Scuola di Scienze del Farmaco e dei Prodotti della Salute, Università di Camerino, via S. Agostino 1, 62032 Camerino, Italy.
Laboratori de Bromatologia i Toxicologia, Facultat de Farmàcia, Universitat de València, Av. Vicent Andrés Estellés s/n, 46100 Burjassot, Spain.
Food Chem. 2012 May 1;132(1):537-43. doi: 10.1016/j.foodchem.2011.10.054. Epub 2011 Nov 4.
Biogenic amines on fish tissue are formed as a result of bacterial contamination and spoilage during storage. A new method based on liquid chromatography (LC) and tandem mass spectrometry (MS/MS) using a triple quadrupole (QqQ) analyser was developed for the analysis of eight biogenic amines (cadaverine, histamine, phenylethylamine, putrescine, spermine, spermidine, tyramine and tryptamine) in fish tissues. Sample preparation was performed by extraction with trichloroacetic acid 5% and solid phase extraction clean up with STRATA X cartridge. The MS/MS method was validated and compared with a method based on the analysis of dansyl derivatives by LC and fluorescence detector (FD). MS/MS achieved higher sensitivity (from 0.02mgkg(-1) for spermidine and phenylethylamine to 0.2mgkg(-1) for spermine) when compared to FD (from 1mgkg(-1) for putrescine and tyramine to 4mgkg(-1) for histamine); MS/MS method showed higher precision too, with intraday relative standard deviations (RSDs) from 1% to 4% with respect to those obtained with FD method (from 3% to 17%). Recovery study was conducted at two different fortification levels and the average ranged from 71% to 93% for all of the studied compounds with RSDs lower than 18%. Matrix-matched standards were used to counteract matrix effect observed in MS/MS determination. The applicability of the method was demonstrated by the analysis of biogenic amines in fish obtained from commercials of Valencia.
鱼组织中的生物胺是在储存过程中细菌污染和变质的结果。本研究建立了一种基于液相色谱-串联质谱（LC-MS/MS）三重四极杆（QqQ）分析的新方法，用于分析鱼组织中的 8 种生物胺（腐胺、组胺、苯乙胺、尸胺、亚精胺、精胺、酪胺和色胺）。样品制备采用 5%三氯乙酸提取，固相萃取 STRATA X 小柱净化。对 LC-FD 方法和 LC-MS/MS 方法进行了验证和比较。与 LC-FD 方法相比（腐胺和酪胺的检测限为 1mgkg(-1)，组胺为 4mgkg(-1)），LC-MS/MS 方法的灵敏度更高（精胺和苯乙胺的检测限为 0.02mgkg(-1)，亚精胺为 0.2mgkg(-1)）；LC-MS/MS 方法的精密度也更高，日内相对标准偏差（RSD）为 1%-4%，而 LC-FD 方法为 3%-17%。在两种不同的加标水平下进行了回收率研究，所有研究化合物的平均回收率为 71%-93%，RSD 低于 18%。采用基质匹配标准来抵消 MS/MS 测定中观察到的基质效应。该方法应用于分析瓦伦西亚市售鱼中的生物胺。
