Evaluation of the genotoxic and antioxidant effects of two novel feed additives (ethoxyquin complexes with flavonoids) by the comet assay and micronucleus test

Abstract

The complexes of antioxidant ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline; EQ) with rutin or quercetin (EQ-R and EQ-Q, respectively) were studied in human lymphocytes for genotoxic and antioxidant activities with the use of the comet assay and micronucleus test. The study was undertaken to search for new potential antioxidants, and was motivated by reports of unfavourable side-effects observed in animals fed with feeds containing EQ, which is allowed up to 150 mg kg⁻¹ (0.015%) in complete animal feed. It was shown that EQ-R induced DNA damage in human lymphocytes when used at all the concentrations studied (1–25 µM), while after EQ-Q treatment, the genotoxic effect was observed mainly after higher doses (10 and 25 µM). An increase in the number of micronuclei was observed only for EQ-Q after a dose of 50 µM. The studied compounds decreased the degree of DNA damage induced by hydrogen peroxide (10 µM) in the comet assay. The results obtained in both tests showed that the antioxidant activity of EQ-Q was comparable with that of EQ, so further detailed studies are necessary to estimate its possible usefulness as a feed preservative.

Keywords:

• Ethoxyquin
• flavonoids
• hydrogen peroxide
• comet assay
• micronucleus test
• human lymphocytes

Acknowledgements

The authors are grateful to Aleksandra Wołczańska and Danuta Paszkiewicz for technical assistance. Work was supported by the State Committee for Scientific Research (KBN, Poland), Grant No. 3 PO4C 021 24.