色谱

色谱 ›› 2022, Vol. 40 ›› Issue (11): 998-1004.DOI: 10.3724/SP.J.1123.2022.07002

• 研究论文 • 上一篇    下一篇

共价三嗪骨架吸附剂-固相萃取-超高效液相色谱-串联质谱法测定牛奶中3种青霉素类残留

李琴1, 代书宇2, 杨媛2, 冯玉敏2, 练鸿振3, 张书胜2, 张文芬2,4,*()   

  1. 1.郑州市食品药品检验所, 河南 郑州 450006
    2.郑州大学化学学院, 河南 郑州 450001
    3.南京大学化学化工学院, 江苏 南京 210023
    4.中国烟草总公司郑州烟草研究院, 河南 郑州 450001
  • 收稿日期:2022-07-04 出版日期:2022-11-08 发布日期:2022-11-10
  • 通讯作者: 张文芬
  • 基金资助:
    国家自然科学基金(22004109);中国博士后科学基金(2022M710167);河南省市场监督管理局科技计划(2022sj134)

Determination of three penicillin residues in milk by solid-phase extraction-ultra performance liquid chromatography-tandem mass spectrometry using a covalent triazine framework sorbent

LI Qin1, DAI Shuyu2, YANG Yuan2, FENG Yumin2, LIAN Hongzhen3, ZHANG Shusheng2, ZHANG Wenfen2,4,*()   

  1. 1. Food and Drug Inspection Institute of Zhengzhou, Zhengzhou 450006, China
    2. College of Chemistry, Zhengzhou University, Zhengzhou 450001, China
    3. College of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China
    4. Zhengzhou Tobacco Research Institute of China National Tobacco Corporation, Zhengzhou 450001, China
  • Received:2022-07-04 Online:2022-11-08 Published:2022-11-10
  • Contact: ZHANG Wenfen
  • Supported by:
    National Natural Science Foundation of China(22004109);China Postdoctoral Science Foundation(2022M710167);Scientific and Technological Project of Administration for Market Regulation Henan Province(2022sj134)

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摘要：

建立了固相萃取-超高效液相色谱-串联质谱法(SPE-UPLC-MS/MS)同时测定牛奶中苄青霉素、邻氯青霉素、氨苄青霉素3种青霉素残留的分析方法。以自制的共价三嗪骨架(CTFs)材料作为固相萃取吸附剂,对影响固相萃取柱效率的吸附剂填充量、洗脱剂种类和用量及上样速率等主要因素进行了优化;同时对样品的提取和净化条件进行了考察。在3 mL/min的样品流速下,采用60 mg CTFs吸附剂和6 mL纯乙腈洗涤液达到了最佳的萃取效果。以0.1%甲酸水溶液-乙腈作为流动相进行梯度洗脱,在Waters ACQUITY UPLC BEH C18色谱柱上分离,电喷雾正离子(ESI+)模式下以动态多反应监测(MRM)采集数据,外标法定量。3种目标分析物的线性回归方程相关系数均大于0.999,检出限(LOD)为0.05~0.10 μg/kg(信噪比S/N=3),定量限(LOQ)为0.1~0.4 μg/kg(S/N=10),加标回收率为84.9%~94.1%,相对标准偏差(RSD, n=5)为1.66%~3.27%。此外,共价三嗪骨架材料与目标物的作用机理研究表明,主客体分子间存在π-π相互作用和氢键作用等多重相互作用,使该吸附剂可成功用于牛奶中青霉素的富集和净化。该方法具有精密度较高、重复性较好、分离度高、分析时间短等优点,可适用于牛奶中青霉素定性定量测定。

关键词: 共价三嗪骨架, 固相萃取, 超高效液相色谱-串联质谱, 青霉素, 牛奶

Abstract:

A method based on solid-phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS) was established for the determination of gpenicillin, cloxacillin, ampicillin residues in milk. Using self-made covalent triazine frameworks (CTFs) as the solid-phase extraction sorbents, the main factors influencing the efficiency of the solid-phase extraction columns, such as the sorbent amount, eluent type, eluent volume, and flow rate, were optimized. The extraction and purification conditions for the samples were also investigated. The optimal extraction effect was achieved at a flow rate of 3 mL/min with 60 mg CTFs and 6 mL eluent solution (acetonitrile). Separation was carried out on a Waters ACQUITY UPLC BEH C18 column, and 0.1% formic acid aqueous solution-acetonitrile was used as the mobile phases for gradient elution. The filtrate was detected by ultra performance liquid chromatography-tandem mass spectrometry, identified by electrospray ionization (ESI) in the positive mode using multiple reaction monitoring, and quantified using external standards. The calibration curves of the three penicillins showed good linearity and the correlation coefficients of the linear regression equations for the three target analytes were all greater than 0.999. The limits of detection (LODs) and limits of quantification (LOQs) were 0.05-0.10 μg/kg and 0.1-0.4 μg/kg, respectively. The average recoveries of the three analytes were 84.9%-94.1%, and the relative standard deviations (RSDs, n=5) were 1.66%-3.27%. Moreover, the mechanism of interaction between the CTFs and the target analytes was analyzed. The results revealed the existence of π-π and hydrogen-bond interactions between the CTFs and analytes. The results further indicated that the CTFs could be successfully used for the enrichment and purification of penicillins in milk. The proposed method has the advantages of high precision, good reproducibility, high resolution, and short analysis time, and it is suitable for the qualitative and quantitative determination of trace targets in complex matrices.

Key words: covalent triazine frameworks (CTFs), solid-phase extraction (SPE), ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), penicillin, milk

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