Derivatization of compounds at the origin of thin-layer plates with fluorescamine.

A method has been developed for the detection of compounds with primary amino groups on thin-layer chromatography (TLC) plates with fluorescamine. The compounds dissolved in buffer were applied to TLC plates and derivatized at the origin by developing with or dipping into an acetone-hexane solution of fluorescamine. Virtually all fluorescamine derivatives stayed at the origin, and they were subsequently separated using appropriate solvent systems. As little as 10 pmoles of fluorescamine-derivatized compound could be detected when viewed under a long-wave (366 nm) ultraviolet lamp. The method was applied to the analysis of peptides, amino acids and amines.