Determination of nitrite/nitrate in human biological material by the simple Griess reaction
Determination of nitrite/nitrate in human biological material by the simple Griess reaction
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DOI：
10.1016/s0009-8981(98)00060-6
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发表时间：
1998-06-22
影响因子：
5
通讯作者：
Szczudlik, A
中科院分区：
文献类型：
--
作者：
Guevara, I;Iwanejko, J;Szczudlik, A
由于许多病理过程如败血性休克、炎症、移植排斥、糖尿病等与一氧化氮（NO）的释放有关，因此感兴趣的是监测NO浓度的快速和准确的方法。用于测量亚硝酸盐和硝酸盐（NO2-，NO3-）-NO的稳定代谢物-的各种方法通常用于此目的。在本文中，我们已经表明，适当的Griess程序测定亚硝酸盐显着增加这种方法的灵敏度。该过程，补充了脱蛋白和还原硝酸盐为亚硝酸盐在NADPH敏感性还原酶的存在下，可以成功地应用于测量人体体液（血清，尿液和CSF）中的NOx水平。需要用甲醇/乙醚对样品进行脱蛋白，并且不影响NO代谢物检测的灵敏度。该方法的回收率为88%+/-6%（n = 30）。通过该程序测量的NOx浓度范围为血液中25.0至39.0 μ mol/l，CSF中4.6至14.6 μ mol/l和尿液中0.37至2.52 mmol/l（根据肌酐浓度调整）。该方法的变异系数在1.3- 2.2%之间。该方法也可推荐用于测量组织细胞培养中细胞产生的NOx。(C)1998年由Elsevier Science B. V.出版，版权所有。
Since a number of pathological processes such as septic shock, inflammation, graft rejection, diabetes, etc. are associated with a release of nitric oxide (NO), rapid and accurate methods of monitoring of NO concentration are of interest. Various methods for measurement of nitrite and nitrate (NO2-, NO3-) - the stable metabolites of NO - are commonly used for this purpose. In this paper we have shown that the proper Griess procedure for nitrite determination significantly increases the sensitivity of this method. This procedure, supplemented with deproteinization and reduction of nitrates to nitrites in the presence of NADPH-sensitive reductase, can be successfully applied for measurement of NOx levels in human body fluids (serum, urine and CSF). Deproteinization of samples with methanol/diethylether is required and does not influence the sensitivity of detection of NO metabolites. The recovery of the method is 88%+/-6% (n = 30). The NOx concentrations measured by this procedure ranged from 25.0 to 39.0 mu mol/l in blood, 4.6 to 14.6 mu mol/l in CSF and 0.37 to 2.52 mmol/l (adjusted to creatinine concentration) in urine. The coefficient of variation for this method was between 1.3-2.2%. This method can also be recommended for measurement of NOx produced by cells in tissue cell culture. (C) 1998 Published by Elsevier Science B.V. All rights reserved.