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Measurement of Human Cytochrome P4501A2 (CYP1A2) Activity In Vitro

Cytochrome P4501A2 (CYP1A2) is responsible for the metabolism of a diverse range of clinically used drugs and dietary and environmental chemicals (including many procarcinogens). CYP1A2 expression is influenced by numerous factors, and hence wide interindividual variability is a characteristic feature of this enzyme in humans. Phenacetin represents a convenient probe for the assessment of human CYP1A2 activity in vitro (hepatic microsomes and recombinant enzyme). It is a relatively high‐turnover substrate that forms only one major primary metabolite, the O‐deethylated derivative acetaminophen. Acetaminophen formation in incubations of phenacetin with a CYP1A2 source is readily measured by HPLC with UV detection. The assay has a low requirement for human liver microsomes or recombinant enzyme, and is both selective and sensitive without the requirement for a solvent extraction step. Overall assay reproducibility is excellent, with coefficients of variation <4%.