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4 protocols using bde 99

1

Thyroid Hormone Metabolism Assay

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2,4,6-TBP (99% pure), T4 (> 99%), T3 (> 99%), rT3 (> 99%), 3,3′-T2 (> 99%), and adenosine 3′-phosphate 5′-phosphosulfate (PAPS) lithium salt hydrate (> 60%) were purchased from Sigma-Aldrich (St. Louis, MO). BDE-47 (> 97%), BDE-99 (> 97%), 3-OH 2,2′,4,4′-BDE-47 (3-OH BDE-47; 99%), and 6-OH 2,2′,4,4′-BDE-47 (6-OH BDE-47; 100%) were purchased as neat standards from AccuStandard (New Haven, CT). [13C6]-3,3′-T2 was purchased from Isotec (Miamisburg, OH). 3,3′-T2 sulfate (3,3′-T2S; 98%) was custom synthesized by the Duke University Small Molecule Synthesis Facility (Durham, NC). T3 sulfate (T3S) was purchased from Toronto Research Chemicals (Toronto, ON; 98.6%).
Ham’s F-12K (Kaighn’s Modification) Medium cell culture media and other cell culture reagents were purchased from Life Technologies (Carlsbad, CA). All solvents and other reagents were purchased from VWR (Radnor, PA). BeWo cells (ATCC® CCL-98) were obtained from the Duke University Cell Culture Facility (Durham, NC). All cells were grown in Tissue Culture treated flasks (75 and 182 cm2) and dishes (36.3 cm2) from Genesee Scientific (San Diego, CA).
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BDE Analysis: Quantification of PBDEs

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Standards of the individual target analytes, including BDE-28, BDE-47, BDE-99, BDE-100, BDE-153, BDE-154, BDE-183, and BDE-209 as well as the internal standard, 13C-BDE-153, were purchased from AccuStandard (New Haven, CT, USA). Silica gel, acetone, n-hexane, dichloromethane, ethyl acetate, and methanol were obtained from Merck Serono Co., Ltd. (Darmstadt Germany). Water was purified using a Milli-Q integral water purification system (Millipore, Milford, MA, USA).
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Comprehensive Analysis of Persistent Organic Pollutants

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Standard solutions of organochlorine pesticides (OCs) and their metabolites, as well as of organophosphate pesticides (Ops) (see Table 1 for the list of OCs and OPs), were purchased from Restek (Bellefonte, PA, USA). Similarly, standard solutions of polycyclic aromatic hydrocarbons (i.e., chrysene, benzo(α)anthracene, benzo(β)fluoranthene, and benzo(α)pyrene) were purchased from Restek (Bellefonte, PA, USA). Mixed solutions of PCB congeners (see Table 2 for the list) and CB-209, which was used as an internal standard (IS) for PCBs and PAHs, as well as mixed solutions of PBDE congeners (BDE-28; BDE-33; BDE-47; BDE-99; BDE-100; BDE-153; and BDE-154) and fluorobromodiphenyl ether (FBDE), which was used as IS for flame retardants, were purchased from AccuStandard (New Haven, CT, USA). The standard solutions of per- and polyfluoroalkyl substances (PFASs; see Table 3 for the list), as well as the two 13C-labeled IS of MPFNA and MPFOS, were purchased from AccuStandard (New Haven, USA). The QuEChERS ‘SupelTM QuE Citrate (EN) tubes for extraction and SupelTM QuE-ZSEP (EN) tubes for the clean-up step used for analyses of OCs, OPs, PCBs, PAHs, and PBDEs were purchased from Supelco (Sigma-Aldrich, St. Louis, MO, USA). The extraction cartridges used for PFASs analyses (Oasis WAX 3 mL, 60 mg) were provided by Waters (Milford, MA, USA).
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Preparation of BDE and BaP Solutions

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BDE-47 (CAS 5436-43-1, 100% purity) and BDE-99 (CAS 60348-60-9, 99.2% purity) were purchased from AccuStandard Inc. (New Haven, CT). Benzo[a]pyrene (BaP, CAS B1760) was purchased from Sigma-Aldrich Co. (St. Louis, U.S.A). Working solutions of BDE-47, BDE-99 (50 and 500 nM) and BaP (500 nM) were diluted from stock solutions prepared in dimethyl sulfoxide with sea salted water (0.0375% sea salt in deionized distilled water). The vehicle control solution contained 0.005% dimethyl sulfoxide. A 4% (w./v.) paraformaldehyde solution was prepared in phosphate-buffered saline solution (PBS) with the pH adjusted to 7.4.
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