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3 protocols using diazepam

1

Quantitative Analysis of Drug Compounds

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Rivaroxaban (purity >99%) and diazepam (purity >98%) were both purchased from J&K Scientific Ltd. (Beijing, China) (Figure 1). Rivaroxaban metabolite M2 (Lot No. 3530-047A1) was purchased from TLC Pharmaceutical Standards Ltd. (Newmarket, Ontario). Ticagrelor (purity > 98%) was purchased from Aladdin (Aladdin, Shanghai, China). Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was obtained from Roche Pharmaceuticals Ltd. (Basel, Switzerland). Acetonitrile and methanol were obtained from Fisher Scientific Co. (Fair Lawn, NJ, USA). Formic acid was got from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was obtained from a Milli-Q water purification system (Millipore, Billerica, MA, USA). All other chemicals were of analytical grade or better.
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2

Analytical Method for Xenobiotic Metabolism

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Phenacetin, bupropion, tolbutamide, dextromethorphan, midazolam, chlorzoxazone, and the diazepam (all purity> 98%) that used as internal standards (ISs) were purchased from J&K Scientific Ltd. (Beijing, China). Vonoprazan was purchased from Beijing Sunflower Scientific Ltd. (Beijing, China). Hydroxybupropion and hydroxymidazolam were purchased from Sigma-Aldrich (St Louis, USA). Dextrorphan, hydroxytolbutamide, hydroxychlorzoxazone, and 4-acetamidophenol were purchased from Toronto Research Chemicals (Toronto, Canada). Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was acquired from Roche Pharmaceuticals Ltd. (Basel, Switzerland). Ultra-pure water was produced by Milli-Q, a reagent-standard water purification system (Millipore, Bedford, USA). Acetonitrile and methanol that of high performance liquid chromatography (HPLC) grade were obtained from Merck Company (Darmstadt, Germany).
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3

Cardiovascular Drugs Metabolism Protocol

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The cardiovascular drugs verapamil hydrochloride, diltiazem hydrochloride, nifedipine, amlodipine besylate, amiodarone hydrochloride, quinidine, propafenone, valsartan, irbesartan, benazepril hydrochloride, and captopril were all purchased from Aladdin (Shanghai, China). The vonoprazan was obtained from Shanghai Send Pharm, and its carboxylic acid metabolite, MI, was provided by Wuxi Apptec Co., Ltd. (Shanghai, China).
Diazepam from J&K Scientific Ltd. (Beijing, China) was selected as the internal standard (IS). Nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Coolaber (Beijing, China). Methanol and acetonitrile of high-performance liquid chromatography (HPLC) grade were obtained from Merck Company (Darmstadt, Germany). Formic acid was purchased from Sigma-Aldrich (St. Louis, United States). Ultra-pure water was produced by the reagent-standard water purification system Milli-Q (Millipore, United States). The RLMs were prepared according to our previously reported method (Wang et al., 2020 (link)), and the HLMs were purchased from Bioruler (Wuhan, China).
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