Pooled human liver cytosol

Commercial SCCP mixtures (Cl degree: 55.5%), MCCP mixtures (Cl degree: 52%), and LCCP mixtures (Cl degree: 49%) were purchased from Dr. Ehrenstorfer (Augsburg, Germany). 13 C-BDE 209 was purchased from Wellington Laboratories Inc (Guelph, ON, Canada). Pooled human liver microsomes (HLM), pooled human liver cytosol (HLCYT), 2,6-uridinediphosphate glucuronic acid (UDPGA), alamethicin (neat, purity > 99%), 3'-phosphoadenosine-5'-phosphosulfate (PAPS; purity > 60%) lithium salt hydrate, nicotinamide adenine dinucleotide phosphate (NADPH; purity > 99%), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (Castle Hill, NSW, Australia). Ultrapure water was obtained from a Milli-Q system (Merck Millipore, MA, USA). Gas chromatography grade acetonitrile (ACN) was purchased from Merck (Darmstadt, Germany).

Ethylphenidate was obtained from LGC Standards (Molsheim, France) as neat powder (purity>99%) and it was dissolved in methanol (purity ≥ 99.9%) purchased from Merck (Darmstadt, Germany). The internal standard, theophylline, was obtained as powder (anhydrous, purity>99%) from Sigma-Aldrich (Diegem, Belgium). Pooled human liver microsomes (HLMs, mix gender, n=200) were purchased from Tebu-Bio (Boechout, Belgium). Pooled human liver cytosol (HLCYT, mix gender, n=50), chemical standards for 2,6-uridinediphosphate glucuronic acid (UDPGA), alamethicin (neat, purity>99%), adenosine 3′-phosphate 5′-phosphosulfate (PAPS; neat, purity>60%) lithium salt hydrate, 4-nitrophenol (4-NP), 4-nitrophenolglucuronide (4-NP-Gluc; neat, purity>99%), 4-nitrophenolsulfate (4-NP-Sulf; neat, purity>99%) and NADPH (neat, purity>99%) were purchased from Sigma-Aldrich.
Baculovirus-insect cell microsomes containing expressed human recombinant CYP enzyme (rCYP1A2, 2B6, 2C9, 2C19, 2D6, 2E1 or 3A4) co-expressed with human CYP oxidoreductase and human cytochrome b5 were purchased from BD Biosciences (Erembodegem, Belgium) and Tebu-Bio. Ultrapure water was prepared using a Purelab flex water system by Elga (Tienen, Belgium). Acetonitrile and formic acid were purchased from Merck. All organic solvents were HPLC grade.