Histamine level determination in fish and fish products is important for human safety, fish quality and food industry. For this reason, a rapid, robust, and precise method is needed. To achieve this objective, an HPLC method with fluorescence detection was developed and validated. Histamine in fish samples was efficiently extracted with perchloric acid, and purified with ion-exchange solid-phase extraction cartridge. A pre-column derivatization was adopted with ortho-phthalaldehyde (OPA) in the presence of the reducing agent 2-mercaptoethanol, and the stability of the histamine-OPA derivatives was achieved with the acidification of the reaction medium. In terms of validation, besides the excellent linear correlations, satisfactory recoveries at all spiking levels ranging between 0 and 200 mg kg −1 were attained, with limit of detection calculated at 1.8 mg kg −1 , whereas limit of quantification determined at 5 mg kg −1 . The proposed method was successfully used in the analysis of reference materials and proficiency tests, and was found to be suitable, accurate, and rapid for detection and quantification of histamine in various fish samples.

中文翻译：

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使用柱前衍生化荧光检测测定鱼样品中组胺的高效液相色谱方法的开发和验证

鱼和鱼制品中组胺水平的测定对人类安全、鱼品质量和食品工业很重要。因此，需要一种快速、稳健且精确的方法。为实现这一目标，开发并验证了具有荧光检测功能的 HPLC 方法。用高氯酸有效提取鱼样品中的组胺，并用离子交换固相萃取柱纯化。在还原剂2-巯基乙醇的存在下，采用邻苯二甲醛(OPA)进行柱前衍生化，通过反应介质的酸化来实现组胺-OPA衍生物的稳定性。在验证方面，除了出色的线性相关性外，在 0 到 200 mg kg -1 之间的所有加标水平下都获得了令人满意的回收率，检测限计算为 1.8 mg kg -1 ，而定量限确定为 5 mg kg -1 。该方法已成功用于标准物质的分析和能力验证，并被证明适用于、准确、快速地检测和定量各种鱼类样品中的组胺。