A continuous-flow detector for cytochrome P-450 and cytochrome P-420

Anal Biochem. 1983 Jul 1;132(1):26-33. doi: 10.1016/0003-2697(83)90421-9.

Abstract

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.

MeSH terms

  • Animals
  • Chromatography
  • Cytochrome P-450 Enzyme System / analysis*
  • Cytochrome P-450 Enzyme System / isolation & purification
  • Cytochromes / analysis*
  • Evaluation Studies as Topic
  • Male
  • Microsomes, Liver / enzymology
  • Rats
  • Rats, Inbred Strains
  • Spectrophotometry / instrumentation*

Substances

  • Cytochromes
  • cytochrome P420
  • Cytochrome P-450 Enzyme System