Liao Benjamin S, Sram Jackie, Cain Teresa T, Halcrow Kenneth R
U.S. Food and Drug Administration, Pacific Regional Laboratory Southwest, General Food Chemistry, Irvine, CA, USA.
J AOAC Int. 2011 Mar-Apr;94(2):565-71.
Aqueous sulfuric acid can be used as the mobile phase in cation ion chromatography to separate the three biogenic amines, putrescine, cadaverine, and histamine, from fish. Various concentrations of aqueous sulfuric acid were investigated to optimize the separation of these three biogenic amines. Aqueous sulfuric acid (5.0 mM) was found to be optimum for the separation and was used to determine the three biogenic amines in fish. The LOQ, defined as the lowest level of the standard calibration curve, was 0.055 ppm (equivalent to 0.55 microg/g sample) for putrescine, 0.05 ppm (equivalent to 0.5 microg/g sample) for cadaverine, and 1.0 ppm (equivalent to 10 microg/g sample) for histamine. From statistical analysis of the LOQ, the method detection limit was 0.003 ppm for putrescine, 0.009 ppm for cadaverine, and 0.16 ppm for histamine. For sample preparation, the fish was composited, homogenized in methanol-water (75 + 25, v/v), incubated for 15 min at 60 degrees C, and centrifuged. The sample solution was micron-filtered before injection. The mobile phase flow rate was 0.8 mL/min under isocratic conditions at room temperature (15-25 degrees C). The three biogenic amines were separated in the order of increasing retention time, i.e., putrescine, cadaverine, and histamine, within 30 min. The chromatograms showed complete peak separation of the three amines regardless of the difference in fish matrixes.
在阳离子离子色谱中，硫酸水溶液可用作流动相，以分离鱼肉中的三种生物胺，即腐胺、尸胺和组胺。研究了不同浓度的硫酸水溶液，以优化这三种生物胺的分离效果。结果发现，5.0 mM的硫酸水溶液最适合分离，并用于测定鱼肉中的三种生物胺。腐胺的定量限（定义为标准校准曲线的最低水平）为0.055 ppm（相当于0.55 μg/g样品），尸胺为0.05 ppm（相当于0.5 μg/g样品），组胺为1.0 ppm（相当于10 μg/g样品）。根据定量限的统计分析，腐胺的方法检出限为0.003 ppm，尸胺为0.009 ppm，组胺为0.16 ppm。样品制备时，将鱼肉混合，在甲醇 - 水（75 + 25，v/v）中匀浆，于60℃孵育15分钟，然后离心。样品溶液在进样前进行微滤。在室温（15 - 25℃）等度条件下，流动相流速为0.8 mL/min。三种生物胺按保留时间增加的顺序分离，即腐胺、尸胺和组胺，在30分钟内完成。色谱图显示，无论鱼肉基质有何差异，三种胺均能实现完全的峰分离。
