China Animal Husbandry and Veterinary Medicine ›› 2020, Vol. 47 ›› Issue (6): 1961-1970.doi: 10.16431/j.cnki.1671-7236.2020.06.036

• Products Quality and Safety • Previous Articles    

Determination of Robenidine and Its Metabolites in Animal Derived Food by UPLC-MS/MS

ZHENG Xingguo, GUO Lingan, ZHAO Shan, LI Xi, LEI Xinyu, ZHONG Lingli   

  1. Analysis and Testing Center of Sichuan Academy of Agricultural Sciences, Chengdu 610066, China
  • Received:2019-12-30 Online:2020-06-20 Published:2020-06-20

Abstract: In order to detect robenidine and its metabolites residues in animal derived food rapidly and effectively,an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of robenidine and its metabolites (4-chlorobenzoic acid,4-chlorobenzoylamino-acetic acid) residues in five kinds of animal derived food,including eggs,chicken,beef,fish and pork.Samples were extracted with 2% (V/V) formic acid-acetonitrile solution,removed water with anhydrous sodium sulfate,fixed volume with methanol,then excepted fat with n-hexane.The purified samples were analyzed by UPLC-MS/MS after freezed and high speed centrifuged.UPLC was achieved using a Waters ACQUITY UPLC BEH C18 column (2.1 mm×100 mm,1.7 μm);the mobile phase consisted of methanol and 0.1% formic acid-water solution.The qualitative analysis was performed with MS/MS under multiple reaction monitoring (MRM) mode with positive and negative electrospray ionization;the matrix matched external standard method was used for quantitation.The results showed that there were good linearities of the three compounds in their respective concentration range (R2>0.999).The limits of detection (LODs) and the limits of quantity (LOQs) for robenidine were 0.5 and 1.0 μg/kg,for 4-chlorobenzoic acid were 2.5 and 5.0 μg/kg,for 4-chlorobenzoylamino-acetic acid were 1.0 and 2.5 μg/kg,respectively.While blank samples spiked four levels (robenidine:1.0,25,50,100 μg/kg;4-chlorobenzoic acid:5.0,25,50,100 μg/kg;4-chlorobenzoylamino-acetic acid:2.5,25,50,100 μg/kg),the recoveries and RSDs (n=6) of the three compounds were 76.0%-95.9% and 2.6%-10.6%,respectively.The matrix effects (|ME|) of the developed method were 0.2%-26.2%.There was a strong matrix effect (|ME|>20%) of robenidine in eggs and 4-Chlorobenzoylamino-acetic acid in chicken,beef and fish;blank pork could be used as representative matrix for quantitative analysis of three compounds.Therefore,the method was simple in pretreatment,high in sensitivity and good in reproducibility,and could be used for the determination of robenidine and its metabolites in five kinds of animal derived food.

Key words: ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS); animal derived food; robenidine; metabolite

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