| Sulfachloropyrazine Sodium-Trimethopriom Soluble Powder is a new compound preparation,which is mainly used for the prevention and control of chicken coccidiosis in veterinary medicine.In this study,the method for the determination of sulfachloropyrazine(SPZ)and trimethoprim(TMP)residues in broiler tissues was established by high performance liquid chromatography(HPLC).The residue elimination of Sulfachloropyrazine Sodium-Trimethopriom Soluble Powder in broiler tissues was studied in order to provide a theoretical basis for the withdrawal time of the new compound preparation in broilers.1.Determination of SPZ Residues in broiler tissues by high performance liquid chromatographySPZ was extracted with ammonia-acetonitrile(5/95,v/v),defatted with n-hexane,the samples were purified by MCX SPE cartridges,and detected by HPLC with UV detection.The mobile phase was acetonitrile-0,02 mol/L potassium dihydrogen phosphate solution(35/65,v/v),the UV detection wavelength was 272 nm,and the external standard method was quantified.The results showed that the standard solution of SPZ had a good linear relationship(r?0.9999)in the range of 0.08?20.00 ?g/mL(0.04?10.00 mg/kg).The limit of detection(LOD)was 0.02 mg/kg and the limit of quantification(LOQ)was 0.04 mg/kg.The average recovery of SPZ in broiler tissues in the range of 0.04?10.00 mg/kg were from 75.46%to 89.40%,the intra-day and inter-day coefficient of variation were less than 9%.The established method of SPZ in this experiment can be used for the residue analysis of SPZ in broiler tissues.2.Determination of TMP Residues in broiler tissues by high performance liquid chromatographyTMP was extracted with acetonitrile,defatted with n-hexane,the samples were purified by MCX SPE cartridges,and determinated by HPLC with UV detection.The mobile phase was acetonitrile-0.01 mol/L potassium dihydrogen phosphate solution(18/82,v/v),the UV detection wavelength was 240 nm,and the external standard method was quantified.The results showed that the standard solution of TMP had a good linear relationship(r?0.9999)in the range of 0.05?8.00 ?g/mL(0.025?4.000 mg/kg).The limit of detection was 0.01 mg/kg and the limit of quantification was 0.025 mg/kg.The average recovery of TMP in broiler tissues in the range of 0.04?4.00 mg/kg were from 77.16%to 90.88%,the intra-day and inter-day coefficient of variation were less than 7%.The established method of TMP in this experiment can be used for the residue analysis of TMP in broiler tissues.3.Study on the residue elimination of Sulfachloropyrazine Sodium-Trimethopriom Soluble Powder in broiler tissuesHealthy commercial broilers were administrated at 300 mg/L(sulfachloropyrazine sodium counted)of the maximum recommended dose of Sulfachloropyrazine Sodium-Trimethopriom Soluble Powder in drinking water for six successive days.Six broilers were respectively slaughtered at 4 h,1 d,3 d,7 d,12 d after the last administration.Concentration of SPZ and TMP in broiler tissues were determined by the established method above.The results showed that the residual of SPZ was the highest in kidney,followed by skin+fat,liver and the lowest in muscle after 4 h withdrawal.The residual of SPZ when lower than the MRL(0.1 mg/kg)in muscle,liver,kidney and skin+fat was 3 d,3 d,7 d,12 d.The residual of TMP was the highest in kidney,followed by the liver,lower in skin+fat and muscle after 4 h withdrawal.The residual of TMP when lower than the MRL(0.05 mg/kg)in muscle,liver,kidney and skin+fat was 1 d,1 d.7 d.3 d.The results showed that SPZ and TMP were all eliminated quickly in muscle and liver,SPZ was eliminated slowly in kidney and skin+fat,TMP was eliminated slowly in kideney,kidney and skin+fat are target tissues of SPZ.and kidney are target tissues of TMP.The raw data were analyzed with software WT1.4 The withdrawal period of SPZ in broiler kidney and skin+fat was 15.47 d,18.18 d,The withdrawal period of TMP in broiler kidney and skin+fat was 9.08 d and 13.69 d.The experiment results showed that Sulfachloropyrazine Sodium-Trimethopriom Soluble Powder according to the recommended dose of water administration,the withdrawal period was 19 d in broiler. |
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