Loperamide, DLOP and midazolam (used as internal standard) were purchased from Shanghai Canspec Scientific & Technology Co., Ltd. (Shanghai, China) Acetonitrile and methanol of analytical grade were bought from Merck (Darmstadt, Germany); formic acid was from Sigma-Aldrich Co. (St. Louis, MO, USA). Ultra-pure water was produced with a Milli-Q reagent system (Millipore, Bedford, MA, USA). The reduced nicotinamide adenine dinucleotide phosphate (NADPH) was purchased from Roche Pharmaceutical Ltd (Basel, Switzerland). Recombinant human CYP3A4 enzymes and purified cytochrome b5 were kind gifts from Beijing Hospital (Beijing, China).24 (link)–26 (link)
Protocol detail
Loperamide and DLOP Metabolism Study
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Acetonitrile
methanol
formic acid
Milli-Q reagent system
reduced nicotinamide adenine dinucleotide phosphate (NADPH)
Acetonitrile
Cyp3a4
Cytochrome b5
Enzymes
Formic acid
Gifts
Human
Loperamide
Methanol
Midazolam
Nicotinamide adenine dinucleotide phosphate
Pharmaceutical
Corresponding Organization : Wenzhou Medical University
Other organizations : First Affiliated Hospital of Wenzhou Medical University, Beijing Hospital
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Variable analysis
independent variables
- Loperamide
- Unspecified
- Midazolam (used as internal standard)
- Acetonitrile and methanol of analytical grade
- Formic acid
- Ultra-pure water produced with a Milli-Q reagent system
- Reduced nicotinamide adenine dinucleotide phosphate (NADPH)
- Recombinant human CYP3A4 enzymes
- Purified cytochrome b5
- Positive control: Recombinant human CYP3A4 enzymes and purified cytochrome b5
- Negative control: Not explicitly mentioned
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