在线衍生-高效液相色谱法测定尿中N-乙酰-S-(2-氨基甲酰乙基)-半胱氨酸  

Determination of N-acetyl-S-( 2-carbamoylethyl) cysteine in urine by on-line derivatization and high performance liquid chromatography

作　　者：吴诗华[1] 丘静静 郭冠浩[1] 吴邦华[2] 刘移民[1] WU Shi-hua;QIU Jing-jing;GUO Guan-hao;WU Bang-hua;LIU Yi-min(Guangzhou Municipal Hospital for Occupational Diseases Prevention and Treatment,Guangzhou,Guangdong 510620,China)

机构地区：[1]广州市职业病防治院,广东广州510620 [2]广东省职业病防治院,广东广州510310

出　　处：《中国卫生检验杂志》2018年第16期1925-1927,1932,共4页Chinese Journal of Health Laboratory Technology

基　　金：广州市民生科技专项(2014Y2-00067);广州市职业环境与健康效应重点实验室建设专项(2014SY000020);广州市卫生局市医药卫生科技项目(20171A011269);广州市医学重点学科建设项目(穗卫科教[2016]27号);广州市产学研协同创新重大专项项目(20170402-0177);广州市"121人才梯队工程"后备人才项目(穗人社发[2011]167号)

摘　　要：目的建立尿样中N-乙酰-S-(2-氨基甲酰乙基)-半胱氨酸的在线衍生-高效液相色谱测定方法。方法尿样中N-乙酰-S-(2-氨基甲酰乙基)-半胱氨酸经酸水解成S-2-羧乙基半胱氨酸,经碱中和后,在高效液相色谱仪自动进样器中使用邻苯二甲醛自动衍生,生成荧光衍生物,经C18色谱柱分离,用荧光检测器在发射波长为340 nm,激发波长为450 nm下测定荧光强度;通过实验比较不同的衍生剂浓度及用量,选择最佳的衍生化条件。结果本法在5.3μmol/L^79.5μmol/L线性相关系数为0.999 2,最低检出浓度为2.12μmol/L(1 ml尿样),批内精密度为6.27%~8.70%,批间精密度为3.63%~8.26%;加标回收率为97.65%~99.06%,初步发现吸烟者尿液中AAMA平均含量明显高于非吸烟者的平均含量。结论该方法操作简单,自动化程度高,灵敏,准确,精密度高,可用于尿中N-乙酰-S-(2-氨基甲酰乙基)-半胱氨酸的测定。Objective To establish a method for the determination of N-acetyl-S-（ 2-carbamoylethyl） cysteine（ AAMA）in urine by high performance liquid chromatography（ HPLC） with online derivatization. Methods The AAMA in sample was hydrolyzed with Hcl and converted to S-2-carboxyethyl cysteine（ CEC） which reacted with o-phthalaldehyde（ OPA） to form fluorescent derivative. The fluorescence intensity was measured by a fluorescence detector at an emission wavelength of 340 nm and an excitation wavelength of 450 nm. The optimum derivatization conditions were selected by comparing the concentration and amount of different derivatizing agents. Results The linear correlation coefficient of this method was 0. 999 2 within0. 99 μmol/L-79. 5 μmol/L,the minimum detectable concentration was 2. 12 μmol/L（ 1 ml urine sample）,the intra-assay precision was within 6. 27%-8. 70%,the batch-between precision was within 3. 63%-8. 26%; the recoveries of spiked samples were within 97. 65%-99. 06%. It was found that the average content of AAMA in the urine of smokers was significantly higher than that of non-smokers. Conclusion The method is automatic,simple,sensitive,accurate and precisive. It can be used for detecting N-acetyl-S-（ 2-carbamoylethyl） cysteine in urine.

关 键 词：丙烯酰胺 N-乙酰-S-(2-氨基甲酰乙基)-半胱氨酸 邻苯二甲醛 在线衍生 高效液相色谱法 

分 类 号：O657.72[理学—分析化学]