生物转化可以极大地影响积累,随后,生物中物质的毒性。尽管传统上这些定量化合物代谢的研究是在体内进行的,目前,正在开发具有非常不同的细胞系的体外测试方法来进行评估。然而,由于具有非常多样化的多个变量,这仍然是一个非常有限的领域。所以,越来越多的分析化学家正在研究非常小的细胞或其他类似的生物样品。这使得有必要解决允许确定其在细胞内部和在其暴露介质中的浓度的分析方法的发展。这项研究的目的是开发一套用于量化多环芳烃的分析方法,PAHs(菲,PHE),和多溴联苯醚,多溴二苯醚(2,2',4,4'-四溴二苯醚,BDE-47),以及它们在细胞中的主要代谢产物和暴露介质。分析方法,基于小型化超声探头辅助提取,气相色谱-质谱-微电子捕获检测器(GC-MS-µECD),和液相色谱-荧光检测器(LC-FL)测定技术,已经过优化,然后在暴露48小时时将其应用于HepG2的生物转化研究。PHE(1-OH,2-OH,3-OH,4-OH-,和9-OH-PHE)和BDE-47(5-MeO-,5-OH-,和3-OH-BDE-47)在细胞内部和暴露培养基中进行检测和定量。这些结果提供了一种测定和改善代谢比率信息的新方法,以便更好地了解代谢途径及其毒性。
Biotransformation can greatly influence the accumulation and, subsequently, toxicity of substances in living beings. Although traditionally these studies to quantify
metabolization of a compound have been carried out with in vivo species, currently, in vitro test methods with very different cell lines are being developed for their evaluation. However, this is still a very limited field due to multiple variables of a very diverse nature. So, an increasing number of analytical chemists are working with cells or other similar biological samples of very small size. This makes it necessary to address the development of analytical methods that allow determining their concentration both inside the cells and in their exposure medium. The aim of this study is to develop a set of analytical methodologies for the quantification of polycyclic aromatic hydrocarbons, PAHs (phenanthrene, PHE), and polybrominated diphenyl ethers, PBDEs (2,2\',4,4\'-tetrabromodiphenyl ether, BDE-47), and their major metabolites in cells and their exposure medium. Analytical methodologies, based on miniaturized ultrasound probe-assisted extraction, gas chromatography-mass spectrometry-microelectron capture detector (GC-MS-µECD), and liquid chromatography-fluorescence detector (LC-FL) determination techniques, have been optimized and then applied to a biotransformation study in HepG2 at 48 h of exposure. Significant concentrations of the major metabolites of PHE (1-OH, 2-OH, 3-OH, 4-OH-, and 9-OH-PHE) and BDE-47 (5-MeO-, 5-OH-, and 3-OH-BDE-47) were detected and quantified inside the cells and in the exposure medium. These results provide a new method for determination and improve information on the
metabolization ratios for a better knowledge of the metabolic pathways and their toxicity.