Zunić G, Spasić S, Jelić-Ivanović Z
Institute for Medical Research, Military Medical Academy, Belgrade, Yugoslavia.
J Chromatogr B Biomed Sci Appl. 1999 Apr 30;727(1-2):73-9. doi: 10.1016/s0378-4347(99)00054-7.
Nitrite and nitrate levels in physiological fluids are commonly used as an index of nitric oxide production. We developed simple and rapid method for the determination of these anions by capillary zone electrophoresis employing borate buffer (pH 10, 100 mmol/l) as running electrolyte. The anions were analyzed in plasma and cerebrospinal fluid (CSF) without deproteinization of the samples. Electrophoresis was carried out in a capillary (36.5 cm x 75 microm) at a potential of 15 kV, with on-column UV detection at 214 nm. Mean retention times for nitrite and nitrates were 4.631 and 5.152 min, respectively. The method was linear (r=0.999) within a 1-500 micromol/l concentration range. Physiological levels of nitrate in plasma (40.2 micromol/l) and CSF (15.3 micromol/l) could be determined with good precision (coefficients of variation <6%) and accuracy (recoveries of added nitrate to plasma and CSF were 97.4 and 104.5%, respectively). Measurements of the physiological levels of nitrite in plasma (6.1 micromol/l) and CSF (0.9 micromol/l) were less precise and accurate.
生理体液中的亚硝酸盐和硝酸盐水平通常用作一氧化氮生成的指标。我们开发了一种简单快速的方法，通过毛细管区带电泳，使用硼酸盐缓冲液（pH 10，100 mmol/L）作为运行电解质来测定这些阴离子。在不对样品进行脱蛋白处理的情况下，对血浆和脑脊液（CSF）中的阴离子进行分析。在毛细管（36.5 cm×75 µm）中，于15 kV的电势下进行电泳，并在214 nm处进行柱上紫外检测。亚硝酸盐和硝酸盐的平均保留时间分别为4.631分钟和5.152分钟。该方法在1 - 500 µmol/L的浓度范围内呈线性（r = 0.999）。血浆（40.2 µmol/L）和脑脊液（15.3 µmol/L）中硝酸盐的生理水平能够以良好的精密度（变异系数<6%）和准确度（添加到血浆和脑脊液中的硝酸盐回收率分别为97.4%和104.5%）进行测定。血浆（6.1 µmol/L）和脑脊液（0.9 µmol/L）中亚硝酸盐生理水平的测量精密度和准确度较低。
