Eine quantitative Bestimmungsmethode des Uracilgehaltes biologischen Materials im Nanogramm-Bereich mit Hilfe der Hochdruck-Flüssigchromatografie / A Method for the Quantitative Estimation of Uracil Content in Biological Materials in the Nanogramme-Range Using High-Pressure Liquid Chromatography
2014
Breter, Hans-Joachim | Schnaus, Marianne | Müller, Werner E. G. | Zahn, Rudolf K.
A method is described for the estimation of the uracil content in biological materials by means of high-pressure liquid chromatography. Hydrolysis of the tissues and total liberation of RNA bases are carried out in 70% perchloric acid. Less than 1 mg of the materials are needed for analysis. A pre-purification of the hydrolyzates is carried out by anion-exchange chromatography. Recoveries are estimated by isotope dilution analysis with [2-¹⁴C] labelled uracil. The method is highly sensitive - about 6000 pmol of uracil content can at least be estimated quantitatively - and analysis time is short. In routine analysis a single sample needs 4 hours to be completed. When preparing several samples at a time an analysis is completed every 45min.
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