Development Of Analytical Methods For Measuring PBDEs And Their Metabolites In Human Blood

Polybrominated diphenyl ethers (PBDEs) are widely used asbrominated flame retardants. Due to their stable chemical properties,PBDEs can be persistent in the environment and bioaccumulating inbiota through food chain. PBDEs present potential risks to the healthof human and animals, such as endcrine disruption, neurodevelopmenttoxicity, immunotoxicity and carcinogenic effects.PBDEs were hydroxylated and methoxylated to Hydroxylated Polyb-rominated diphenyl ethers(OH-PBDEs)and Methoxylated polybrominateddiphenyl ethers (MeO-PBDEs). OH-PBDEs and MeO-PBDEs weredetected in a variety of substrates or biotic, even human. OH-PBDEspresent potential risks to the health of human and animals such asbiologicalaccumulating,neurotoxicity, cytotoxicity and so on. Hydroxyl-ated metabolites with thyroid hormone transport protein has a highaffinity, they are easier to combined with thyroid hormone receptorTRα1and TRβ1. The toxicity of OH-PBDEs are higher than the PBDEsitself.There are few reportsand less analysis related to PBDEs andtheir metabolites. Some researchers used some derivatization reagents(diazomethane, methylchloformate, and dansylchloride) to deriveatizedtoxicsubstances, and then detected OH-PBDEs by using gas chromatogr-aphy mass spectrometry (GC-MS). However, these methods were notconductive to environmmental protection.Researches about transformati-on and toxicity of OH-PBDEs are even less. In the condition of laborato-ry long time engaring in the study of persistent organic pollutants(POPs). This subject was able to carry out a simultaneous analysis ofdetections of PBDEs their metabolites. There are four parts：Ⅰ.This section describeds the hazards of polybrominated diphenylethers and their metabolites, such as endocrine, disruption, Neurodevlo-pmental toxicity, immuneotoxicity and carcinogenic effects and so on.The correspponding source, mutual conversionexposurelevels, analyticalmethods were introduced.Ⅱ.A simplified analytical method comprised of solid-phaseextraction(SPE) and determination using gas chromatography coupled with negative chemical ionisation mass spectrometry (GC-NCI-MS) weredeveloped for the determination of10kinds of PolybrominatedDiphenyls ethers (PBDEs) in human serum. The analyte was decomposedby treatment with concentrated sulfuric acid directly on the SPE column.The solvent for denaturing protein and the SPE conditions, such aselution solvent and its volume were optimized during the experiment.The method has been validation by spiking fetal bovine serum at threelevels. The mean accuracy given as recovery relative to internalstandards was78.5%-109.7%for all PBDEs congeners. Therepeatability given as inter-RSDs was0.3%-7.4%, while thereproductivity given as intra-RSDs was1.42%-4.1%. Estimated limit ofdetection (LOD, S/N=3) and limit of quantification (LOQ, S/N=10) werein the range0.104ng/L～0.272ng/L and0.346ng/L～0.906ng/L serumfor TriHepta-BDE. LOQ (blank concentration value×3) for BDE-209was7.914ng/L serums. The method was verified by accurate analysis oforganic contaminants according to standard reference material in1957;and1958. This fast, simple method possesseds high sensitiveity andwere accuracy and precision, and consumed low amounts solvent, and itwas fit for determinati-on of PBDEs.Ⅲ.A simplified analytical method comprised of solid-phaseextraction (SPE) and ultra performance liquid chromatography coupledwith negative electrospray ion source tandem mass spectrometry (UPLC-ESI--MS-MS) has been developed for the determination of eleven kindsof hydroxylated polybrominated diphenyl ethers (OH-PBDEs) in humanblood． The solvent for the mobile phase and the SPE conditions wereoptimized． The recoveries of the OH-PBDEs spiked in human bloodrelative to internal standards were in the range of53.2%～117.9%atthree spiked levels. The relative standard deviations (RSDs) werebetween4.71%and18.85%. The limits of detection(LOD, S/N=3) wasin the range of0.00886～0.0589ng/ml. The proposed method wassensitive, accurate, fast, simple, low solvent consumption and suitablefor the determination of OH-PBDEs in human serum withoutderivatization.Ⅳ.Liquid-liquid extraction (LLE) combined with SPE extractionand separation of PBDEs, MeO-PBDEs and OH-PBDEs. PBDEs andMeO-PBDEs were cleaned-up through silica column. Gaschromatography coupled with electrospray ion mass spectrometry has been developed for the determination of ten kinds of polybrominateddiphenyl ethers and twelve kinds of Methoxylated polybrominateddiphenyl ethers in human blood. Ultra performance liquidchromatography coupled with negative electrospray ion source tandemmass spectrometry (UPLC-ESI--MS-MS) has been developed for thedetermination of eleven kinds of hydroxylated polybrominated diphenylethers (OH-PBDEs) in human blood. Kinds of extracting solvent, SPEcolumn, and the SPE conditions were optimized. Also, the solvent forthe Chromatography Parameter and Mass spectrum parameters of theGC-MS and UPLC-MS/MS were optimized. The recoveries of thePBDEs、OH-PBDEs and MeO-PBDEs spiked in human blood relative tointernal standards were in the range of48.90%~115.9%at three spikedlevels.The relative standard deviations (RSDs) were between5.23%and20.88%. The limits of detection(LOD, S/N=3) was in the range ofPBDEs、 MeO-PBDEs and OH-PBDEs were0.00102ng/ml～0.00734ng/ml and0.0082ng/ml～0.0542ng/ml. The proposed method issensitive, accurate, fast, simple, low solvent consumption and suitablefor the determination of PBDEs, MeO-PBDEs and OH-PBDEs in humanserum without derivatization.