The Isolation, Characterisation, And Evaluation Of Bioactivities Of Secondary Metabolites From Leaf Extracts Of Karamunting  (Rhodomyrtus Tomentosa (Aiton) Hassk)

Marwati Marwati; Yusnita  Rifai; Gemini   alam; Risfah  Yulianty

doi:10.22146/ijp.10363

Marwati Marwati 1.Department of Pharmaceutical Science and Technology, Faculty of Pharmacy, Hasanuudin University, Makassar, 90245, Indonesia and 2.Department of Biology , Faculty of Pharmacy , University of Almarisah Madani, 90242, Indonesian
Yusnita Rifai Department of Pharmaceutical Science and Technology, Faculty of Pharmacy, Hasanuudin University, Makassar, Jalan Perintis Kemerdekaan Km.10, Tamalanrea Indah, Kec. Tamalanrea, Kota Makassar, Sulawesi Selatan 90245, Indonesia
Gemini alam 3Department of Phytochemistry, Faculty of Pharmacy, Hasanuddin University, Makassar, Jalan Perintis Kemerdekaan Km.10, Tamalanrea Indah, Kec. Tamalanrea, Kota Makassar, Sulawesi Selatan 90245, Indonesia and Drug Discovery and Development Centre, Institue of Research and Community Service, Hasanuddin University, Jalan Perintis Kemerdekaan Km.10, Tamalanrea Indah, Kec. Tamalanrea, Kota Makassar, Sulawesi Selatan 90245, Indonesia
Risfah Yulianty

Keywords: Rhodomyrtus tomentosa (Aiton) Hassk, Isolation, cytotoxic, Anticancer

Abstract

Cancer is a condition of abnormal cell development with one of the p53 gene mutation pathways that can molecularly and biologically lead to the formation of cancer cells, and abnormalities in the p53 gene occur in 60% of cancer cases. Under normal conditions, p53 has an important role in cell division. One of the plants that can be developed as an anticancer agent is leaves R. tomentosa Which has chemical compounds such as phenolic compounds, flavonoids, tannins, saponins, and terpenoids. This study aims to isolate, identify, and determine the bioactivity of secondary metabolite compounds in extracts, fractions, and methanol isolates of R. tomentosa leaves against cancer cells. The samples used were from Sulu Sopai Village, Tana Toraja, South Sulawesi Province. Fine powdered leaves of R. tomentosa leaves were macerated with 70% ethanol solvent. The ethanol extract obtained was partitioned with n-hexane, ethyl acetate, and ethanol-water n-hexane and then fractionated by the isolation of compounds carried out by chromatographic techniques, including KLT, The spectra of the isolated compounds were determined using FT-IR, and then the extracts, fractions, and isolates were tested for cytotoxic properties and selectivity against MCF-7, T47D, WiDr, HeLa, and normal cells (Vero Cell). by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolim bromide (MTT) method. The results showed that the extracts and fractions contained flavonoids, tannins, saponins, alkaloids, steroid compounds, and isolates from R. tomentosa leaves. Identification with the KLT Three Eluent System showed a single yellow stain on the KLT plate, which is a typical color of flavanoid compounds, with Rfs of 0.35, 0.67, and 0.72. The AlCl3 reagent test also showed that the isolate obtained was a flavanoid compound group. As well as FTIR spectrum data showing the presence of C-O, C=O, and O-H primary groups, the isolate obtained is a flavanoid compound group with a melting point of 288–290 °C. The bioactivity test results of extracts, fractions, and isolates of R. tomentosa leaves show that the results obtained in ethanol-water extracts have activity as anticancer in inhibiting cancer cell proliferation with IC₅₀ values obtained in MCF-7 cells of 9.66 µg/mL (strong), T47D 59.47 µg/mL (medium), WiDr 39.21 µg/mL (strong), HeLa 199.6 µg/mL (medium), and on normal cells (Vero Cell) has a weak toxic effect with an IC₅₀ value of 1292.50 µg/mL. And doxorubicin, 3.2753 µg/mL. Based on the results obtained from this study, it can be concluded that the ethanol-water fraction of R. tomentosa leaves has anticancer activity on MCF-7, T47D, WiDr, and HeLa cancer cells. and has a weak toxic effect on normal cells (vero cells) compared to ethanol extract, ethyl acetate fraction, n-hexane fraction, and isolates. The ethanol extract and ethanol-water fraction of R. tomentosa leaves are active and have active selectivity.

References

Aisyah, A., Lukitaningsih, E., Rumiyati, R., Marwati, M., Sapra, A., Khairi, N., Fadri, A., Nur, S.,2022. In Vitro antioxidant and cytotoxic evaluation of ethyl acetate fraction of Angiopteris ferox Copel tuber against HTB lung cancer cell. Egypt. J. Chem. 0, 0-0 https://doi.org/10.21608/ejchem.2022.91843.4364
Akinmoladun, A.C., Falaiye, O.E., Ojo, O.B., Adeoti, A., Amoo, Z.A., Olaleye, M.T., 2022. Effect of extraction technique, solvent polarity, and plant matrix on the antioxidant properties of Chrysophyllum albidum G. Don (African Star Apple). Bull Natl Res Cent 46, 40. https://doi.org/10.1186/s42269-022-00718-y
Chae, S.W., Sohn, J.H., Kim, D.-H., Choi, Y.J., Park, Y.L., Kim, K., Cho, Y.H., Pyo, J.-S., Kim, J.H., 2011. Overexpression of Cyclin B1, cdc2, p16 and p53 in Human Breast Cancer: The Clinicopathological Correlations and Prognostic Implications. Yonsei Med J 52, 445. https://doi.org/10.3349/ymj.2011.52.3.445.
Dai, J., Mumper, R.J., 2010. Plant Phenolics: Extraction, Analysis and Their Antioxidant and Anticancer Properties. Molecules 15,7313-7352. https://doi.org/10.3390/molecules15107313.
Devlin, E.J., Denson, L.A., Whitford, H.S., 2017. Cancer Treatment Side Effects: A Meta-analysis of the Relationship Between Response Expectancies and Experience. Journal of Pain and Symptom Management 54, 245-258.e2. https://doi.org/10.1016/j.jpainsymman.2017.03.017
Emens, L.A., Middleton, G., 2015. The Interplay of Immunotherapy and Chemotherapy: Harnessing Potential Synergies. Cancer Immunol Res 3, 436-443. https://doi.org/10.1158/2326-6066.CIR-15-0064
Fischer, K.M., 2022. Cancer Treatment: Past, Present And Future. Journal Of Cancer Research, Treatment & Prevention. https://doi.org/10.37191/Mapsci-JCRTP-1(1)-1
Gayathri, V., Kiruba, D., 2014. Phytochemical Analysis Of Leaf Powder Extracts Of Rhodomyrtus Tomentose. International Journal Of Current Research 6, 6527-6530.
Globocan, 2020a. Cancer today. International Agency for Research on Cancer 1-2.
Globocan, 2020b. Indonesia. International Agency for Research on Cancer.
Hamid, H.A., Mutazah, S.S.Z.R., Mashitah M Yusoff, 2016. Rhodomyrtus Tomentosa: Phytochemical And Pharmacological Review. Asian J Pharm Clin Res 10, 10. https://doi.org/10.22159/ajpcr.2017.v10i1.12773.
Hassanpour, S.H., Dehghani, M., 2017. Review of cancer from a molecular perspective. Journal of Cancer Research and Practice 4, 127-129. https://doi.org/10.1016/j.jcrpr.2017.07.001.
Khairani, S., Keban, S.A., Afrianty, M., 2019. Evaluation of Drug Side Effects Chemotherapy on Quality of Life (QOL) Breast Cancer Patients at Hospital X in Jakarta. jifi 17, 9. https://doi.org/10.35814/jifi.v17i1.705
Kuete, V., Karaosmanoglu, O., Sivas, H., 2017. Chapter 10 - Anticancer Activities of African Medicinal Spices and Vegetables. Medicinal Spices and Vegetables from Africa 27.
Kusuma, I.W., Ainiyati, N., Suwinarti, W., 2016. Search for biological activities from an invasive shrub species rose myrtle (Rhodomyrtus tomentosa). Nusantara Biosci 8, 55-59. https://doi.org/10.13057/nusbiosci/n080110.
Liu, H.-X., Tan, H.-B., Qiu, S.-X., 2016a. Antimicrobial acylphloroglucinols from the leaves of (Rhodomyrtus tomentosa). Journal of Asian Natural Products Research 18, 535-541. https://doi.org/10.1080/10286020.2015.1121997.
Liu, H.-X., Zhang, W.-M., Xu, Z.-F., Chen, Y.-C., Tan, H.-B., Qiu, S.-X., 2016b. Isolation, synthesis, and biological activity of tomentosenol A from the leaves of Rhodomyrtus tomentosa. Royal Society of Chemistry 6, 25882-25886. https://doi.org/10.1039/c6ra01594h.
Luliana, S., Riza, H., Indriyani, E.N., 2019. The Effect of the Extraction Method on Total Phenolic Content and Antioxidant Activity of Salam Leaves (Syzygium polyanthum) using DPPH (1,1-Diphenyl-2-Picrylhydrazyl). Trad.Med.J 24, 72. https://doi.org/10.22146/mot.33955
Marwati, M., Anggriani, A., Burhan, A., Awaluddin, A., Nur, S., Dharmayanti, R., Lilingan, E., Tiboyong, M.D., 2021. Antioxidant Activity and Cytotoxicity Against WiDR Cell and Vero Cell of The Karamunting (Rhonomyrtus tomentosa L.) Leaves Ethanol Extract. IJPST 8, 111. https://doi.org/10.24198/ijpst.v8i3.26769.
Marwati, Rifai, Y., Alam, G., Yulianty, R., Nursamsiar, 2022. Toxicity Test of Karamunitng Leaf (Rhodomyrtus tomentosa (Aiton) Hassk.) Extract with Finder Liquid Variation Using the Brine Shrimp Lethality Test (BSLT) Method. Presented at the Tapanuli International Health Conference 2022, Atlantis Press, pp. 103-109. https://doi.org/doi.org/10.2991/978-94-6463-032-9_16.
Marwati, Salampe, M., Burhan, A., Megawati, Khairuddin, Naneng, A.A.A.M., Oktaviani, N., 2020. Antioxidant And Anticancer Screening Of Ethanol Extract Of Karamunting (Rhonomyrtus Tomentosa L.) Leaves As An Alternative Medicine. Scientific Journal of Manutung 6, 6.
Na-Phatthalung, P., Teles, M., Voravuthikunchai, S.P., LluísTort, Fierro-Castro, C., 2018. Immunomodulatory effects of Rhodomyrtus tomentosa leaf extract and its derivative compound, rhodomyrtone on head kidney macrophages of rainbow trout (Oncorhynchus mykiss). Fish physiology and biochemistry 44, 543-555. https://doi.org/10.1007/s10695-017-0452-2.
Nur, S., Aisyah, A.N., Lukitaningsih, E., Rumiyati, Juhardi, R.I., Rezkiawati Andirah, Hajar, A.S., 2021. Evaluation of antioxidant and cytotoxic effect against cancer cells line of Angiopteris ferox Copel tuber and its compounds by LC-MS analysis. J Appl Pharm Sci. https://doi.org/10.7324/JAPS.2021.110808.
Nur, S., Aisyah, A.N., Lukitaningsih, E., Rumiyati, R., Fadri, A., Marwati, M., 2022. Cytotoxic Effect of the Paku Atai Merah (Angiopteris ferox Copel) Fraction on MCF-7 and HeLa Cells and its Compound Profile by GC-MS. Borneo J Pharm 5, 48-55. https://doi.org/10.33084/bjop.v5i1.2943.
Nur, S., Aisyah, A.N., Sami, F.J., Fadri, A., Khairi N., Sapra, A., 2023. Standardisation and GC-MS Analysis of Kersen (Muntingia calabura L.) Fruit Ethanol Extract As An Herbal Raw Material. Bulletin of Pharmaceutical Sciences. Assiut 46, 173-187. https://doi.org/10.21608/bfsa.2023.300915.
Saising, J.,Voravuthikunchai, S.P., 2012. Anti Propionibacterium acnes activity of rhodomyrtone, an effective compound from Rhodomyrtus tomentose (Aiton) Hassk. leaves. Elsevier Ltd Anaerobe 18, 400-404. https://doi.org/10.1016/j.anaerobe.2012.05.003.
Salni, S., Marisa, H., 2020. Antibacterial Activity of Essential Oil from Rosemytle Leaves (Rhodomyrtus tomentosa (Ait.) Hassk). Molecule 15, 158. https://doi.org/10.20884/1.jm.2020.15.3.601.
Salsabila, S., Hartati, R., Al Muqarrabun, L.M.R., Qomaladewi, N.P., Haniffadli, A., Rosandy, A.R., Chahyadi, A., Elfahmi, E., 2022. Isolation of bioactive compounds with tyrosinase inhibitory activity from the methanol extract of meniran herb (Phyllanthus niruri 401 Linn.).crbb 3, 196-201. https://doi.org/10.5614/crbb.2022.3.2/GOO6ZULV.
Shah, R., Rosso, K., Nathanson, S.D., 2014. Pathogenesis, prevention, diagnosis, and treatment of breast cancer. WJCO 5, 283. https://doi.org/10.5306/wjco.v5.i3.283.
Vo, T., Ngo, D., 2019. The Health-Beneficial Properties of Rhodomyrtus Tomentose as Potential Functional Food. Biomolecules 9, 76. https://doi.org/10.3390/biom9020076