申请人:KYOWA MEDEX CO., LTD.
公开号:EP0926244A1
公开(公告)日:1999-06-30
The present invention relates to a method for the quantitative determination of ascorbic acid in a sample using ascorbate oxidase which catalyzes the reaction of reduced form of ascorbic acid with oxygen to form oxidized ascorbic acid and hydrogen peroxide (hereinafter the enzyme is referred to as ASOD), wherein the reaction of reduced ascorbic acid with oxygen in the presence of ASOD to form oxidized ascorbic acid and hydrogen peroxide and the reaction of the formed hydrogen peroxide with a chromogen in the presence of peroxidase to form a pigment are carried out in an aqueous medium in the same reaction system, and then the formed pigment is determined.
The present invention also relates to a method for the determination of total ascorbic acid, wherein the enzyme reactions are carried out in the presence of a reducing agent which is capable of converting oxidized ascorbic acid into reduced ascorbic acid in said aqueous medium. The method enables the determination of total ascorbic acid at a low concentration.
The present invention further relates to a method for the determination of total ascorbic acid, wherein oxidized ascorbic acid in said sample is reduced by the use of a reducing agent which is capable of converting oxidized ascorbic acid into reduced ascorbic acid, and then the enzyme reactions are carried out in the presence of a compound which is capable of deactivating the reducing agent.
Furthermore, the present invention relates to a reagent and a kit for the determination of ascorbic acid suitable for use in the above methods.
The method of the present invention is a simple method for the determination of ascorbic acid which is applicable to screening in clinical assays and which is capable of analyzing a large number of samples at the same time.
本发明涉及一种利用抗坏血酸氧化酶定量测定样品中抗坏血酸的方法,抗坏血酸氧化酶可催化还原型抗坏血酸与氧反应生成氧化型抗坏血酸和过氧化氢(以下将该酶称为 ASOD)、在同一反应体系中,还原型抗坏血酸在 ASOD 的存在下与氧反应生成氧化型抗坏血酸和过氧化氢,生成的过氧化氢在过氧化物酶的存在下与发色剂反应生成色素,然后测定生成的色素。
本发明还涉及一种测定总抗坏血酸的方法,其中酶反应是在还原剂存在下进行的,还原剂能够在所述水介质中将氧化的抗坏血酸转化为还原的抗坏血酸。该方法可以测定低浓度的总抗坏血酸。
本发明还涉及一种测定总抗坏血酸的方法,其中通过使用能够将氧化抗坏血酸转化为还原抗坏血酸的还原剂来还原所述样品中的氧化抗坏血酸,然后在能够使还原剂失活的化合物存在下进行酶反应。
此外,本发明还涉及一种适用于上述方法的测定抗坏血酸的试剂和试剂盒。
本发明的方法是一种测定抗坏血酸的简单方法,适用于临床检测中的筛选,能够同时分析大量样品。