rdf:type |
|
lifeskim:mentions |
|
pubmed:issue |
1
|
pubmed:dateCreated |
1993-10-21
|
pubmed:abstractText |
A high-performance liquid chromatographic method which measures formation of product 4-nitrocatechol (4NC) has been developed and applied to the study of human liver microsomal 4-nitrophenol (4NP) hydroxylation. Following diethyl ether extraction, 4NC and the assay internal standard (salicylamide) were separated by reversed-phase (C18) liquid chromatography. Extraction efficiencies of 4NC and internal standard were both > 90%. The assay, which has a limit of detection of 15 pmol injected (or an incubation 4NC concentration of 0.25 microM), is accurate, reproducible and straightforward. With a chromatographic time of 12 min, 40-50 samples may be analyzed per day. Rates of 4NC formation were linear with time and protein concentration to 50 min and 0.5 mg/ml, respectively. Preliminary studies of 4NP hydroxylation showed that this reaction followed single enzyme Michaelis-Menten kinetics in human liver microsomes.
|
pubmed:language |
eng
|
pubmed:journal |
|
pubmed:citationSubset |
IM
|
pubmed:chemical |
|
pubmed:status |
MEDLINE
|
pubmed:month |
Jun
|
pubmed:issn |
0021-9673
|
pubmed:author |
|
pubmed:issnType |
Print
|
pubmed:day |
23
|
pubmed:volume |
616
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
73-8
|
pubmed:dateRevised |
2006-11-15
|
pubmed:meshHeading |
pubmed-meshheading:8376495-Catechols,
pubmed-meshheading:8376495-Chromatography, High Pressure Liquid,
pubmed-meshheading:8376495-Cytochrome P-450 CYP2E1,
pubmed-meshheading:8376495-Cytochrome P-450 Enzyme System,
pubmed-meshheading:8376495-Glycerol,
pubmed-meshheading:8376495-Humans,
pubmed-meshheading:8376495-Hydroxylation,
pubmed-meshheading:8376495-Microsomes, Liver,
pubmed-meshheading:8376495-Nitrophenols,
pubmed-meshheading:8376495-Oxidoreductases, N-Demethylating,
pubmed-meshheading:8376495-Quality Control,
pubmed-meshheading:8376495-Solvents,
pubmed-meshheading:8376495-Spectrophotometry, Ultraviolet
|
pubmed:year |
1993
|
pubmed:articleTitle |
High-performance liquid chromatographic assay for 4-nitrophenol hydroxylation, a putative cytochrome P-4502E1 activity, in human liver microsomes.
|
pubmed:affiliation |
Department of Clinical Pharmacology, Flinders Medical Centre, Bedford Park, Adelaide, Australia.
|
pubmed:publicationType |
Journal Article,
In Vitro,
Research Support, Non-U.S. Gov't
|