Preliminary Bioactivity Profiling of Methanol Crude Extract from Clinacanthus nutans Leaves: Antioxidant, Antimicrobial, and Apoptotic Properties
DOI:
https://doi.org/10.54987/jobimb.v13i1.1085Keywords:
C. nutans, Antioxidant, Antimicrobial, Apoptosis, Anti-cancerAbstract
Clinacanthus nutans (C. nutans), or “Belalai gajah,” is traditionally used to treat microbial infections, cancer, inflammation, and diabetes due to its rich phytochemical profile. This study aimed to extract bioactive compounds from C. nutans leaves using varying methanolic concentrations (100%, 70%, and 60% v/v) and evaluate their total phenolic content (TPC), antioxidant, antimicrobial, and apoptotic activities. Extraction yields were 8.50%, 15.80%, and 14.70% for 100%, 70%, and 60% methanol extracts, respectively. The 100% methanol extract showed the highest TPC (23.40 mg GAE/g), followed by 70% (20.48 mg GAE/g) and 60% (20.32 mg GAE/g). Antioxidant activity assessed by DPPH assay showed the strongest radical scavenging in the 100% extract (IC₅₀ = 1.13 mg/mL), followed by 70% (1.87 mg/mL) and 60% (2.82 mg/mL). Only the 60% and 70% extracts exhibited antimicrobial activity with a minimum inhibitory concentration (MIC) of 10 mg/mL. Cytotoxicity assessed by MTT assay on CT26 cells revealed the lowest IC₅₀ for the 100% extract (0.18 ± 0.071 mg/mL), followed by 70% (0.20 ± 0.062 mg/mL) and 60% (0.49 ± 0.042 mg/mL). Apoptotic populations in CT26 cells, determined via Trypan Blue assay, were 47.34% ± 8.89, 44.37% ± 2.34, and 43.00% ± 0.95 for the 100%, 70%, and 60% extracts, respectively. These findings suggest that C. nutans exhibits promising antioxidant, antimicrobial, and anticancer properties, supporting its potential as a natural therapeutic agent. Further studies are needed to explore its mechanisms, dosage optimization, and synergistic interactions with other compounds.
References
Azemi AK, Mokhtar SS, Rasool AH. Clinacanthus nutans: Its potential against diabetic vascular diseases. Braz J Pharm Sci. 2020;56:e18838.
Fazil FN, Azzimi NS, Yahaya BH, Kamalaldin NA, Zubairi SI. Kinetics extraction modelling and antiproliferative activity of Clinacanthus nutans water extract. Sci World J. 2016;2016(1):7370536.
Manzitto?Tripp EA, Darbyshire I, Daniel TF, Kiel CA, McDade LA. Revised classification of Acanthaceae and worldwide dichotomous keys. Taxon. 2022 Feb;71(1):103-53.
Alam A, Ferdosh S, Ghafoor K, Hakim A, Juraimi AS, Khatib A, Sarker ZI. Clinacanthus nutans: A review of the medicinal uses, pharmacology and phytochemistry. Asian Pac J Trop Med. 2016 Apr 1;9(4):402-9.
Kai Xiang T & Shu Xian C. Phytochemical Screening and Antibacterial Properties of Methanol, Hexane and Water Extracts of Clinacanthus nutans. Malays J Chem. 2023;25(4):199-206.
Turkan I. ROS and RNS: key signalling molecules in plants. J Exp Bot. 2018 Jun 19;69(14):3313-5.
Xu DP, Li Y, Meng X, Zhou T, Zhou Y, Zheng J, Zhang JJ, Li HB. Natural antioxidants in foods and medicinal plants: Extraction, assessment and resources. Int J Mol Sci. 2017 Jan 5;18(1):96.
Arumugam A, Wong NK. Comparison study of antioxidant, antimicrobial and cytotoxic properties of secondary metabolite and protein extracts from Clinacanthus nutans. Int J Pharm Pharm Sci. 2020;12:98-102.
Yong YK, Tan JJ, Teh SS, Mah SH, Ee GC, Chiong HS, Ahmad Z. Clinacanthus nutans extracts are antioxidant with antiproliferative effect on cultured human cancer cell lines. Evid Based Complement Alternat Med. 2013;2013(1):462751.
Raya KB, Ahmad SH, Farhana SF, Mohammad M, Tajidin NE, Parvez A. Changes in phytochemical contents in different parts of Clinacanthus nutans (Burm. f.) lindau due to storage duration. Bragantia. 2015 Aug 21;74(4):445-52.
Syed Najmuddin SU, Romli MF, Hamid M, Alitheen NB, Nik Abd Rahman NM. Anti-cancer effect of Annona Muricata Linn Leaves Crude Extract (AMCE) on breast cancer cell line. BMC Complement Altern Med. 2016 Dec;16:1-20.
Akinduti PA, et al. Suitability of spectrophotometric assay for determination of honey microbial inhibition. J Phys Conf Ser. 2019 Aug 1;1299(1):012131.
Riyadi PH, Susanto E, Anggo AD, Arifin MH, Rizki L. Effect of methanol solvent concentration on the extraction of bioactive compounds using ultrasonic-assisted extraction (UAE) from Spirulina platensis. Food Res. 2023;7(3):59-66.
Speight JG. Environmental organic chemistry for engineers. Butterworth-Heinemann. 2016 Nov 2.
Brini E, et al. How water's properties are encoded in its molecular structure and energies. Chem Rev. 2017;117(19):12385-12414.
Borges A, et al. Comparison of Techniques and Solvents on the Antimicrobial and Antioxidant Potential of Extracts from Acacia dealbata and Olea europaea. Antibiotics (Basel). 2020 Feb;9(2):48.
Mustapa AN, et al. Extraction of phytocompounds from the medicinal plant Clinacanthus nutans Lindau by microwave-assisted extraction and supercritical carbon dioxide extraction. Ind Crops Prod. 2015 Nov 15;74:83-94.
Ghasemzadeh A, Jaafar HZ, Rahmat A. Antioxidant activities, total phenolics and flavonoids content in two varieties of Malaysia young ginger (Zingiber officinale Roscoe). Molecules. 2010 Jun 14;15(6):4324-33.
Yee HS, Eng CB, Eng HS. Evaluation of antioxidant activity of Clinacanthus nutans (Acanthaceae). Short Commun Biotechnol. 2017;4:63-73.
Bakar MF, et al. Phytochemicals and antioxidant activity of different parts of bambangan (Mangifera pajang) and tarap (Artocarpus odoratissimus). Food Chem. 2009 Mar 15;113(2):479-83.
Lee SY, et al. Antioxidant and alpha.-glucosidase inhibitory activities of the leaf and stem of selected traditional medicinal plants. Int Food Res J. 2014;21(1):379.
Chiangchin S, Thongyim S, Pandith H, Kaewkod T, Tragoolpua Y, Inta A, et al. Clinacanthus nutans genetic diversity and its association with anti-apoptotic, antioxidant, and anti-bacterial activities. Sci Rep. 2023 Nov 10;13(1):19566.
Ismail NZ, et al. Antioxidant effects, antiproliferative effects, and molecular docking of Clinacanthus nutans leaf extracts. Molecules. 2020 Apr 29;25(9):2067.
Susanti N, et al. Phytochemical, metabolite compound, and antioxidant activity of Clinacanthus nutans leaf extract from Indonesia. Sci Technol Indones. 2023;8(1):38-44.
Kowalska-Krochmal B, Dudek-Wicher R. The minimum inhibitory concentration of antibiotics: methods, interpretation, clinical relevance. Pathogens. 2021 Feb 4;10(2):165.
Arullappan S, Rajamanickam P, Thevar N, Kodimani CC. In vitro screening of cytotoxic, antimicrobial and antioxidant activities of Clinacanthus nutans (Acanthaceae) leaf extracts. Trop J Pharm Res. 2014 Oct 10;13(9):1455-61.
Aligiannis N, et al. Composition and antimicrobial activity of the essential oils of two Origanum species. J Agric Food Chem. 2001 Sep 17;49(9):4168-70.
Mogana R, et al. Antibacterial activities of the extracts, fractions and isolated compounds from Canarium patentinervium Miq. against bacterial clinical isolates. BMC Complement Med Ther. 2020 Dec;20:1-1.
Lim SH, et al. Antibacterial activity of Clinacanthus nutans polar and non-polar leaves and stem extracts. Biomed Pharmacol J. 2020 Sep 1;13:1169-74.
Dehelean CA, et al. Plant-derived anticancer compounds as new perspectives in drug discovery and alternative therapy. Molecules. 2021 Feb 19;26(4):1109.
Gong G, et al. Unveiling the neuroprotective properties of isoflavones: current evidence, molecular mechanisms and future perspectives. Crit Rev Food Sci Nutr. 2024 May 18:1-37.
Al-Yousef HM, et al. UPLC?ESI?MS/MS profile and antioxidant, cytotoxic, antidiabetic, and antiobesity activities of the aqueous extracts of three different hibiscus species. J Chem. 2020;2020(1):6749176.
Phung JH, Fong IL, Khong HY, Ban WK. In vitro antioxidant and anticancer activities of Clinacanthus nutans extracts. Sci Eng Health Stud. 2021 Dec 27:21050014.
Fong SY, Piva T, Dekiwadia C, Urban S, Huynh T. Comparison of cytotoxicity between extracts of Clinacanthus nutans (Burm. f.) Lindau leaves from different locations and the induction of apoptosis by the crude methanol leaf extract in D24 human melanoma cells. BMC Complement Altern Med. 2016 Sep 20;16(1):368.
Artun T, et al. In vitro anticancer and cytotoxic activities of some plant extracts on HeLa and Vero cell lines. J BUON. 2016 May 1;21(3):720-5.
Rashidi M, et al. Selective cytotoxicity and apoptosis-induction of Cyrtopodion scabrum extract against digestive cancer cell lines. Int J Cancer Manag. 2017 May 1;10(5):e8633.
Bendale Y, et al. Evaluation of cytotoxic activity of platinum nanoparticles against normal and cancer cells and its anticancer potential through induction of apoptosis. Integr Med Res. 2017 Jun 1;6(2):141-8.
Indrayanto G. The importance of method validation in herbal drug research. J Pharm Biomed Anal. 2022 May 30;214:114735.
Sun D, et al. Why 90% of clinical drug development fails and how to improve it?. Acta Pharm Sin B. 2022 Jul 1;12(7):3049-62.
Hussein HA, et al. Cytotoxicity and 1H NMR metabolomics analyses of microalgal extracts for synergistic application with Tamoxifen on breast cancer cells with reduced toxicity against Vero cells. Heliyon. 2022 Mar 1;8(3).
Riss T, et al. Cytotoxicity assays: in vitro methods to measure dead cells. Assay Guid Man. 2019 May 1.
Maioli E, et al. Critical appraisal of the MTT assay in the presence of rottlerin and uncouplers. Biol Proced Online. 2009 Dec;11:227-40.
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Copyright (c) 2025 Syed Umar Faruq Syed Najmuddin, Rasidah Abdul Rajak, Nur Aina Aqilah Arlihamdi, Ahmad Muqri Akhiroman, Nurul Fadhilah Amirah Abdul Zim, Andrea Sheena Albert Switzer Chia, Nur Adlina Syuhada Anurul Hiraq, Mohd Khalizan Sabullah
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