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Samples are extracted following JIS (Japanese Industrial Standard) methods or by a sonication extraction method. The crude extract is concentrated, resuspended in hexane and subjected to the XDS patented clean-up method. The sample is then applied to acid silica column and XCARB (activated carbon) columns and PCDD/Fs and coplanar PCBs are separated using appropriate solvents (separation of the PCDD/Fs and coplanar PCBs is not always necessary). The solvent is replaced with DMSO and added to cultures of recombinant mouse hepatoma (H1L6.1c3) CALUX cells grown in 96-well microplates. A dilution series of 2,3,7,8-TCDD is included in each plate as the positive control and to generate a standard curve of luciferase induction. After 24 hours, luciferase activity in each sample extract well is measured and compared to the 2,3,7,8-TCDD standard curve to allow calculation of total CALUX TEQ. |
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