total phenolics of clove (Syzygium aromaticum L.) seed extracts. Pakistan Journal of Pharmaceutical Sciences, 27(4), 893-899.
- Tinh, T.H. Nuidate, T., Vuddhakul, V., & Rodkhum, C. (2016). Antibacterial Activity of Pyrogallol, a Polyphenol Compound against Vibrio parahaemolyticus Isolated from The Central Region of Thailand. Procedia Chemistry, 18, 162 168. https://doi.org/10.1016/j.proche.2016.01.025
- Ulanowska, M., & Olas, B. (2021). Biological Properties and Prospects for the Application of Eugenol - A Review. International Journal of Molecular Sciences, 22(7), 3671. https://doi.org/0.3390/ijms22073671
- Vijisaral Elezabeth, D., & Arumugam, S. (2014). GC-MS analysis of bioactive constituents of Indigofera suffruticosa leaves. Journal of Chemical and Pharmaceutical Research, 6(8), 294-300.
- Wichtl, M., & Anton, R. (1999). Plantes thérapeutiques. 2nd Ed. Lavoisier, Paris, 106-108.
- Xu, Q., Li, Y.H., & Lu, X.Y. (2007). Investigation on influencing factors of 5-HMF content in Schisandra. Journal of Zhejiang University Science B., 8(6), 439 345. https://doi.org/10.1631/jzus.2007.B0439
- Yakubu Bello, R., Umar, S., Wakil, S., & Bello, A. (2022). Antibacterial activity of clove extracts (Syzygium aromaticum) against Bacillus cereus. International Journal of Advanced Academic Research, 8(8), 33-46. https://doi.org/10.46654
- Zolkowska, D., Dhir, A., Krishnan, K., Covey, D.F., & Rogawski, M.A. (2014). Anticonvulsant potencies of the enantiomers of the neurosteroids androsterone and etiocholanolone exceed those of the natural forms. Psychopharmacology, 231 (17), 3325 3332. https://doi.org/10.1007/s00213-014-3546-x