title: Validation of the CALUX bioassay as a screening and semi-quantitative method for PCDD/F levels in cow's milk.
abstract: The objectives of this study were first to set up and validate the quality criteria of the Xenobiotic Detection Systems-chemical activated luciferase gene expression (CALUX) bioassay for the analysis of cow's milk samples spiked with polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs). The application of this bioassay was then tested by analyzing 28 commercially available pasteurized milk samples and comparing the data with the reference method for PCDD/F analysis. The CALUX criteria from the U.S. EPA and the European Union (EU) were calculated from 16 replaced spiked milk (SM) samples and 8 performance evaluation (PE) samples to validate the CALUX bioassay system. The CALUX bioassay criteria included control chart for quality control (QC) standards, recovery efficiency, and data comparability. The control chart for QC standards were both within the micro+/-2sigma range. The recovery efficiencies ranged from 60.4% to 106% with an average of 79.9% (relative standard deviation (RSD): 20.7%). The mean of data comparability (i.e., relative percent difference, RPD) between CALUX bioassay and high-resolution gas chromatography and high-resolution mass spectrometry (HRGC/HRMS) was 19.0% for SM samples. For the PE samples, Pearson's correlation coefficient between CALUX and the HRGC/HRMS method was 0.953. The high correlation shows that the CALUX system is suitable as a screening method and a semi-quantitative method to analyze the PCDD/F concentration in milk samples. Next, the validated CALUX bioassay was applied to measure 28 commercially available pasteurized milk samples. These milk samples were also analyzed with the HRGC/HRMS method to compare the analysis data from two different methods. There is no false negative sample when applying the bioassay to pasteurized milk and PE samples as a screening method and a semi-quantitative method. The present study indicates that CALUX is a powerful bioassay method for screening a large number of milk samples.
authors.affiliation: Department of Environmental Engineering, National Cheng Kung University, Tainan 70101, Taiwan.
authors.forename: I-Cheng
authors.initials: IC
authors.lastname: Chou
authors.forename: Wen-Jhy
authors.initials: WJ
authors.lastname: Lee
authors.forename: Lin-Chi
authors.initials: LC
authors.lastname: Wang
authors.forename: Guo-Ping
authors.initials: GP
authors.lastname: Chang-Chien
authors.forename: Wei-Shan
authors.initials: WS
authors.lastname: Lee
authors.forename: Hsinyu
authors.initials: H
authors.lastname: Lee
mesh_terms: D000818:Animals; D001572:Benzofurans / Q000032:analysis*; D001681:Biological Assay; D002417:Cattle; D045744:Cell Line, Tumor; D000072338:Dibenzofurans, Polychlorinated; D004785:Environmental Pollutants / Q000032:analysis*; D005260:Female; D015870:Gene Expression; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008892:Milk / Q000737:chemistry*; D000072317:Polychlorinated Dibenzodioxins / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D001572:Benzofurans; D000072338:Dibenzofurans, Polychlorinated; D004785:Environmental Pollutants; D000072317:Polychlorinated Dibenzodioxins; D008156:Luciferases