Simple and rapid determination of biogenic amines in fish and fish products by liquid chromatography-tandem mass spectrometry using 2,4,6-triethyl-3,5-dimethyl pyrylium trifluoromethanesulfonate as a derivatization reagent.

A simple and rapid analytical method was developed for determination of four biogenic amines [histamine (Him), cadaverine (Cad), tyramine (Tym), 2-phenylethylamine (Pea)] in fish and fish products. This method uses a new derivatization reagent, 2,4,6-triethyl-3,5-dimethyl pyrylium trifluoromethanesulfonate (Py-Tag). The four biogenic amines in the samples were extracted with trichloroacetic acid. The diluted extract was derivatized with Py-Tag (15 min at 50°C) and then subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS). The limits of quantification for the method were 2 mg/kg for Him, Tym, and Pea and 10 mg/kg for Cad. The matrix effects derived from the tested fish and fish products were negligible in the LC-MS/MS analysis. The impact of the sample matrices on the Py-Tag derivatization was also negligible. The trueness and repeatability of the method were assessed by performing replicate analyses (n = 5) of five samples of fish and fish products, each spiked with the four biogenic amines at three different concentration levels. Analysis of the samples found 87%-104% of the spiked concentrations and the relative standard deviations were <6.1%. A reference sample and quality control canned fish samples were analyzed by the method, and the concentrations of the Him were within acceptable limits. The developed method was successfully used to determine concentrations of the four biogenic amines in 48 fish and fish products on the Japanese market. The developed method does not require cleanup using a solid-phase extraction column or similar, and the derivatization reaction time was only 15 min. The results suggested that the present method is reliable and suitable for rapid analysis of the four biogenic amines in fish and fish products.