Optimization of a DNA Nicking Assay to Evaluate Oenocarpus bataua and Camellia sinensis Antioxidant Capacity

International Journal of Molecular Sciences

Louis-Jérôme Leba²  Christel Brunschwig²  Mona Saout²  Karine Martial²  Emmanuelle Vulcain²  Didier Bereau²  Jean-Charles Robinson¹ 

[1] Université des Antilles et de la Guyane, UMR QUALITROP, campus universitaire de Troubiran, P.O. Box 792, 97337 Cayenne Cedex, French Guiana, France;

关键词: pUC18;    antioxidant;    prooxidant;    DNA nicking assay;    Fenton;    Amazonian palm;    Oenocarpus bataua;    Camellia sinensis;   

DOI  :  10.3390/ijms151018023

来源: mdpi

【 摘 要 】

This study was aimed at assessing the DNA damage protective activity of different types of extracts (aqueous, methanolic and acetonic) using an in vitro DNA nicking assay. Several parameters were optimized using the pUC18 plasmid, especially FeSO₄, EDTA, solvent concentrations and incubation time. Special attention has been paid to removing the protective and damaging effect of the solvent and FeSO₄ respectively, as well as to identifying the relevant positive and negative controls. For each solvent, the optimal conditions were determined: (i) for aqueous extracts, 0.33 mM of FeSO₄ and 0.62 mM of EDTA were incubated for 20 min at 37 °C; (ii) for acetone extracts, 1.16% solvent were incubated for 15 min at 37 °C with 1.3 mM of FeSO₄ and 2.5 mM of EDTA and (iii) for methanol extracts, 0.16% solvent, were incubated for 1.5 h at 37 °C with 0.33 mM of FeSO₄ and 0.62 mM of EDTA. Using the optimized conditions, the DNA damage protective activity of aqueous, methanolic and acetonic extracts of an Amazonian palm berry (Oenocarpus bataua) and green tea (Camellia sinensis) was assessed. Aqueous and acetonic Oenocarpus bataua extracts were protective against DNA damage, whereas aqueous, methanolic and acetonic extracts of Camellia sinensis extracts induced DNA damage.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.