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Food Chem. 2020 Mar 16;320:126625. doi: 10.1016/j.foodchem.2020.126625. [Epub ahead of print]

Influences of calcium and magnesium ions on cellular antioxidant activity (CAA) determination.

Author information

1
SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310012, China.
2
SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310012, China. Electronic address: lijingke@zjgsu.edu.cn.
3
Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
4
Hunan Salt Industry Co., Ltd., Changsha 410004, China.

Abstract

The cellular antioxidant activity (CAA) assay is wildly used for quantifying antioxidant activities of foods and dietary supplements in vitro. Among various incubation and handling buffers used in different laboratories, the inconsistence in concentrations of ions, particularly calcium and magnesium, has somehow been neglected. We hired the Hank's balanced salt solution with or without calcium and magnesium to perform CAA assay in Caco-2 cells and HepG2 cells, evaluating the impacts of these cations. The absence of calcium and magnesium reduced intracellular ROS level and underestimated the CAA of quercetin, Trolox and catechin. The abnormally high extracellular calcium and magnesium can also produce inaccurate results. Hank's buffer is recommended to ensure the accuracy and reproducibility. It elucidates precautions must be taken on these cations' concentrations of the buffers while conducting CAA determinations on different types of cells and when comparing foods and beverages with various calcium/magnesium contents.

KEYWORDS:

Calcium; Cellular antioxidant activity (CAA); Hank’s balanced salt solution (HBSS); Magnesium; Quantitative assay

Conflict of interest statement

Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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