Bibliographic Details
| Title: |
Vinyl chloride-induced DNA adducts I: Quantitative determination of N,3-ethenoguanine based on electrophore labeling. |
| Authors: |
Fedtke, N., Boucheron, J.A., Turner, M. J., Swenberg, J. A. |
| Superior Title: |
Carcinogenesis; 1990, Vol. 11 Issue 8, p1279-1285, 7p |
| Abstract: |
A sensitive assay for quantitative determination of the vinyl chloride (VC)-induced cyclic DNA adduct N,3-ethenoguanine (EG) was developed. The method is based on the detection of EG as its di-pentafluorobenzyl derivative (3,5-PFB-EG) This compound exhibited good gas chromatographic properties and was detected with high sensitivity by gas chromatography with electron capture detection (limit of detection 300 amol/µl injected solution) or with negative ion chemical ionization mass spectrometry monitoring the [M-181] fragment ion at m/z 354 (GC-NICI MS, limit of detection 190 amol/µl injected solution).EG, its C-labeled analog {C]-EG and 3,5-PFB-EG were synthesized and characterized by UV and fluorescence spectrophotometry, H- and C-NMR spectroscopy and mass spectrometry. The standards were used to optimize the isolation of EG and its derivatization with pentafluorobenzyl bromide (electrophore labeling) at fmol quantities. DNA solutions were spiked with EG, the DNA was depurinated by mild acid hydrolysis, and EG was isolated from the hydrolysates by low-pressure strong cation exchange chromatography with subsequent C solid-phase extraction. The extracted EG was electrophore labeled and 3,5-PFB EG was detected using GC-NICI-MS. [C] EG served as internal standard. 3,5-PFB-EG was quantitated relative to its C-labeied analog by measuring the ion ratio m/z 354/358. The limit of detection for the complete method was 60 fmol EG/µmol guanine. The method was applied to liver DNA from young Sprague-Dawley rats exposed to 600 p.p.m. VC from day 10 through day 14 after birth. The EG concentration in these samples was 1.8 ± 0.3 pmo/µmol guanine. [ABSTRACT FROM PUBLISHER] |
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| Database: |
Complementary Index |
