Wang Sha, Kong Wei-jun, Yang Mei-hua
Yao Xue Xue Bao. 2016 Jan;51(1):110-5.
A suitable ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of 11 mycotoxins with isotope internal standard in malt. The mycotoxins in malt were extracted and purified by one-step ultrasonic extraction procedure using acetonitrile/water/acetic acid (80 : 19 : 1), and then detected and confirmed by UPLC-MS/MS, and quantified by isotope labeled AFB1 ([13C17]-AFB1) and ZEN ([13C18]-ZEN) internal standards. Rapid separation of the 11 mycotoxins was successfully achieved on a Phenomenex Kinetex C18 column (100 mm x 2.1 mm, 2.6 μm) with gradient elution using the mobile phase of methanol containing 0.1% formic acid and 2 mmol x L(-1) ammonium acetate in water. Simultaneous acquisition was performed in multiple reaction monitoring (MRM) mode with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The established method provided a good linearity for the 11 mycotoxins within their respective linear ranges with correlation coefficients all higher than 0.999 1. The average recoveries ranged from 75.0% to 117.0% with relative standard deviations (RSDs) below 5.1%. The limits of detection (LODs) and quantitation (LOQs) ranged from 0.05 to 30 μg x kg(-1) and 0.15 to 87.5 μg x kg(-1), respectively, which were below the maximum residue levels (MRLs) set by the European Union. Twenty malt samples were analyzed and nine samples were detected with mycotoxins, which were confirmed according to the same fragment ions found in positive samples and the standards at the same retention time. This study has demonstrated that the one-step extraction procedure of mycotoxins from complex matrices coupled to UPLC-MS/MS method is simple, quick, accurate and sensitive for quantitative and qualitative analysis of multiple mycotoxins in malt.
建立了一种合适的超高效液相色谱-串联质谱法（UPLC-MS/MS），用于测定麦芽中11种带同位素内标的霉菌毒素。麦芽中的霉菌毒素采用乙腈/水/乙酸（80:19:1）通过一步超声提取程序进行提取和净化，然后通过UPLC-MS/MS进行检测和确证，并采用同位素标记的AFB1（[13C17]-AFB1）和ZEN（[13C18]-ZEN）内标进行定量。在Phenomenex Kinetex C18柱（100 mm×2.1 mm，2.6μm）上，以含0.1%甲酸的甲醇和含2 mmol·L(-1)乙酸铵的水溶液为流动相进行梯度洗脱，成功实现了11种霉菌毒素的快速分离。在电喷雾电离（ESI）源的正、负离子模式下，采用多反应监测（MRM）模式进行同时采集。所建立的方法在各自线性范围内对11种霉菌毒素具有良好的线性，相关系数均高于0.999 1。平均回收率为75.0%至117.0%，相对标准偏差（RSDs）低于5.1%。检测限（LODs）和定量限（LOQs）分别为0.05至30μg·kg(-1)和0.15至87.5μg·kg(-1)，均低于欧盟设定的最大残留限量（MRLs）。对20个麦芽样品进行了分析，9个样品检测到霉菌毒素，根据阳性样品和标准品在相同保留时间发现的相同碎片离子进行确证。本研究表明，从复杂基质中一步提取霉菌毒素并结合UPLC-MS/MS方法，对于麦芽中多种霉菌毒素的定量和定性分析简单、快速、准确且灵敏。
