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2 protocols using uridine 5 diphospho glucuronosyltransferase udpga

1

In Vitro Metabolic Profiling of Isobavachin

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Isobavachin (purity above 98%) was purchased from Shanghai Winherb Medical Technology Co., Ltd (Shanghai, China). Alamethicin, D-saccharic-1, 4-lactone monohydrate, magnesium chloride (MgCl2), nicotinamide adenine dinucleotide phosphate (NADPH) and uridine 5′-diphospho-glucuronosyltransferase (UDPGA) were all obtained from Sigma-Aldrich (St. Louis, MO, USA). Specific substrates and their corresponding metabolites including phenacetin, paracetamol, bupropion, hydroxybupropion, paclitaxel, 6α-hydroxy-paclitaxel, tolbutamide, 4-hydroxytolbutamide, mephenytoin, 4-hydroxymephenytoin, chlorzoxazone, 6-hydroxychlorzoxazone, nifedipine, oxidized nifedipine, β-estradiol, propofol and zidovudine were acquired from Aladdin Chemicals (Shanghai, China). β-estradiol-3-O-glucuronide, propofol-O-glucuronide and AZT-N-glucuronide used in this study were from Toronto Research Chemicals (North York, ON, Canada).
Pooled HLM, individual HLM (iHLM), recombinant human CYP isozymes (rCYP1A2, 2B6, 2C8, 2C9, 2C19, 2E1, 3A4) and UGT isoforms (rUGT1A1, 1A9, 2B7) were all provided by Corning Biosciences (Corning, NY, USA). Mice liver microsomes (MLM) and mice intestine microsomes (MIM) were prepared in our laboratory using the differential centrifugation method as described previously.[16 (link)] Other chemicals and reagents were of analytical grade or better.
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2

Eurycomanone-rich Standardised Herbal Extract

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Standardised water extract of E. longifolia used conformed to the MS: 2409:201 and contained eurycomanone (1.36%), total protein (30.5%), total polysaccharide (37.8%), and glycosaponin (52%). Eurycomanone (94.8% purity) was obtained from ChromaDex Inc. (Irvine, CA, USA). Propranolol, estriol, metoprolol tartrate, diethylstil bestrol, erythromycin, atenolol, carbamazepine, propantheline bromide, enalapril, dasatinib, midazolam, terfinadine, alamethicin, uridine 5′-diphospho-glucuronosyltransferase (UDPGA), nicotinamide adenine dinucleotide phosphate (NADPH), N-methyl-2-pyrrolidone (NMP), 2-hydroxypropyl-beta-cyclodextrin (HPCD), and chemicals used in reagents and buffer preparations were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Plasma and microsomes from mice and rats were prepared in-house, whilst plasma and microsomes from dog, monkey, and human were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Solvents such as methanol, formic acid, acetonitrile (ACN), and DMSO used were of LC-MS and HPLC grade and were purchased from Fisher Chemicals (Waltham, MA, USA).
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