Rapid Determination of Clopidol in Animal Feed by Solid-Phase Extraction Coupled with Liquid Chromatography-Tandem Mass Spectrometry (SPE-LC-MS/MS)

PAN Fei; LIN Xuexian; LIU Zhenni; DING Chenhong; WANG Weili

doi:10.11924/j.issn.1000-6850.casb2025-0373

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Chin Agric Sci Bull ›› 2026, Vol. 42 ›› Issue (9) : 163-170. DOI: 10.11924/j.issn.1000-6850.casb2025-0373

Rapid Determination of Clopidol in Animal Feed by Solid-Phase Extraction Coupled with Liquid Chromatography-Tandem Mass Spectrometry (SPE-LC-MS/MS)

PAN Fei ¹^,² ,
LIN Xuexian ³ ,
LIU Zhenni ²^,⁴ ,
DING Chenhong ²^,⁴ ,
WANG Weili ¹^,²

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Abstract

To address the problems of low sensitivity, cumbersome pretreatment, large matrix interference, and easy missed detection of low concentration of clopidol in feed, this study aimed to establish a solid-phase extraction- liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) method for the rapid determination of clopidol content in feed, in order to meet the requirements of accurate quantification and safety supervision. Clopidol in feed samples was extracted using a 0.2% ammonia-methanol solution, purified through an alkaline alumina solid-phase extraction (SPE) cartridge, and separated on a C₁₈ column with gradient elution using ammonium formate solution and methanol as the mobile phase. Detection was performed by LC-MS/MS in positive electrospray ionization (ESI⁺) mode, and quantification was conducted using matrix-matched external standard calibration. The results showed that clopidol exhibited a good linearity within the concentration range of 0.2-100 μg/L, with a correlation coefficient (r²) greater than 0.999. The method detection limits (LOD) and quantification limits (LOQ) were 0.05 mg/kg and 0.1 mg/kg, respectively. Method accuracy was evaluated in various types of feed, yielding recoveries of 90%-102%, with a relative standard deviation (RSD) below 7%. This analytical method is simple to perform, highly sensitive, accurate, and reproducible, and significantly improves the detection limit and qualitative reliability compared with the national standard method. It meets the methodological requirements for the determination of clopidol in feed and provides a reliable technical approach for its precise quantification.

Key words

feeds / clopidol / liquid chromatography-tandem mass spectrometry / solid-phase extraction / anticoccidial drugs / matrix effect

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PAN Fei , LIN Xuexian , LIU Zhenni , et al . Rapid Determination of Clopidol in Animal Feed by Solid-Phase Extraction Coupled with Liquid Chromatography-Tandem Mass Spectrometry (SPE-LC-MS/MS)[J]. Chinese Agricultural Science Bulletin. 2026, 42(9): 163-170 https://doi.org/10.11924/j.issn.1000-6850.casb2025-0373

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