Vanderperren H, Van Wouwe N, Behets S, Windal I, Van Overmeire I, Fontaine A
Federal Agency for the Safety of the Food Chain, 17 Leuvensesteenweg, B-3080 Tervuren, Belgium.
Talanta. 2004 Aug 8;63(5):1277-80. doi: 10.1016/j.talanta.2004.05.027.
Polyhalogenated aromatic hydrocarbons, such as polychlorinated dibenzo-p-dioxins are a large and diverse group of environmental pollutants. Their tendency to accumulate in the food chain and their toxicity make monitoring necessary. The reference analysis method is laborious and very expensive, therefore cheap and rapid bioassays have been developed. The chemical-activated luciferase bioassay (CALUX) bioassay uses a recombinant cell line, which responds to dioxins and dioxin-like molecules with Ah receptor (AhR)-dependent induction of firefly luciferase in a dose related response. The CALUX was tested for its use in the screening of feed. Aliquots of 20g of enriched feed were extracted with a toluene:methanol mixture (20:4 v/v) and extracts were defatted on 33% H(2)SO(4) silica columns and purified on carbon columns. Only the dioxin and furan fraction was analysed, the PCB fraction was discarded. The precision of the method is acceptable and in compliance with an R.S.D. <30% as suggested for cell-based bioassays in the Commission Directive 2002/70/EC of July 2002. The results evidence good agreement between TEQ-values obtained by either CALUX or GC-HRMS. The method is now routinely in use for a feed screening programme designed by the Federal Agency for the Safety of the Food chain. Approximately, 25 samples are analysed weekly. From the obtained results approximately 10% was confirmed by GC-HRMS. The false positive ratio is 1% and no false negatives were found, making the use of the CALUX technology advantageous.
多卤代芳烃，如多氯二苯并 - 对 - 二噁英，是一大类多样的环境污染物。它们在食物链中积累的趋势及其毒性使得监测成为必要。参考分析方法既费力又非常昂贵，因此已开发出廉价且快速的生物测定法。化学激活荧光素酶生物测定法（CALUX）使用一种重组细胞系，该细胞系对二噁英和二噁英类分子作出反应，通过芳烃受体（AhR）依赖性诱导萤火虫荧光素酶，呈现剂量相关反应。对CALUX在饲料筛选中的应用进行了测试。取20克富集饲料的等分试样，用甲苯：甲醇混合物（20:4 v/v）萃取，萃取液在33%硫酸硅胶柱上脱脂，并在碳柱上纯化。仅分析二噁英和呋喃部分，多氯联苯部分被丢弃。该方法的精密度是可接受的，符合2002年7月欧盟委员会指令2002/70/EC中针对基于细胞的生物测定法建议的相对标准偏差<30%。结果表明通过CALUX或气相色谱 - 高分辨质谱法（GC - HRMS）获得的毒性当量（TEQ）值之间具有良好的一致性。该方法目前常规用于由联邦食物链安全局设计的饲料筛选计划。每周大约分析25个样品。从获得的结果来看，约10%通过GC - HRMS得到确认。假阳性率为1%，未发现假阴性，这使得CALUX技术的应用具有优势。
