固相萃取-分光光度法测定人体尿液中亚硝酸盐的含量  

Determination of Nitrite Content in Human Urine by Spectrophotometry with Solid-phase Extraction

作　　者：王瑞萍 陈纠[1] 王致[1] WANG Ruiping;CHEN Jiu;WANG Zhi(Key Laboratory of Occupational Environment and Health,Guangzhou Twelfth People's Hospital,Guangzhou,Guangdong 510620,China)

机构地区：[1]广州市第十二人民医院,职业环境与健康重点实验室,广州510620

出　　处：《中国无机分析化学》2024年第7期875-880,共6页Chinese Journal of Inorganic Analytical Chemistry

基　　金：广州市科技计划项目重点研发计划项目(202206010061);广州市医学重点学科建设项目(2021)。

摘　　要：亚硝酸盐中毒发生时,为了快速准确测定人体尿液中亚硝酸盐的含量,建立了固相萃取-分光光度的分析方法。基于亚硝酸盐的重氮偶合反应,采用固相萃取技术对尿液进行净化处理后,消除了基质干扰,提高了测试灵敏度,通过对比考察了固相萃取柱类型、重氮化剂种类、重氮化剂质量浓度、反应酸度、偶合剂质量浓度及反应时间等实验参数对测试结果的影响,得出了最佳实验条件。结果显示,亚硝酸盐在0.033~0.986μg/mL线性关系良好,相关系数为0.9998,尿液在0.246~2.054μg/mL三种加标水平范围内亚硝酸盐的平均回收率为96.3%~106%,表明HLB固相萃取柱有效去除了影响重氮偶合反应的干扰物质,达到了满意的净化效果。日内和日间测定值的相对标准偏差(RSD)分别为2.0%~3.6%和2.2%~4.8%,方法检出限为0.010μg/mL,定量下限为0.033μg/mL。方法操作简单、快速,精密度和准确度良好,且检出限低,适用于人体尿液中亚硝酸盐含量的快速检测。In order to achieve rapid and accurate determination of nitrite content in human urine when nitrite poisoning occurs,a method of spectrophotometry coupled with solid-phase extraction(SPE)was established.The method was based on the diazo-coupling reaction of nitrite.Urine was purified using SPE technique to eliminate the interference of matrix and improve the testing sensitivity.The influences of different operating parameters such as type of SPE columns,type of diazotization reagents,concentration of diazotization reagent,reaction acidity,concentration of coupling reagent,reaction time were compared and studied,and the optimum experimental conditions were obtained,and the testing method was determined.The results showed that nitrite had a good linear relationship in the range of 0.033—0.986μg/mL and the correlation coefficient was 0.9998.The spiked recoveries of nitrite in urine samples were ranged from 96.3% to 106% at three spiked levels from 0.246-2.054μg/mL,indicating that the interfering substances that affect the diazo-coupling reaction were successfully removed by the HLB SPE column,and the purification effect was satisfied.The relative standard deviations(RSD)for intra-day and inter-day were ranged from 2.0% to 3.6% and from 2.2% to 4.8%,respectively,and the limit of detection and quantification were 0.010μg/mL and 0.033μg/mL,respectively.The method has the advantages of simple operation,fast detection speed,good precision,high accuracy and low detection limit,so it can be used for the rapid determination of nitrite in human urine.

关 键 词：分光光度法 固相萃取 亚硝酸盐 尿液 

分 类 号：O657.32[理学—分析化学]