LC-MS／MS法分析尿液中丙烯醛、巴豆醛代谢产物--3-HPMA和3-HMPMA

Determination of Metabolites of Acrolein and Crotonaldehyde in Human Urine by Liquid Chromatography with Tandem Mass Spectrometry

建立了一种同时检测人尿样中丙烯醛和巴豆醛巯基尿酸类代谢物——N－乙酰基－S－（3－羟基丙基）半胱氨酸（3－HPMA）、N－乙酰基－S－（3－羟基丙基－1－甲基）半胱氨酸（3－HMPMA）的液相色谱－三重四级杆串联质谱（HPLC－MS／MS）方法。该方法以3－HPMA—D3、3－HMPMA—D，作内标，采用IsoluteENV．4-固相萃取柱对样品进行净化，Agilent Extend C18柱分离，多离子监测模式检测3－HPMA和3－HMPMA，并采用该方法测定了8名吸烟者和8名非吸烟者尿样中3－HPMA和3－HMPMA的含量。结果表明：①3－HPMA和3－HMPMA的精密度为2．41％～11．22％，回收率为91．11％～114．66％，检测限分别为1．0和1．3ng／mL；②8名吸烟者尿样中3－HPMA含量为943．05～6303．54ng／mL，3－HMPMA含量为1061．73～6373．74ng／mL。8名非吸烟者尿样中3－HPMA含量为14．13—640．93ng／mL，3－HMPMA含量为23．36～119．40ng／mL。

A method based on liquid chromatography with tandem mass spectrometry （HPLC-MS/MS） was developed for the simultaneous determination of 3 - hydroxypropyl mercapturic acid （3 - HPMA） and 3 - hydroxy - 1 - methyl - propylmercapturic acid （3 - HMPMA）, the mercapturic acid metabolites of acrolein and crotonaldehyde in human urine. Using 3 - HPMA - D3 and 3 - HMPMA - D3 as internal standards, the urine samples were cleaned up by Isolute ENV ＋ solid phase extraction （SPE） cartridge and separated by an Angilent Extend C18 column, and then 3 -HPMA and 3 -HMPMA in the samples were detected by the tandem mass detector under multi - reaction monitoring scan mode. The contents of 3 - HPMA and 3 - HMPMA in urine samples from eight smokers and eight nonsmokers were determined, the results showed that： 1 ） the recoveries of 3 -HPMA and 3 -HMPMA ranged from 91.11% to 114.66% with the relative standard deviations （RSDs） of 2. 41% to 11.22%, and the limit of detection was of 1. 0 and 1. 3 ng/mL, respectively; 2） the contents of 3 - HPMA and 3 - HMPMA in urine samples from eight smokers ranged from 943.05 to 6303.54 and from 1061.73 to 6373. 74 ng/mL, and from eight nonsmokers ranged from 14.13 to 640.93 and from 23.36 to 119.40 ng/mL, respectively.