Skip to Content
Merck

Safety assessment of potential food ingredients in canine hepatocytes.

Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association (2015-02-11)
Leshuai W Zhang, Juraj Koci, Brett Jeffery, Jim E Riviere, Nancy A Monteiro-Riviere
ABSTRACT

This research aimed to develop in vitro methods to assess hazard of canine food ingredients. Canine hepatocytes were harvested and cell viability of clove-leaf oil (CLO), eugenol (EUG), lemongrass oil (LGO), guanosine monophosphate (GMP), inosine monophosphate (IMP), sorbose, ginger-root extract (GRE), cinnamon-bark oil (CBO), cinnamaldehyde (CINA), thymol oil (TO), thymol (THYM), and citric acid were assessed with positive controls: acetaminophen (APAP), aflatoxin B1 and xylitol. Molecular Toxicology PathwayFinder array (MTPF) analyzed toxicity mechanisms for LGO. LC50 for APAP was similar among human (3.45), rat (2.35), dog (4.26 mg/ml). Aflatoxin B1 had an LC50 of 4.43 (human), 5.78 (rat) and 6.05 (dog) µg/ml; xylitol did not decrease viability. LC50 of CLO (0.185 ± 0.075(SD)), EUG (0.165 ± 0.112), LGO (0.220 ± 0.012), GRE (1.54 ± 0.31) mg/ml; GMP (166.03 ± 41.83), GMP + IMP (208.67 ± 15.27) mM; CBO (0.08 ± 0.03), CINA (0.11 ± 0.01), TO (0.21 ± 0.03), THYM (0.05 ± 0.01), citric acid (1.58 ± 0.08) mg/ml, while sorbose was non-toxic. LGO induced upregulation of 16 and down-regulation of 24 genes, which CYP and heat shock most affected. These results suggest that in vitro assays such as this may be useful for hazard assessment of food ingredients for altered hepatic function.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Xylitol, ≥99% (GC)
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, for molecular biology, ≥97.0%
Sigma-Aldrich
D-(+)-Sorbose, ≥99%
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, BioXtra, ≥97 .0%
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, ≥97.0%
Supelco
Xylitol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Xylitol
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, BioUltra, for molecular biology, ≥99.0% (T)
Paracetamol, European Pharmacopoeia (EP) Reference Standard
USP
Xylitol, United States Pharmacopeia (USP) Reference Standard
Xylitol, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Citric acid, ≥99.5%, FCC, FG
Sigma-Aldrich
Citric acid, BioUltra, anhydrous, ≥99.5% (T)
Sigma-Aldrich
2-Hydroxybutyric acid sodium salt, 97%
Sigma-Aldrich
Citric acid, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetaminophen, meets USP testing specifications, 98.0-102.0%, powder
Sigma-Aldrich
Acetaminophen, analytical standard
Sigma-Aldrich
Citric acid, anhydrous, suitable for cell culture, suitable for plant cell culture
Sigma-Aldrich
Citric acid, 99%
Sigma-Aldrich
Acetaminophen, BioXtra, ≥99.0%
Supelco
Citric acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
USP
Citric acid, United States Pharmacopeia (USP) Reference Standard
Supelco
Citric acid, Anhydrous, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Acetaminophen, Pharmaceutical Secondary Standard; Certified Reference Material
Citric acid, anhydrous, European Pharmacopoeia (EP) Reference Standard
Supelco
Acetaminophen solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
Sigma-Aldrich
Citric acid, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
USP
Acetaminophen, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Citric acid, meets analytical specification of Ph. Eur., BP, USP, E330, anhydrous, 99.5-100.5% (based on anhydrous substance)