Antioxidant Activity of 96% Ethanol Extract of Pepaya Jepang Leaves (Cnidoscolus aconitifolius (Mill.) I. M. Johnst) Using DPPH Method (1,1-diphenyl-2-picrylhydrazyl)

Riskianto Riskianto (1) , Melieta Windi (2) , Karnelasatri Karnelasatri (3) , Maroloan Aruan (4)
(1) Universitas Pelita Harapan , Indonesia
(2) Universitas Pelita Harapan , Indonesia
(3) Universitas Pelita Harapan , Indonesia
(4) Universitas Pelita Harapan , Indonesia


Free radicals in the human body can be helped neutralized by natural antioxidants derived from plants. Pepaya Jepang (Cnidoscolus aconitifolius (Mill.) I.M.Johnst) contains antioxidant compounds such as flavonoids and polyphenols. This study examined the antioxidant activity of 96% ethanol extract of C. aconitifolius leaves. Maceration of C. aconitifolius leaves using 96% ethanol solvent, phytochemical screening, and antioxidant activity assay was carried out with 1,1-Diphenyl-2-Picryl Hydrazil (DPPH) on 96% ethanol extract of C. aconitifolius. Extraction obtained a yield of 16.1834%, and phytochemical screening on the 96% ethanol extract of C. aconitifolius contained flavonoids, tannins, saponins, phenols, alkaloids, and steroids. The antioxidant activity assay on 96% ethanol extract of C. aconitifolius had an IC50 value of 145.3855 ppm and an IC50 value of vitamin C of 7.0806 ppm. The Antioxidant Activity Index (AAI) of 96% ethanol extract of C. aconitifolius was 0.4127, and the AAI of vitamin C was 8.4739. Based on the results obtained, although in a weak classification, a 96% ethanol extract of C. aconitifolius leaves has antioxidant activity.

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1. Phaniendra A, Jestadi DB, Periyasamy L. Free radicals: properties, sources, targets, and their implication in various diseases. Indian J Clin Biochem. 2015;30(1):11-26. doi:10.1007/s12291-014-0446-0
2. Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Pharmacogn Rev. 2010;4(8):118-26. doi:10.4103/0973-7847.70902
3. Kurutas EB. The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: current state. Nutr J. 2016;15(1):71. doi:10.1186/s12937-016-0186-5
4. Lourenço SC, Moldão-Martins M, Alves VD. Antioxidants of Natural Plant Origins: From Sources to Food Industry Applications. Molecules. 2019;24(22):4132. doi:10.3390/molecules24224132
5. Pem D, Jeewon R. Fruit and Vegetable Intake: Benefits and Progress of Nutrition Education Interventions- Narrative Review Article. Iran J Public Health. 2015;44(10):1309-21.
6. Riskianto, Kamal SE, Aris M. Aktivitas Antioksidan Ekstrak Etanol 70% Daun Kelor (Moringa oleifera LAM) terhadap DPPH. J Pro-Life. 2021; 8(2):168-77. doi:10.33541/jpvol6Iss2pp102
7. Kasote DM, Katyare SS, Hedge MV, Bae H. Significance of antioxidant potential of plants and its relevance to therapeutic applications. Int J Biol Sci. 2015;11(8):982-91. doi:10.7150/ijbs.12096
8. Orji OU, Ibiam UA, Aja PM, Okechukwu PCU, Uraku AJ, Aloke C, et al. Evaluation of the Phytochemical and Nutritional Profiles of Cnidoscolus aconitifolius Leaf Collected in Abakaliki South East Nigeria. World J Med Sci. 2016;13(3):213-7. doi:10.5829/idosi.wjms.2016.213.217
9. Obichi EA, Monago CC, Belonwu DC. Effect of Cnidoscolus aconitifolius (Family Euphorbiaceae) Aqueous Leaf Extract on Some Antioxidant Enzymes and Haematological Parameters of High Fat Diet and Streptozotocin Induced Diabetic Wistar Albino Rats. J Appl Sci Environ Manag. 2015;19(2):201-9. doi:10.4314/jasem.v19i2.5
10. Oyeyemi MO, Ajani OS. Haematological parameters and serum testosterone of West African dwarf rams treated with aqueous extract of Cnidoscolus aconitifolius (Chaya). J Med Plants Res. 2014;8(14):571–5. doi:10.5897/jmpr2013.5185
11. Mordi JC. Antidiabetic potential of the aqueous leaf extract of Cnidoscolus aconitifolius on streptozotocin Induced Diabetes in Wistar rat hepatocytes. Curr Res J Biol Sci. 2012;4(2):164-7.
12. Azeez OI, Oyagbemi AA, Oyeyemi MO, Odetola AA. Ameliorative effects of Cnidoscolus aconitifolius on alloxan toxicity in Wistar rats. Afr Health Sci. 2010;10(3):283–91.
13. Victor M, Abbey PA, Joseph Y, Jonathan Z, Bobai YK, Maria O. An Underexploited Tropical Plant with Promising Economic Value and the Window of Opportunities for Researchers: Cnidoscolus aconitifolius. Am J Food Sci Nutr Res. 2016;3(6):177-87.
14. Donkoh A, Atuahene CC, Poku-Prempeh YB, Twum IG. The nutritive values of chaya leaf meal (Cnidoscolus aconitifolius) studies with broilers chickens. Anim Feed Sci Technol. 1999;77(1-2):163–72. doi:10.1016/S0377-8401(98)00231-4
15. Achi NK, Ohaeri OC, Ijeh II, Eleazu C. Modulation of the lipid profile and insulin levels of streptozotocin induced diabetic rats by ethanol extract of Cnidoscolus aconitifolius leaves and some fractions: Effect on the oral glucose tolerance of normoglycemic rats. Biomed Pharmacother. 2017;86:562–9. doi:10.1016/j.biopha.2016.11.133
16. Oboh G. Effect of Some Post-Harvest Treatment on the Nutritional Properties of Cnidoscolus aconitifolus leaf. Pak J Nutr. 2005;4(4):226-30. doi:10.3923/pjn.2005.226.230
17. Oyagbemi AA, Odetola AA, Azeez OI. Ameliorative effects of Cnidoscolus aconitifolius on anaemia and osmotic fragility induced by protein energy malnutrition. Afr J Biotechnol. 2008;7(11):1721-6. doi:10.5897/AJB08.247
18. Oladeinde FO, Kinyua AM, Laditan AA, Michelin R, Bryant JL, Denaro F, et al. Effect of Cnidoscolus aconitifolius leaf extract on blood glucose and insulin levels of inbred type 2 diabetic mice. Cell Mol Biol (Noisy-le-grand). 2007;53(3):34-41.
19. Sarmiento-Franco L, Sandoval-Castro CA, McNab JM, Quijano–Cervera R, Reyes-Ramirez RR. Effect of age of regrowth on chemical composition of chaya (Cnidoscolus aconitifolius) leaves. J Sci Food Agric. 2003;83(6):609-12. doi:10.1002/jsfa.1372
20. Awoyinka AO, Balogun IO, Ogunnowo AA. Phytochemical screening and in vitrobioactivity of Cnidoscolus aconitifolius (Euphorbiaceae). J Med Plant Res. 2007;1(3):63-5.
21. Ross-Ibarra J, Molina-Cruz A. The Ehnobotany of Chaya (Cnidoscolus aconofifolius): A nutritious maya vegetable. Econ Bot. 2002;56:350-64. doi:10.1663/0013-0001(2002)056[0350:TEOCCA]2.0.CO;2
22. Chukwu EC, Osuocha KU, Iwueke AV. Phytochemical profiling, body weight effect and anti-hypercholesterolemia potentials of Cnidoscolus aconitifolius leaf extracts in male albino rat. J Pharmacogn Phytother. 2020;12(2):19–27. doi:10.5897/jpp2016.0436
23. Akar Z, Küçük M, Doğan H. A new colorimetric DPPH• scavenging activity method with no need for a spectrophotometer applied on synthetic and natural antioxidants and medicinal herbs. J Enzyme Inhib Med Chem. 2017;32(1):640-7. doi:10.1080/14756366.2017.1284068
24. Altemimi A, Lakhssassi N, Baharlouei A, Watson DG, Lightfoot DA. Phytochemicals: Extraction, Isolation, and Identification of Bioactive Compounds from Plant Extracts. Plants. 2017;6(4):42. doi:10.3390/plants6040042
25. Gangwar M, Gautam MK, Sharma AK, Tripathi YB, Goel RK, Nath G. Antioxidant capacity and radical scavenging effect of polyphenol rich Mallotus philippenensis fruit extract on human erythrocytes: an in vitro study. ScientificWorldJournal. 2014;2014:279451. doi:10.1155/2014/279451
26. Scherer R, Godoy HT. Antioxidant activity index (AAI) by the 2,2-diphenyl-1-picrylhydrazyl method. Food Chem. 2009;112(3):654-8. doi:10.1016/j.foodchem.2008.06.026
27. Riskianto, Soemardji AA, Tan MI. Cytotoxic Effects of Kirinyuh Herb (Austroeupatorium inulaefolium (Kunth) R. d. King & H. Robinson) Extracts and Fractions on BSLT, MCF-7 Cells and T-47D Cells. Pharmacogn J. 2022;14(2):374-8. doi:10.5530/pj.2022.14.48
28. Abubakar AR, Haque M. Preparation of medicinal plants: Basic extraction and fractionation procedures for experimental purposes. J Pharm Bioallied Sci. 2020;12(1):1-10. doi:10.4103/jpbs.JPBS_175_19
29. Ullah A, Munir S, Badshah SL, Khan N, Ghani L, Poulson BG, et al. Important Flavonoids and Their Role as a Therapeutic Agent. Molecules. 2020;25(22):5243. doi:10.3390/molecules25225243
30. Unuofin JO, Lebelo SL. Antioxidant Effects and Mechanisms of Medicinal Plants and Their Bioactive Compounds for the Prevention and Treatment of Type 2 Diabetes: An Updated Review. Oxid Med Cell Longev. 2020;2020:1356893. doi:10.1155/2020/1356893
31. Kumar N, Goel N. Phenolic acids: Natural versatile molecules with promising therapeutic applications. Biotechnol Rep. 2019;24:e00370. doi:10.1016/j.btre.2019.e00370
32. Soldado D, Bessa RJB, Jerónimo E. Condensed Tannins as Antioxidants in Ruminants-Effectiveness and Action Mechanisms to Improve Animal Antioxidant Status and Oxidative Stability of Products. Animals. 2021;11(11):3243. doi:10.3390/ani11113243
33. Thawabteh A, Juma S, Bader M, Karaman D, Scranno L, Bufo SA, et al. The Biological Activity of Natural Alkaloids against Herbivores, Cancerous Cells and Pathogens. Toxins. 2019;11(11):656. doi:10.3390/toxins11110656
34. Kotta S, Ansari SH, Ali J. Exploring scientifically proven herbal aphrodisiacs. Pharmacogn Rev. 2013;7(13):1-10. doi:10.4103/0973-7847.112832
35. Kedare SB, Singh RP. Genesis and development of DPPH method of antioxidant assay. J Food Sci Technol. 2011;48(4):412-22. doi:10.1007/s13197-011-0251-1
36. Macan AM, Kraljević TG, Raić-Malić S. Therapeutic Perspective of Vitamin C and Its Derivatives. Antioxidants. 2019;8(8):247. doi:10.3390/antiox8080247
37. Phongpaichit S, Nikom J, Rungjindamai N, Sakayaroj J, Hutadilok-Towatana N, Rukachaisirikul V, et al. Biological activities of extracts from endophytic fungi isolated from garcinia plants. FEMS Immunol Med Microbiol. 2007;51(3):517-25. doi:10.1111/j.1574-695x.2007.00331.x
38. Ajiboye BO, Oyinloye BE, Agboinghale PE, Ojo OA. Cnidoscolus aconitifolius (Mill.) I. M. Johnst leaf extract prevents oxidative hepatic injury and improves muscle glucose uptake ex vivo. J Food Biochem. 2019;43(12):e13065. doi:10.1111/jfbc.13065
39. Hamid AA, Oguntoye SO, Negi AS, Ajao A, Owolabi NO. Chemical constituents, antibacterial, antifungal and Antioxidant activities of the aerial parts of Cnidoscolus aconitifolius. Ife J Sci. 2016;18(2):561-71.
40. Ajiboye BO, Ojo OA, Okesola MA, Oyinloye BE, Kappo AP. Ethyl acetate leaf fraction of Cnidoscolus aconitifolius (Mill.) I. M. Johnst: antioxidant potential, inhibitory activities of key enzymes on carbohydrate metabolism, cholinergic, monoaminergic, purinergic, and chemical fingerprinting. Int J Food Prop. 2018;21(1):1697-715. doi:10.1080/10942912.2018.1504787


Riskianto Riskianto (Primary Contact)
Melieta Windi
Karnelasatri Karnelasatri
Maroloan Aruan
Author Biographies

Riskianto Riskianto, Universitas Pelita Harapan

Pharmacy Diploma Study Program, Universitas Pelita Harapan, Tangerang, Banten, Indonesia

Melieta Windi, Universitas Pelita Harapan

Pharmacy Diploma Study Program, Universitas Pelita Harapan, Tangerang, Banten, Indonesia

Karnelasatri Karnelasatri, Universitas Pelita Harapan

Pharmacy Diploma Study Program, Universitas Pelita Harapan, Tangerang, Banten, Indonesia

Maroloan Aruan, Universitas Pelita Harapan

Medical Laboratory Technology Study Program, Universitas Pelita Harapan, Tangerang, Banten, Indonesia

Riskianto R, Windi M, Karnelasatri K, Aruan M. Antioxidant Activity of 96% Ethanol Extract of Pepaya Jepang Leaves (Cnidoscolus aconitifolius (Mill.) I. M. Johnst) Using DPPH Method (1,1-diphenyl-2-picrylhydrazyl). Borneo J Pharm [Internet]. 2022Nov.30 [cited 2023Jan.31];5(4):315-24. Available from:

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