ABSTRACT
A limited overview is given of the separation and detection of specific cytochrome P450 enzymes of the rat. Separation methods include group-specific chromatographic separation and electrophoretic separation in and elution from polyacrylamide gels. Detection methods that are considered include enzymatic analysis with and without chromatographic step using liquid chromatography and immunochemical methods following separation of the cytochrome P450 enzymes by polyacrylamide gel electrophoresis (Western blotting). The advantages and limitations of the various methods have been compared and discussed.
Subject(s)
Cytochrome P-450 Enzyme System/analysis , Electrophoresis, Polyacrylamide Gel/methods , Animals , Blotting, Western , Chromatography, High Pressure Liquid , Colorimetry , Cytochrome P-450 Enzyme System/isolation & purification , Fluorescence , Luminescent Measurements , Rats , Spectrophotometry, UltravioletABSTRACT
The reactions catalysed by the various cytochrome P-450 enzymes are reviewed with respect to the analysis of products by high-performance liquid chromatography (HPLC). Especially biotransformation reactions of purified cytochrome P-450 enzymes in a reconstituted system and in microsomes mainly of rat liver origin are considered. Emphasis is put on the specificity of product formation due to the individual isozymes of cytochrome P-450. It is shown that the presence of eight cytochrome P-450 isozymes can be monitored and determined by specific product formation after HPLC analysis, which is an important parameter in toxicological studies.