Aplicación del bioensayo EROD-H4IIE para la determinación de dioxinas en carnes de pollos broiler: un estudio de equivalencia con la cromatografía de gases de alta resolución acoplada a espectrometría de masas de alta resolución

Abstract

ARTÍCULO ORIGINALAplicación del bioensayo EROD-H4IIE para la determinación de dioxinas en carnes de pollos broiler: un estudio de equivalencia con la cromatografía de gases de alta resolución acoplada a espectrometría de masas de alta resolución # � Application of the EROD-H4IIE bioassay for the determination of dioxins in broiler chicken meat: an equivalence study with high resolution gas chromatography coupled to high resolution mass spectrometry SUMMARYThe EROD bioassay with H4IIE cell line was applied in this study to determine the equivalence of the results for chicken meat between the EROD H4IIE Bioassay in pg TCDD-EQ/g of tissue, and the results of the gas chromatography coupled to high resolution spectrometry (HRGC/HRMS) in pg WHO-TEQ/g of fat of TEQs. 41 compound samples of chicken drumsticks were used. The samples were obtained in slaughtering plants of 4 different production facilities of Chile, during the sacrifice of animals (38 and 43 days old) between 2004 and 2007. Each sample was analysed with both analytical techniques. A regression model for the equivalence of both techniques was determined from the results. The model obtained was: HRGC/ HRMS = 0.481 + 0.051[EROD-H4IIE] 2 , R 2 = 0.885, therefore for a value in EROD-H4IIE of 2.2 pg TCDD-EQ/g of tissue it is estimated that it will correspond to 0.73 pg WHO-TEQ/g of fat in HRGC/HRMS. Also, for the same value of EROD-H4IIE with a 95% confidence, an estimate as an upper limit equal to 0.83 pg WHO-TEQ/g of fat in HRGC/HRMS will be obtained. An upper limit equal to 0.88 pg WHO-TEQ/g of fat in HRGC/HRMS is estimated by taking the same value of EROD-H4IIE with 99% confidence. It is concluded that de EROD-H4IIE bioassay can be applied as a screening method in animal production systems and specifically in broiler chicken production.Palabras clave: dioxina, bioensayo H4IIE, HRGC/HRMS, EROD.

Application of the EROD-H4IIE bioassay for the determination of dioxins in pork in comparison to high resolution gas chromatography coupled to high resolution mass spectrometry

“…These samples were analysed by HRGC/HRMS, effectively confirming 36 as positive. Overestimates have also been reported in the analysis of poultry samples (Schoffer et al 2011) and ash (Till et al 1997). The difference between the two methods may be due to compounds that can trigger the aryl hydrocarbon receptor (AhR) pathway, such as polycyclic aromatic hydrocarbons (Chaloupka et al 1993), polybrominated compounds (Baston and Denison 2011), indol-3-carbinol (13C), which is present in cruciferous plants and has been described as a moderate activity binder with the AhR receptor (Chen et al 1996), in addition to DL-PCBs which in this study work were not analysed by HRGC/HRMS.…”

“…In this study the bioassay lasted an average of 12 days from the beginning of the extraction until the reporting of the results and it showed a high sensitivity (LOD = 0.142 ± 0.1208) and a high reproducibility considering the quality control criteria of the assay. Schoffer et al (2011) demonstrated the equivalence between the bioassay and HRCG/HRMS for the detection of dioxins in broiler chicken meat, reporting that it can be a useful technique to biomonitor these xenobiotics in animal production. However, in this study it was not possible to estimate a significant equivalence between the results obtained from both methods in pork samples, because there was not significant correlation between the data produced by the two analytical methods, this precluded the use of a model to estimate the expected value of PCDD/Fs contamination, which would be obtained by HRCG/HRMS, only if the EROD-H4IIE bioassay is used.…”

“…Therefore, a number of screening assays has been developed, such as the 7-ethoxyresorufin-O-deethylase (EROD)-bioassay, the aryl hydrocarbon hydroxylase (AHH) bioassay, the enzyme immunoassay (EIA), the chemical-activated luciferase gene expression (CALUX), the gel retardation of AhR DNA binding (GRAB) assay, the recAhr DELFIA assay kit, the Ah receptor (AhR) (or filtration) assay with radiolabeled dioxins, the Ah-immunoassay (AhIA) (Behnisch et al 2001), the chemical-activated fluorescent expression (CAFLUX) (Zhao et al 2010) and DR-EcoScreen® bioassay (Anezaki et al 2009, Kojima et al 2010. A viable alternative is to use in vitro bioassay cell line H4IIE rat hepatoma as screening method in pork production, considering that Schoffer et al (2011) determined the equivalence between the results of the bioassay of meat from broiler chickens with HRGC/HRMS, showing that the first one provides very accurate estimates (R 2 =0.885), and is therefore considered a useful technique for biomonitoring, as a screening method, in broiler chicken production and can be extended to other animal production systems. Therefore, the objective of this study is to determine the presence of dioxins and furans in pork produced in Chile by HRGC/HRMS in pg WHO-TEQ/g fat and H4IIE-EROD bioassay in pg TCDD-EQ 2 /g of sampling tissue, and to determine the equivalence of the results generated by the bioassay as a screening method and HRGC/HRMS as identity confirmatory method.…”

Comparison of a Biosensor and a Commercial UV-Visible Method for Measuring Hexavalent Chromium in Liquid Medium

“…These chemical analyzes require pretreatment to preserve the sample during transfer to the laboratory, to avoid altering the results. This occurs, for example, in gas chromatography and mass detection analyzes; expensive and large equipment that requires its use in the laboratory (Schoffer et al, 2011).…”

Application of a model to estimate total concentrations of dioxins and DL-PCBs in pork production in Chile

“…The modified protocol of Tillitt et al (1991) was used, as described by Schoffer et al (2011) and Samsing et al (2011), and incorporated into the standard procedure of the CIE Mayor P-TEC-001. The results of the analysis were expressed in pg TCDD-EQ/g of sample.…”