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Main Title Short Communication: Improved Thin-Layer Chromatographic Separation of 32P-Postlabeled DNA Adducts.
Author Spencer, G. G. ; Beach, A. C. ; Gupta, R. C. ;
CORP Author Kentucky Univ., Lexington.;Health Effects Research Lab., Research Triangle Park, NC.;National Inst. of Environmental Health Sciences, Research Triangle Park, NC.
Publisher c1993
Year Published 1993
Report Number EPA-R-816185 ;NIEHS-ES-07266-02; EPA/600/J-93/343;
Stock Number PB93-229615
Additional Subjects DNA damage ; Deoxyribonucleic acids ; Thin layer chromatography ; Phosphorus radioisotopes ; Reagents ; Solvents ; Reprints ;
Library Call Number Additional Info Location Last
NTIS  PB93-229615 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 9p
DNA adducts represent the putative initiating event in the chemical carcinogenesis process. (32)P-Postlabeling is one of several assays which have been developed for the sensitive detection of DNA adducts. An integral part of the (32)P-postlabeling assay is the separation of adducted nucleotides by multidirectional, multisolvent, anion-exchange polyethyleneimine-cellulose thin-layer chromatography. In the study the use of a dilute ammonium hydroxide solvent for D4 chromatography is introduced and compared 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. 0.2 M ammonium hydroxide worked well for the recovery, separation, and resolution of a wide array of adducts derived from highly lipophilic polycyclic aromatic hydrocarbons and aromatic amines. In addition, this solvent required much less time (< 1/4) as compared to the other solvents and more importantly allowed the separation of adducts which otherwise comigrated and were not visible when using the other three D4 solvents. Background radioactivity per sq cm of the thin layers was also reduced two- to three-fold with the ammonia solvent versus the urea-based solvents. Thus, the use of this dilute ammonia system provides powerful resolution in a time-efficient manner.