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    同位素内标-超高效液相色谱-串联质谱法测定水果和蔬菜中4种杀菌剂残留量

    UPLC-MS/MS Determination of Residual Amounts of 4 Fungicides in Fruits and Vegetables with Quantification by Using Isotope-Labelled Internal Standards

    • 摘要: 所分析的水果、蔬菜按照标准GB 2763-2016制备分析样品,并在-20℃下冷冻保存。称取样品5.000 g,加入乙腈10 mL及同位素内标混合溶液500 μL,超声提取30 min,高速离心5 min,取上清液,于-40℃冷冻不少于30 min,待乙腈与水相分层后,迅速取出上清液1.0 mL,用N-丙基乙二胺50 mg、十八烷基键合硅胶50 mg、石墨化炭黑7.5 mg和无水硫酸镁150 mg作为混合吸附剂与上清液混匀并高速离心5 min,进行净化处理。取上清液通过0.22 μm滤膜过滤,滤液作为测定溶液供超高效液相色谱-串联质谱法分析。色谱分离中用Waters AcquityTM UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7 μm)为固定相,进样体积为2 μL,流动相由0.1%(体积分数)甲酸溶液和乙腈以不同比例混合组成。按梯度洗脱程序对样品中的4种杀菌剂(甲基硫菌灵、腈菌唑、异菌脲和吡唑醚菌酯以及它们的同位系内标)进行分离,并用质谱法测定。质谱分析中采用电喷雾离子源正离子模式和多反应监测模式。方法中采用同位素内标法定量解决了基质效应问题。结果表明:4种杀菌剂的质量浓度均在0.1~50.0 μg·L-1内与其对应的峰面积之间呈线性关系,检出限(3S/N)在0.1~0.2 μg·kg-1之间。按标准加入法进行回收试验,测得回收率在79.1%~112%之间,测定值的相对标准偏差(n=6)均小于15%。

       

      Abstract: Analytical samples of fruits and vegetables were prepared in accordance with the National Standard GB 2763-2016 and cryopreserve at -20℃. 5.000 g of the sample, after adding 500 μL of the mixed internal standard solution of the 4 isotope-labelled fungicides, was extracted ultrasonically with 10 mL of acetonitrile for 30 min, and then the mixture was centrifuged for 5 min at high speed. The supernatant was collected and frozen at -40℃ for at least 30 min, until phase separation between CH3CN and H2O occured. An aliquot of 1.0 mL of the supernatant was taken rapidly, mixed with an adsorbent mixture containing 50 mg of PSA, 50 mg of C18, 7.5 mg of GCB and 150 mg of anly. MgSO4 for 1 min and centrifuged at high speed for 5 min for purification. The supernatant was taken and filtered through 0.22 μm filtering membrane, the filtrate was used as testing solution for UPLC-MS/MS analysis. Waters AcquityTM UPLC BEH C18 chromatographic column (2.1 mm×100 mm, 1.7 μm) was used as the stationary phase, and 2 μL of the testing solution were introduced to pass through the column. Mixtures with various ratios of 0.1% (φ) formic acid solution and acetonitrile were used as mobile phases in the programed gradient elution for separation of the 4 fungcides (thiophanate-methyl, myclobutanil, pyraclostrobin and iprodione together with their isotope-labelled internal standards) and their determinations by MS/MS. Modes of ESI+ and MRM were adopted in MS analysis. It was shown that linear relationships between values of peak areas of the fungicides and their respective mass concentrations were found in the same range of 0.1-50.0 μg·L-1, and that their detection limits (3S/N) were in the range of 0.1-0.2 μg·kg-1. Test for recovery was made by standard addition method, giving results of recovery in the range of 79.1%-112%, and RSDs (n=6) all less than 15%.

       

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