Highly Sensitive Densitometry for Sugarphosphoeters and Nucleotides Using Enzymatic Hydrolysis on TLC Plate

概要

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Sensitive TLC for the determination of sugarphosphoesters and nucleotides was established. The individual separation of four sugarphosphoesters and three nucleotides on a silica gel plate was carried out in 1-butanol-acetic acid-water (1 : 1 : 1,v/v). Following the development of phophate esters, alkaline phophatase solution was sprayed on the plate at 45℃ to librate phosphate from each phophate ester. After spraying a mixture of molybdic acid and Melachite Green on the plate, the liberated phophate appeared as blue-green spots of molybdophosphate-Malachite Green aggregates on a yellow-brown back ground. The absorbance of each spot on the plate was measured at 620 nm with a densitometer to obtain a chromatogram. Fructose 1-phophate, fructose 1,6-phophate, glucose 6-phosphate, mannose 6-phophate, adenosine 5'-triphosphate, adenosine 5'-diphosphate and adenosne 5'-monophosphate could be detected separately at a sensitivy of as little as 0.5 pmol/spot. The values of relative standard deviation for the esters was less than 3.9% (n=7) at 20 pmo/spot.
公益社団法人日本薬学会の論文
1996-06-15