Brezina Ulrike, Valenta Hana, Rempe Inga, Kersten Susanne, Humpf Hans-Ulrich, Dänicke Sven
Institute of Animal Nutrition, Federal Research Institute for Animal Health, Friedrich-Loeffler-Institut, Braunschweig, Germany.
Mycotoxin Res. 2014 Aug;30(3):171-86. doi: 10.1007/s12550-014-0200-8. Epub 2014 Jun 13.
A sensitive and selective liquid chromatography tandem mass spectrometry method using negative electrospray ionisation (LC-ESI-MS/MS) was developed for the simultaneous determination of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites α-zearalenol, β-zearalenol, zearalanone, α-zearalanol, β-zearalanol and de-epoxy-deoxynivalenol in pig serum. For method development, different sample preparation columns were tested for their suitability for extraction and clean up. Finally, preparation of serum samples was carried out using Oasis™ HLB solid-phase extraction (SPE) columns. The analyte concentrations were determined by the use of isotopically labelled internal standards (IS). The method was in-house validated for all analytes. Calibration graphs (0.3-480 ng/ml) were prepared and high degree of linearity was achieved (r ≥ 0.99). Results for method precision ranged between 2.7 and 21.5 % for inter-day and between 1.1 and 11.1 % for intra-day. The recoveries were in the range of 82-131 %. Limits of detection and quantification ranged 0.03-0.71 and 0.08-2.37 ng/ml, respectively. The method has been successfully used for quantitative determination of ZEN, DON and their metabolites in pig serum from a feeding trial with practically relevant ZEN and DON concentrations. This method is precise and reproducible and can be used as a multi-biomarker method to assess animal exposure to these mycotoxins and for diagnosis of intoxications.
建立了一种灵敏且具选择性的液相色谱串联质谱法，采用负电喷雾电离（LC - ESI - MS/MS）同时测定猪血清中的玉米赤霉烯酮（ZEN）、脱氧雪腐镰刀菌烯醇（DON）及其代谢产物α - 玉米赤霉醇、β - 玉米赤霉醇、玉米赤霉酮、α - 玉米赤霉烯醇、β - 玉米赤霉烯醇和脱环氧脱氧雪腐镰刀菌烯醇。为了方法开发，测试了不同的样品制备柱对提取和净化的适用性。最后，使用Oasis™ HLB固相萃取（SPE）柱进行血清样品的制备。通过使用同位素标记的内标（IS）来测定分析物浓度。该方法对所有分析物进行了内部验证。制备了校准曲线（0.3 - 480 ng/ml）并实现了高度线性（r≥0.99）。方法精密度结果日间为2.7%至21.5%，日内为1.1%至11.1%。回收率在82%至131%范围内。检测限和定量限分别为0.03 - 0.71 ng/ml和0.08 - 2.37 ng/ml。该方法已成功用于定量测定来自实际相关ZEN和DON浓度的饲养试验猪血清中的ZEN、DON及其代谢产物。该方法精确且可重复，可作为一种多生物标志物方法用于评估动物对这些霉菌毒素的暴露情况以及中毒诊断。
