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中国精品科技期刊2020
刘彤彤,梁瑞强,韩伟娜,等. 超高效液相色谱-串联质谱法测定肉制品中的虾过敏原[J]. 食品工业科技,2023,44(6):292−299. doi: 10.13386/j.issn1002-0306.2022050152.
引用本文: 刘彤彤,梁瑞强,韩伟娜,等. 超高效液相色谱-串联质谱法测定肉制品中的虾过敏原[J]. 食品工业科技,2023,44(6):292−299. doi: 10.13386/j.issn1002-0306.2022050152.
LIU Tongtong, LIANG Ruiqiang, HAN Weina, et al. Determination of Shrimp Allergens in Meat Products by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry[J]. Science and Technology of Food Industry, 2023, 44(6): 292−299. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022050152.
Citation: LIU Tongtong, LIANG Ruiqiang, HAN Weina, et al. Determination of Shrimp Allergens in Meat Products by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry[J]. Science and Technology of Food Industry, 2023, 44(6): 292−299. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022050152.

超高效液相色谱-串联质谱法测定肉制品中的虾过敏原

Determination of Shrimp Allergens in Meat Products by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

  • 摘要: 目的:建立超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)法检测肉制品中虾过敏原的定量方法。方法:选取基质较为复杂的肉制品(西式火腿、香肠和肉丸)作为研究对象。样品经磷酸盐缓冲液(Phosphate buffer solution,PBS)超声提取30 min,离心(10000 r/min,15 ℃,10 min)后加入乙腈去除脂肪,取样液加入内标肽段(2.5 μmol/L,40 μL)和胰蛋白酶(1 mg/mL,10 μL)在37 ℃下酶解16 h后上液质进行分析,样品经T3柱进行分离,0.1%甲酸-水溶液和乙腈梯度洗脱,多反应监测(multiple reaction monitoring,MRM)正离子模式采集数据,内标法定量。结果:采用该方法测定肉制品中虾过敏原蛋白含量,其中定量肽段在0.001~2.0 μmol/L范围内,线性关系良好,决定系数R2为1.0000,检出限为0.67 mg/kg,定量限为2.00 mg/kg;在三个加标浓度水平下,回收率为83.2%~104.5%,精密度为2.63%~7.92%。结论:该方法具有特异性强、灵敏度高和回收率好等优势,适用于肉制品中虾过敏原的筛查定量。

     

    Abstract: Objective: To establish a method for the determination of shrimp allergens in meat products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods: Meat products (cooked ham, sausages and meatballs) with complex matrix components were selected as the detection target. The samples were extracted by ultrasonic in phosphate buffer solution (PBS) for 30 min, and acetonitrile was added to remove fat after centrifugation (10000 r/min, 15 ℃, 10 min). Finally, trypsin (1 mg/mL, 10 μL) was added to the sample diluents which had been mixed with internal peptide (2.5 μmol/L, 40 μL), and the enzymolysis process lasted 16 h at 37 ℃ before UPLC-MS/MS analysis. The samples were separated on a T3 column, and eluted by gradient elution with 0.1% formic acid aqueous solution and acetonitrile. The data was collected by multiple reaction monitoring (MRM) in positive ion mode, and quantified by internal standard method. Results: The method was used to determine the content of shrimp allergen in meat products, and the quantitative peptide had a good linear relationship with the determination coefficients 1.0000 in the range of 0.001~2.0 μmol/L. The limits of detection was 0.67 mg/kg, and the limits of quantification was 2.00 mg/kg. The recovery was 83.2%~104.5% and the precision was 2.63%~7.92% at three spiked levels. Conclusion: The method has the advantages of strong specificity, high sensitivity and good recovery. It was suitable for the screening and quantification of shrimp allergens in meat products.

     

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