色谱

色谱 ›› 2021, Vol. 39 ›› Issue (1): 96-103.DOI: 10.3724/SP.J.1123.2020.06010

• 研究论文 • 上一篇    

QuEChERS-同位素稀释-气相色谱-串联质谱法测定动物源性食品中9种N-亚硝胺类化合物

孔祥一1,2, 庄丽丽2, 方恩华2, 林鹏1, 郑子龙3, 郑向华2, 徐敦明1,2,*()   

  1. 1.集美大学水产学院, 福建 厦门 361021
    2.厦门海关技术中心, 福建 厦门 361000
    3.福建省市场监督管理局, 福建 福州 350003
  • 收稿日期:2020-07-03 出版日期:2021-01-08 发布日期:2020-12-20
  • 通讯作者: 徐敦明
  • 作者简介:徐敦明:厦门海关技术中心研究员,受聘第二届食品安全国家标准审评委员会委员,海关总署科技委专业委委员,海关技术规范食品检验专业技术委员会委员、国家燕窝市场专业委员会专家委员、厦门市食品安全委员会专家委员会副主任委员、福建农林大学/集美大学食品安全检测与控制方向硕士生导师。长期从事食品安全研究与检测、食品安全科普,专业于有毒有害物的分析。近年,主持参与35项与食品安全相关的国家及省部级科技项目,主持参与28项国家标准、行业标准的制修订。发表学术论文60余篇,撰写专著6部,授权专利10项,获各类科技进步奖17项、省标准贡献奖4项。*Tel:(0592)6800818,E-mail:Dunmingxu@163.com.
  • 基金资助:
    国家重点研发计划(2019YFC1605400);食品安全国家标准项目(spaq-2019-40);福建省自然科学基金项目(2017J01020)

Determination of nine N-nitrosamines in animal derived foods by QuEChERS-isotope dilution combined with gas chromatography-tandem mass spectrometry

KONG Xiangyi1,2, ZHUANG Lili2, FANG Enhua2, LIN Peng1, ZHENG Zilong3, ZHENG Xianghua2, XU Dunming1,2,*()   

  1. 1. Fisheries College of Jimei University, Xiamen 361021, China
    2. Xiamen Customs Technology Center, Xiamen 361000, China
    3. Fujian Administration for Market Regulation, Fuzhou 350003, China
  • Received:2020-07-03 Online:2021-01-08 Published:2020-12-20
  • Contact: XU Dunming
  • Supported by:
    National Key Research and Development Program of China(2019YFC1605400);National Food Safety Standard Project(spaq-2019-40);Natural Science Foundation of Fujian Province(2017J01020)

RichHTML

54

PDF

161

摘要：

建立了同时测定动物源性食品中9种N-亚硝胺类化合物的气相色谱-串联质谱分析方法。当下动物源性食品中N-亚硝胺类化合物污染种类较多,对人体危害较大,但国标GB 5009.26-2016仅针对N-二甲基亚硝胺的检测,且存在样品前处理复杂、标准方法回收率低、再现性差等问题,因此建立同时快速检测多种N-亚硝胺类化合物的方法有一定现实意义。称取10.0 g样品,置于50 mL离心管中,加入200 μL内标工作液和10 mL乙腈,冷冻30 min后,加入4 g硫酸镁和1 g氯化钠进行脱水,以9000 r/min离心5 min。取5 mL上清液使用150 mg聚苯乙烯二乙烯苯(PLS-A)粉末净化,再使用1.6 g MgSO4和0.4 g NaCl脱水,过0.22 μm滤膜,上机分析。在初始温度为50 ℃时采用程序升温模式,0.16 min后,以900 ℃/min的速率将温度升至220 ℃。采用毛细管气相色谱柱HP-Innowax(30 m×0.25 mm×0.25 μm)分离,使用电子轰击电离(EI)源检测,在多反应监测模式下,以保留时间和特征离子对信息进行定性和定量分析,使用内标法定量N-亚硝胺类化合物。结果表明,N-亚硝胺类化合物在0.1~50.0 μg/L范围内具有良好的线性关系,方法的检出限(S/N=3)和定量限(S/N=10)分别为0.03~0.30 μg/kg和0.10~1.00 μg/kg。对不同样品基质进行0.5、1.0、3.0 μg/kg3个水平的加标回收试验,9种N-亚硝胺类化合物的回收率为80.4%~98.5%, RSD(n=6)为2.41%~12.50%。应用建立的方法检测市面上常见的动物源性食品,除N-亚硝基乙胺、N-亚硝基吗啡胆碱外,其他7种N-亚硝胺类化合物均有不同程度检出。检测结果表明,腌制水产品中N-亚硝胺类化合物含量普遍高于其他样品。研究建立的方法操作简单,不需要长时间蒸馏提取,可快速对动物源性食品中N-亚硝胺类化合物进行定性和定量分析,且样品和试剂的消耗量更少,节省成本,对环境污染小。该法的建立对我国动物源性食品中N-亚硝胺类化合物残留水平的控制、检测标准的制定和采取相应的管理措施具有一定的理论和现实意义。

关键词: 气相色谱-串联质谱, QuEChERS, N-亚硝胺类化合物, 动物源性食品

Abstract:

In this study, a comprehensive analytical method based on gas chromatography-tandem mass spectrometry (GC-MS/MS) was developed for the determination of nine N-nitrosamines in animal derived foods. There are many kinds of N-nitrosamines in foods that are harmful to human health. However, the national standard GB 5009.26-2016 pertains only to the detection of N-dimethylnitrosamine; there are many drawbacks of this method, such as complicated sample preparation, low recovery rate, and poor reproducibility. Hence, it is of practical significance to establish a method for the simultaneous determination of a variety of N-nitrosamines. The optimal extraction conditions for the developed method were as follows: 10.0 g aliquots of the sample were placed in a 50 mL centrifuge tube, followed by the addition of 10 mL acetonitrile and 200 μL internal working standard solutions. After 30 min of freezing treatment, 4 g magnesium sulfate and 1 g sodium chloride were added for dehydration, and the tube was centrifuged at 9000 r/min for 5 min. After vortex centrifugation, 5 mL of the clear supernatant was purified using 150 mg polystyrene divinylbenzene (PLS-A). The purified extracts were dewatered using 1.6 g MgSO4 and 0.4 g NaCl, and then filtered through a 0.22 μm membrane filter unit prior to GC-MS/MS analysis. Temperature-programmed was applied at an initial temperature of 50 ℃. After 0.16 min, the temperature was raised to 220 ℃ at the rate of 900 ℃/min for 5 min. N-Nitrosamines were separated on an HP-Innowax column (30 m×0.25 mm×0.25 μm). Identification and quantification were achieved using an electron impact ion (EI) source in positive ion mode with multiple reaction monitoring (MRM). The internal standard method was used to quantify the N-nitrosamines. Under the optimal conditions, the correlation coefficients of the standard calibration curves were not less than 0.99 in the range of 0.1-50.0 μg/L. The limits of detection were 0.03-0.30 μg/kg (S/N=3), and limits of quantification were 0.15-1.00 μg/kg (S/N=10). At spiked levels of 0.5, 1.0, and 3.0 μg/kg, the average recoveries of N-nitrosamines in spiked samples ranged from 80.4% to 98.5%, with relative standard deviations between 2.41% and 12.50%. This method was used to determine animal derived food products, except N-itrosomethylethylamine and N-nitrosomorpholine, others were founded. The results showed that N-nitrosamines levels in salted aquatic products were generally higher than those of the other samples. The method established in this study is simple to operate, and it does not require any time-consuming distillation extraction. Furthermore, there is minimal consumption of samples and reagents; consequently, the experiment cost is reduced, and the method is environmentally friendly. This method has theoretical and practical significance for the control of N-nitrosamines residues in animal derived foods, establishment of detection standards, and corresponding management measures.

Key words: gas chromatography-tandem mass spectrometry (GC-MS/MS), QuEChERS, N-nitrosamines, animal derived foods

中图分类号: