高效液相色谱法测定水体中的痕量一氧化氮

High performance liquid chromatography method for the determination of trace nitric oxide in water

基于4,5-二氨基荧光素与一氧化氮(NO)反应生成高荧光三唑荧光素的原理,建立了一种测定水体中溶解NO的高效液相色谱方法。通过不同流动相(乙腈∶磷酸盐缓冲溶液)配比实验和色谱柱温度梯度实验,确定了该方法测定水体中NO的最佳实验条件:流动相配比为乙腈∶磷酸盐缓冲溶液=8∶92(V/V);柱温为25℃。在优化条件下,测得NO浓度与荧光强度在0.33~3.32 nmol/L和13.3~266 nmol/L范围内具有良好的线性关系,检出限为90 pmol/L,相对标准偏差为5.1%,平均加标回收率为122.4%。方法可用于测定可口可乐、植物饮料、海水和日本星杆藻藻液中的NO浓度。

Based on the principle that the reaction between 4,5-diaminofluorescein and NO could produce high fluorescent triazolofluorescein,a high performance liquid chromatography method for the determination of dissolved NO in water was proposed.The effects of different mobile phase(acetonitrile:phosphate buffer solution)ratio and chromatographic column temperature gradient were investigated.The experimental results indicated that the optimum experimental conditions for the determination of NO in water were as follows:the mobile phase of acetonitrile∶phosphate buffer of 8∶92(V/V),and the column temperature of 25℃.There were good linear relationships between the NO concentration and the fluorescence intensity in the ranges of 0.33-3.32 and 13.3-266 nmol/L.The detection limit was 90 pmol/L,the relative standard deviation was 5.1%,and the average standard recovery was122.4%.The method worked well in determining the concentration of NO in the Coca Cola,plant drink,Qingdao Shilaoren seawater and the culture medium of Stellaria japonicum.