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一氧化氮在癌症中的作用。通过高效离子色谱法测量亚硝酸盐和硝酸盐的改进方法。

The role of nitric oxide in cancer. Improved methods for measurement of nitrite and nitrate by high-performance ion chromatography.

作者信息

Stratford M R, Dennis M F, Cochrane R, Parkins C S, Everett S A

机构信息

Gray Laboratory Cancer Research Trust, Mount Vernon Hospital, Northwood, Middlesex, UK.

出版信息

J Chromatogr A. 1997 May 16;770(1-2):151-5. doi: 10.1016/s0021-9673(96)01074-6.

Abstract

The short lifetime of nitric oxide (NO) in vivo impedes its quantitation directly; however, the determination of nitrite and nitrate ions as the end-products of NO oxidation has proven a more practical approach. High-performance ion chromatographic analysis of nitrite in biological fluids is hampered by the large amount of chloride ion (up to approximately 100 mmol/l) which results in insufficient peak resolution when utilizing conductimetric detection. Analysis of both anions in small sample volumes is also constrained by the need to minimise sample handling to avoid contamination by environmental nitrate. We report a means to remove Cl- ions from small sample volumes using Ag+ resin which facilitates quantitation of either nitrite and nitrate anions in biological samples, using silica or polymer based ion-exchange resins with conductimetric or electrochemical and spectrophotometric detection. Including a reversed-phase guard column before the anion-exchange guard and analytical column also greatly extends column lifetime.

摘要

一氧化氮(NO)在体内的短寿命阻碍了其直接定量;然而,将亚硝酸盐和硝酸盐离子作为NO氧化的终产物进行测定已被证明是一种更实用的方法。生物体液中亚硝酸盐的高效离子色谱分析受到大量氯离子(高达约100 mmol/L)的阻碍,这在使用电导检测时会导致峰分辨率不足。小样本体积中两种阴离子的分析也受到需要尽量减少样本处理以避免环境硝酸盐污染的限制。我们报告了一种使用Ag+树脂从小样本体积中去除Cl-离子的方法,该方法有助于使用基于硅胶或聚合物的离子交换树脂,通过电导、电化学和分光光度检测对生物样品中的亚硝酸盐和硝酸盐阴离子进行定量。在阴离子交换保护柱和分析柱之前加入反相保护柱也大大延长了柱寿命。

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