Studies on micromethod for the determination of blood ingredents by enzymatic cycling
酶循环测定血液成分的微量方法研究
基本信息
- 批准号:10672189
- 负责人:
- 金额:$ 1.98万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:1998
- 资助国家:日本
- 起止时间:1998 至 1999
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
We developed a new and highly sensitive method for determination of lactate and pyruvate by the enzymatic cycling reaction with lactate dehydrogenase (LD) and lactate oxidase (LOX). The method involves a new enzymatic cycling technique with NADH, LD and LOX, and measures decrease of absorbance at 340nm of NADH oxidized at 37℃ during the reaction. The reagents for present method were designed to be used automated clinical analyzers. Lactate and pyruvate concentration was calculated by comparison with the reaction rate obtained with 10mmol/L of standard solution. The linearity ranged from 1 to 20mmol/L. The blood or the serum quantity per test are equivalent to 0.16μL, and absorbance for 1mmol/L standard was 0.0045/min. The CV values in within-run precision (n=10) were 2.3% (2.51mmol/L) and 1.4% (7.28mmol\L).This method was not affected by bilirubin (300 mg/L), hemoglobin (5g/L) and ascorbate (1g/L). The coefficient of correlation with LOX-POD-TOOS method was 0.987 (n=93). Thus the simple and highly sensitive kinetic assay method of lactate and pyruvate, less affected by interferents, was established.On the other hand, we developed a highly sensitive method for determination of ketone bodies (acetoacetate and 3-hydroxybutylate) by the enzymatic cycling reaction with 3-hydroxy butyrate dehydrogenase (3-HBAD) and Thio-NAD. The method measures increase of absorbance at 405nm of Thio-NADH deducted at 37℃ during the reaction.
建立了一种新的、高灵敏度的测定乳酸和丙酮酸的方法,该方法利用乳酸脱氢酶(LD)和乳酸氧化酶(LOX)的酶循环反应。该方法采用了一种新的酶循环技术,以NADH、LD和LOX为底物,在37℃下氧化NADH,测定其在340 nm处吸光度的下降。本方法的试剂设计用于自动化临床分析仪。通过与用10 mmol/L标准品溶液获得的反应速率进行比较,计算乳酸盐和丙酮酸盐浓度。线性范围为1 ~ 20 mmol/L。方法的批内精密度CV值(n=10)分别为2.3%(2.51mmol/L)和1.4%(7.28mmol/L),不受胆红素(300 mg/L)、血红蛋白(5g/L)和抗坏血酸(1g/L)的影响。与LOX-POD-TOOS法的相关系数为0.987(n=93)。建立了一种简便、高灵敏、受干扰物影响小的乳酸和丙酮酸的动力学测定方法,并以3-羟基丁酸脱氢酶(3-HBAD)和Thio-NAD为底物,建立了一种高灵敏的酮体(乙酰乙酸和3-羟基丁酸)测定方法。该方法测量了反应期间在37℃下扣除的硫代-NADH在405 nm处的吸光度增加。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
NISHINA Toshihiro其他文献
NISHINA Toshihiro的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}