4 nitrophenol sulfate 4 np sulf

Chemical standards for PMA, PMMA, 4-MTA, MBDB, BDB, Gluc; neat, purity>99%), 4-nitrophenol-sulfate (4-NP-Sulf; neat, purity>99%), naphtholglucuronide (Naphth-Gluc; neat, purity>99%), 5-bromo-4-chloro-3-indolyl sulfate (Indolyl-Sulf; neat, purity>99%) and NADPH (neat, purity>99%) were purchased from Sigma-Aldrich (Diegem, Belgium). Ultrapure water was prepared using a Purelab flex water system by Elga (Tienen, Belgium). Acetonitrile, methanol, ammonium hydroxide, hydrochloric acid and ammonium acetate were purchased from Merck (Darmstadt, Germany). All organic solvents were HPLC grade or higher.

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.