Kirschbaum J, Rebscher K, Brückner H
Department of Food Sciences, Institute of Nutritional Science, Interdisciplinary Research Center, Justus Liebig University, Giessen, Germany.
J Chromatogr A. 2000 Jun 9;881(1-2):517-30. doi: 10.1016/s0021-9673(00)00257-0.
The reagent 3,5-dinitrobenzoyl chloride (DNBZ-Cl) was tested for pre-column derivatization of biogenic amines (BAs). Samples were derivatized within 3 min in 1 M NaOH at ambient temperature by adding 2-propanole and 50 mM DNBZ-Cl in acetonitrile. The reaction was terminated by addition of 2 M HCl. For high-performance liquid chromatography an encapsulated stationary reversed-phase and gradient elution using a ternary gradient system were used. The DNBZ derivatives were quantified by their UV-absorption at 260 nm. The structures of the derivatives were elucidated using coupling of HPLC with electrospray ionization mass spectrometry. Detection limits of BAs were approximately 124-864 microg l(-1) (injected amounts 203-1410 pg) at a signal-to-noise ratio of 3:1. The coefficients of determination were 0.989-0.996, with the exceptions of cadaverine (0.976) and serotonin (0.965). The method was applied to the quantitative determination of agmatine, cadaverine, histamine, octopamine, 2-phenylethylamine, putrescine, serotonin, spermidine, spermine, tryptamine and tyramine, in fermented cabbage juices, soy sauces, Misos (soy pastes), fermented fish sauces, and anchovy paste.
对试剂3,5 - 二硝基苯甲酰氯(DNBZ - Cl)进行了生物胺(BAs)柱前衍生化测试。在室温下,于1 M氢氧化钠中,通过加入2 - 丙醇和乙腈中的50 mM DNBZ - Cl,样品在3分钟内完成衍生化。通过加入2 M盐酸终止反应。对于高效液相色谱,使用了封装的固定反相柱和采用三元梯度系统的梯度洗脱。通过在260 nm处的紫外吸收对DNBZ衍生物进行定量。利用高效液相色谱与电喷雾电离质谱联用阐明衍生物的结构。在信噪比为3:1时,生物胺的检测限约为124 - 864 μg l⁻¹(进样量203 - 1410 pg)。除尸胺(0.976)和血清素(0.965)外,测定系数为0.989 - 0.996。该方法应用于发酵白菜汁、酱油、味噌(豆酱)、发酵鱼露和凤尾鱼酱中胍丁胺、尸胺、组胺、章鱼胺、2 - 苯乙胺、腐胺、血清素、亚精胺、精胺、色胺和酪胺的定量测定。