Chinese Journal of Chromatography

Chinese Journal of Chromatography

   

Simultaneous determination of antibiotics and psychoactive substances in mollusks by QuEChERS and ultra-high performance liquid chromatography-tandem mass spectrometry

GU Xiaorong1,2, FU Jie3,*, LIN Feng4, WU Yajin3, YANG Dan3, CHEN Linhai3, ZHANG Aiqian1,2,3, FU Jianjie1,2,3,*   

  1. 1. State Key Laboratory of Environmental Chemistry and Toxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China;
    2. College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China;
    3. Zhejiang Key Laboratory of Environment and Health of New Pollutants, School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou 310024, China;
    4. Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China
  • Received:2026-02-12
  • Supported by:
    Youth Fund of the National Natural Science Foundation of China (No. 22306040); China Postdoctoral Science Foundation (No. 2023M744123); the Research Funds of the Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences (No. 2023HIAS-Y019); the Central Guiding Local Science and Technology Development Fund Projects (No. 2025ZY01044); Research Project of Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs (No. ZJK202516).

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Abstract: The widespread detection of antibiotics and psychoactive substances (PSs) in aquatic products has raised significant concerns. However, sensitive methods for detecting both compound classes simultaneously are lacking. To address this analytical gap, this study developed a novel analytical method for the determination of mollusks by QuEChERS-ultra-high performance liquid chromatography- triple quadrupole mass spectrometry (UHPLC-MS/MS). This method simultaneously determines 28 antibiotics and 10 PSs in shellfish and gastropods. During the extraction phase, target compounds were extracted using methanol-acetonitrile (1:1, V/V) containing 0.1% ammonia solution. This optimized solvent system ensured high extraction efficiency while significantly reducing matrix effects. The subsequent purification step was performed with a mixed sorbent (C18 100 mg, PSA 100 mg, GCB 100 mg, anhydrous MgSO4 300 mg). This specific combination effectively removed lipids, proteins, and pigment interferences from the sample extracts. Mass spectrometry was performed in positive electrospray ionization (ESI+) mode, with selected reaction monitoring (SRM) as the acquisition mode. Results demonstrated that all target compounds exhibited good linearity in the range of 0.1‒50 μg/L, with correlation coefficients (r2) exceeding 0.995. The instrument limits of detection (LODs) ranged from 0.002 to 0.1 ng/g, and the limits of quantification (LOQs) were between 0.005 and 0.3 ng/g. The method reporting limits (MRLs) varied from 0.5 to 2.9 ng/g. Accuracy and precision were comprehensively evaluated at three distinct spiked levels (0.5, 2.5, and 10 ng/g). The average recoveries for the targeted analytes ranged from 70.7% to 124.9%, with relative standard deviations (RSDs, n = 6) of 1.0%‒18.6%. Furthermore, the intra-day and inter-day precision values were 1.4%‒14.7% and 0.7%‒12.4%, respectively, indicating high method stability and reproducibility. The optimized method was applied to analyze antibiotics and PSs in 7 shellfish and 3 gastropod samples collected from coastal markets in Zhejiang Province, revealing the presence of 15 antibiotics and 9 PSs. Total antibiotic concentrations ranged from not detected (nd) to 3β251.3 ng/g dw(dry weight). The mean antibiotic content was 325.1 ng/g dw. Notably, sulfamethoxypyridazine emerged as the predominant antibiotic residue, presenting an average concentration of 111.7 ng/g dw and accounting for 34.5% of the total antibiotic load. The total concentrations of PSs ranged from nd to 4β909.2 ng/g dw. The mean PSs content was 490.9 ng/g dw. Ketamine was identified as the primary psychoactive contaminant, with a mean concentration of 199.2 ng/g dw, representing 40.5% of the total PSs load. In conclusion, this method is sensitive and efficient, provides satisfactory recovery and reproducibility, and is suitable for the simultaneous rapid determination of trace levels of antibiotics and PSs in shellfish and gastropod mollusks.

Key words: shellfish and gastropods mollusks, antibiotics, psychoactive substances, QuEChERS, ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)

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