Research on highly sensitive analysis of trace metals in biological fluid using enzymes as analytical reagent.

以酶为分析试剂高灵敏分析生物体液中痕量金属的研究

• 批准号: 61571111
• 负责人: MASAYOSHI Tabata
• 金额: $ 1.28万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61571111/
• 关键词: enzymatic analysis; magnesium; calcium ion; zink; hexokinase; glucokinase; phospholipase D; アルカリホスファターゼ; マグネシウムイオン【Mg^(2+)】; カルシウムイオン【Ca^(2+)】; グルコキナーゼ

I have made " Research on highly sensitive analysis of trace metals in biological fluid using enzymes as analytical reagent". The main points of this research can be classified into the following three methods, and these results are as follows.1. Research on the development of enzymatic analysis of magnesium in serum and urine.I have developed a novel enzymatic method for measuring magnesium in serum and urine using hexokinase (HK) or glucokinase (GK) and glucose-6-phosphate dehydrogenase (G6PDH). Both HK-G6PDH and GK-G6PDH methods are relatively simple and do not require expensive instrumentation, but the results correlate satisfactorily with those obtained by atomic absorption method. I, furthermore, applied this method to chemiluminescence determination with high sensitivity, and obtained the excellent results.2. Research on the development of enzymatic analysis of calcium ion in serum.I have developed a new enzymatic method for determining calcium ion in serum using phospholipase D, choline oxidase and peroxidase. The present method is highly Ca^<2+> -specific, accurate, and reproducible, giving results in reasonably good agreement with those obtained by the Ca^<2+> selective electrode method.3. Research on the development of enzymatic analysis of zink in serum.I tried to develope a novel method for determining zink in serum by measuring the degree of activation by zink present in a given solution of the zink enzyme, alkaline phosphatase, which was removed from by chelating agents. It was found the conditions giving linearity between zink amounts (zink standard solution) and the acivity of the enzyme. However, there was a few troubles in the case of the determination of zink in serum.

我做了“用酶作为分析试剂高灵敏度分析生物体液中痕量金属的研究”。本研究的主要观点可归纳为以下三种方法，研究结果如下.血清和尿中镁的酶法分析进展研究本人开发了一种新的酶法测定血清和尿中镁的方法，该方法利用己糖激酶（HK）或葡萄糖激酶（GK）和葡萄糖-6-磷酸脱氢酶（G6 PDH）。HK-G6 PDH和GK-G6 PDH方法都相对简单，不需要昂贵的仪器，但结果与原子吸收法所得结果令人满意。并将该方法应用于化学发光分析，获得了良好的结果.血清中钙离子酶法分析的研究利用磷脂酶D、胆碱氧化酶和过氧化物酶建立了一种新的酶法测定血清中钙离子。本方法具有高度的Ca^<2+>特异性、准确性和重现性，所得结果与Ca^<2+>选择性电极法所得结果相当一致.血清中锌的酶分析方法的发展研究：我试图发展一种新的方法来测定血清中的锌，通过测定锌酶，碱性磷酸酶，这是从螯合剂去除给定的溶液中存在的锌的激活程度。结果表明，锌的加入量（锌标准溶液）与酶的活性呈线性关系。但在血清锌的测定中存在一些问题。

项目成果

Masayoshi Tabata: Agricultural and Biological Chemistry. 50. 1909-1910 (1986)

Masayoshi Tabata：农业和生物化学。

Takahiro Kido: "Enzymatic method for the determination of magnesium in urine using hexokinase or glucokinase and glucose-6-phosphate dehydrogenase." Japanese Journal of Clinical Chemistry. 15. 146-151 (1986)

Takahiro Kido：“使用己糖激酶或葡萄糖激酶和葡萄糖-6-磷酸脱氢酶测定尿液中镁的酶法。”

松下泰子: 日本臨床検査自動化学会会誌. (1988)

Yasuko Matsushita：日本临床实验室自动化学会杂志（1988 年）。

田畑勝好: マグネシウム. 5. 45-51 (1986)

田端胜义：镁。5. 45-51 (1986)