Hokkaido Institute of Public Health, Kita-19, Nishi-12, Kita-ku, Sapporo 060-0819, Japan.
Chemosphere. 2011 Apr;83(6):753-9. doi: 10.1016/j.chemosphere.2011.02.062. Epub 2011 Mar 30.
There is a strong need for the development of relatively rapid and low-cost bioassays for the determination of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in environmental and food samples. In this study, we applied a reporter gene assay using DR-EcoScreen cells (DR-cell assay), which is highly sensitive to dioxins, to the determination of PCDD/Fs and dl-PCBs in fish and seafood samples. The PCDD/Fs and dl-PCBs were extracted from homogenated samples (10 g) of 30 fish and shellfish, purified by clean-up procedure using a multilayered silica gel column and an alumina column, and applied to DR-cell assay. Interestingly, the bioanalytical equivalent (BEQ) values obtained from the DR-cell assay [<0.1∼5.4 pg BEQ g(-1) wet weight (ww)] were closely correlated with the toxicity equivalent (TEQ) values from conventional high-resolution gas chromatography/high-resolution mass spectrometry (HRGC-HRMS) analysis (r(2)=0.912), and the slope of regression line was 0.913. Therefore, we multiplied the BEQ values from the DR-cell assay by a conversion coefficient (1.095, the reciprocal of 0.913) to approximate the TEQ values from the HRGC-HRMS analysis. Furthermore, we used this DR-cell assay to perform a prescreening test of PCDD/Fs and dl-PCBs in 16 fish and seafood samples purchased from a supermarket, revealing that a sample from the fatty flesh of a bluefin tuna exceeded 8 pg TEQ g(-1)ww (the European Union-tolerance limit). Taken together, these results suggest that the DR-cell assay might be applicable as a rapid and low-cost prescreening method to determine dioxin levels in fish and seafood samples.
对于环境和食品样品中多氯二苯并对二恶英和二苯并呋喃(PCDD/Fs)以及类似二恶英的多氯联苯(dl-PCBs)的测定,人们强烈需要开发相对快速且低成本的生物测定法。在这项研究中,我们应用了一种高灵敏度的二恶英检测报告基因检测法(DR-EcoScreen 细胞法,DR-cell assay),用于测定鱼类和贝类样品中的 PCDD/Fs 和 dl-PCBs。将 30 种鱼和贝类的均质样品(10 g)中的 PCDD/Fs 和 dl-PCBs 提取出来,通过使用多层硅胶柱和氧化铝柱的净化程序进行纯化,然后应用于 DR-cell assay。有趣的是,从 DR-cell assay 获得的生物分析等效值(BEQ)[<0.1∼5.4 pg BEQ g(-1)湿重(ww)]与传统高分辨率气相色谱/高分辨率质谱(HRGC-HRMS)分析的毒性等效值(TEQ)密切相关(r(2)=0.912),回归线的斜率为 0.913。因此,我们将 DR-cell assay 中的 BEQ 值乘以转换系数(1.095,即 0.913 的倒数),以近似 HRGC-HRMS 分析中的 TEQ 值。此外,我们使用这种 DR-cell assay 对从超市购买的 16 种鱼类和贝类样品中的 PCDD/Fs 和 dl-PCBs 进行了预筛选测试,结果表明,来自蓝鳍金枪鱼的脂肪肉的一个样品超过了 8 pg TEQ g(-1)ww(欧盟容忍限度)。综上所述,这些结果表明,DR-cell assay 可能适用于快速、低成本的鱼类和贝类样品中二恶英水平的预筛选方法。