Piskorska-Pliszczyńska Jadwiga, Małagocki Paweł, Furga Beata, Gembal Magdalena, Cebulska Joanna
Radiobiology Department, National Veterinary Research Institute, 24-100 Pulawy, Poland.
J Vet Res. 2017 Dec 27;61(4):473-481. doi: 10.1515/jvetres-2017-0066. eCollection 2017 Dec.
Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DL-PCBs) belong to a well-known group of pollutants. Present in feedstuffs, they bioaccumulate in tissues of food-producing animals. Food is the source of over 90% of human PCDD/Fs and DL-PCBs intake. Thus, feed control is one of the pillars of the EU strategy and a mean of reducing human exposure. The article presents AhR based reporter gene bioassay method for PCDD/Fs and DL-PCBs analysis in feed and its validation results.
Analytes were extracted from samples with fat. Subsequently, fat and other interferences were removed from extract using sulphuric acid modified silica. Extract was further cleaned and PCDD/Fs separated from DL-PCBs using carbon column. Contaminants detection was performed using H1L6.1c3 cell line, which produces luciferase in response to AhR ligands present in extract.
Performance characteristics (repeatability, reproducibility, and apparent recovery) fulfil the requirements of Regulation 2017/771/EU. The positive correlation between bioassay and reference HRGC-HRMS method was confirmed. Moreover, the role of screening method used in connection with the confirmatory HRGC-HRMS method in providing feed and food safety has been discussed.
Bioassay is a useful method for dioxin and DL-PCBs analysis, allowing cost reduction of monitoring programmes with minimal risk of false negative results.
多氯代二苯并 - 对 - 二噁英、多氯代二苯并呋喃（PCDD/Fs）和类二噁英多氯联苯（DL - PCBs）属于一类广为人知的污染物。它们存在于饲料中，会在产肉动物组织中生物累积。食物是人类摄入PCDD/Fs和DL - PCBs的90%以上的来源。因此，饲料控制是欧盟战略的支柱之一，也是减少人类接触的一种手段。本文介绍了基于芳烃受体（AhR）的报告基因生物测定法用于饲料中PCDD/Fs和DL - PCBs的分析及其验证结果。
用脂肪从样品中提取分析物。随后，使用硫酸改性硅胶从提取物中去除脂肪和其他干扰物。提取物进一步净化，并用碳柱将PCDD/Fs与DL - PCBs分离。使用H1L6.1c3细胞系进行污染物检测，该细胞系会因提取物中存在的AhR配体而产生荧光素酶。
性能特征（重复性、再现性和表观回收率）符合欧盟法规2017/771/EU的要求。证实了生物测定法与参考高分辨气相色谱 - 高分辨质谱（HRGC - HRMS）方法之间存在正相关。此外，还讨论了与确证性HRGC - HRMS方法结合使用的筛查方法在保障饲料和食品安全方面的作用。
生物测定法是一种用于二噁英和DL - PCBs分析的有用方法，可降低监测计划的成本，同时将假阴性结果的风险降至最低。
