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Updated: Jan 31, 2026

A General Method for Detecting Nitrosamide Formation in the In Vitro Metabolism of Nitrosamines by Cytochrome P450s
Published on: September 25, 2017
A New Method for Quantitative Determination of Steroid Metabolites of Cytochrome P450-Dependent Reactions Using
A V Kuzikov1,2, R A Masamrekh1,2, V V Shumyantseva3,4
1Institute of Biomedical Chemistry, ul. Pogodinskaya 10, str. 8, Moscow, 119121, Russia.
Abstract:
A method for determination of hydroxylase activity of cytochrome P450 3A4 (CYP3A4) towards its substrate hydrocortisone using fluorescent analysis of the product was developed. 6β-hydroxycortisol, formed during CYP3A4-dependent electrocatalysis, has a characteristic fluorescent peak at λ = 427 ± 2 nm after treating with the sulfuric acid : ethanol (3 : 1) mixture and excitation at λ = 365 nm, which is different from the substrate (hydrocortisone) fluorescence (λ = 525 ± 2 nm). The limit of detection of 6β-hydroxycortisol was 0.32 μM. The developed analytical approach was used to determine the kinetic parameters of CYP3A4-dependent hydrocortisone hydroxylation.
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