Simultaneous Determination Of Albendazole And Its Three Metabolites Residues In Pork,Poultry Muscle And Poultry Eggs By UPLC-FLD

In this experiment,three-way cross hybrid pig(Duroc×Landrace×Large White),Haiyang yellow chicken,Jinghai yellow chicken,Gaoyou duck and Yangzhou goose were selected as the experimental animals.The sample preparation technique of liquid-liquid extraction combined with solid phase extraction(LLE-SPE)was used to extract the targets substance from the sample matrix.The purpose of this study was to establish a method for the simultaneous determination of albendazole(ABZ)and its three metabolites(albendazole sulfone,ABZ-SO2;albendazole sulfoxide,ABZ-SO and albendazol-2-amino sulfone,ABZ-2NH2-SO2)residues in pork,poultry muscle(chicken muscle,duck muscle,goose muscle)and poultry eggs(chicken eggs,duck eggs,goose eggs)by ultra-high performance liquid chromatography-fluorescence detection(UPLC-FLD)method.The main results of this study are as follows:1.A method for the simultaneous extraction of ABZ,ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2 residues in pork,poultry muscle and poultry eggs by liquid-liquid extraction was established and optimized.ABZ and its three metabolites were extracted with ethylacetate in pork and poultry muscle,and were extracted with 0.2%formic acid-acetonitrile in poultry eggs.After that nitrogen was used to dry the samples and mobile phase was used to reconstitute the samples.Then acetonitrile saturated n-hexane was used to degrease the samples,and HLB solid phase extraction column was used to clean up the samples.After that the samples were dried by nitrogen and reconstituted by mobile phase before tested on the machine.The extraction method has a simple process and high recovery rates and the recovery rates of the four drugs were all above 80.37%.2.An UPLC-FLD method for simultaneous determination of ABZ,ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2 residues in pork and poultry muscle was developed and optimized.The peak area of each drug had good linear relationship with its added concentration when the concentration range of the mixed standard working solutions of albendazole and its three metabolites were at the limit of quantitation(LOQ)～400 μg/kg,and the coefficients of determination R2 were greater than 0.999 0.When the concentration of albendazole and its three metabolites in pork and poultry muscle were LOQ,0.5 MRL,1.0 MRL,2.0 MRL,respectively,the average recovery rates of ABZ,ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2 in pork were 80.37%～92.07%,91.69%～97.08%,84.65%～97.93%and 84.56%～97.18%,respectively;the intra-day relative standard deviations(RSDs)were 2.54%～4.67%,2.15%～4.53%,2.70%～4.34%and 2.52%～4.71%,respectively;the inter-day RSDs were 3.56%～6.20%,3.54%～4.84%,2.93%～5.03%and 4.35%～6.29%,respectively;the limits of detection(LODs)were 2.8,0.4,3.8 and 0.9 μg/kg,respectively;the LOQs were 10.0,1.5,9.7 and 3.0 μg/kg,respectively.The average recovery rates of ABZ,ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2 in poultry muscle were 84.09%～98.39%,83.13%～97.68%,87.37%～97.03%and 83.53%～97.25%,respectively;the intra-day RSDs were 2.18%～4.85%,1.33%～5.11%,1.68%～4.10%and 2.04%-4.25%,respectively;the inter-day RSDs were 2.23%～5.36%,2.10%～5.10%,2.72%～4.77%and 2.50%～4.65%,respectively;the LODs were 3.1～3.6,0.2～0.3,2.4～3.0 and 0.5～0.6 μg/kg,respectively;the LOQs were 10.0～10.9,1.0,8.0～8.5 and 1.5～1.9 μg/kg,respectively.3.An UPLC-FLD method for simultaneous determination of ABZ,ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2 residues in poultry eggs was developed and optimized.The peak area of each drug had good linear relationship with its added concentration when the concentration range of the mixed standard working solutions of albendazole and its three metabolites were at LOQ～400 μg/kg,and the coefficients of determination R2 were greater than 0.999 3.When the concentration of albendazole and its three metabolites in poultry eggs were LOQ,0.5 MRL,1.0 MRL,2.0 MRL,respectively,the average recovery rates of ABZ,ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2 in poultry eggs were 80.50%～92.42%,85.22%～98.60%,80.78%～96.80%and 84.41%～96.51%,respectively;the intra-day RSDs were 2.11%～4.91%,2.05%～4.53%,2.16%～4.86%and 2.00%～4.96%,respectively;the inter-day RSDs were 3.01%～5.49%,2.58%～5.13%,2.37%～5.42%and 2.41%～5.32%,respectively;the LODs were 3.4～4.6,0.3～0.9,2.7～3.2 and 0.6～1.1 μg/kg,respectively;the LOQs were 11.0～12.9,1.0～2.3,9.0～10.0 and 2.0～2.8 μg/kg,respectively.In conclusion,the UPLC-FLD method established in this research was efficient,rapid,accurate,sensitive and was suitable for simultaneous detection of albendazole and its three metabolites(ABZ-SO2,ABZ-SO and ABZ-2NH2-SO2)residues in pork,poultry meat and poultry eggs.