Abstract：
Headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME-GC-MS) offers an alternative analysis method for isoeugenol (an active ingredient in fish sedatives) that avoids the use of organic solvents, simplifies sample preparation, and can be fully automated. This work focuses on developing and evaluating an HS-SPME-GC-MS method for isoeugenol in aquaculture samples and testing the stability of isoeugenol itself. Because of isoeugenol\'s relatively low volatility, more polar SPME fiber coatings (polyacrylate and polydimethylsiloxane/divinylbenzene) had better performance and the headspace extractions took over 30 min to reach equilibrium. Additionally, it was found that isoeugenol was relatively unstable compared to a deuterated standard (d3-eugenol) in the presence of water. To address this, after the fish samples were homogenized with water, they were heated at 50 °C for 1 h prior to analysis for equilibration. By using the method developed in this work, isoeugenol\'s detection limits in multiple aquaculture matrices (shrimp, tilapia, and salmon) were in the low ng/g range (<15 ng/g), well below the target testing level (200 ng/g). Additionally, by adding d3-eugenol as an internal standard, excellent linearity (R2 > 0.98), accuracy (97-99% recoveries), and precision (5-13% RSDs) were all achieved.
摘要：
顶空固相微萃取与气相色谱-质谱联用（HS-SPME-GC-MS）提供了一种避免使用有机溶剂的异丁香酚（鱼类镇静剂中的活性成分）的替代分析方法，简化样品制备，并且可以完全自动化。这项工作的重点是开发和评估水产养殖样品中异丁香酚的HS-SPME-GC-MS方法，并测试异丁香酚本身的稳定性。由于异丁香酚的挥发性相对较低，极性更强的SPME纤维涂层（聚丙烯酸酯和聚二甲基硅氧烷/二乙烯基苯）具有更好的性能，顶空萃取需要30分钟才能达到平衡。此外,发现在水存在下,与氘代标准品(d3-丁香酚)相比,异丁香酚相对不稳定。为了解决这个问题，鱼样品用水匀浆后，在分析平衡之前,将它们在50°C加热1小时。通过使用这项工作中开发的方法，异丁香酚在多种水产养殖基质中的检测限(虾，罗非鱼,和鲑鱼)在低ng/g范围内(<15ng/g)，远低于目标测试水平(200纳克/克)。此外,通过添加d3-丁香酚作为内标，良好的线性度（R2>0.98），准确度(97-99%回收率)，和精度（5-13%RSD）都达到了。
