Evaluation of Clastogenic Potential of Ethanolic Extract of Leaf of <i>Couroupita guianensis</i> using Micronucleus Test in Mice


  • Department of Toxicology (Mutagenicity), Jai Research Foundation, Valvada – 396105, Gujarat
  • Department of Toxicology (Mutagenicity), Jai Research Foundation, Valvada – 396105, Gujarat
  • Department of Toxicology (Mutagenicity), Jai Research Foundation, Valvada – 396105, Gujarat
  • Department of Toxicology (Mutagenicity), Jai Research Foundation, Valvada – 396105, Gujarat
  • Department of Ecotoxicology, Jai Research Foundation, Valvada – 396105, Gujarat
  • C.G. Bhakta Institute of Biotechnology, Uka Tarsadia University (UTU), Maliba Campus, Bardoli – 394350, Gujarat
  • Department of Toxicology (Mutagenicity), Jai Research Foundation, Valvada – 396105, Gujarat
  • Department of Toxicology (Mutagenicity), Jai Research Foundation, Valvada – 396105, Gujarat
  • C.G. Bhakta Institute of Biotechnology, Uka Tarsadia University (UTU), Maliba Campus, Bardoli – 394350, Gujarat




<i>Couroupita guianensis</i>, Ethanolic Leaf Extract, Genotoxicity, Medicinal Plant, Micronucleus Test


Medicinal plants are being used since ancient times in different tribes in India for numerous medicinal purposes. The essential facets of plant-based medicines, such as safety, quality, and efficiency, must be evaluated. Couroupita guianensis is a medicinally important plant, being used conventionally for curing varieties of medical ailments. From the safety point of view, limited information exists for the C. guianensis plant. Considering the facts mentioned above, this study evaluates the genotoxic potential of ethanolic leaf extract of C. guianensis, through an in vivo micronucleus test , which was conducted on mice (Strain: Hsd: ICR (CD1)). The main study consisted of three groups (5 male mice per group): vehicle control (corn oil), a treatment group (2000 mg/kg body weight), and positive control (Mitomycin-C, 1.0 mg/kg body weight). The methodology followed was the standard in vivo procedure as per the internationally approved OECD TG 474 guidelines with a two-day dosing schedule separated by 24 hours. Dose-volume used was 10 mL/kg body weight. Mice in each group exhibited normal behavior and were found clinically normal after dosing. Toxicity to bone marrow, as well as any statistically significant rise in the percent micronucleated polychromatic erythrocytes (% MNPCE), was not detected in mice that were treated with C. guianensis at 2000 mg/kg body weight when compared with the data of concurrent vehicle control. Statistically, a significant rise in the % MNPCE was obtained in mice treated with Mitomycin-C (1.0 mg/kg body weight), which confirmed the test system's suitability, the procedures, and efficiency of conditions employed in the test. Our study concluded that the plant is not showing any genotoxicity and can be used as a medicine to cure various reported ailments.


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Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy by CK Kokate. 16th ed.; 2001.

Trease G, Evans WJP. The Pharmacological Action of Plant Drugs. 12th ed.; 1983.

Kavitha K. Effect of Courouptia guianensis on n-diethylnitrosamine induced oxidative stress in wistar rats. JKK Nattraja Coll Pharmacy, Komar. 2012.

Idu M, Onyibe HI, Timothy O, Erhabor JO. Ethnomedicinal flora of Otuo people of Edo State, Nigeria. Asian J Plant Sci. 2008; 7(1):8–12. https://doi.org/10.3923/ajps.2008.8.12

Sheba LA, Anuradha V. An updated review on Couroupita guianensis Aubl: A sacred plant of India with myriad medicinal properties. J Herbmed Pharmacol. 2020; 9(1):1–11. https://doi.org/10.15171/jhp.2020.01

Priyadarshini N, Veeramani A. Antitubercular compounds isolated and characterized in Tithonia diversifolia and Couroupita guianensis. Int J Myco bacteriology. 2020; 9(2):195–9.

Ilangovan S, Thavasumani P. Preliminary screening of phytochemical constituents, antioxidant and antimicrobicidal activities in the methanolic leaf extract of Couroupita guianensis. Asian J Pharm Clin Res. 2021:203–6. https://doi.org/10.22159/ajpcr.2021. v14i1.39923

Jiang M, Cui BW, Wu YL, Nan JX, Lian LH. Genus Gentiana: A review on phytochemistry, pharmacology and molecular mechanism. J Ethnopharmacol. 2021; 264. https://doi.org/10.1016/j.jep.2020.113391. PM id:32931880

Elumalai A, Eswaraiah M, Didala A. Investigations on anti-oxidant, anti-arthritic and anti-platelet studies in Couroupita guianensis aubl leaves by in-vitro methods. Pharma Sci Monit. 2012; 3(3):2262–9.

Umachigi SP, Jayaveera KN, Kumar CKA, Kumar GS. Antimicrobial, wound healing and antioxidant potential of Couroupita guianensis in rats. Pharmacologyonline. 2007; 3:269–81.

Khan MR, Kihara M, Omoloso AD. Antibiotic Activity of Couroupita guianensis. J Herbs, Spices Med Plants. 2003; 10(3):95–108. https://doi.org/10.1300/J044v10n03_10

Kavitha R, Kamalakannan P, Deepa T, Elamathi R, Sridhar S, Kumar Suresh J. In vitro antimicrobial activity and phytochemical analysis of Indian medicinal plant Couroupita guianensis aubl. J Chem Pharm Res. 2011; 3(6):115–21.

Sivakumar T, Shankar T, Geetha G. Efficacy of Couroupita guianensis against selected human pathogens. Adv Biol Res (Rennes). 2012; 6(2):59–63.

Prabhu V, Ravi S. Quantification of quercetin and stigmasterol of Couroupita guianensis aubl by HPTLC method and in-vitro cytototoxic activity by MTT assay of the methanol extract against HeLa, NIH 3T3 and HepG2 cancer cell lines. Int J Pharm Pharm Sci. 2012; 4(4):126–30.

Begum K, Motobayashi T, Hasan N, Appiah KS, Shammi M, Fujii Y. Indigo as a plant growth inhibitory chemical from the fruit pulp of Couroupita guianensis Aubl. Agronomy. 2020; 10(9). https://doi.org/10.3390/agronomy10091388

Mattana CM, Cangiano MA, Alcaráz LE, et al. Evaluation of cytotoxicity and genotoxicity of acacia aroma leaf extracts. Sci World J. 2014; 2014. https://doi.org/10.1155/2014/380850. PMid:25530999. PMC id:PMC4228826

Plewa MJ, Wagner ED. Activation of promutagens by green plants. Annu Rev Genet. 1993; 27:93–113. https://doi.org/10.1146/annurev.ge.27.120193.000521. PMid:8122914

Gothai S, Vijayarathna S, Chen Y, et al. In vitro and in vivo-scientific evaluation on cytotoxicity and genotoxicity of traditional medicinal plant Couroupita guianensis aubl. flower. Pharmacologyonline. 2019; 2:24–38.

Hadi NSA, Bankoglu EE, Schott L, et al. Genotoxicity of selected pyrrolizidine alkaloids in human hepatoma cell lines HepG2 and Huh6. Mutat Res - Genet Toxicol Environ Mutagen. 2021:861–2. https://doi.org/10.1016/j.mrgentox.2020.503305. PMid:33551105

Cha SB, Kim SS, Oh JJ, et al. Evaluation of the in vitro and in vivo genotoxicity of a Dioscorea Rhizome water extract. Toxicol Res. 2021. https://doi.org/10.1007/s43188-020-00077-8

Lee S-B, Lee J-S, Wang J-H, et al. Genotoxicity of Water extract from bark-removed rhus verniciflua stokes. molecules. 2021; 26(4):896. https://doi.org/10.3390/molecules26040896. PMid:33567750. PMCid:PMC7914431

Shokrzadeh M, Habibi E, Shadboorestan A, Chabra A, Ahmadi A. The protective effects of Origanum vulgare L. extract on genetic damage of cyclophosphamide in mice blood lymphocytes using micronucleus test. Pharm Biomed Res. 2021. https://doi.org/10.18502/pbr.v6i4.5116

Ganaie AH. Review of the active principles of medicinal and aromatic plants and their disease fighting properties. In: Medicinal and Aromatic Plants. 2021:1–36. https://doi.org/10.1016/B978-0-12-819590-1.00001-X

George SA, Bhadran S, Sudhakar M, Harini BP. Comprehensive in vitro evaluation of pharmacological activities of selected mass spectrometry profiling of Flacourtia jangomas flower extract. Asian J Pharm Clin Res. 2017; 10(5):237–44. https://doi.org/10.22159/ajpcr.2017.v10i5.17419

OECD. Test No. 474: Mammalian Erythrocyte Micronucleus Test. OECD; 2016. https://doi.org/10.1787/9789264264762-en

EFSA. Guidance for submission for food additive evaluations. EFSA J. 2012; 10(7). https://doi.org/10.2903/j.efsa.2012.2760

Hardy A, Benford D, Halldorsson T, et al. Clarification of some aspects related to genotoxicity assessment. EFSA J. 2017; 15(12):5113. https://doi.org/10.2903/j.efsa.2017.5113. PMid:32625393

Asanami S, Shimono K. Hypothermia induces micronuclei in mouse bone marrow cells. Mutat Res - Genet Toxicol Environ Mutagen. 1997; 393(1–2):91–8. https://doi.org/10.1016/S1383-5718(97)00089-2

Asanami S, Shimono K. High body temperature induces micronuclei in mouse bone marrow. Mutat Res - Genet Toxicol Environ Mutagen. 1997; 390(1–2):79–83. https://doi.org/10.1016/S0165-1218(97)00002-5

Asanami S, Shimono K, Kaneda S. Transient hypothermia induces micronuclei in mice. Mutat Res - Genet Toxicol Environ Mutagen. 1998; 413(1):7–14. https://doi.org/10.1016/S1383-5718(98)00004-7

MacGregor JT, Heddle JA, Hite M, et al. Guidelines for the conduct of micronucleus assays in mammalian bone marrow erythrocytes. Mutat Res Toxicol. 1987; 189(2):103–12. https://doi.org/10.1016/0165-1218(87) 90016-4

Krishna G, Hayashi M. In vivo rodent micronucleus assay: Protocol, conduct and data interpretation. Mutat Res - Fundam Mol Mech Mutagen. 2000; 455(1– 2):155–66. https://doi.org/10.1016/S0027-5107(00)00 117-2

Heddle JA, Stuart E, Salamone MF. The Bone Marrow Micronucleus Test. In: Handbook of Mutagenicity Test Procedures. 1984:441–57. https://doi.org/10.1016/B978-0-444-80519-5.50025-0

Gad SC, Weil C. Statistics for toxicologists. In: Principles and methods of toxicology. 3rd Editio. Raven press Ltd., New York; 1994:221–74.

Jasmine FS, Moorthi VP. Couroupita guianensis: The reservoir of medicinal compounds of human welfare. Asian J Pharm Clin Res. 2017; 10(3):50–2. https://doi.org/10.22159/ajpcr.2017.v10i3.16174

Landge L. Ameliorative role of flower extract of Couroupita guianensis on chloramphenicol induced hematological changes in Mus musculus. Int J Res Anal Rev. 2018; 5:22–6.

Yadav A, Mendhulkar V. Repellency and toxicity of Couroupita guianensis leaf extract against Silverleaf Whitefly (Bemisia tabaci). Int J Sci Res Publ. 2014; 5(4):1–4.

de Queiroz FM, de Oliveira Matias KW, da Cunha MMF, Schwarz A. Evaluation of (anti)genotoxic activities of Phyllanthus niruri L. in rat bone marrow using the micronucleus test. Brazilian J Pharm Sci. 2013; 49(1):137–48. https://doi.org/10.1590/S1984- 82502013000100015

Sousa HG, Uchí´a VT, Cavalcanti SMG, et al. Phytochemical screening, phenolic and flavonoid contents, antioxidant and cytogenotoxicity activities of Combretum leprosum Mart. (Combretaceae). J Toxicol Environ Heal - Part A Curr Issues. 2021. https://doi.org/10.1080/15287394.2021.1875345. PMid:33494643

Boriollo MFG, Resende MR, da Silva TA, et al. Evaluation of the mutagenicity and antimutagenicity of Ziziphus joazeiro Mart. Bark in the micronucleus assay. Genet Mol Biol. 2014; 37(2):428–38. https://doi.org/10.1590/S1415-47572014000300016. PMid:25071 409. PMCid:PMC4094613

Lee JS, Cho JH, Lee DS, Son CG. Genotoxicity evaluation of an ethanol extract mixture of astragali radix and salviae miltiorrhizae radix. Evidence-based Complement Altern Med. 2018; 2018. https://doi.org/10.1155/2018/5684805. PMid:30402128. PMCid: PMC6198562

Mohamed HM, Aly MS. Evaluation of genotoxicity of Euphorbia triaculeata Forssk. extract on mice bone marrow cells in vivo. Toxicol Reports. 2018; 5(April):625–31. https://doi.org/10.1016/j.toxrep.2018.05.007. PM id:29854632. PMCid:PMC5977870

Ji KY, Kim KM, Oh JJ, et al. Assessment of the 4-week repeated-dose oral toxicity and genotoxicity of GHX02. J Appl Toxicol. 2020; 40(2):270–84. https://doi.org/10.1002/jat.3902. PMid:31515828. PMCid:P MC7004199

Dodda S, Alluri VK, Golakoti T, Sengupta K. Acute, subacute, and genotoxicity assessments of a proprietary blend of Garcinia mangostana fruit rind and Cinnamomum tamala leaf extracts (CinDura®). J Toxicol. 2020; 2020. https://doi.org/10.1155/2020/1435891. PMid:32802054. PMCid: PMC7414347

Food. International Conference on Harmonisation; guidance on S2(R1) Genotoxicity testing and data interpretation for pharmaceuticals intended for human use; availability. Notice. Fed Regist. 2012.

Sahu R, Divakar G, Divakar K. In vivo rodent micronucleus assay of Gmelina arborea Roxb (Gambhari) extract. J Adv Pharm Technol Res. 2010; 1(1):22–9.

Gupta VH, Gunjal MA, Wankhede SS, Deshmukh VS. Neuropharmacological evaluation of the methanolic extract of Couroupita guianensis Aubl. flower in mice. Int J Res Stud Biosci. 2012; 3(10):115–21.

Elumalai A, Bargavi K, Krishna S, Eswaraiah MC. Evaluation of anti-oxidant and hepatoprotective activity of Couroupita guianensis Leaves. J Cell Tissue Res. 2013; 13(2):3745–8.

Han EH, Lim MK, Lee SH, Rahman MM, Lim YH. An oral toxicity test in rats and a genotoxicity study of extracts from the stems of Opuntia ficus-indica var. saboten. BMC Complement Altern Med. 2019; 19(1). https://doi.org/10.1186/s12906-019-2442-7. PMid:306 91445. PMCid:PMC6350306

Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Investig Suppl. 1968; 97:77–89.

Pradhan D, Panda PK, Tripathy G. Evaluation of the immunomodulatory activity of the methanolic extract of Couroupita guianensis aubl. Flowers in rats. Nat Prod Radiance. 2009; 8(1):37–42.



How to Cite

Nagane, R. M., Desai, K. R., Barad, I. M., Patel, M. V., Rana, J. R., Sheth, U. V., Patel, V. M., Patel, K. A., & Krishnamurthy, R. (2021). Evaluation of Clastogenic Potential of Ethanolic Extract of Leaf of <i>Couroupita guianensis</i> using Micronucleus Test in Mice. Toxicology International, 28(2), 187–198. https://doi.org/10.18311/ti/2021/v28i2/27433



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