尿中8-羟基脱氧尿苷及反-反式黏糠酸和苯巯基尿酸测定的液相色谱-质谱法  

Determination of urinary 8-hydroxy-2＇-deoxyguanosine, trans,trans-muconic acid, and S-phenylmercapturic acid by liquid chromatography-mass spectrometry

作　　者：刘华良[1] 朱宝立[1] 史俊文[2] 

机构地区：[1]江苏省疾病预防控制中心理化检测所,南京210009 [2]苏州工业园区疾病防治中心

出　　处：《中华劳动卫生职业病杂志》2014年第2期142-144,共3页Chinese Journal of Industrial Hygiene and Occupational Diseases

摘　　要：目的 建立苯作业工人尿中3种生物标志物8-羟基脱氧尿苷、反-反式黏糠酸和苯巯基尿酸同时测定的方法.方法 尿样经固相萃取柱净化后,用液相色谱-质谱仪进行测定,用外标法定量,色谱柱采用C18反相色谱柱,流动相为甲醇与酸性甲酸铵溶液梯度淋洗,质谱采集方式为多反应监测.结果 反-反式粘糠酸测定的线性范围为10～1000μg/L,当添加浓度为50μg/L和500μg/L时,回收率均大于90％,相对标准偏差小于3％；苯巯基尿酸与8-羟基脱氧尿苷测定的线性范围为1～100 μg/L,当添加浓度为5μg/L和50μg/L时,回收率均大于85％,相对标准偏差小于5％.结论 本检测方法各项指标均符合WS/T 68-1996《研制生物样品监测检验方法指南》的要求,适用于尿中苯的3种生物标志物的同时测定.Objective To establish a method for simultaneously determining the urinary concentrations of 8-hydroxy-2＇-deoxyguanosine （8-OHdG）,trans,trans-muconic acid （tt-MA）,and S-phenylmercapturic acid （S-PMA） in subjects exposed to benzene.Methods After being purified by a solid-phase extraction column,the urine samples were transferred to a liquid chromatography-mass spectrometry system,and the concentrations of 8-OHdG,tt-MA,and S-PMA were determined by external standard method.A C18 reversed-phase column was used as the chromatographic column,and methanol/acidic ammonium formate solution was used as the mobile phase for gradient elution.The mass spectrometer was operated in a multi-reaction monitoring mode.Results For tt-MA,the calibration curves were linear in the range of 10-1000 μg/L,and the recovery rates were over 90％ （relative standard deviation （RSD） ＜3％） at spiked levels of 50 μg/L and 500 μg/L.For S-PMA and 8-OHdG,the calibration curves were linear in the range of 1-100 μg/L,and the recovery rates were over 85％ （RSD ＜5％） at spiked levels of 5 μg/L and 50 μg/L.Conclusion This determination method meets the requirement of Biological materials-Methods of monitoring-Guide of development （WS/T 68-1996） and can be used for simultaneous determination of 8-OHdG,tt-MA,and S-PMA in urine.

关 键 词：反-反式黏糠酸 苯巯基尿酸 8-羟基脱氧尿苷 色谱法 液相色谱-质谱 

分 类 号：R446.12[医药卫生—诊断学] O657.63[医药卫生—临床医学]