Method for the simultaneous determination of free/protein malondialdehyde and lipid/protein hydroperoxides

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• 作者：Konstantinos Grintzalis^a ; Dimitrios Zisimopoulos^a ; Tilman Grune^b ; Daniela Weber^b ; Christos D. Georgiou^a ; ^c ; ^{c.georgiou@upatras.gr} ; ^{christos.georgiou@nasa.gov}
• 关键词：Lipid/protein malondialdehyde ; Thiobarbituric acid ; Lipid/protein hydroperoxides ; Oxidative stress ; Xylenol orange ; Free radicals
• 刊名：Free Radical Biology and Medicine
• 出版年：2013
• 出版时间：June, 2013
• 年：2013
• 卷：59
• 期：Complete
• 页码：27-35
• 全文大小：454 K

文摘

A simple and sensitive method is presented for the simultaneous quantification (spectrophotometric and spectrofluorimetric) of the main lipid and protein peroxidation products after their initial fractionation: free malondialdehyde (FrMDA), protein-bound malondialdehyde (PrMDA), total hydroperoxides (LOOH), and protein hydroperoxides (PrOOH). FrMDA and PrMDA (released from proteins by alkaline hydrolysis) are measured after the reaction of MDA with thiobarbituric acid (TBA) under acidic conditions, by the specific fluorimetric quantification of the resulting MDA-(TBA)₂ adduct chromophore. The measurement of LOOH and PrOOH is based on the reaction of Fe³⁺ (resulting from the reaction of LOOH and PrOOH with Fe²⁺) with xylenol orange (XO) and the photometric quantification of the resulting XO-Fe complex. The sensitivity of the assays for FrMDA/PrMDA and LOOH/PrOOH is 20 and 100 pmol, respectively. The method was applied successfully on human plasma and can be used for the evaluation of oxidative stress in both basic and clinical research.