University of Valencia, Department of Food Science, Av. Vicent Andrés Estellés s/n, 46100 Burjassot, Spain.
J Chromatogr A. 2012 Dec 28;1270:28-40. doi: 10.1016/j.chroma.2012.10.061. Epub 2012 Nov 5.
A new analytical method for the rapid and simultaneous determination of ten mycotoxins including patulin, zearalenone and eight trichothecenes (nivalenol, fusarenon-X, diacetoxyscirpenol, 3-acetyl-deoxynivalenol, neosolaniol, deoxynivalenol, T-2 and HT-2) in wheat semolina has been developed and optimized. Sample extraction and purification were performed with a modified QuEChERS-based (acronym of Quick, Easy, Cheap, Effective, Rugged and Safe) procedure and determined by gas chromatography (GC) coupled to triple quadrupole instrument (QqQ). This is the first paper on the application of GC-QqQ-MS/MS to analysis of mycotoxins. Careful optimization of the gas chromatography-tandem mass spectrometry parameters was achieved in order to attain a fast separation with the best sensitivity allowing a total run time of 16 min. The validation was performed by analyzing recovery samples at three different spiked concentrations, 20, 40 and 80 μg kg(-1), with four replicates (n=4) at each concentration. Recoveries ranged from 74% to 124% and the intra-day precision and inter-day precision, expressed as relative standard deviation, were lower than 13% and 17%, respectively for all studied compounds, except for zearalenone. Limits of quantification (LOQ) were lower than 10 μg kg(-1) for all studied mycotoxins. Eight concentration levels were used for constructing the calibration curves which showed good linearity between LOQ and 100 times LOQ concentration levels (linear range). Matrix-matched calibration for applying the method in routine analysis is recommended for reliable quantitative results. The method validated was successfully applied to fifteen wheat semolina samples detecting occurrence of mycotoxins at concentrations below the maximum permissible level.
一种新的分析方法，用于快速和同时测定十种真菌毒素，包括棒曲霉素、玉米赤霉烯酮和八种单端孢霉烯族化合物（玉米赤霉醇、伏马菌素-X、二乙酰基脱氧雪腐镰刀菌烯醇、3-乙酰基-脱氧雪腐镰刀菌烯醇、新茄病镰刀菌醇、脱氧雪腐镰刀菌烯醇、T-2 和 HT-2）在小麦粗粉中的含量。采用改良的基于 QuEChERS（Quick，Easy，Cheap，Effective，Rugged and Safe 的缩写）的方法进行样品提取和净化，并通过气相色谱（GC）与三重四极杆仪器（QqQ）联用进行测定。这是第一篇应用 GC-QqQ-MS/MS 分析真菌毒素的论文。仔细优化了气相色谱-串联质谱参数，以实现快速分离，获得最佳灵敏度，总运行时间为 16 分钟。通过在三个不同的加标浓度（20、40 和 80 μg kg(-1)）下分析回收率样品，并在每个浓度下进行四个重复（n=4）来进行验证。回收率范围为 74%至 124%，日内精密度和日间精密度（表示为相对标准偏差）分别低于 13%和 17%，除玉米赤霉烯酮外，所有研究化合物均如此。所有研究的真菌毒素的定量限（LOQ）均低于 10 μg kg(-1)。使用八个浓度水平来构建校准曲线，结果表明在 LOQ 和 100 倍 LOQ 浓度水平之间具有良好的线性关系（线性范围）。建议使用基质匹配校准来进行常规分析，以获得可靠的定量结果。该方法经验证后成功应用于 15 个小麦粗粉样品中，检测到真菌毒素的浓度低于最大允许水平。
