Fluorometric determination of a 1,3-diethyl-2-thiobarbituric acid-malondialdehyde adduct as an index of lipid peroxidation in plant materials

• Published in: Bioscience, biotechnology, and biochemistry Vol. 58; no. 1; pp. 14 - 17
• Main Authors: Suda, I. (Kyushu National Agricultural Experiment Station, Chikugo, Fukuoka (Japan)), Furuta, S, Nishiba, Y
• Format: Journal Article
• Language: English
• Published: Tokyo Taylor & Francis 01-01-1994; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: ALDEHIDOS; ALDEHYDE; BARBITURICOS; BARBITURIQUE; Biological and medical sciences; FLUORIMETRIA; FLUORIMETRIE; Food industries; Fundamental and applied biological sciences. Psychology; General aspects; LIPIDE; LIPIDOS; Methods of analysis, processing and quality control, regulation, standards; OXIDACION; OXYDATION; PLANTAS; PLANTE; TECHNIQUE ANALYTIQUE; TECNICAS ANALITICAS; Performance evaluation; Fruit; Reaction product; Analytical method; Lipids; Vegetable; Fluorescence spectrometry; Peroxidation
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.58.14

A fluorometric method has been developed to measure a 1,3-diethyl-2-thiobarbituric acid (DETBA)-malondialdehyde (MDA) adduct as an index of lipid peroxidation in plant materials. Plant tissue samples were prepared under nitrogen gas and then added to an assay system containing butylated hydroxytoluene. Following the reaction between DETBA and the plant tissue samples, the DETBA-MDA adduct was extracted with ethyl acetate and measured by spectrofluorometry or high-performance liquid chromatography (HPLC) with a fluorescence detector. The species of influencing substances with spectrofluorometry were fewer and their interfering concentration was higher than that by traditional colorimetry. When this method was applied to plant materials, the detection limits for the DETBA-MDA adduct were 2.5 nmol/g of fresh weight and 0.0625 nmol/g of fresh weight by spectrofluorometry and HPLC with a fluorescence detector, respectively. Using this sensitive, specific and simple fluorometric method, DETBA-MDA adducts ranging from 0.8 to 18.0 pmol/g of fresh weight could easily be detected from vegetables, fruits, and potatoes