Cytochrome P450 bio-affinity detection coupled to gradient HPLC: On-line screening of affinities to cytochrome P4501A2 and 2D6

Here we describe novel on-line human CYP1A2 and CYP2D6 Enzyme Affinity Detection (EAD) systems coupled to gradient HPLC. The use of the systems lies in the detection of individual inhibitory ligands in mixtures (e.g. metabolic mixtures or herbal extracts) towards two relevant drug metabolizing human...

Full description

Saved in:
Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-10, Vol.858 (1), p.49-58
Main Authors: Kool, Jeroen, van Liempd, Sebastiaan M., Harmsen, Stefan, Beckman, Joran, van Elswijk, Danny, Commandeur, Jan. N.M., Irth, Hubertus, Vermeulen, Nico P.E.
Format: Article
Language:English
Subjects:
Analysis
Analytical, structural and metabolic biochemistry
Bio-affinity detection
Biological and medical sciences
Chromatography, High Pressure Liquid - methods
Cytochrome P-450 CYP1A2 - analysis
Cytochrome P-450 CYP1A2 - chemistry
Cytochrome P-450 CYP2D6 - analysis
Cytochrome P-450 CYP2D6 - chemistry
Cytochrome P-450 Enzyme System - analysis
Cytochrome P-450 Enzyme System - chemistry
Cytochrome P450
Fundamental and applied biological sciences. Psychology
General pharmacology
HPLC
Humans
Medical sciences
On-line
Pharmacology. Drug treatments
Stereoisomerism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Here we describe novel on-line human CYP1A2 and CYP2D6 Enzyme Affinity Detection (EAD) systems coupled to gradient HPLC. The use of the systems lies in the detection of individual inhibitory ligands in mixtures (e.g. metabolic mixtures or herbal extracts) towards two relevant drug metabolizing human CYPs. The systems can rapidly detect individual compounds in mixtures with affinities to CYP1A2 or 2D6. The HPLC–EAD systems were first evaluated and validated in flow injection analysis mode. IC50 values of known ligands for both CYPs, tested both in flow injection and in HPLC mode, were well comparable with those measured in microplate reader formats. Both EAD systems were also connected to gradient HPLC and used to screen known compound mixtures for the presence of CYP1A2 and 2D6 inhibitors. Finally, the on-line CYP2D6 EAD system was used to screen for the inhibitory activities of stereoisomers of a mixture of five methylenedioxy-alkylamphetamines (XTC analogs) on a chiral analytical column.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2007.08.005