Grova Nathalie, Salquèbre Guillaume, Hardy Emilie M, Schroeder Henri, Appenzeller Brice M R
Laboratory of Analytical Human Biomonitoring, CRP-Santé, Luxembourg, 1511, Luxembourg France.
Laboratory of Analytical Human Biomonitoring, CRP-Santé, Luxembourg, 1511, Luxembourg France.
J Chromatogr A. 2014 Oct 17;1364:183-91. doi: 10.1016/j.chroma.2014.08.082. Epub 2014 Sep 6.
Since exposure to benzo[a]pyrene is suspected to be associated with several health issues, significant efforts have been made to develop efficient strategies for the assessment of human exposure to this ubiquitous compound. In this context, a method was developed for the analysis of four tetrahydroxylated-benzo[a]pyrene isomers resulting from the hydrolysis of their respective diol-epoxide precursors which are involved in DNA-adduct formation. The analytical sensitivity necessary to reach environmental levels of concentration was obtained by using gas chromatography-tandem mass spectrometry. The recovery determined at the four concentration levels were estimated in average at 83% for benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol(±), 29% for benzo[a]pyrene-r-7,t-8,t-9,t-10-tetrahydrotetrol(±), and 82% for benzo[a]pyrene-r-7,t-8,C-9,c-10-tetrahydrotetrol(±). The coefficient of determination of the calibration curve was above 0.997 for all the analytes investigated and the limit of quantification ranged from 0.5 to 2 adduct/10(8) nucleotides. The precision was between 5.3% and 22.3%. The suitability of the method was firstly evaluated by the analysis of DNA isolated from white blood cells of rats submitted after controlled exposure to benzo[a]pyrene. The four targeted tetra-OH-benzo[a]pyrenes as well as two unknown isomers were detected in all the treated animals. Benzo[a]pyrene-r-7,t-8,c-9,c-10-tetrahydrotetrol(±) appeared as the most abundant isomer in both treated and control animals followed by benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol(±). The method was afterwards applied to the analysis of DNA isolated from white blood cells of human volunteers. The results confirmed that this method was sufficiently sensitive to monitor environmental levels of exposure since all the specimens analyzed were above the limit of quantification for benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydrotetrol(±) and two of them were positive for benzo[a]pyrene-r-7,t-8,c-9,c-10-tetrahydrotetrol(±), thereby highlighting interspecies differences in the nature of the tetrahydroxylated-benzo[a]pyrene isomers formed. This study confirms the necessity to focus on all the tetrahydroxylated-benzo[a]pyrene isomers, which could be indicators of benzo[a]pyrene-associated toxicity related to an individual's own metabolism, rather than limit to a single form.
由于怀疑接触苯并[a]芘与多种健康问题有关，人们已做出巨大努力来开发有效的策略，以评估人类对这种普遍存在的化合物的接触情况。在此背景下，开发了一种方法，用于分析四种四羟基化苯并[a]芘异构体，这些异构体是由其各自参与DNA加合物形成的二醇环氧化物前体水解产生的。通过使用气相色谱 - 串联质谱法获得了达到环境浓度水平所需的分析灵敏度。在四个浓度水平下测定的回收率平均估计为：苯并[a]芘 - r - 7,t - 8,t - 9,c - 10 - 四氢四醇（±）为83%，苯并[a]芘 - r - 7,t - 8,t - 9,t - 10 - 四氢四醇（±）为29%，苯并[a]芘 - r - 7,t - 8,C - 9,c - 10 - 四氢四醇（±）为82%。所研究的所有分析物的校准曲线的决定系数均高于0.997，定量限范围为0.5至2个加合物/10(8)个核苷酸。精密度在5.3%至22.3%之间。该方法的适用性首先通过分析从经控制暴露于苯并[a]芘后的大鼠白细胞中分离的DNA进行评估。在所有处理过的动物中均检测到了四种目标四羟基化苯并[a]芘以及两种未知异构体。苯并[a]芘 - r - 7,t - 8,c - 9,c - 10 - 四氢四醇（±）在处理组和对照组动物中均表现为最丰富的异构体，其次是苯并[a]芘 - r - 7,t - 8,t - 9,c - 10 - 四氢四醇（±）。该方法随后应用于分析从人类志愿者白细胞中分离的DNA。结果证实，该方法灵敏度足以监测环境暴露水平，因为所有分析的样本均高于苯并[a]芘 - r - 7,t - 8,t - 9,c - 10 - 四氢四醇（±）的定量限，其中两个样本对苯并[a]芘 - r - 7,t - 8,c - 9,c - 10 - 四氢四醇（±）呈阳性，从而突出了所形成的四羟基化苯并[a]芘异构体性质的种间差异。这项研究证实了有必要关注所有四羟基化苯并[a]芘异构体，它们可能是与个体自身代谢相关的苯并[a]芘相关毒性的指标，而不是局限于单一形式。
