Abstract

A simple, practical technique is presented for the selective determination and measurement of histamine (HA) levels in fermented food. The method involved a solid-phase extraction cleanup using a Sep-Pak Plus C-18 cartridge and ion-paired reversed-phase high-performance liquid chromatographic (IP-RP-HPLC) separation, followed by detection of HA at its UV absorbance wavelength of 220 nm. After evaporating a methanolic extract from the food sample, the resulting residue was reconstituted with 0.2 M phosphate buffer (pH 3.0), and subsequently passed through the cartridge. The aliquot of the solution which came out of the cartridge was chromatographed in IP-RP mode on a C-18 column, as the stationary phase, and with a solution of 0.2 M phosphate buffer (pH 3.0)–acetonitrile–water (1:24:166, v/v) containing 2 mM sodium 1-octane sulfonic acid, as the mobile phase. When this method was applied to a mixture of HA, Cadaverine (Cad), Putrescine (Put), Serotonin (5HT), and Tyramine (Tyr), only HA was detected at 16.4 min of retention time. The method was fully validated and validation parameters were: linearity range 2–1000 ppm; correlation coefficient >0.991; mean recovery >99.5%; limit of quantification 2 ppm and limit of detection 0.5 ppm. The method was next applied to 12 brands of Miso (fermented soybean paste), 9 brands of Sake (rice wine), and 5 brands of Shouyu (Japanese soy sauce) to verify its ability to detect the presence of HA in a variety of fermented foods. The method proved to be both rapid and accurate and is therefore recommended for use in HA pollution surveys and in the routine practice of food-quality control.

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