Pinchuk Ilya, Lichtenberg Dov
Department of Physiology and Pharmacology, Sackler School of Medicine, Tel-Aviv University, Tel-Aviv, 69987, Israel,
Methods Mol Biol. 2015;1208:171-80. doi: 10.1007/978-1-4939-1441-8_13.
Antioxidants inhibit oxidation processes and by this affect many biological processes. This, in turn, promotes continuing efforts to synthesize new efficient antioxidants and discover compounds of natural origin capable of preventing peroxidation. Although many assays have been developed to evaluate antioxidants, the search for improved protocols is still actual. The presented protocol is based on the effect of antioxidant on the kinetics of peroxidation of lipids in human blood serum. Specifically, we evaluate the capacity of antioxidant by the relative prolongation of lag phase (delay) of copper-induced peroxidation of lipids in unfractionated serum. The main advantage of the assay is that it implements inhibition of peroxidation in physiologically relevant system. We propose expressing the results of the assay either in terms of the relative prolongation of the lag per 1 μM of antioxidant or as the concentration of antioxidant required to double the lag. To allow for comparing the results with those of other assays, these results may be normalized and expressed in terms of the unitless "TROLOX equivalents."
抗氧化剂可抑制氧化过程,从而影响许多生物过程。这反过来又促使人们不断努力合成新的高效抗氧化剂,并发现能够防止过氧化的天然来源化合物。尽管已经开发了许多测定抗氧化剂的方法,但寻找改进的方案仍然很有必要。本文提出的方案基于抗氧化剂对人血清中脂质过氧化动力学的影响。具体而言,我们通过未分级血清中铜诱导的脂质过氧化的滞后相(延迟)的相对延长来评估抗氧化剂的能力。该测定方法的主要优点是它在生理相关系统中实现了对过氧化的抑制。我们建议以每1μM抗氧化剂滞后相的相对延长或使滞后相加倍所需的抗氧化剂浓度来表示测定结果。为了便于将结果与其他测定方法的结果进行比较,这些结果可以进行归一化处理,并以无量纲的“Trolox当量”表示。