Propofol glucuronide

R(+)-NAF (>99.5% ee) and S(-)-NAF (>99.5% ee) (Figure 1) were synthesized according to previous methods (Shivani et al., 2007 (link)). The internal standard (IS) Z10 (CN201110148322.7) was obtained from our laboratory. HPLC-grade methanol, acetonitrile, and acetic acid were obtained from Merck (Darmstadt, Germany). Uridine diphosphate glucuronic acid (UDPGA), alamethicin, Tris, magnesium chloride, methylum-belliferone (MU), 4-methylumbelliferone (4-MU), 4-methylum-belliferyl-β-D-glucuronide hydrate (4-MU-G), propofol, propofol glucuronide, zidovudine, zidovudine glucuronide, fluconazole, and niflumic acid, trifluoperazine dihydrochloride (TFP), and TFP-glucuronide (TFP-G) were purchased from Sigma–Aldrich (St. Louis, MO, United States). Recombinant human UGTs (UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B15, and 2B17) and the commercially available microsomes from human liver (HLMs), intestine (HIMs), and kidney (HKMs), as well as those from rat liver (RLMs), intestine (RIMs), and kidney (RKMs), were purchased from BD Gentest (Woburn, MA, United States). Protein contents of the microsomes were used according to the manufacturers’ instructions. Ultrapure water was used in all experiments (Millipore, Billerica, MA, United States).

Ammonium bicarbonate (AB), dithiothreitol (DTT), iodoacetamide (IAA), trifluoroacetic acid (TFA) and phenylmethanesulfonyl fluoride (PMSF) were bought from Sigma-Aldrich, USA. Sequencing grade modified trypsin was obtained from Promega (Madison, WI). All peptides and internal standard (purity > 95%) were got from Your R&D Partner. HPLC-grade methanol, formic acid and acetonitrile were acquired from Merck (Darmstadt, Germany).
NADPH solution A and NADPH solution B were got from BD Bioscience, USA. Alamethicin, Tetracycline, Glucosyl monophosphate, Uridine diphosphate glucuronic acid (UDPGA), 3′-phosphoadenosine-5′-phosphosulfate (PAPS), MgCl2, Chlorzoxazone, Testosterone, 6β-hydroxyTestosterone, Propofol, Dopamine, 6-hydroxy Chlorzoxazone, Propofol-glucuronide and 4-Nitrophenyl sulfate metabolite were acquired from Sigma-Aldrich, USA. Dopamine 3-O-sulfate and Dopamine 4-O-sulfate were got from TRC, Canada. P-Nitrophenol was bought from Aladdin, China. Genistein and ammonium acetate were got from Chengdu Mansite Biotechnology Co., Ltd. and Dalian Meilun Biotechnology Co., Ltd., respectively. Coomassie brilliant blue, providing for protein measurement, was bought from Bio-Rad (Hercules, California, USA).