血浆氨酶促终点法测定的实验研究
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Enzymatic End-Point Assay for Determination of Ammonia in Plasma with GLDH-NADH Reaction System
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    摘要:

    介绍一种用谷氨酸脱氢酶-还原辅酶Ⅰ(GLDH-NADH)为反应系统的血浆氨酶促终点比色测定法.反应体系中加入1.9 kU/L的乳酸脱氢酶(LDH),可排除内源性丙酮酸盐引起NADH消耗所致的吸光度持续下降对测定的干扰.此法测定线性范围为6~200 μmol/L,回收率为90%~105%,批内变异系数小于5%,常规条件下的变异系数小于10%.30例健康志愿者清晨空腹静脉血浆氨水平为10~70 μmol/L.此法操作简单、迅速、精确,适合于常规分析.

    Abstract:

    An improved enzymatic end-point method for determination of plasma ammonia was reported. Semi-micromethod were used with a total incubation volume 1 165 μl. Mix 125 μl EDTA Na2-anticoagulation plasma and 1 000 μl PBS-substrate-solution containing 1.9 kU/L LDH (pH 8.0), then add 40 μl GLDH (≥12×104 U/L) to start the reaction. Incubation periods were shortened to 5 min. The limited detection for ammonia in plasma is 4 μmol/L, and this method is linear from 6 to 200 μmol/L. The method is accurate and reliable. The analytical recovery is from 90% to 105%. The within-run CV% was≤5%. The routine condition variance of the method ranged from 5% to 10%. It was found that 1.9 kU/L LDH was sufficient to eliminate the decreace of NADH by endogenouse pyruvate and LDH in plasma. The normal fasting ammonia in plasma is 10~70 μmol/L.

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徐国宾,朱立华,洪健美,夏铁安.血浆氨酶促终点法测定的实验研究[J].生物化学与生物物理进展,1997,24(2):173-176

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  • 收稿日期:1996-04-16
  • 最后修改日期:1996-07-29
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