Abstract Histamine (HA) is an important biochemical for human health and food quality. It is produced due to the decarboxylation of histidine. High level of HA was reported in a large number of fish tissues in Scombridae family. The presence of a low concentration of histamine is the indicator of the freshness of fish. Therefore, this study aims to develop a simple novel method for the quantification of histamine by revers-phase high-performance liquid chromatography (RP-HPLC-UV) followed by conventional UV detector at the wavelength of 205 nm. The extraction of the fish sample was optimized without derivatization, 2-5 dihydroxybenzoic acid (2-5 DHBA) and acetonitrile (ACN) was used in the mobile phase for the first time. Here we thoroughly investigate key experimental conditions such as UV absorbing probe, pH and mobile phase. We found that concentration of 6 mmol/L of 2,5-DHBA in the mobile phase, pH 2.65 and 50:50% ratios of 2,5-dihydroxybenzoic acid and acetonitrile were the suitable conditions for determination of histamine in the fish sample. By applying our new method, we determine the HA concentration within 5 min. This novel method was validated by using different parameters like linearity, limit of detection (LOD), limit of quantification (LOQ), repeatability, matric effect, specificity, trueness, ruggedness, accuracy, precision, and specificity. The LOD and LOQ were 1 and 3 mg/kg respectively, and the correlation coefficient was >0.9995. We applied this method to the fish sample. Which were obtained from a local market here in Shanghai. Recovery was determined after spiking the fish sample; good recovery was observed with average recovery >82%. Thus, the proposed method provides a better reference for the successful analysis of fish safety, quality and trade.

中文翻译：

---

间接反相-HPLC法测定未经衍生化处理的鱼类中的组胺

摘要 组胺（HA）是影响人类健康和食品质量的重要生化物质。它是由于组氨酸脱羧而产生的。在鲭科的大量鱼类组织中报道了高水平的 HA。低浓度组胺的存在是鱼新鲜度的指标。因此，本研究旨在开发一种通过反相高效液相色谱 (RP-HPLC-UV) 和常规紫外检测器在 205 nm 波长下定量组胺的简单新方法。鱼样品的萃取优化无衍生化，首次在流动相中使用2-5二羟基苯甲酸（2-5 DHBA）和乙腈（ACN）。在这里，我们彻底研究了关键实验条件，例如紫外线吸收探针、pH 值和流动相。我们发现流动相中 6 mmol/L 的 2,5-DHBA 浓度、pH 2.65 和 50:50% 的 2,5-二羟基苯甲酸和乙腈是测定鱼样品中组胺的合适条件。通过应用我们的新方法，我们可以在 5 分钟内确定 HA 浓度。这种新方法通过使用线性、检测限 (LOD)、定量限 (LOQ)、重复性、基质效应、特异性、真实性、耐用性、准确度、精密度和特异性等不同参数进行了验证。LOD 和 LOQ 分别为 1 和 3 mg/kg，相关系数 >0.9995。我们将此方法应用于鱼类样本。这些是从上海当地市场获得的。加标鱼样品后确定回收率；观察到良好的回收率，平均回收率 > 82%。因此，