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    QuEChERS结合超高效液相色谱-串联质谱法测定果汁和果酒中5种真菌毒素的含量

    Determination of 5 Mycotoxins in Fruit Juices and Fruit Wines by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry Combined with QuEChERS

    • 摘要: 取样品2 g,加入15 mL 10%(体积分数)乙腈溶液,涡旋提取30 min,加入QuEChERS盐包(内含4 g无水硫酸镁和1 g氯化钠),涡旋1 min,离心5 min,分取上清液500 μL,加入500 μL 10%(体积分数)甲醇溶液,涡旋混匀后进入超高效色相色谱-串联质谱仪,其中的霉酚酸(MPA)、青霉酸(PA)、桔青霉素(CIT)、黄绿青霉素(CTV)、异烟棒曲霉素C(ROC)等5种真菌毒素在Thermo Acclaim RSLC C18色谱柱上用不同体积比的甲醇和0.1%(体积分数)甲酸溶液的混合溶液进行梯度洗脱分离,以电喷雾离子源正离子(ESI+)模式电离,多反应监测(MRM)模式检测,基质匹配法定量。结果表明,5种真菌毒素的质量浓度均在一定范围内与对应的峰面积呈线性关系,检出限(3S/N)为0.3~5.0 μg·kg-1。按标准加入法进行回收试验,回收率为81.1%~105%,测定值的相对标准偏差(n=6)为0.63%~5.9%。方法用于市售41批果汁和59批果酒样品的分析,在果汁样品中检出了PA(2批)、MPA(1批)和CTV(1批),检出量分别为137,52.6 μg·kg-1(PA)、2.92 μg·kg-1(MPA)和2.81 μg·kg-1(CTV);在果酒样品中检出了PA(1批)和MPA(2批),检出量分别为34.3 μg·kg-1(PA)和3.51,2.94 μg·kg-1(MPA)。

       

      Abstract: An aliquot (2 g) of the sample was taken, and 15 mL of 10% (volume fraction) acetonitrile solution was added. The mixture was extracted for 30 min by vortex, and QuEChERS salt package (containing 4 g of anhydrous magnesium sulfate and 1 g of sodium chloride) was added. The mixture was vortexed for 1 min, and centrifuged for 5 min. An aliquot (500 μL) of the supernatant was taken, and mixed with 500 μL of 10% (volume fraction) methanol solution by vortex. The solution was introduced into the ultra-high performance chromatograph-tandem mass spectrometer. Five mycotoxins, including mycophenolic acid (MPA), penicillic acid (PA), citrinin (CIT), penicillium citreoviride (CTV) and roquefortine C (ROC), were separated on the Thermo Acclaim RSLC C18 column by gradient elution with mixed solutions composed of methanol and 0.1% (volume fraction) formic acid solution at different volume ratios, ionized by the ESI+ mode, detected by the MRM mode, and quantified by the matrix matching method. It was shown that linear relationships between values of the mass concentration of the 5 mycotoxins and the corresponding peak area were kept in definite ranges, with detection limits (3S/N) in the range of 0.3-5.0 μg·kg-1. Test for recovery was made according to standard addition method, giving recoveries in the range of 81.1%-105%, and RSDs (n=6) of the determined values ranged from 0.63% to 5.9%. The proposed method was applied to the analysis of 41 batches of fruit juice samples and 59 batches of fruit wine samples from the market. PA (2 batches), MPA (1 batch) and CTV (1 batch) were detected in fruit juice samples, with detection amounts of 137, 52.6 μg·kg-1 for PA, 2.92 μg·kg-1 for MPA and 2.81 μg·kg-1 for CTV; and PA (1 batch) and MPA (2 batches) were detected in fruit wine samples, with detection amounts of 34.3 μg·kg-1 for PA and 3.51, 2.94 μg·kg-1 for MPA.

       

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