Safety and Toxicity Evaluation of Nutraceuticals in Animal Models

  • Nikolay Goncharov
  • Vladislav Sobolev
  • Maxim Terpilowski
  • Ekaterina Korf
  • Richard Jenkins


Nutraceuticals are derived from various natural sources such as medicinal plants, marine organisms, vegetables, and fruits. Most of them possess antioxidant or anti-inflammatory properties and are claimed to provide protection against many diseases if taken regularly. At the same time, toxicological studies of nutraceuticals have been limited, so the safety of many of them cannot be guaranteed. Animals share many genetic, anatomical, and physiological similarities with humans, and they continue to be widely used in preclinical studies of drugs, in spite of a lack of their validity which is due to the great phenotypic differences. The absence of toxicity in animals provides little probability that adverse reactions will also be absent in humans. There are currently thousands of researchers involved in the development of alternatives to animal use in the life sciences. Statistical machine-learning tools, once developed, might become a powerful means to explain the complex physiological effects of nutraceuticals. The use of different models and algorithms can provide a more scientific basis for risk assessment of nutraceuticals for humans.


Preclinical studies Biomarkers System analysis Alternative models Cytotoxic power 



Acceptable daily intake


Adverse drug reactions


α-linolenic acid


Animals in Research: Reporting In Vivo Experiments


Benchmark dose


Benchmark dose lower bound


Chlorinated dibenzo-p-dioxins


Docosahexaenoic acid


Derived no-effect level


Dietary Supplement Health and Education Act


Estimated daily intake




Eicosapentaenoic acid




Generally recognized as safe


Green tea extract


Harm-benefit analysis


Lowest-observed-adverse-effect level


Likelihood ratios


Mode of action


Margin of exposure


Nonhuman primates


No-observed-adverse-effect level


Organosulfur compounds


Particular nutritional uses


Pyrrolizidine alkaloids


Plant food supplement


Points of departure


Reference concentration


Reference dose


Reactive oxygen species


Relative potency factors


Red yeast rice


Toxic equivalency factors


Threshold of toxicological concern



This work has been supported by the Russian Foundation for Basic Research Grant 18-015-00304 and by the Russian FASO Programme АААА-А18-118012290142-9.


  1. Akingbemi BT, Braden TD, Kemppainen BW et al (2007) Exposure to phytoestrogens in the perinatal period affects androgen secretion by testicular Leydig cells in the adult rat. Endocrinology 148:4475–4488PubMedGoogle Scholar
  2. Al-Malahmeh AJ, Al-Ajlouni AM, Wesseling S et al (2016) Determination and risk assessment of naturally occurring genotoxic and carcinogenic alkenylbenzenes in basil-containing sauce of pesto. Toxicol Rep 4:1–8PubMedPubMedCentralGoogle Scholar
  3. Al-Malahmeh AJ, Alajlouni AM, Ning J et al (2017) Determination and risk assessment of naturally occurring genotoxic and carcinogenic alkenylbenzenes in nutmeg-based plant food supplements. J Appl Toxicol 37(10):1254–1264PubMedGoogle Scholar
  4. Alzoubi K, Calabrò S, Faggio C et al (2015) Stimulation of suicidal erythrocyte death by sulforaphane. Basic Clin Pharmacol Toxicol 116(3):229–235PubMedGoogle Scholar
  5. Anon (2018b) 5th international conference of the Basel Declaration Society openness and transparency: building trust in animal research, 14th–15th Feb 2018.
  6. Augustin MA, Sanguansri L, Lockett T (2013) Nano- and microencapsulated systems for enhancing the delivery of resveratrol. Ann N YAcad Sci 1290:107–112Google Scholar
  7. Bailey J, Thew M, Balls M (2014) An analysis of the use of animal models in predicting human toxicology and drug safety. Altern Lab Anim 42(3):181–199PubMedGoogle Scholar
  8. Bayan L, Koulivand PH, Gorji A (2014) Garlic: a review of potential therapeutic effects. Avicenna J Phytomed 4(1):1–14PubMedPubMedCentralGoogle Scholar
  9. Bunchorntavakul C, Reddy KR (2013) Review article: herbal and dietary supplement hepatotoxicity. Aliment Pharmacol Ther 37(1):3–17PubMedGoogle Scholar
  10. Cerella C, Dicato M, Jacob C et al (2011) Chemical properties and mechanisms determining the anti-cancer action of garlic-derived organic sulfur compounds. Anti Cancer Agents Med Chem 11(3):267–271Google Scholar
  11. Chen L, Mulder PPJ, Louisse J et al (2017) Risk assessment for pyrrolizidine alkaloids detected in (herbal) teas and plant food supplements. Regul Toxicol Pharmacol 86:292–302PubMedGoogle Scholar
  12. Das L, Bhaumik E, Raychaudhuri U et al (2012) Role of nutraceuticals in human health. J Food Sci Technol 49(2):173–183PubMedGoogle Scholar
  13. Desbrow B, McCormack J, Burke LM (2014) Sports dietitians Australia position statement: sports nutrition for the adolescent athlete. Int J Sport Nutr Exerc Metab 24(5):570–584PubMedGoogle Scholar
  14. Directive 89/398/EEC (1989.) Accessed Jun 2018
  15. Doke SK, Dhawale SC (2015) Alternatives to animal testing: a review. Saudi Pharm J 23(3):223–229PubMedGoogle Scholar
  16. Eggel M, Grimm H (2018) Necessary, but not sufficient. The benefit concept in the project evaluation of animal research in the context of directive 2010/63/EU. Animals (Basel) 8(3):E34Google Scholar
  17. Elgawish RAR, Rahman HGA, Abdelrazek HMA (2015) Green tea extract attenuates CCl4-induced hepatic injury in male hamsters via inhibition of lipid peroxidation and p53-mediated apoptosis. Toxicol Rep 2:1149–1156PubMedPubMedCentralGoogle Scholar
  18. Espín JC, García-Conesa MT, Tomás-Barberán FA (2007) Nutraceuticals: facts and fiction. Phytochemistry 68(22–24):2986–3008PubMedGoogle Scholar
  19. European Commission (2010) Sixth report on the statistics on the number of animals used for experimental and other scientific purposes in the member states of the European Union, BrusselsGoogle Scholar
  20. Ferdowsian HR, Beck N (2011) Ethical and scientific considerations regarding animal testing and research. PLoS One 6(9):e24059PubMedPubMedCentralGoogle Scholar
  21. Filippich LJ, Zhu J, Oelrichs P et al (1991) Hepatotoxic and nephrotoxic principles in Terminalia oblongata. Res Vet Sci 50(2):170–177PubMedGoogle Scholar
  22. Fischer K, Kettunen J, Würtz P et al (2014) Biomarker profiling by nuclear magnetic resonance spectroscopy for the prediction of all-cause mortality: an observational study of 17,345 persons. PLoS Med 11(2):e1001606PubMedPubMedCentralGoogle Scholar
  23. Franco NH (2013) Animal experiments in biomedical research: a historical perspective. Animals (Basel) 3(1):238–273Google Scholar
  24. Gerhauser C (2018) Impact of dietary gut microbial metabolites on the epigenome. Philos Trans R Soc Lond Ser B Biol Sci 373(1748):20170359Google Scholar
  25. Ghezzi P, Davies K, Delaney A et al (2018) Theory of signs and statistical approach to big data in assessing the relevance of clinical biomarkers of inflammation and oxidative stress. Proc Natl Acad Sci USA 115(10):2473–2477PubMedGoogle Scholar
  26. Goncharov NV, Ukolov AI, Orlova TI et al (2015) Metabolomics: on the way to an integration of biochemistry, analytical chemistry, and informatics. Biol Bull Rev 5(4):296–307Google Scholar
  27. Goncharov N, Maevsky E, Voitenko N et al (2016a) Nutraceuticals in sports activities and fatigue. In: Gupta RC (ed) Nutraceuticals: efficacy, safety and toxicity. Academic Press/Elsevier, Amsterdam, pp 177–188Google Scholar
  28. Goncharov N, Orekhov A, Voitenko N et al (2016b) Organosulfur compounds as nutraceuticals. In: Gupta RC (ed) Nutraceuticals: efficacy, safety and toxicity. Academic Press/Elsevier, Amsterdam, pp 555–568Google Scholar
  29. Goncharov NV, Belinskaia DA, Shmurak VI et al (2017a) Serum albumin binding and esterase activity: mechanistic interactions with organophosphates. Molecules 22(7):E1201PubMedGoogle Scholar
  30. Goncharov NV, Nadeev AD, Jenkins RO, Avdonin PV (2017b) Markers and biomarkers of endothelium: when something is rotten in the state. Oxidative Med Cell Longev 2017:9759735, 27 ppGoogle Scholar
  31. Goncharov NV, Terpilovskii MA, Shmurak VI et al (2017c) Comparative analysis of esterase and paraoxonase activities of different serum albumin species. J Evol Biochem Physiol 53(4):271–281Google Scholar
  32. Goncharov NV, Terpilowski MA, Nadeev AD et al (2018) Cytotoxic power of hydrogen peroxide effect on endothelial cells in vitro. Biochemistry (Moscow), Supplement Series A: Membr Cell Biol 12(2):180–188Google Scholar
  33. Greek R, Menache A (2013) Systematic reviews of animal models: methodology versus epistemology. Int J Med Sci 10(3):206–221PubMedPubMedCentralGoogle Scholar
  34. Grimm H, Eggel M, Deplazes-Zemp A et al (2017) The road to hell is paved with good intentions: why harm-benefit analysis and its emphasis on practical benefit jeopardizes the credibility of research. Animals (Basel) 7(9):E34Google Scholar
  35. Gupta RC (2016) Nutraceuticals: efficacy, safety and toxicity. Academic Press/Elsevier, Amsterdam, 1040 ppGoogle Scholar
  36. Habs M, Binder K, Krauss S et al (2017) A balanced risk-benefit analysis to determine human risks associated with pyrrolizidine alkaloids (PA)—the case of tea and herbal infusions. Nutrients 9(7):E717PubMedGoogle Scholar
  37. Hassanin LA, Salama AM, Essa EA et al (2017) Potential role of some nutraceuticals in neurotoxicity induced by aluminum oxide in experimental animal model. Int J Adv Res Biol Sci 4(11):72–89Google Scholar
  38. Hui L, Qigui L, Sashuang R et al (2014) Nonspecific changes in clinical laboratory indicators in unselected terminally ill patients and a model to predict survival time based on a prospective observational study. J Transl Med 12:78PubMedPubMedCentralGoogle Scholar
  39. Ismail T, Calcabrini C, Diaz AR et al (2016) Ellagitannins in cancer chemoprevention and therapy. Toxins (Basel) 8(5):E151Google Scholar
  40. James KD, Kennett MJ, Lambert JD (2018) Potential role of the mitochondria as a target for the hepatotoxic effects of (-)-epigallocatechin-3-gallate in mice. Food Chem Toxicol 111:302–309PubMedGoogle Scholar
  41. Jurgens TM, Whelan AM, Killian L et al (2012) Green tea for weight loss and weight maintenance in overweight or obese adults. Cochrane Database Syst Rev 12:CD008650PubMedGoogle Scholar
  42. Kilkenny C, Browne WJ, Cuthill IC et al (2010) Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol 8:e1000412PubMedPubMedCentralGoogle Scholar
  43. Kim KB, Nam YA, Kim HS et al (2014) α-Linolenic acid: nutraceutical, pharmacological and toxicological evaluation. Food Chem Toxicol 70:163–178PubMedGoogle Scholar
  44. Knight A (2008) Systematic reviews of animal experiments demonstrate poor contributions toward human healthcare. Rev Recent Clin Trials 3(2):89–96PubMedGoogle Scholar
  45. Konar D, Devarasetty M, Yildiz DV et al (2016) Lung-on-a-chip technologies for disease modeling and drug development. Biomed Eng Comput Biol 7(Suppl 1):17–27PubMedPubMedCentralGoogle Scholar
  46. Korf EA, Kubasov IV, Vonsky MS et al (2017) Green tea extract increases the expression of genes responsible for regulation of calcium balance in rat slow-twitch muscles under conditions of exhausting exercise. Bull Exp Biol Med 164(1):6–9PubMedGoogle Scholar
  47. Kruger CL, Mann SW (2003) Safety evaluation of functional ingredients. Food Chem Toxicol 41(6):793–805PubMedGoogle Scholar
  48. Mahady G, Parrot J, Lee C et al (2003) Botanical dietary supplement use in peri- and postmenopausal women. Menopause 10(1):65–72PubMedGoogle Scholar
  49. Manteiga R, Park DL, Ali SS (1997) Risks associated with consumption of herbal teas. Rev Environ Contam Toxicol 150:1–30PubMedGoogle Scholar
  50. Mason BC, Lavallee ME (2012) Emerging supplements in sports. Sports Health 4(2):142–146PubMedPubMedCentralGoogle Scholar
  51. Mazzanti G, Menniti-Ippolito F, Moro PA et al (2009) Hepatotoxicity from green tea: a review of the literature and two unpublished cases. Eur J Clin Pharmacol 65(4):331–341PubMedGoogle Scholar
  52. Mazzanti G, Di Sotto A, Vitalone A (2015) Hepatotoxicity of green tea: an update. Arch Toxicol 89(8):1175–1191PubMedGoogle Scholar
  53. Mazzanti G, Moro PA, Raschi E et al (2017) Adverse reactions to dietary supplements containing red yeast rice: assessment of cases from the Italian surveillance system. Br J Clin Pharmacol 83:894–908PubMedPubMedCentralGoogle Scholar
  54. Merz KH, Schrenk D (2016) Interim relative potency factors for the toxicological risk assessment of pyrrolizidine alkaloids in food and herbal medicines. Toxicol Lett 263:44–57PubMedGoogle Scholar
  55. Mindukshev I, Kudryavtsev I, Serebriakova M et al (2016) Flow cytometry and light scattering technique in evaluation of nutraceuticals. In: Gupta RC (ed) Nutraceuticals: efficacy, safety and toxicity. Academic Press/Elsevier, Amsterdam, pp 319–332Google Scholar
  56. Mueller C (1999) The regulatory status of medical foods and dietary supplements in the United States. Nutrition 15:249–251PubMedGoogle Scholar
  57. Munday R (2012) Harmful and beneficial effects of organic monosulfides, disulfides, and polysulfides in animals and humans. Chem Res Toxicol 25(1):47–60PubMedGoogle Scholar
  58. Navarro SL, Li F, Lampe JW (2011) Mechanisms of action of isothiocyanates in cancer chemoprevention: an update. Food Funct 2(10):579–587PubMedPubMedCentralGoogle Scholar
  59. Ning J, Cui X, Kong X et al (2018) Risk assessment of genotoxic and carcinogenic alkenylbenzenes in botanical containing products present on the Chinese market. Food Chem Toxicol 115:344–357PubMedGoogle Scholar
  60. Novozhilov AV, Tavrovskaya TV, Voitenko NG et al (2015) Efficacy of green tea extract in two exercise models. Bull Exp Biol Med 158(3):342–345PubMedGoogle Scholar
  61. Oelrichs PB, Pearce CM, Zhu J et al (1994) Isolation and structure determination of terminalin A toxic condensed tannin from Terminalia oblongata. Nat Toxins 2(3):144–150PubMedGoogle Scholar
  62. Olson H, Betton G, Robinson D et al (2000) Concordance of the toxicity of pharmaceuticals in humans and in animals. Regul Toxicol Pharmacol 32:56–67PubMedGoogle Scholar
  63. Päivärinta E, Pajari AM, Törrönen R et al (2006) Ellagic acid and natural sources of ellagitannins as possible chemopreventive agents against intestinal tumorigenesis in the Min mouse. Nutr Cancer 54(1):79–83PubMedGoogle Scholar
  64. Patel S (2016) Functional food red yeast rice (RYR) for metabolic syndrome amelioration: a review on pros and cons. World J Microbiol Biotechnol 32:32–87Google Scholar
  65. Pound P, Nicol CJ (2018) Retrospective harm benefit analysis of pre-clinical animal research for six treatment interventions. PLoS One 13(3):e0193758PubMedPubMedCentralGoogle Scholar
  66. Pound P, Ebrahim S, Sandercock P et al (2004) Where is the evidence that animal research benefits humans? BMJ 328:514–517PubMedPubMedCentralGoogle Scholar
  67. Prakash M, Shetty JK, Rao L et al (2008) Serum paraoxonase activity and protein thiols in chronic renal failure patients. Indian J Nephrol 18(1):13–16PubMedPubMedCentralGoogle Scholar
  68. Prokofieva DS, Goncharov NV (2014) Effects of biogenic and abiogenic disulphides upon endothelial cells in culture: comparison of three methods of viability assessment. Tsitologiya 56(6):410–418Google Scholar
  69. Ried K, Fakler P (2014) Potential of garlic (Allium sativum) in lowering high blood pressure: mechanisms of action and clinical relevance. Integr Blood Press Control 7:71–82PubMedPubMedCentralGoogle Scholar
  70. Ronis M, Hennings L, Gomez-Acevedo H et al (2014) Different responses to soy and estradiol in the reproductive system of prepubertal male rats and neonatal male pigs. FASEB J 28:373.5Google Scholar
  71. Ronis MJ, Gomez-Acevedo H, Blackburn ML et al (2016) Uterine responses to feeding soy protein isolate and treatment with 17β-estradiol differ in ovariectomized female rats. Toxicol Appl Pharmacol 297:68–80PubMedGoogle Scholar
  72. Ronis MJJ, Pedersen KB, Watt J (2018) Adverse effects of nutraceuticals and dietary supplements. Annu Rev Pharmacol Toxicol 58:583–601PubMedGoogle Scholar
  73. Santini A, Cammarata SM, Capone G et al (2018) Nutraceuticals: opening the debate for a regulatory framework. Br J Clin Pharmacol 84(4):659–672PubMedPubMedCentralGoogle Scholar
  74. Sauer S, Luge T (2015) Nutriproteomics: facts, concepts, and perspectives. Proteomics 15(5–6):997–1013PubMedGoogle Scholar
  75. Sauer S, Plauth A (2017) Health-beneficial nutraceuticals-myth or reality? Appl Microbiol Biotechnol 101(3):951–961PubMedGoogle Scholar
  76. Shimshoni JA, Duebecke A, Mulder PP et al (2015) Pyrrolizidine and tropane alkaloids in teas and the herbal teas peppermint, rooibos and chamomile in the Israeli market. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 32(12):2058–2067PubMedGoogle Scholar
  77. Stahl BU, Kettrup A, Rozman K (1992) Comparative toxicity of four chlorinated dibenzo-p-dioxins (CDDs) and their mixture. Part I: acute toxicity and toxic equivalency factors (TEFs). Arch Toxicol 66(7):471–477PubMedGoogle Scholar
  78. Stoner GD, Chen T, Kresty LA et al (2006) Protection against esophageal cancer in rodents with lyophilized berries: potential mechanisms. Nutr Cancer 54(1):33–46PubMedPubMedCentralGoogle Scholar
  79. Sugihara G, May R, Ye H et al (2012) Detecting causality in complex ecosystems. Science 338(6106):496–500PubMedGoogle Scholar
  80. Tan KAL, Walker M, Morris K et al (2006) Infant feeding with soy formula milk: effects on puberty progression, reproductive function and testicular cell numbers in marmoset monkeys in adulthood. Hum Reprod 21:896–904PubMedGoogle Scholar
  81. Terpilowski MA, Korf EA, Jenkins RO, Goncharov NV (2018) An algorithm for deriving combinatorial biomarkers based on ridge regression. J Bioinform Genom 1(6).
  82. Terry C, Rasoulpour RJ, Knowles S et al (2015) Utilizing relative potency factors (RPF) and threshold of toxicological concern (TTC) concepts to assess hazard and human risk assessment profiles of environmental metabolites: a case study. Regul Toxicol Pharmacol 71(2):301–317PubMedGoogle Scholar
  83. Toyokuni S (2014) Iron and thiols as two major players in carcinogenesis: friends or foes? Front Pharmacol 5:200PubMedPubMedCentralGoogle Scholar
  84. Ukolov AI, Kessenikh ED, Radilov AS, Goncharov NV (2017) Toxicometabolomics: identification of markers of chronic exposure to low doses of aliphatic hydrocarbons. J Evol Biochem Physiol 53(1):25–36Google Scholar
  85. Vanacloig-Pedros E, Proft M, Pascual-Ahuir A (2016) Different toxicity mechanisms for citrinin and ochratoxin A revealed by transcriptomic analysis in yeast. Toxins 8:273PubMedCentralGoogle Scholar
  86. Vaughan RA, Conn CA, Mermier CM (2014) Effects of commercially available dietary supplements on resting energy expenditure: a brief report. ISRN Nutr 2014:650264PubMedPubMedCentralGoogle Scholar
  87. Weinberg R (2010) Point: hypotheses first. Nature 464(7289):678PubMedGoogle Scholar
  88. Wiedenfeld H, Edgar J (2011) Toxicity of pyrrolizidine alkaloids to humans and ruminants. Phytochem Rev 10:137–151Google Scholar
  89. Zhang CL, Zeng T, Zhao XL, Xie KQ (2013) Garlic oil attenuated nitrosodiethylamine-induced hepatocarcinogenesis by modulating the metabolic activation and detoxification enzymes. Int J Biol Sci 9(3):237–245PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Nikolay Goncharov
    • 1
    • 2
  • Vladislav Sobolev
    • 1
  • Maxim Terpilowski
    • 2
  • Ekaterina Korf
    • 2
  • Richard Jenkins
    • 3
  1. 1.Research Institute of Hygiene, Occupational Pathology and Human EcologyLeningrad RegionRussia
  2. 2.Sechenov Institute of Evolutionary Physiology and BiochemistrySt. PetersburgRussia
  3. 3.School of Allied Health SciencesDe Montfort UniversityLeicesterUK

Personalised recommendations