PREPARATIVE CAPILLARY-ZONE-ELECTROPHORESIS IN COMBINATION WITH OFF-LINE GRAPHITE-FURNACE ATOMIC-ABSORPTION FOR THE ANALYSIS OF DNA COMPLEXES FORMED BY A NEW AMINOCOUMARINE PLATINUM (II) COMPOUND
Dld. Deforce et al., PREPARATIVE CAPILLARY-ZONE-ELECTROPHORESIS IN COMBINATION WITH OFF-LINE GRAPHITE-FURNACE ATOMIC-ABSORPTION FOR THE ANALYSIS OF DNA COMPLEXES FORMED BY A NEW AMINOCOUMARINE PLATINUM (II) COMPOUND, Electrophoresis, 19(14), 1998, pp. 2454-2458
Citations number
20
Categorie Soggetti
Biochemical Research Methods","Chemistry Analytical
Calf thymus DNA was incubated in vitro with a new aminocoumarin platin
um(II) complex in order to study its interaction with DNA. The platina
ted DNA was hydrolyzed enzymatically to the 5'-mononucleotide level us
ing DNAase I and nuclease P1. Analysis of the DNA hydrolysate with cap
illary zone electrophoresis (CZE), using sample stacking, revealed the
presence of unhydrolyzed oligonucleotides in the platinated DNn. A ho
memade system, using only some plastic pipet tips, was constructed to
collect the oligonucleotide fraction during CZE analysis. The platinum
content of this fraction was determined using graphite furnace atomic
absorption with Zeeman background correction. This system proved to b
e a useful tool to detect platinated DNA species (with a quantifiable
detection limit for the detection of platinum of 0.78 ng). Subsequent
gel filtration experiments confirmed the presence of high molecular we
ight oligonucleotides that were platinated. This was proven by reversa
l of the platination using thiourea and subsequent enzymatic hydrolysi
s to 5'-mononucleotides.