Suda I, Furuta S, Nishiba Y
a Kyushu National Agricultural Experiment Station, Ministry of Agriculture, Forestry and Fisheries , Nishigoshi , Kumamoto 861-11 , Japan.
Biosci Biotechnol Biochem. 1994 Jan;58(1):14-7. 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.
已开发出一种荧光法来测量1,3 - 二乙基 - 2 - 硫代巴比妥酸（DETBA） - 丙二醛（MDA）加合物，以此作为植物材料中脂质过氧化的指标。植物组织样品在氮气保护下制备，然后添加到含有丁基化羟基甲苯的测定系统中。在DETBA与植物组织样品反应后，用乙酸乙酯萃取DETBA - MDA加合物，并通过荧光光谱法或配备荧光检测器的高效液相色谱法（HPLC）进行测定。与传统比色法相比，荧光光谱法中影响物质的种类较少，其干扰浓度较高。当该方法应用于植物材料时，通过荧光光谱法和配备荧光检测器的HPLC测定DETBA - MDA加合物的检测限分别为2.5 nmol/g鲜重和0.0625 nmol/g鲜重。使用这种灵敏、特异且简便的荧光法，可以轻松地从蔬菜、水果和土豆中检测出浓度范围为0.8至18.0 pmol/g鲜重的DETBA - MDA加合物。
