<p>This work reports the development and validation of a fast and selective high-performance thin-layer chromatography (HPTLC) method for determining the most relevant biogenic amines (BAs) present in beers, namely, spermine, spermidine, putrescine, cadaverine, tryptamine, histamine, tyramine, and 2-phenylethylamine. Dansyl-BAs were separated on dipotassium hydrogen phosphate-treated HPTLC silica gel 60 plates, using a mobile phase composed of <i>tert</i>-butyl methyl ether‒toluene‒triethylamine (2:2.5:6, <i>V/V</i>). Densitometric quantification was carried out by fluorescence at 366/&gt; 400&#xa0;nm. Validation was established according to the International Conference on Harmonisation (ICH) guidelines. Each calibration data (in matrix) fitted a polynomial regression model with <i>R</i>² &gt; 0.996. Repeatability (<i>n</i> = 6) and intermediate precision (<i>n</i> = 6) in matrix showed relative standard deviation (RSD) values &lt; 3.57 and &lt; 2.58%, respectively. Selectivity was evaluated by mass spectrometry in full scan mode (<i>m/z</i> 100‒2000), comparing sample and standards mass spectra. The limits of detection and limits of quantification ranged from 0.30 to 1.91&#xa0;ng/band and from 1.00 to 6.36&#xa0;ng/band, respectively. This method was applied to determine BA content in beer samples, finding a total BA content from 2.00 to 9.00&#xa0;mg L^‒1, levels lower than those considered a human health risk. This analytical method proved to be fast, accurate, and precise alternative for monitoring beer quality and safety.</p>