色谱 ›› 2021, Vol. 39 ›› Issue (6): 624-632.DOI: 10.3724/SP.J.1123.2020.12026

• 研究论文 • 上一篇    下一篇

聚多巴胺纳米纤维膜固相萃取-超高效液相色谱-串联质谱检测淡水鱼中四环素类和氟喹诺酮类药物残留

梁思慧1, 戴海蓉1, 张铧尹1, 李建2, 张秋萍2, 许茜1,3,*(), 王春民2,*()   

  1. 1.东南大学公共卫生学院营养与食品卫生学系, 江苏 南京 210009
    2.苏州市疾病预防控制中心, 江苏 苏州 215100
    3.东南大学环境与医学工程教育部重点实验室, 江苏 南京 210009
  • 收稿日期:2020-12-24 出版日期:2021-06-08 发布日期:2021-04-13
  • 通讯作者: 许茜,王春民
  • 作者简介:E-mail: chunminwang@163.com(王春民).
    * E-mail: q_xu68@163.com(许茜);
  • 基金资助:
    国家自然科学基金(81973098)

Determination of tetracycline and fluoroquinolone residues in fish by polydopamine nanofiber mat based solid phase extraction combined with ultra performance liquid chromatography-tandem mass spectrometry

LIANG Sihui1, DAI Hairong1, ZHANG Huayin1, LI Jian2, ZHANG Qiuping2, XU Qian1,3,*(), WANG Chunmin2,*()   

  1. 1. Department of Nutrition and Food Hygiene, School of Public Health, Southeast University, Nanjing 210009, China
    2. Suzhou Center for Disease Control and Prevention, Suzhou 215100, China
    3. Key Laboratory of Ministry of Education of Environment and Medical Engineering, Southeast University, Nanjing 210009, China
  • Received:2020-12-24 Online:2021-06-08 Published:2021-04-13
  • Contact: XU Qian,WANG Chunmin
  • Supported by:
    National Nature Science Foundation of China(81973098)

摘要:

制备聚多巴胺(PDA)修饰的聚苯乙烯纳米纤维膜(PS NFsM)作为固相萃取吸附介质,可快速提取淡水鱼中3种四环素类(四环素、金霉素、土霉素)和3种氟喹诺酮类(恩诺沙星、环丙沙星、诺氟沙星)药物残留,结合超高效液相色谱-串联质谱(UPLC-MS/MS),建立了药物残留检测的新方法。利用静电纺丝法制备了聚苯乙烯纳米纤维膜,将其作为模板,通过自聚合作用,进行聚多巴胺功能化修饰,得到PDA-PS NFsM材料。对制得的PS NFsM和PDA-PS NFsM材料进行傅里叶红外光谱和场发射扫描电镜表征,证明PDA的成功修饰,修饰后的纳米纤维表面粗糙,呈现核-壳形貌,纤维内部为蜂窝状多孔结构。以空白加标样品的回收率为指标,对PDA-PS NFsM材料的用量、离子强度、样品溶液的流速、洗脱液和突破体积等影响SPE的因素进行考察及条件优化,确定了最佳的SPE条件。该方法对6种目标物的检出限为0.3~1.5 μg/kg,定量限为1.0~5.0 μg/kg,低于国家标准和行业标准;在各目标物的线性范围内均有良好的线性关系,决定系数(R2)大于0.999,方法的回收率为94.37%~102.82%,日间和日内的相对标准偏差(RSD)均小于10%,与国家标准和行业标准相当。通过固相萃取前后的基质效应对比,表明PDA-PS NFsM具有优秀的净化能力。最后,通过实际样品分析验证了方法的实际应用可行性。该文建立的基于PDA-PS NFsM材料的SPE方法是一种高效环保的方法,可为淡水鱼中药物残留的常规监测提供技术支持。

关键词: 超高效液相色谱-串联质谱, 固相萃取, 聚多巴胺, 纳米纤维膜, 药物残留, 淡水鱼

Abstract:

Tetracyclines and fluoroquinolones are common antibacterial drugs used in aquaculture, and their residues may pose a risk to human health. The low concentration of drug residues and complex matrixes such as fats and proteins in aquatic products necessitate the urgent development of efficient sample pretreatment methods. Solid phase extraction (SPE) is the most common sample pretreatment method, in which the core is an adsorbent. Compared with traditional SPE adsorbents, nanofiber mat (NFsM) has more interaction sites because of their large specific surface area. Furthermore, NFsMs modified with specific functional groups can significantly improve the extraction efficiency of tetracyclines and fluoroquinolones. Polydopamine (PDA) is spontaneously synthesized by the oxidative self-polymerization of dopamine-hydrochloride in alkaline solutions (pH>7.5). Because of its rich amino and catechol groups, PDA can form π-π stacking, electrostatic attraction, hydrophobic interaction, and hydrogen bonding interactions with target molecules. By exploiting the above advantages, polystyrene (PS) NFsM, as a template, was prepared by the electrostatic spinning method, and PDA-PS NFsM was obtained by functional modification of PDA through self-polymerization. Fourier transform infrared spectroscopy (FT-IR) and field-emission scanning electron microscopy (FESEM) were used to characterize the synthesized PS NFsM and PDA-PS NFsM. It was proved that PDA was successfully modified on the PS NFsM, with the SEM images revealing a rough outer core shell structure and an inner honeycomb structure. Subsequently, the handmade SPE column with PDA-PS NFsM was completed. A novel and efficient screening analytical method based on PDA-PS NFsM for the simultaneous determination of three tetracyclines (tetracycline (TET), chlortetracycline (CTC), and oxytetracycline (OTC)) and three fluoroquinolones (enrofloxacin (ENR), ciprofloxacin (CIP), and norfloxacin (NOR)) in fish by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. The SPE procedure was optimized to develop an efficient method for sample preparation. Evaluate parameters including the amount of NFsM usage, ionic strength, flow rate of the sample solution, composition of eluent, and breakthrough volume were investigated. Only (20±0.1) mg of PDA-PS NFsM was sufficient to completely adsorb the targets, and the analytes retained on NFsM could be eluted by 1 mL of formic acid-ethyl acetate (containing 20% methanol) (1∶99, v/v). The residues were redissolved in 0.1 mL 10% methanol aqueous solution containing 0.2% formic acid. In addition, no adjustment of the pH and ionic strength of the sample solutions was required, and the breakthrough volume was 50 mL. The limits of detection (LODs) and limits of quantification (LOQs) of the six target compounds were measured at 3 times and 10 times the signal-to-noise ratio (S/N), respectively. The LODs and LOQs were 0.3-1.5 μg/kg and 1.0-5.0 μg/kg, respectively. The linear ranges of the six target compounds were LOQ-1000 μg/kg, and the coefficient of determination (R2) was greater than 0.999. To evaluate the accuracy and precision, blank spiked samples at three levels (low, medium, and high) were prepared for the recovery experiments, and each level with six parallel samples (n=6). The recoveries ranged from 94.37% to 102.82%, with intra-day and inter-day relative standard deviations of 2.38% to 8.06% and 4.10% to 9.10%, respectively. To evaluate the purification capacity of PDA-PS NFsM, the matrix effects before and after SPE were calculated and compared. Matrix effects before SPE were -12.98% to -38.68%. After the completion of SPEbased on PDA-PS NFsM, the matrix effect of each target analyte was significantly reduced to -2.15% to -7.36%, which proved the significant matrix removal capacity of PDA-PS NFsM. Finally, the practicality of this method was evaluated by using it to analyze real samples. This SPE method based on PDA-PS NFsM is efficient, practical, and environmentally friendly, and it has great potential for use in the routine monitoring of drug residues in fish.

Key words: ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), solid phase extraction (SPE), polyaniline (PDA), nanofiber mat (NFsM), drug residues, fish

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