A simplified method for the estimation of individual amino acid radioactivity in plasma samples.
A method is presented for the quantitative estimation of the individual amino acid radioactivity in biological samples. The material is deproteinized with cold acetone, and, after acetone evaporation, is passed through a column containing 1 g of Amberlite XAD-2, then eluted with 10% ethanol. The samples are derivatized with Sanger's reagent (alkaline 1-fluoro-2,4-dinitrobenzene) and passed again through the Amberlite XAD-2 column; the 10% ethanol eluate is now discarded and the DNP-amino acids eluted with acetone. Aliquots are used for TLC chromatography on Silicagel plates; the spots are identified, cut away and their radioactivity estimated. The actual recovery of radioactivity in the spots is about 86-92% of the initial radioactivity. No contamination with radioactive glucose, lactate, pyruvate or glycerol has been observed.
D000596:Amino Acids / Q000097:blood*; D000818:Animals; D001786:Blood Glucose / Q000032:analysis; D002855:Chromatography, Thin Layer / Q000379:methods; D005990:Glycerol / Q000097:blood; D007773:Lactates / Q000097:blood; D019344:Lactic Acid; D011773:Pyruvates / Q000097:blood; D019289:Pyruvic Acid; D051381:Rats
D000596:Amino Acids; D001786:Blood Glucose; D007773:Lactates; D011773:Pyruvates; D019344:Lactic Acid; D019289:Pyruvic Acid; D005990:Glycerol