New solvent systems for thin-layer chromatographic determination of nine biogenic amines in fish and squid.

Different solvent systems were evaluated for their ability to separate biogenic amines by thin-layer chromatography (TLC). Dansyl derivatives of agmatine, putrescine, tryptamine, cadaverine, spermidine, histamine, spermine, tyramine and beta-phenylethylamine were separated using the solvent system chloroform-diethyl ether-triethylamine (6:4:1), followed by chloroform-triethylamine (6:1). After separation dansyl amines were quantified by fluorescence densitometry at 330 nm. Correlation coefficients of linear regressions were higher than 0.99 for all amines, except for agmatine (0.976). Detection limits were 10ng for tryptamine, tyramine, histamine and beta-phenylethylamine, and 5 ng for the other amines. The overall repeatability of the chromatography was 1.82% when including agmatine and barely 1.02% for the other amines. The accuracy ranged from 105.97% (agmatine) to 49.92% (tryptamine). This thin-layer chromatography method was found to be an effective and precise analytical procedure to separate and determine biogenic amines. Its main advantages compared to previous procedures are that it uses less harmful solvent (diethyl ether instead of benzene) and can separate a larger group of biogenic amines.