Scientific article

A cocktail approach for assessing the in vitro activity of human cytochrome P450s: An overview of current methodologies

Published inJournal of pharmaceutical and biomedical analysis, vol. 101, p. 221-237
Publication date2014

An assessment of cytochrome P450 (CYP) enzyme activity is essential for characterizing the phase I metabolism of biological systems or to evaluate the inhibition/induction properties of xenobiotics. CYPs have generally been investigated individually by single probes, and metabolite formation has been monitored by liquid chromatography-mass spectrometry (LC–MS). To increase the throughput, many probes have been applied to assess multiple CYP activities simultaneously within a single experiment. This strategy is called the cocktail approach, and it has already been reviewed for in vivo applications, but never for in vitro ones. This review focuses for the first time on an in vitro cocktail approach, and it references the most notable articles on this topic. The advantages and limitations of applying cocktails for the in vitro activity assessment of major human CYPs, namely, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and subfamily CYP3A, are discussed. This article considers the probe reaction selections for each CYP according to regulatory recommendations, probe metabolic properties (i.e., specificity and turnover), probe concentrations and analytical sensitivity, but it also highlights a challenge specific to cocktail design, which is probe-probe interaction. The last part of the review reports some methodologies for incubating these cocktails and discusses some important issues regarding the incubation time,enzyme concentrations and sample preparation.

  • Cocktail
  • CYP phenotyping
  • Human liver microsomes
  • Drug–drug interactions
  • In vitro methodology
Citation (ISO format)
SPAGGIARI, Dany et al. A cocktail approach for assessing the in vitro activity of human cytochrome P450s: An overview of current methodologies. In: Journal of pharmaceutical and biomedical analysis, 2014, vol. 101, p. 221–237. doi: 10.1016/j.jpba.2014.03.018
Main files (1)
Article (Published version)
ISSN of the journal0731-7085

Technical informations

Creation11/02/2016 3:35:00 PM
First validation11/02/2016 3:35:00 PM
Update time03/15/2023 12:54:15 AM
Status update03/15/2023 12:54:15 AM
Last indexation10/19/2023 2:21:24 AM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack