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    QuEChERS-超高效液相色谱-串联质谱法测定糌粑中15种真菌毒素的含量

    Determination of 15 Mycotoxins in Zanba by Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry with QuEChERS

    • 摘要: 取1.000 g均质过的糌粑粉末,加入5 mL水和10 mL含10%(体积分数)甲酸的乙腈溶液,涡旋振荡提取10 min,加入4 g硫酸钠和1 g氯化钠,涡旋振荡、离心。取1.5 mL上清液,置于装有150 mg硫酸镁、100 mg C18、100 mg N-丙基乙二胺(PSA)的2 mL净化管中,涡旋振荡、离心。取0.8 mL上清液,氮吹至近干,用0.4 mL 20%(体积分数)乙腈溶液复溶,涡旋混合5 min,经0.22 μm滤膜过滤后,采用超高效液相色谱-串联质谱法测定滤液中15种真菌毒素的含量。以ACQUITY UPLC HSS T3色谱柱为固定相,以不同体积比的0.2%(体积分数)甲酸溶液和乙腈混合液为流动相进行梯度洗脱,质谱分析采用电喷雾离子(ESI)源,正负离子模式同时扫描,以多反应监测模式检测,采用基质校正标准曲线进行准确定量。结果表明,15种真菌毒素的质量浓度均在一定范围内与峰面积呈线性关系,检出限(3S/N)为0.1~5.8 μg·kg-1。按照标准加入法进行回收试验,回收率为75.8%~105%,测定值的相对标准偏差(n=6)均小于10%。方法用于分析10批实际糌粑样品,发现两批糌粑中赭曲霉毒素A(OTA)含量超标。

       

      Abstract: Homogenized zanba powders (1.000 g) was taken, and 5 mL of water and 10 mL of acetonitrile solution containing 10% (volume fraction) formic acid were added. After extracting for 10 min by vortex, 4 g of sodium sulfate and 1 g of sodium chloride were added. The mixture was vortexed and centrifuged, and 1.5 mL of supernatant was placed in a 2 mL-purification tube filled with 150 mg of magnesium sulfate, 100 mg of C18, 100 mg of N-propyl ethylenediamine (PSA). After vortex and centrifugation, 0.8 mL of supernatant was blown to near dryness by nitrogen, and then the residue was redissolved with 0.4 mL of 20% (volume fraction) acetonitrile solution. The solution was vortexed for 5 min, and filtered by 0.22 μm filter membrane. 15 mycotoxins in the filtrate were determined by ultra-high performance liquid chromatography tandem mass spectrometry, using ACQUITY UPLC HSS T3 column as stationary phase and mixtures of 0.2% (volume fraction) formic acid solution and acetonitrile at different volume ratios as mobile phase for gradient elution. Electrospray ion (ESI) source was used for mass spectrometry. The target compounds were analyzed by multiple reaction monitoring (MRM) mode in positive and negative scanning mode. Matrix calibration standard curves were used for accurate quantification. As shown by the results, linear relationships between values of peak area and mass concentration of 15 mycotoxins were found in definite ranges, with detection limits (3S/N) in the range of 0.1-5.8 μg·kg-1. Test for recovery was made by standard addition method, giving results in the range of 75.8%-105%, with RSDs (n=6) of the determined values less than 10%. This method was applied to the analysis of 10 batches of actual zanba samples, and it was shown that ochratoxin A (OTA) content in 2 batches of zanba samples exceeded the standard.

       

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