Fluorescence detection of benzo[a]pyrene-DNA adducts in human lung

Improved techniques are described for the specific identification of benzo[a]pyrene-diolepoxide (BPDE)-DNA adducts in human tissues. Immunoaffinity chromatography, synchronous fluorescence spectroscopy and second-derivative synchronous fluorescence spectroscopy have previously been used to detect BP...

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Bibliographic Details
Published in:Carcinogenesis (New York) 1991-08, Vol.12 (8), p.1445-1449
Main Authors: Weston, Ainsley, Bowman, Elise D.
Format: Article
Language:English
Subjects:
Adolescent
Adult
Aged
Aged, 80 and over
Benzo(a)pyrene - analysis
Biological and medical sciences
Carcinogenesis, carcinogens and anticarcinogens
Chemical agents
Chromatography, Affinity
Chromatography, High Pressure Liquid
DNA - analysis
DNA Adducts
Female
Humans
Infant
Infant, Newborn
Lung - chemistry
Male
Medical sciences
Middle Aged
Tumors
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Description
Summary:Improved techniques are described for the specific identification of benzo[a]pyrene-diolepoxide (BPDE)-DNA adducts in human tissues. Immunoaffinity chromatography, synchronous fluorescence spectroscopy and second-derivative synchronous fluorescence spectroscopy have previously been used to detect BPDE-DNA adducts in human placenta. Here we report how these methods, together with HPLC and the generation of complete fluorescence excitation—emission matrices, have been used to identify unequivocally BPDE-DNA adducts in samples of human lung. BPDE nucleotide adducts were isolated with immunoaffinity chromatography columns bearing antibodies raised against the (±)anti-7,8-diol-9,10-epoxide-deoxyguanosine adduct of betizo[a]pyrene. These adducts were hydrolyzed to tetrahydrotetrols and the hydrolysis products subjected to HPLC. The major product isolated by HPLC, benzo[a]-pyrene-7,10/8,9-tetrahydrotetrol, was determined by fluorescence spectroscopy. Using this method, levels of BPDE-DNA adducts in the range of 1–40 in 108 nucleotides were measured in 6 out of 25 samples, with a lower detection limit of one adduct in 108 nucleotides. The data may also indicate that adduct levels show regional variation in different parts of the same lung.
ISSN:0143-3334
1460-2180
DOI:10.1093/carcin/12.8.1445