Improved sample preparation for the testosterone hydroxylation assay using disposable extraction columns

The preparation of samples for injection into a high-performance liquid chromatography from assay mixtures for the determination of cytochrome P-450-dependent testosterone hydroxylation has been substantially facilitated. By replacing the multiple cumbersome extraction steps of the conventional meth...

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Bibliographic Details
Published in:Journal of chromatography. Biomedical applications 1992-11, Vol.582 (1), p.232-235
Main Authors: Oesch, F., Wagner, H., Platt, K.L., Arand, M.
Format: Article
Language:English
Subjects:
Analysis of complex biological substances
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Blood and biological fluids
Fundamental and applied biological sciences. Psychology
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Summary:The preparation of samples for injection into a high-performance liquid chromatography from assay mixtures for the determination of cytochrome P-450-dependent testosterone hydroxylation has been substantially facilitated. By replacing the multiple cumbersome extraction steps of the conventional method with a single column extraction the time for sample preparation was reduced from hours to minutes. The new procedure also yields better recoveries for most of the testosterone metabolites than the original protocol. The use of extraction columns for sample preparation allows the simultaneous treatment of a large number of samples or even the automation of the whole assay procedure. The modified procedure is a straightforward, easy-to-perform method that should greatly facilitate the implementation of the testosterone hydroxylation assay for sharply discriminating between many individual cytochrome P-450 species in routine enzyme diagnostics.
ISSN:0378-4347
DOI:10.1016/0378-4347(92)80324-J