色谱

色谱 ›› 2025, Vol. 43 ›› Issue (7): 767-778.DOI: 10.3724/SP.J.1123.2024.11017

• 研究论文 • 上一篇    下一篇

基于复合净化柱-超高效液相色谱-串联质谱法同时测定鱼、虾、蟹中50种兽药残留

邓惠丹1,2, 吉小凤2,*(), 肖英平2, 夏强1,*(), 杨华2,*()   

  1. 1.宁波大学食品科学与工程学院，浙江 宁波 315832
    2.浙江省农业科学院农产品质量安全与营养研究所，浙江 杭州 310021
  • 收稿日期:2024-11-18 出版日期:2025-07-08 发布日期:2025-07-04
  • 通讯作者: *Tel：（0574）87604388，E-mail：xiaqiang@nbu.edu.cn（夏强）；Tel：（0571）86415208，E-mail：jixiaofeng@zaas.ac.cn（吉小凤）；Tel：（0571）86404005，E-mail： yanghua@zaas.ac.cn（杨华）.
  • 基金资助:
    浙江省农业农村厅风险评估项目(2022R19T60D18);浙江省农业农村厅风险评估项目(2023R19T60D14)

Simultaneous determination of 50 veterinary drug residues in fish， shrimp， and crabs using a composite purification column-ultra-performance liquid chromatography-tandem mass spectrometry

DENG Huidan1,2, JI Xiaofeng2,*(), XIAO Yingping2, XIA Qiang1,*(), YANG Hua2,*()   

  1. 1. School of Food Science and Engineering，Ningbo University，Ningbo 315832，China
    2. Institute of Agro-product Safety and Nutrition，Zhejiang Academy of Agricultural Sciences，Hangzhou 310021，China
  • Received:2024-11-18 Online:2025-07-08 Published:2025-07-04
  • Supported by:
    Risk Assessment Project of the Department of Agriculture and Rural Affairs of Zhejiang Province(2022R19T60D18);Risk Assessment Project of the Department of Agriculture and Rural Affairs of Zhejiang Province(2023R19T60D14)

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摘要：

建立了一步式复合净化柱样品前处理方法-超高效液相色谱-串联质谱（UPLC-MS/MS）测定鱼、虾、蟹中喹诺酮类、大环内酯类、硝基咪唑类、磺胺类、酰胺醇类、苯并咪唑类及敌百虫、金刚烷胺、尼卡巴嗪共50种兽药残留的方法。5 g样品经1.0%乙酸乙腈溶液提取，无水硫酸钠盐包去除水分，FAVEX净化柱净化，净化液氮吹至近干，0.2%甲酸水溶液-甲醇（9∶1，v/v）复溶残渣至1.0 mL，过滤膜后待测物经Waters ACQUITY UPLC® BEH C18色谱柱（100 mm×2.1 mm，1.7 μm）分离，在正、负离子模式下以UPLC-MS/MS多反应监测模式采集数据，分别用同位素内标法、基质溶液外标法定量。在最优的仪器参数及前处理条件下进行方法验证，结果表明，目标化合物在各自相应的质量浓度范围内线性关系良好，决定系数（R2）均大于0.990 0，检出限（LOD）为0.2～6.0 μg/kg，定量限（LOQ）为0.5～20.0 μg/kg。取鱼、虾、蟹的空白样品进行3个水平的加标回收试验（n=6），50种兽药的平均回收率为60.1%～119.7%，相对标准偏差（RSD）为1.11%～15.6%。将该方法应用于150批次实际样品的测定，兽药残留检出率为47.3%，超标率为7.3%，说明水产品中确实存在兽药残留的现象，后续应加强对水产品中兽药残留的风险预警和评估。该方法具有前处理过程简单、检测效率高、有机试剂消耗少、成本低等优点，适用于同时测定鱼、虾、蟹中多种兽药残留，可为水产品风险监测、风险评估和预警提供强有力的技术支持。

关键词: 水产品, 兽药残留, 食品安全, 超高效液相色谱-串联质谱法

Abstract:

China is the world’s largest producer and consumer of aquatic products. Insecticides， as well as veterinary drugs， such as quinolones， macrolides， nitroimidazoles， sulfonamides， and amphenicols， are widely used in aquaculture. These compounds play crucial roles in preventing， controlling， and treating diseases in aquatic organisms， improving feed conversion rates， and promoting the healthy growth of farmed species. However， the illegal use of veterinary drugs， their overuse， and noncompliance with drug withdrawal periods by farmers can lead to aquatic products containing veterinary drug residues. This study focused on fish， shrimp， and crabs with the aim of meeting the green-development needs of China’s aquaculture industry， strengthening risk warnings and preventing veterinary drug residues in aquatic products， enhancing the quality and competitiveness of farmed aquatic products， and promoting the sustainable development of the aquaculture industry. Instrumental working conditions were optimized， and the extraction solvent， extraction method， purification materials， and the redissolution solvent used in the sample-pretreatment process were determined. These efforts led to the establishment of a protocol for the detection of 50 veterinary drug residues， including quinolones， macrolides， nitroimidazoles， sulfonamides， amphenicols， and insecticides， in fish， shrimp， and crabs， which involved one-step pretreatment with a composite purification column in combination with ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）. A 5-g sample was extracted with a 1.0% acetic acid solution， dehydrated with anhydrous sodium sulfate， and purified using a FAVEX purification column， after which the purified solution was nitrogen-blown to near dryness. The residue was re-dissolved in 1.0 mL of 0.2% formic acid aqueous solution-methanol （9∶1， v/v） ， filtered， and then separated using a Waters ACQUITY UPLC® BEH C18 column （100 mm × 2.1 mm， 1.7 μm）. Data were collected in positive- and negative-ion multiple reaction monitoring modes， and quantified using isotopic internal and external standards. All target compounds exhibited good linear relationships within their respective concentration ranges， with coefficients of determination （R²） greater than 0.990 0. Average recovery rates of 60.1%–117.8% were determined for the 50 veterinary drugs in the fish matrix， with RSDs of between 1.89% and 15.0%， while the shrimp matrix delivered average recovery rates of 60.2%–119.7%， with RSDs of 1.11%–15.6%， and the crab matrix exhibited average values of 60.7%–119.7% and 2.94%–15.0%， respectively. The developed method is advantageous owing to its simple pretreatment step， high throughput， low organic-solvent consumption， good accuracy， high sensitivity， and cost-effectiveness. One hundred and fifty batches of aquatic products， including 50 batches of fish， 50 batches of shrimp， and 50 batches of crab， were risk-screened for 50 veterinary drug residues using the established detection method. Veterinary drug residues were detected at a rate of 47.3%， with an over-standard rate of 7.3%. Specifically， the fish samples exhibited a detection rate of 66.0%， with contamination concentrations of 1.20–257.1 μg/kg. In contrast， the shrimp samples exhibited a detection rate of 18.0%， with contamination concentrations of 1.10–8.60 μg/kg， while values of 22.0% and 3.00–7.30 μg/kg， respectively， were determined for the crab samples. This study identified a total of seven veterinary drug residues， namely pefloxacin， norfloxacin， ciprofloxacin， enrofloxacin， sarafloxacin， sulfadimethoxine， and sulfamethoxazole， with detection rates of 0.67%， 5.3%， 12.7%， 20.7%， 4.6%， 2.00%， and 1.33%， respectively. The detection method established in this study provides strong technical support for monitoring and assessing aquatic-product risks in a timely manner， and is expected play an important role in ensuring food safety， protecting consumer health， and promoting the healthy development of the aquatic-product industry.

Key words: aquatic products, veterinary drug residues, food safety, ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）

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