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The determination of ADP and ATP in picomole amounts by steady-state mixed zones in isotachophoresis

✍ Scribed by J.P.M. Wielders; J.L.M. Muller

Publisher: Elsevier Science
Year: 1980
Tongue: English
Weight: 672 KB
Volume: 103
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(80)90628-4

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✦ Synopsis

A new electrophoretic method for the determination of adenine nucleotides in picomole amounts is described. The method depends upon the careful balancing of the effective electrophoretic mobilities of the component to be analyzed, e.g., ADP, and an inert carrier component (e.g., acetate) during an isotachophoretic analysis, whereby a steady-state mixed zone of ADP and its carrier is generated. This makes it possible spectrophotometrically to quantify traces of nucleotides at a concentration level several orders of magnitude below that of other sample constituents. A comparison was made between this new method and the luciferase assay using firefly lantern extract. Although more time-consuming than the luciferase assay and not quite as sensitive, the electrophoretic method is more precise and has the advantage that other anions in the sample are also measured. The method may also be used for trace analysis of other nucleotides, e.g., AMP, NAD(H), and FAD.

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