{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T17:42:57Z","timestamp":1770745377210,"version":"3.49.0"},"reference-count":50,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,10,13]],"date-time":"2020-10-13T00:00:00Z","timestamp":1602547200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Agriculture, Malaysia","award":["05-03-20-SF1001"],"award-info":[{"award-number":["05-03-20-SF1001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Coffee infused with the additive Eurycoma longifolia, also known as Tongkat ali (TA), has become widely available in the Malaysian market. Safety evaluations for consumption of the products have been called for due to the herbal addition. This study investigates the acute, subacute and chronic effects of a commercial TA coffee in Sprague Dawley rats when given in a single, repeated and prolonged dosage. The dosages of 0.005, 0.05, 0.30 and 2 g\/kg body weight (BW) were used in the acute study and 0.14, 0.29 and 1 g\/kg BW were used in the repeated dose studies. The in-life parameters measured were food and water intake, body weight and clinical observations. Blood were collected for hematology and clinical biochemistry analyses. All animals were subjected to full necropsies. Non-toxicity-related changes were observed in the food and water consumption parameters. Body weight showed normal increments and none of the animals had any clinical signs of toxicity. Microscopically assessed organ tissues did not reveal any abnormalities. There was significant decrease of platelet count in all the chronic study male treated groups. Significant elevation of renal profile parameters in both gender groups given 0.29 g\/kg BW, along with liver and lipid profile elevation in some female groups of the chronic study were noted. No dose-dependent relationship was apparent in the dosage range tested, though these changes may suggest an initial safety indication to the TA coffee. The study concludes that the no observed adverse effect level (NOAEL) for this commercial TA coffee was 1 g\/kg BW.<\/jats:p>","DOI":"10.3390\/nu12103125","type":"journal-article","created":{"date-parts":[[2020,10,13]],"date-time":"2020-10-13T22:02:02Z","timestamp":1602626522000},"page":"3125","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Eurycoma longifolia\u2014Infused Coffee\u2014An Oral Toxicity Study"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6549-0552","authenticated-orcid":false,"given":"Norzahirah","family":"Ahmad","sequence":"first","affiliation":[{"name":"Herbal Medicine Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Shah Alam 40170, Malaysia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0789-170X","authenticated-orcid":false,"given":"Bee Ping","family":"Teh","sequence":"additional","affiliation":[{"name":"Herbal Medicine Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Shah Alam 40170, Malaysia"}]},{"given":"Siti Zaleha","family":"Halim","sequence":"additional","affiliation":[{"name":"Herbal Medicine Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Shah Alam 40170, Malaysia"}]},{"given":"Nor Azlina","family":"Zolkifli","sequence":"additional","affiliation":[{"name":"Herbal Medicine Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Shah Alam 40170, Malaysia"}]},{"given":"Nurulfariza","family":"Ramli","sequence":"additional","affiliation":[{"name":"Herbal Medicine Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Shah Alam 40170, Malaysia"}]},{"given":"Hussin","family":"Muhammad","sequence":"additional","affiliation":[{"name":"Herbal Medicine Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Setia Alam, Shah Alam 40170, Malaysia"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rehman, S.U., Choe, K., and 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.","DOI":"10.3390\/molecules21030331"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1016\/j.ctim.2015.07.009","article-title":"Efficacy of Tongkat Ali (Eurycoma longifolia) on erectile function improvement: Systematic review and meta-analysis of randomized controlled trials","volume":"23","author":"Kotirum","year":"2015","journal-title":"Complement. Ther. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1002\/ptr.5017","article-title":"Tongkat Ali as a potential herbal supplement for physically active male and female seniors\u2014A pilot study","volume":"28","author":"Henkel","year":"2014","journal-title":"Phytother. Res. PTR"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1111\/and.12082","article-title":"In vivo effects of Eurycoma longifolia Jack (Tongkat Ali) extract on reproductive functions in the rat","volume":"46","author":"Solomon","year":"2014","journal-title":"Andrologia"},{"key":"ref_5","first-page":"872406","article-title":"Combined effects of Eurycoma longifolia and testosterone on androgen-deficient osteoporosis in a male rat model","volume":"2012","author":"Shuid","year":"2012","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Shuid, A.N., El-arabi, E., Effendy, N.M., Abdul Razak, H.M., Muhammad, N., Mohamed, N., and Soelaiman, I.M. (2012). Eurycoma longifolia upregulates osteoprotegerin gene expression in androgen-deficient osteoporosis rat model. BMC Complement. Altern. Med., 12.","DOI":"10.1186\/1472-6882-12-152"},{"key":"ref_7","first-page":"337","article-title":"Physico-chemical effects of the major quassinoids in a standardized Eurycoma longifolia extract (Fr 2) on the bioavailability and pharmacokinetic properties, and their implications for oral antimalarial activity","volume":"6","author":"Low","year":"2011","journal-title":"Nat. Prod. Commun."},{"key":"ref_8","first-page":"155","article-title":"Effect of Eurycoma longifolia extract on the glutathione level in Plasmodium falciparum infected erythrocytes in vitro","volume":"22","author":"Abdullah","year":"2005","journal-title":"Trop. Biomed."},{"key":"ref_9","first-page":"14","article-title":"Toxicological and antimalaria activity of Eurycoma longifolia Jack extracts in mice","volume":"5","author":"Satayavivad","year":"1998","journal-title":"Thai. J. Phytopharm."},{"key":"ref_10","first-page":"232","article-title":"Acute, subacute and subchronic 90-days toxicity of Eurycoma longifolia aqueous extract (Physta) in Wistar rats","volume":"4","author":"Choudhary","year":"2012","journal-title":"Int. J. Pharm. Pharm. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"641","DOI":"10.3923\/ijp.2011.641.646","article-title":"Acute and subacute toxicity studies of Eurycoma longifolia in male rats","volume":"7","author":"Shuid","year":"2011","journal-title":"Int. J. Pharmacol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"102987","DOI":"10.1155\/2013\/102987","article-title":"Evaluation of acute 13-week subchronic toxicity and genotoxicity of the powdered root of Tongkat Ali (Eurycoma longifolia jack)","volume":"2013","author":"Li","year":"2013","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"910","DOI":"10.1111\/1541-4337.12446","article-title":"International perspectives on substantiating the efficacy of herbal dietary supplements and herbal medicines through evidence on traditional use","volume":"18","author":"Lenssen","year":"2019","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_14","unstructured":"Ministry of Health (1983). Food Act."},{"key":"ref_15","unstructured":"Ministry of Health (1985). Food Regulation."},{"key":"ref_16","unstructured":"World Health Organization (2004). WHO Guidelines on Safety Monitoring of Herbal Medicines in Pharmacovigilance Systems, World Health Organization."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1055\/s-0043-123764","article-title":"Do we need plant food supplements? A critical examination of quality, safety, efficacy, and necessity for a new regulatory framework","volume":"84","year":"2018","journal-title":"Planta Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.phanu.2017.10.002","article-title":"Food or pharmaceuticals? Consumers\u2019 perception of health-related borderline products","volume":"5","author":"Khedkar","year":"2017","journal-title":"PharmaNutrition"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1016\/j.yrtph.2015.10.009","article-title":"Adverse food-drug interactions","volume":"73","author":"Bast","year":"2015","journal-title":"Regul. Toxicol. Pharmacol. RTP"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.phymed.2011.10.002","article-title":"European regulation of herbal medicinal products on the border area to the food sector","volume":"19","author":"Quintus","year":"2012","journal-title":"Phytomedicine"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.taap.2009.12.005","article-title":"Assessment of herbal medicinal products: Challenges, and opportunities to increase the knowledge base for safety assessment","volume":"243","author":"Jordan","year":"2010","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_22","unstructured":"OECD (2001). OECD Guideline for Testing of Chemicals No. 420: Acute Oral Toxicity\u2014Fixed Dose Procedure, OECD."},{"key":"ref_23","unstructured":"OECD (1995). OECD Guideline for Testing of Chemicals No. 407: Repeated Dose 28-Day Oral Toxicity Study in Rodents, OECD."},{"key":"ref_24","unstructured":"OECD (2009). OECD Guideline for Testing of Chemicals No. 452: Chronic Toxcity Studies, OECD."},{"key":"ref_25","unstructured":"Ministry of Health (2002). Principles and Guide to Ethical Use of Laboratory Animals."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1177\/0192623310388431","article-title":"Gavage-related reflux in rats: Identification, pathogenesis, and toxicological implications","volume":"39","author":"Damsch","year":"2011","journal-title":"Toxicol. Pathol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Whishaw, I.Q., and Kolb, B. (2004). Eating. The Behaviour of the Laboratory Rat. A Handbook with Tests, Oxford University Press.","DOI":"10.1093\/acprof:oso\/9780195162851.001.0001"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Whishaw, I.Q., and Kolb, B. (2004). Drinking. The Behaviour of the Laboratory Rat. A Handbook with Tests, Oxford University Press.","DOI":"10.1093\/acprof:oso\/9780195162851.001.0001"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/0041-008X(81)90421-X","article-title":"The effect of brewed and instant coffee on reproduction and teratogenesis in the rat","volume":"58","author":"Nolen","year":"1981","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1080\/13685538.2017.1350156","article-title":"Reference database of hematological parameters for growing and aging rats","volume":"21","author":"Lima","year":"2018","journal-title":"Aging Male"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"He, Q., Su, G., Liu, K., Zhang, F., Jiang, Y., Gao, J., Liu, L., Jiang, Z., Jin, M., and Xie, H. (2017). Sex-specific reference intervals of hematologic and biochemical analytes in Sprague-Dawley rats using the nonparametric rank percentile method. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0189837"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1177\/096032719601500802","article-title":"Reference values for young normal Sprague-Dawley rats: Weight gain, hematology and clinical chemistry","volume":"15","author":"Lillie","year":"1996","journal-title":"Hum. Exp. Toxicol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.etp.2005.10.002","article-title":"Clinical chemistry and haematology historical data in control Sprague-Dawley rats from pre-clinical toxicity studies","volume":"57","author":"Petterino","year":"2006","journal-title":"Exp. Toxicol. Pathol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/S0278-6915(99)00155-6","article-title":"Characteristics of rat platelets and relative contributions of platelets and blood coagulation to haemostasis","volume":"38","author":"Takahashi","year":"2000","journal-title":"Food Chem. Toxicol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.fct.2019.02.039","article-title":"Effects of coffee, energy drinks and their components on hemostasis: The hypothetical mechanisms of their action","volume":"127","author":"Olas","year":"2019","journal-title":"Food Chem. Toxicol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1002\/ame2.12041","article-title":"Reference values for selected hematological, biochemical and physiological parameters of Sprague-Dawley rats at the animal house, Faculty of Medicine, University of Colombo, Sri Lanka","volume":"1","author":"Delwatta","year":"2018","journal-title":"Anim. Models Exp. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"231","DOI":"10.31344\/ijhhs.v3i4.107","article-title":"The comparison effect of energy drinks and coffee on creatinin level in rats","volume":"3","author":"Riza","year":"2019","journal-title":"Int. J. Hum. Health Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"140","DOI":"10.2991\/efood.k.190802.001","article-title":"In vivo modulatory effect of coffee (Coffea canephora var. Robusta) on the expression levels of murine microRNA-124-3p associated with antioxidant defenses","volume":"1","author":"Curti","year":"2020","journal-title":"eFood"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Kolb, H., Kempf, K., and Martin, S. (2020). Health effects of coffee: Mechanism unraveled?. Nutrients, 12.","DOI":"10.3390\/nu12061842"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2552","DOI":"10.1016\/j.clnu.2018.11.030","article-title":"A systematic review and a dose-response meta-analysis of coffee dose and nonalcoholic fatty liver disease","volume":"38","author":"Chen","year":"2019","journal-title":"Clin. Nutr."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1186\/s42826-019-0024-y","article-title":"Investigation of the effect of coffee on body weight, serum glucose, uric acid and lipid profile levels in male albino Wistar rats feeding on high-fructose diet","volume":"35","author":"Feyisa","year":"2019","journal-title":"Lab. Anim. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.yclnex.2019.10.005","article-title":"Sub-chronic administration of brewed coffee on rat behavior and cognition and oxidative stress Alzheimer\u2019s disease model","volume":"28","author":"Sedaghat","year":"2019","journal-title":"Clin. Nutr. Exp."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/0278-6915(95)80242-8","article-title":"Analysis of the potential carcinogenicity of coffee and its related compounds in a medium-term liver bioassay of rats","volume":"33","author":"Hasegawa","year":"1995","journal-title":"Food Chem. Toxicol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"172880","DOI":"10.1016\/j.pbb.2020.172880","article-title":"Chronic voluntary caffeine intake in male Wistar rats reveals individual differences in addiction-like behavior","volume":"191","author":"Lee","year":"2020","journal-title":"Pharmacol. Biochem. Behav."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1016\/j.fct.2017.04.002","article-title":"Systematic review of the potential adverse effects of caffeine consumption in healthy adults, pregnant women, adolescents, and children","volume":"109","author":"Wikoff","year":"2017","journal-title":"Food Chem. Toxicol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1016\/j.yrtph.2016.07.011","article-title":"The acute lethal dose 50 (LD50) of caffeine in albino rats","volume":"80","author":"Adamson","year":"2016","journal-title":"Regul. Toxicol. Pharmacol. RTP"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Ahmad, N., Samiulla, D.S., Teh, B.P., Zainol, M., Zolkifli, N.A., Muhammad, A., Matom, E., Zulkapli, A., Abdullah, N.R., and Ismail, Z. (2018). Bioavailability of eurycomanone in its pure form and in a standardised Eurycoma longifolia water extract. Pharmaceutics, 10.","DOI":"10.3390\/pharmaceutics10030090"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1055\/s-2005-871259","article-title":"Bioavailability and pharmacokinetic studies of eurycomanone from Eurycoma longifolia","volume":"71","author":"Low","year":"2005","journal-title":"Planta Med."},{"key":"ref_49","first-page":"1091","article-title":"Coffee modify pharmacokinetics of acetaminophen","volume":"7","author":"Win","year":"2019","journal-title":"EC Pharmacol. Toxicol."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Hayes, A.W., Wang, T., and Dixon, D. (2020). Chapter 13\u2014Toxicologic testing methods. Loomis\u2019s Essentials of Toxicology, Academic Press. [5th ed.].","DOI":"10.1016\/B978-0-12-815921-7.00013-2"}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/12\/10\/3125\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:20:23Z","timestamp":1760178023000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/12\/10\/3125"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,13]]},"references-count":50,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["nu12103125"],"URL":"https:\/\/doi.org\/10.3390\/nu12103125","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,13]]}}}