Document Detail


HPLC analysis of benzo[a]pyrene-albumin adducts in benzo[a]pyrene exposed rats. Detection of cis-tetrols arising from hydrolysis of adducts of anti- and syn-BPDE-III with proteins.
MedLine Citation:
PMID:  10597906     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Quantitation of protein-benzo[a]pyrene adducts represent a more sensitive analysis method than quantitation of benzo[a]pyrene-DNA adducts. By accurate analysis of benzo[a]pyrene-protein adducts several different molecular adduct forms can be studied. Male Wistar rats were injected i.p. with benzo[a]pyrene, and serum albumin was isolated and subjected to acid hydrolysis at 90 degrees C for 3 h. The hydrolysate was analyzed by HPLC with fluorescence detection. The HPLC profiles obtained after albumin hydrolysis from benzo[a]pyrene exposed animals were compared to similar HPLC profiles from in vitro adducted bovine serum albumin (BSA) and direct hydrolysis of both r-10,t-9-dihydrodiol-c-7,8-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (syn-BPDE-III) and r-10,t-9-t-dihydrodiol-t-7,8-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE-III). After acid hydrolysis of albumin from benzo[a]pyrene exposed rats, 6 fluorescent peaks were separated. Four of the peaks were isomers of benzo[a]pyrene-tetrahydrotetrols, (+/-)-benzo[a]pyrene-r-7,t-8,9,10-tetrahydrotetrol, (+/-)-benzo[a]pyrene-r-7,t-8,9,c-10-tetrahydrotetrol, (+/-)-benzo[a]pyrene-r-7,t-8,c-9,t-10-tetrahydrotetrol and (+/-)-benzo[a]pyrene-r-7,t-8,c-9,10-tetrahydrotetrol. In addition we found two fluorescent peaks, named X1 and X2 with retention times similar to the benzo[a]pyrene-tetrols. The unknown fluorescent peaks reacted similar to the four known tetrols in both dose response experiments and time course experiments. Fluorescent material with retention times equal to X1 and X2 were found after acid hydrolysis of syn-BPDE-III and anti-BPDE-III in acid and in hydrolysates from BSA treated in vitro with syn-BPDE-III and anti-BPDE-III. The ratio X1/X2 was relatively constant indicating epimerization equilibrium between these to species. Synchronous fluorescence analysis of fractions containing X1 or X2 from both in vivo and in vitro experiments showed fluorescence spectra characteristic of benzo[a]pyrene tetrols using a wavelength difference of 34 nm.
Authors:
G A Islam; T Greibrokk; R G Harvey; S Ovrebø
Related Documents :
12204426 - An enzymatic method for the measurement of glycated albumin in biological samples.
8246146 - Facilitated dissociation of albumin-fatty acid complexes by rat hepatocytes.
638196 - Influence of fatty acid and time of focusing on the isoelectric focusing of human plasm...
6656536 - Oleic acid transfer from microsomes to egg lecithin liposomes: participation of fatty a...
6651286 - Growth of candida famata and trichosporon cutaneum on uric acid as the sole source of c...
1377456 - Enzymatically amplified time-resolved fluorescence immunoassay with terbium chelates.
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Chemico-biological interactions     Volume:  123     ISSN:  0009-2797     ISO Abbreviation:  Chem. Biol. Interact.     Publication Date:  1999 Nov 
Date Detail:
Created Date:  2000-01-05     Completed Date:  2000-01-05     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  0227276     Medline TA:  Chem Biol Interact     Country:  IRELAND    
Other Details:
Languages:  eng     Pagination:  133-48     Citation Subset:  IM    
Affiliation:
Department of Toxicology, National Institute of Occupational Health, Oslo, Norway.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Benzo(a)pyrene / analysis*,  metabolism,  toxicity
Benzopyrenes / analysis,  metabolism*,  toxicity
Biotransformation
Chromatography, High Pressure Liquid
Hydrolysis
Male
Rats
Rats, Wistar
Serum Albumin, Bovine / analysis*,  metabolism
Spectrometry, Fluorescence
Stereoisomerism
Chemical
Reg. No./Substance:
0/Benzopyrenes; 0/Serum Albumin, Bovine; 50-32-8/Benzo(a)pyrene; 66212-61-1/BPDE-III

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Chromium(VI)-mediated DNA damage: oxidative pathways resulting in the formation of DNA breaks and ab...
Next Document:  Captan impairs CYP-catalyzed drug metabolism in the mouse.