Objective: A gas chromatography-tandem mass spectrometry detection method for benzene and its metabolites was established to provide methodological support and theoretical basis for the study of benzene toxicity mechanism. Methods: In August 2019 to March 2020, the animal model of containing high concentration of benzene by inhalation of poison through the respiratory tract of mice was established, taken the blood of mice after dyeing the poison, and the HLB solid phase extraction method was used to extract and purify the samples. The gas chromatography-tandem mass spectrometry detection method was used to qualitative and quantitative analysis of the target substances. After separated by HP-17MS capillary chromatographic column, the compounds were ionized with EI ion source, mass spectrometry detection was carried out by selective ion scanning method (SIM) , and quantification was carried out by external standard curve method. Results: Benzene and its metabolites (phenol, catechol, hydroquinone and m-trihydroxybenzene) in blood could be effectively separated and quasi deterministic and quantitative by this method. The regression equations and correlation coefficients of this method for detecting benzene and its metabolites were: benzene: y=3252.1x+1540, r=0.9993; phenol: y=2046.5x+1423, r=0.9991; catechol: y=1853.9x+945, r=0.9993; hydroquinone: y=1891.5x+840, r=0.9992; m-trihydroxybenzene: y=1052.4x+655, r=0.9991. The detection limits for benzene, phenol, catechol, hydroquinone and m-trihydroxybenzene were 0.03, 0.03, 0.05, 0.05 and 0.10 μg/g, respectively. And the lower limits of quantification were 0.10, 0.10, 0.15, 0.15 and 0.30 μg/g, respectively. The intra-assay precision interval was 2.64%-10.06%, the inter-assay precision interval was 1.37%-10.17%, and the spike recovery rate was 89.8%-102.3%. This method could be used to quantitatively detect benzene, phenol, catechol, hydroquinone and m-trihydroxybenzene in the blood of benzene-infected mice. Conclusion: Solid phase extraction-gas chromatography-mass spectrometry can be used for qualitative and quantitative detection of benzene and its metabolites (phenol, catechol, hydroquinone and m-trihydroxybenzene) accurately.
目的： 建立苯及其部分代谢物的气相色谱-串联质谱检测方法，为苯毒性机制研究提供方法支持和理论依据。 方法： 于2019年8月至2020年3月，用小鼠经呼吸道吸入染毒的方法建立苯高浓度接触的动物模型，取染毒后小鼠血液标本，采用HLB固相萃取的方法进行样品提取和净化，气相色谱质谱-串联法进行目标物质定性、定量分析，化合物经HP-17MS型毛细管柱分离后，EI离子源进行离子化，选择离子扫描法（SIM）进行质谱检测，采用外标标准曲线法定量。 结果： 该方法可实现血液中苯及其代谢物苯酚、邻苯二酚、对苯二酚、间苯三酚的有效分离和准确定性、定量。该方法检测苯及其代谢物回归方程分别是：苯：y=3 252.1x+1 540，r=0.999 3；苯酚：y=2 046.5x+1 423，r=0.999 1；邻苯二酚：y= 1 853.9x+945，r=0.999 3；对苯二酚：y=1 891.5x+840，r=0.999 2；间苯三酚：y=1 052.4x+655，r=0.999 1。苯、苯酚、邻苯二酚、对苯二酚和间苯三酚的检出限分别为0.03、0.03、0.05、0.05和0.10 μg/g，定量下限分别为0.10、0.10、0.15、0.15和0.30 μg/g，该方法批内精密度为2.64%~10.06%，批间精密度为1.37%~10.17%，加标回收率为89.8%~102.3%。采用该方法可对苯染毒小鼠血液中苯、苯酚、邻苯二酚、对苯二酚和间苯三酚进行定量检测。 结论： 固相萃取-气相色谱串联质谱法可对苯及其代谢物（苯酚、邻苯二酚、对苯二酚和间苯三酚）进行准确定性、定量检测。.
Keywords: Benzene; Biomarkers; Catechol; Hydroquinone; M-trihydroxybenzene; Phenol.