Abstract
2,4-Dinitro-5-fluoroaniline was reacted with various amino acids and proteins. The N-2,4-dinitro-5-aminophenyl (DNAP) derivatives of twenty amino acids were prepared and three solvent systems which can effect their separation on thin-layer chromatograms were developed. DNAP insulin was prepared and it was shown that N-terminal amino acids can be determined starting with 50 μg of this derivative or 100 μg of insulin. The color of DNAP amino acids can be intensified on thin-layer chromatograms by first exposing the chromatograms to nitrogen dioxide fumes and then spraying with a solution of β-naphthol and triethylamine in benzene; this permits the detection of as little as 0.1 μg (about one nanomole) of DNAP amino acid.
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