PMID: 8468388Feb 26, 1993Paper

Improved thin-layer chromatographic separation of 32P-postlabeled DNA adducts

Journal of Chromatography
G G SpencerR C Gupta

Abstract

DNA adducts represent the putative initiating event in the chemical carcinogenesis process. 32P-Postlabeling is one of several assays which have been developed for the sensitive detection of DNA adducts. An integral part of the 32P-postlabeling assay is the separation of adducted nucleotides by multidirectional, multisolvent, anion-exchange polyethyleneimine-cellulose thin-layer chromatography. Standard since the introduction of this assay has been the use of high-salt, high-urea solvents for the resolution of adducts during the D3 and D4 phases of the chromatography. Urea solvents are able to separate adducts resulting from a number of chemicals, however, they are time-consuming, retain a lot of background noise, may push adducts into inadequately resolved diagonal radioactive zones, and may not separate adducts of similar structure. In this study we introduce the use of a dilute ammonium hydroxide solvent for D4 chromatography and compare it to other standard solvents such as lithium chloride-Tris.HCl-urea, sodium phosphate-Tris.HCl-urea, and isopropanol-4 M ammonium hydroxide for adduct separation, resolution, recovery, retention of background noise, and chromatography development time. We found that 0.2 M ammonium hydroxide...Continue Reading

References

Jul 1, 1992·Carcinogenesis·A C Beach, R C Gupta
Oct 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·K RanderathR C Gupta
Oct 1, 1964·Journal of Chromatography·K RANDERATH, E RANDERATH

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Citations

Mar 3, 1996·Journal of Chromatography. B, Biomedical Applications·G G Spencer-BeachR C Gupta
Feb 1, 1995·Current Opinion in Biotechnology·G Keith, G Dirheimer
Sep 1, 1996·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·C M Dale, R C Garner
Oct 16, 2007·The Science of the Total Environment·Marco PelusoPaolo Boffetta
Jan 1, 1997·Biomarkers : Biochemical Indicators of Exposure, Response, and Susceptibility to Chemicals· Shu-Xin Qu Cheng-Long Bai And Neill H Stacey

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