engleski

Use of small diameter column particles to enhance HPLC determination of histamine and other biogenic amines in seafood

Pre-column and post-column HPLC derivatization methods were modified and evaluated for the identification and quantification of nine biogenic amines in seafood. Two HPLC methods with column particles of 1.8 µm or 3 µm in diameter were modified and compared to classical methods using 5 µm column particles. Both pre-column derivatization with dansyl chloride and post-column derivatization with O-phthalaldehyde were studied. The HPLC methods were compared with respect to the time of elution, eluent consumption, back-pressure as well as separation, sensitivity, recovery and repeatability for determination of biogenic amines in lean canned tuna and fatty frozen herring. The modified methods using smaller column particles of 1.8 µm or 3 µm allowed biogenic amines to be separated and quantified faster (23-59%) and with less eluent consumption (59-62%) than classical HPLC methods. Back-pressures were below 170 bar and this allowed the use of classical HPLC systems rather than dedicated and costly ultra-high-pressure liquid chromatography (UHPLC) equipment. Biogenic amine separation, sensitivity, recovery and repeatability for the modified methods were similar to or performed better than for the classical HPLC methods. Due to reduced analyses time and eluent consumption the HPLC methods would improve biogenic amine analysis.

histamine; biogenic amines; HPLC columns with small particle-size

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