Document Detail


Determination of 3-oxo-delta4- and 3-oxo-delta4,6-bile acids and related compounds in biological fluids of infants with cholestasis by gas chromatography-mass spectrometry.
MedLine Citation:
PMID:  9200514     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A method has been developed for the determination of 3-oxo-delta4- and 3-oxo-delta4,6-bile acids and related bile acids in biological fluids of infants by gas chromatography-mass spectrometry (GC-MS) of the methyl ester-dimethylethylsilyl ether-methoxime derivatives. The 7alpha-hydroxylated 3-oxo-delta4-bile acids were partially dehydrated to give the 3-oxo-delta4,6-bile acids by trimethylsilyl or dimethylethylsilyl derivatization and other pretreatments under acidic or alkaline conditions for GC-MS analysis. To prevent dehydration, the 3-oxo-delta4-bile acids were derivatized to the oximes by treatment with O-methylhydroxylamine prior to pretreatments such as solid-phase extraction, enzymatic solvolysis and hydrolysis of the conjugates, and silylation with dimethylethylsilylimidazole. Calibration curves for the bile acids were linear over a range of 5-250 ng and the detection limit was 100 pg for each 3-oxo-delta4-bile acid. Recoveries of the bile acids and their glycine and taurine conjugates from bile acid-free urine and serum ranged from 94.2 to 105.9% of their added amounts. The bile acids in urine and serum of four patients with severe cholestatic liver disease were measured by the analytical method, and the 3-oxo-delta4-bile acids were determined to be the major bile acids (59-68%) in the urines associated with 3-oxo-delta4-steroid 5beta-reductase deficiency or dysfunction.
Authors:
M Suzuki; T Murai; T Yoshimura; A Kimura; T Kurosawa; M Tohma
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of chromatography. B, Biomedical sciences and applications     Volume:  693     ISSN:  1387-2273     ISO Abbreviation:  J. Chromatogr. B Biomed. Sci. Appl.     Publication Date:  1997 May 
Date Detail:
Created Date:  1997-08-20     Completed Date:  1997-08-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9714109     Medline TA:  J Chromatogr B Biomed Sci Appl     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  11-21     Citation Subset:  IM    
Affiliation:
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan.
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MeSH Terms
Descriptor/Qualifier:
Bile Acids and Salts / blood*,  urine*
Cholestasis / blood*,  urine*
Gas Chromatography-Mass Spectrometry
Humans
Infant
Chemical
Reg. No./Substance:
0/Bile Acids and Salts

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