REQUIMTE, Laboratory of Bromatology and Hydrology, Faculty of Pharmacy, University of Porto, Rua Aníbal Cunha 164, 4099-030 Porto, Portugal.
Food Chem Toxicol. 2012 Mar;50(3-4):1019-26. doi: 10.1016/j.fct.2011.12.028. Epub 2011 Dec 28.
The determination of deoxynivalenol (DON) and its metabolites such as deepoxy-deoxynivalenol (DOM-1) in human urine is complicated due its low levels (ng/mL) and the complexity of the matrix. A gas chromatography-mass spectrometry method was optimized and validated for the confirmation analysis of DON and its metabolites in urine samples using 13C isotopic-labeled DON as internal standard. In the sample preparation the type and amount of β-glucuronidase for enzymatic hydrolysis was investigated as well as the cleanup procedure, being compared the immunoaffinity column with solid-phase extraction (SPE). As far as we know, SPE C18 cleanup procedure was applied for the first time in the analysis of DON and its metabolites in human urine. Using this analytical methodology the detection and quantification limits achieved ranged from 0.06 to 0.30 ng/mL and from 0.2 to 1.0 ng/mL, respectively. Recoveries were higher than 73% for fortification levels between 25 and 100 ng/mL and repeatability were lower than 13%. The natural occurrence of DON and its metabolites in human urine samples from the north zone of Portugal was studied. Free DON was detected in 15% of the samples whereas total (free+conjugated) DON was detected in 69% of the samples. Deepoxy-deoxynivalenol, 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol were not detected in any of the samples analyzed.
由于脱氧雪腐镰刀菌烯醇（DON）及其代谢物（如去氧雪腐镰刀菌烯醇-1（DOM-1））的含量较低（ng/mL）且基质复杂，因此在人尿液中测定 DON 及其代谢物较为复杂。本研究采用 13C 同位素标记 DON 作为内标，优化并验证了气相色谱-质谱法用于尿液中 DON 及其代谢物的确证分析。在样品制备过程中，研究了酶水解用β-葡萄糖醛酸酶的类型和用量，以及净化程序，比较了免疫亲和柱和固相萃取（SPE）。据我们所知，SPE C18 净化程序首次应用于人尿液中 DON 及其代谢物的分析。采用该分析方法，检测限和定量限分别为 0.06-0.30ng/mL 和 0.2-1.0ng/mL。在 25-100ng/mL 的加标水平下，回收率高于 73%，重复性低于 13%。本研究还研究了葡萄牙北部地区人尿液中 DON 及其代谢物的自然发生情况。15%的样品中检测到游离 DON，69%的样品中检测到总（游离+结合）DON。未在任何分析样品中检测到去氧雪腐镰刀菌烯醇-1、3-乙酰脱氧雪腐镰刀菌烯醇和 15-乙酰脱氧雪腐镰刀菌烯醇。
