A continuous-flow detector for cytochrome P-450 and cytochrome P-420.
A spectrophotometric detector for automatically monitoring levels of cytochrome P-450 and cytochrome P-420 in chromatographic column effluents is described. Levels of cytochrome P-450 and P-420 as low as 30 pmol/ml can be detected above an absorbance background noise equivalent to 10 pmol/ml of these heme proteins. Spectra and the concentrations of cytochrome P-450 and cytochrome P-420 are determined every 5 sec. The automatic flow reactor and detector offer the following advantages over existing manual methods: (i) greatly reduced analysis time, (ii) measurement of a larger number of independent samples than is practical manually, (iii) simultaneous measurement of cytochrome P-450 and its degradation product cytochrome P-420 immediately after elution from the column, thus avoiding further sample degradation, and (iv) greatly increased accuracy and threshold resolution due to highly reproducible reaction conditions and constant optics.
D000818:Animals; D002845:Chromatography; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000302:isolation & purification; D003580:Cytochromes / Q000032:analysis*; D005069:Evaluation Studies as Topic; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D013053:Spectrophotometry / Q000295:instrumentation*
D003580:Cytochromes; C013464:cytochrome P420; D003577:Cytochrome P-450 Enzyme System