Linked Life Data

12906903

Source:http://linkedlifedata.com/resource/pubmed/id/12906903

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2003-8-8
pubmed:abstractText
A sensitive assay for the determination of (-)-verbenone 10-hydroxylation catalyzed by rat liver microsomes was developed using high-performance liquid chromatography. Verbenone was incubated in vitro with liver microsomes of untreated rats and rats treated with phenobarbital and the products thus formed were extracted with CH(2)Cl(2) and the extracts were separated by HPLC with a C(18) 5-microm analytical column. Elution was conducted with 40% methanol containing 20 mM NaClO(4) and the detection of UV absorbance was done at 251 nm. Product formation was dependent on the incubation time at least up to 30 min and the microsomal protein concentration between 0.01 and 0.1 mg protein/ml. The limit of detection of (-)-10-hydroxyverbenone with the HPLC was found to be about 40 pg, indicating that this method is about 100-fold sensitive than the GC-MS method. Optimized pH for the reaction was at 7.4 when examined with 100 mM potassium phosphate buffer in different pHs. Kinetic analysis showed that K(m) values for liver microsomes of untreated and phenobarbital-treated rats were 206 and 41 microM and V(max) values were 5.8 and 44 nmol/min/mg protein, respectively. Thus the present results provided a sensitive and useful method for the determination of verbenone 10-hydroxylation catalyzed by rat liver microsomes.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1570-0232
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
793
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
291-6
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
2003
pubmed:articleTitle
High-performance liquid chromatographic method for the determination of (-)-verbenone 10-hydroxylation catalyzed by rat liver microsomes.
pubmed:affiliation
Department of Applied Chemistry, Faculty of Science and Engineering, Kinki University, Kowakae, Higashiosaka, Osaka 577-8502, Japan. miyazawa@apch.kindai.ac.jp
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't