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

✍ Scribed by R.F. Bonner; P.M. Bungay; L.R. Pohl; R.V. Branchflower

Publisher
Elsevier Science
Year
1983
Tongue
English
Weight
629 KB
Volume
132
Category
Article
ISSN
0003-2697

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✦ Synopsis

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.

KEY WORDS: cytochrome P-450; cytochrome P-420; chromatography; instrumentationspectrophotometric; continuous detector.

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In two cases on p. 707, the word "potentially" should have been removed. The sentences should read: "CYP450, another heme-containing protein, is one of the most versatile enzymes in nature and could efficiently catalyze hundreds of different substrates through a variety of difficult biotransformatio