Development Of Methods For Determination Of Robenidine And Levamisole Residues In Animal Derive Food

Robenidine is a common anticoccidial drug used in the poultry,and it has good curative effect in prevent acute and chronic coccidiosis in chicken and rabbit as feeding stuff.Levamisole is a potent broad-spectrum anthelmintic drug,which is widely used in veterinary medicine for the control of gastrointestinal parasites in cattle,sheep and pigs. Incorrect use of robenidine and levamisole may leave residues in edible tissues threat the health of mankind.This study used a external standard method to quantitate,developed method to analyse robenidine residue in chicken and rabbit,to detect levamisole residue in chicken and pig by liquid chromatography,respectively.In this paper,a rapid and effective analytical procedure for the determination of robenidine in chicken and rabbit tissues by high-performance liquid chromatorgraphy was developed.Robenidine was extracted from samples with ethyl acetate,centrifuged,the upper organic layer was evaporated to nearly dryness,then the residues was dissolved with hexane and clean with florsil SPE cartridge,10%dimethylketone in methanol eluted, evaporated,dissolved with lmL methanol.Aliquots of 20uL of the resulting sample were injected into the HPLC system set at 353nm.Retention time of robenidine was 5min. Linearity of the detector response was verified with robenidine standard solution in the range of 5~500μg /L,the linear regression was y=0.00715x+1.386(r=0.9999).The quantitative methed indicated that,the LOD was 5μg/kg and the LOQ was lOμg/kg. Recoveries of robenidine from spiked samples at different concentrations was between 70%-105%,the RSD for intra- and inter-day data were below 15%and 20%, respectively.Another high-performance liquid chromatography method was optimised and validated for the determination of levamisole in pig and chicken tissues.A liquid-liquid extraction procedure in alkaline medium,using ethyl acetate as extraction solvent.The ethyl acetate extracts were re-extracted with O.lmol/L HC1,then the solution was followed by a solid-phase extractaction procedure using an MCX column to clean up, eluted by 4%ammonia in methanol,the eluate was evaporated to dryness under a gentle stream of nitrogen gas.The dry residue was redissolved in lmL mobile phase.The reconstituted sample was transferred to HPLC system to detect.Retention time of levamisole was 12min.Linearity of the detector response was verified with robenidine standard solution in the range of 10-1000μg/L,the linear regression was y=196.37x+81.599(r=0.9999).The quantitative methed indicated that,the LOD was 2μg/kg and the LOQ was 5μg/kg.Recoveries of robenidine from spiked samples at different concentrations was between 70%-110%,the RSD for intra- and inter-day data were below 10%and 15%,respectively.The conditions of sample preparation and detection by HPLC were optimized in this study.These two methods were successfully validate in compliance with veterinary residues analysis requirements.The development of the methods provide technical support for monitoring of residues of these drugs in food derived from Animals.