Reyns Tim, Belpaire Claude, Geeraerts Caroline, Van Loco Joris
Scientific Institute of Public Health, Food, Medicines and Consumer Safety, Chemical Residues and Contaminants, Juliette Wytsmanstraat 14, 1050 Brussel, Belgium.
Research Institute for Nature and Forest, Duboislaan 14, 1560 Groenendaal-Hoeilaart, Belgium.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Mar 15;953-954:92-101. doi: 10.1016/j.jchromb.2014.02.002. Epub 2014 Feb 10.
Beside the possible illegal use of malachite green in aquaculture, other familiar dyes could also been applied by fraudulent producers due to their antiseptic and antibacterial activity. In this contribution, a new sensitive multi-residue method was developed to determine triarylmethane, xanthene, phenothiazine and phenoxazine dyes in fish by ultra-performance liquid chromatography-tandem mass spectrometry. Samples were extracted with acetonitrile, followed by an oxidation step using 2,3-dichloro-5,6-dicyanobenzoquinone. Further clean-up was performed by tandem solid phase extraction using weak and strong cation exchange cartridges. Extracts were analysed by UPLC-MS(n) operating in the positive electrospray ionisation mode (ESI+). The fourteen dyes were separated within only 12min on a C18 BEH column using 1mM ammonium acetate in water at pH 4.5 and acetonitrile as mobile phases at a flowrate of 0.4mLmin(-1). The presented method was validated as defined by the European Union and scientific literature. Good linearity (R ≥0.99 and goodness-of-fit (g) ≤10%) was achieved over the tested concentration range (0.25-2ngg(-1)). Limit of quantification was 0.25ngg(-1) for all dyes, with a signal-to-noise ratio of at least 10/1. This is at least 5 to 10 times lower than previous published methods. Limits of detection were all <0.1ngg(-1). Precision and trueness fell within the criteria requested by the EC requirements for this concentration range. Decision limit (CCα) and detection capability (CCβ) were all <1 and <0.25ngg(-1), respectively. Due to background levels of the xanthene dyes, the two rhodamine dyes could only be determined above 0.75ngg(-1). For these dyes, the method can only be used for screening purposes. To show the applicability of the method, a monitoring study was performed to investigate the occurrence of artificial dyes in wildlife European eel in Flemish rivers.
除了水产养殖中可能非法使用孔雀石绿外，其他常见染料因其防腐和抗菌活性也可能被不法生产商使用。在本研究中，开发了一种新的灵敏多残留方法，用于通过超高效液相色谱 - 串联质谱法测定鱼类中的三芳基甲烷、呫吨、吩噻嗪和吩恶嗪染料。样品用乙腈提取，然后使用2,3 - 二氯 - 5,6 - 二氰基苯醌进行氧化步骤。通过使用弱阳离子交换柱和强阳离子交换柱的串联固相萃取进行进一步净化。提取物在正电喷雾电离模式（ESI +）下通过UPLC - MS(n)进行分析。在C18 BEH柱上，使用pH为4.5的1mM醋酸铵水溶液和乙腈作为流动相，流速为0.4mLmin(-1)，仅在12分钟内就分离出了14种染料。该方法按照欧盟和科学文献的定义进行了验证。在测试浓度范围（0.25 - 2ngg(-1)）内实现了良好的线性（R≥0.99且拟合优度（g）≤10%）。所有染料的定量限为0.25ngg(-1)，信噪比至少为10/1。这比以前发表的方法低至少5至10倍。检测限均<0.1ngg(-1)。精密度和准确性在欧盟针对该浓度范围要求的标准内。决策限（CCα）和检测能力（CCβ）分别均<1和<0.25ngg(-1)。由于呫吨染料的背景水平，两种罗丹明染料只能在0.75ngg(-1)以上测定。对于这些染料，该方法仅可用于筛查目的。为了展示该方法的适用性，进行了一项监测研究，以调查佛兰芒河流域野生欧洲鳗鱼中人工染料的存在情况。
