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Flavorings used in cookies, electronic cigarettes, popcorn, and breads contain approximately 30 chemical compounds, which makes it difficult to determine and correlate signs and symptoms of acute, subacute or chronic toxicity. The aim of this study was to characterize a butter flavoring chemically and subsequently examine the in vitro and in vivo toxicological profile using cellular techniques, invertebrates, and lab mammals. For the first time, the ethyl butanoate was found as the main compound of a butter flavoring (97.75%) and 24 h-toxicity assay employing Artemia salina larvae revealed a linear effect and LC50 value of 14.7 (13.7-15.7) mg/ml (R2 = 0.9448). Previous reports about higher oral doses of ethyl butanoate were not found. Observational screening with doses between 150-1000 mg/kg by gavage displayed increased amount of defecation, palpebral ptosis, and grip strength reduction, predominantly at higher doses. The flavoring also produced clinical signs of toxicity and diazepam-like behavioral changes in mice, including loss of motor coordination, muscle relaxation, increase of locomotor activity and intestinal motility, and induction of diarrhea, with deaths occurring after 48 h exposure. This substance fits into category 3 of the Globally Harmonized System. Data demonstrated that butter flavoring altered the emotional state in Swiss mice and disrupted intestinal motility, which may be a result of neurochemical changes or direct lesions in the central/peripheral nervous systems.


Nárcia Mariana Fonseca Nunes, Jurandy do Nascimento Silva, Micaely Lorrana Pereira Conceição, Joaquim Soares da Costa Júnior, Edymilais da Silva Sousa, Maria das Dores Alves de Oliveira, Antonia Maria das Graças Lopes Citó, Dalton Dittz, Ana Paula Peron, Paulo Michel Pinheiro Ferreira. In vitro and in vivo acute toxicity of an artificial butter flavoring. Journal of toxicology and environmental health. Part A. 2023 Mar 19;86(6):181-197

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PMID: 36794368

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