Cytotoxic Evaluation of Eurycoma longifolia Jack Root Extract on Chromosome Aberrations in Human Lymphocytes In vitro

Somkid Chaiphech(1), Isara Patawang(2), Sumalee Phimphan(3), Sukhonthip Ditcharoen(4), Chatmongkon Suwannapoom(5), Alongklod Tanomtong(6*)

(1) Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group on Toxic Substances in Livestock and Aquatic Animals, Khon Kaen University, Khon Kaen 40002, Thailand
(2) Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50000; Center of Excellence in Bioresources for Agriculture, Industry and Medicine, Chiang Mai University, Chiang Mai, 50000 Thailand
(3) Biology program, Faculty of Science and Technology, Phetchabun Rajabhat University, Phetchabun 67000, Thailand
(4) Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi Khlong 6, Khlong Luang, Pathum Thani 12110, Thailand
(5) Department of Fishery, School of Agriculture and Natural Resources, University of Phayao, 56000,
(6) Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen 40002; Research Group on Toxic Substances in Livestock and Aquatic Animals, Khon Kaen University, Khon Kaen 40002, Thailand
(*) Corresponding Author


This study aimed to investigate chromosomal aberrations of Eurycoma longifolia Jack (EL) root extract in human lymphocytes in vitro. Human whole blood was cultured in medium solution that treated with distilled water, 20% DMSO, extract of EL roots at the concentration of 2.5, 5, 10, 20, 40, 80 µg/mL (extracted with distilled water and ethanol), and nontreated (blank: only culture medium and whole blood). All experiments were cultured for 72 hours in the 37°C incubator. The effects of EL roots extract on cytotoxicity were compared with the control groups including the blank, distilled water, and 20% DMSO. This study found that EL root extract significantly decreased metaphase cell number and increased chromosome aberrations dose dependent manner (p<0.01). The 7 types of chromosome aberration that were observed consisted of dicentric chromosome, single chromatid breaks, isochromatid break, isochromatid gap, single chromatid gap, fragmentation, and deletion. The dicentric chromosome was the most common chromosomal aberrations type that was treated with EL root extract both distilled water and ethanol. Moreover, the ethanolic extract of EL root was more effective to stimulate chromosome aberrations compared to the water extract of EL root (the deletion and fragmentation were not found in the water extract of EL root). This study demonstrated that the phytochemicals of EL root extract had cytotoxicity effect (decreased metaphase cells and increase cells death) and genotoxic effect (increased chromosomal aberrations. The use of EL root crude extract with distilled water is therefore safer for cells. However, when EL is used at high levels, it may lead to the inhibition of cell division process and cause side effects (toxicity). EL extracts consist of various phytochemicals with different properties and dosages, thus more studies should be conducted on the effect of those substances on cytotoxicity, especially their effects on genotoxicity humans.



biotechnology; Cell biology

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Aida, E.M.I. et al., 2016. Assessment of genetic and reproductive toxicity of Tongkat Ali Plus®. International Journal of Pharma and Bio Sciences, 7(2), pp.492–499.

Ang, H.H., Cheang, H.S. & Yusof, A.P., 2000. Effects of Eurycoma longifolia Jack (Tongkat Ali) on the initiation of sexual performance of inexperienced castrated male rats. Experimental Animals, 49(2), pp.35–38. doi: 10.1538/expanim.49.35.

Ang, H.H. & Cheang, H.S., 2001. Effects of Eurycoma longifolia jack on laevator ani muscle in both uncastrated and testosterone-stimulated castrated intact male rats. Archives of Pharmacal Research, 24(5), pp.437–440. doi: 10.1007/BF02975191.

Askin C.T. & Aslantürk, O.S., 2018. Cytotoxic and Genotoxic Effects of Aqueous Extracts of Rosmarinus officinalis L., Lavandula stoechas L. and Tilia cordata Mill. on in vitro Human Peripheral Blood Lymphocytes. Cumhuriyet Science Journal, 39(1), pp.127–143. doi: 10.17776/csj.405629.

Bhat et al., 2017. Toxicological evaluation and effective dose selection of eurycomanone, a quassinoid of Eurycoma longifolia plant in fishes. Aquaculture, 481, pp.94–102. doi: 10.1016/j.aquaculture.2017.08.030.

Bhat, R. & Karim, A.A., 2010. Tongkat Ali (Eurycoma longifolia Jack): a review on its ethnobotany and pharmacological importance. Fitoterapia, 81(7), pp.669–679. doi: 10.1016/j.fitote.2010.04.006.

Choo, C.Y. & Chan, K.L., 2002. High performance liquid chromatography analysis of canthinone alkaloids from Eurycoma longifolia. Planta Medica, 68, pp.382–384.

Freshney, R.I., 2005. Culture of animal cell: a manual of basic technique. 5thed. John Wiley and Sons, Inc., Hoboken, New Jersey.

Galloway, S.M., 2000. Cytotoxicity and chromosome aberrations in vitro: experience in industry and the case for an upper limit on toxicity in the aberration assay. Environmental and Molecular Mutagenesis, 35(3), pp.191–201.

George, A., Kawasaki, Y., & Abas, A.B., 2016. Eurycoma Longifolia Extract and Its Use in Enhancing and/or Stimulating Immune System Available Source: e82f3/ WO2016036232A2.pdf. August 20, 2019.

Han, S.N. et al., 2016. Vitamin E supplementation increases T helper 1 cytokine production in old mice infected with influenza virus. Journal of Immunology, 100(4), pp.487–493. doi: 10.1046/j.1365-2567.2000.00070.x.

Hien, D.T. et al., 2019. Anti-inflammatory effects of alkaloid enriched extract from roots of Eurycoma longifolia Jack. Asian Pacific Journal Tropical Biomedicine, 9, pp.18–23. doi: 10.4103/2221-1691.250265.

Kavitha, N. et al., 2010. Cytotoxicity activity of root extract/fractions of Eurycoma longifolia jack root against Vero and Hs27cells. Journal of Medicinal Plants Research, 4(22), pp.2383–2387. doi: 10.5897/JMPR10.568.

Khanam, Z., Wen, C.S. & Bhat, I.U.H., 2015. Phytochemical screening and antimicrobial activity of root and stem extracts of wild Eurycoma longifolia Jack (Tongkat Ali). Journal of King Saud University - Computer and Information Sciences, 27(1), pp.23–30. doi: 10.1016/j.jksus.2014.04.006.

Kuo, P.C. et al., 2003. Cytotoxic and antimalarial β-carboline alkaloids from the roots of Eurycoma longifolia. Journal of Natural Products, 66, pp.1324–1327.

Kuo, P.C. et al., 2004. Cytotoxic and antimalarial constituents from the roots of Eurycoma longifolia. Bioorganic & Medicinal Chemistry, 12, pp.537–544. doi: 10.1016/j.bmc.2003.11.017.

Li, C.H. et al., 2013. Evaluation of acute 13-week subchronic toxicity and genotoxicity of the powdered root of Tongkat Ali (Eurycoma longifolia Jack). Evidence-Based Complementary and Alternative Medicine, 102987. doi: 10.1155/2013/102987.

Low, B.S., Das, P.K & Chan, K.L., 2013. Standardized quassinoid-rich Eurycoma longifolia extract improved spermatogenesis and fertility in male rats via the hypothalamic-pituitary-gonadal axis. Journal of Ethnopharmacology, 145(3), pp.706–714. doi: 10.1016/j.jep.2012.11.013.

Mohd, E.N, et al., 2012. Eurycoma longifolia: Medicinal plant in the prevention and treatment of male osteoporosis due to androgen deficiency. Evidence-Based Complementary and Alternative Medicine, 2012: doi: 10.1155/2012/125761.

Ming, K.Y. et al., 2014. Evaluation of the Genotoxicity of Eurycoma longifolia Aqueous Extract (Physta®) Using in Vivo Mammalian Micronucleus Test. International Journal of Pharmacy and Pharmaceutical Sciences, 6(4), pp.652–657.

Qari, S.H. & El-Assouli, S.M., 2019. Evaluation of cytological and genetic effects of Tribulus terrestris fruit aqueous extract on cultured human lymphocytes. Saudi Juanal of Biological Sciences, 26(1), pp. 91-95. doi: 10.1016/j.sjbs.2017.01.012.

Razak, M.F.A. & Aidoo, K.E., 2011. Toxicity studies of Eurycoma longifolia (jack)-based remedial products. Asian Journal of Pharmaceutical and Clinical Research, 4(3), pp.23–27.

Rehman, S.U., Choe, K. & Yoo, H.H., 2016. Review on a traditional herbal medicine, Eurycoma longifolia Jack (Tongkat Ali): its traditional uses, chemistry, evidence-based pharmacology and toxicology. Molecules, 21(3), pp.331. doi: 10.3390/molecules21030331.

Rooney, D.E., 2001. Human cytogenetics: constitutional analysis. Oxford University Press, Oxford.

Sangpakdee, W. et al., 2016. Characterization of chromosomal rearrangements in pileated gibbon (Hylobates pileatus) using multiplex-FISH technique. The Nucleus, 59, pp.131– 135. doi: 10.1007/s13237-016-0171-6.

Segaran, A. et al., 2021. A narrative review on pharmacological significance of Eurycoma longifolia jack roots. Longhua Chinese Medicine. doi: 10.21037/lcm-21-32.

Shuid, A.N. et al., 2011. Acute and subacute toxicity studies of Eurycoma longifolia in male rates. International Journal of Pharmacology, 7(5), pp.641–646. doi: 10.3923/ijp.2011.641.646.

Talbott, S.M. et al., 2013. Effect of Tongkat Ali on stress hormones and psychological mood state in moderately stressed subject. Journal of the International Society of Sports Nutrition, 10(1), pp.1–7. doi: 10.1186/1550-2783-10-28.

Tambi, M.I., 2005. Standardized water-soluble extract of Eurycoma longifolia (LJ100) on men’s health. International Journal of Andrology, 28(1), pp.25–44. doi: 10.1111/j.1439-0272.2011.01168.x.

Turck, et al., 2021. Safety of Eurycoma longifolia (Tongkat Ali) root extract as a novel food pursuant to Regulation (EU) 2015/2283. EFSA Journal, 19(12), pp.6937. doi: 10.2903/j.efsa.2021.6937.


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