| The purpose of determination Ampicillin residue in swine,HPLC and CE methods was established.It was prefer HPLC to CE to detect Ampicillin residue in swine.The protein in plasma,muscle and liver were precipitated by treichoroacetic, derivatized at 100℃in water for 30 Min with formaldehyde.With the condition of Agilent C18column(4.6*250mm,Φ5μm)detection.The ampicillin fluorescence derivative was analyzed by HPLC with fluorescence detection at an excitation wavelength of 346nm and an emission wavelength of 422nm,mobile phase was PBS(PH3.5)/acetonitrile:76/24(v/v),at 1mL/Min flow rate.The limit of quantitation was 0.01μg/mL in plasma and tissue.The calibration curve was linear in rage of 0.01-10μg/mL.The recoveries of Ampicillin added to plasma at levels at 0.02,0.1,2,20μg/mL were 93.4±2.06%,93.5±3.81%,99.0±0.98%,93.3±1.06%.The average was 95.23±1.98%in plasma.The recoveries of ampicillin added to tissue at levels 0.1,0.5,10,100μg/mL were 76.6±3.65%,77.7±2.96%,71.0±3.81%,72.3±3.60%.The average was 74.40±3.50%.The recoveries of ampicillin added to liver at levels 0.1,0.5,10,100μg/mL were 75.6±2.18%,75.8±0.52%,76.0±2.40%,76.0±1.20%, The average was 75.9±1.60%in liver.At 0.02,0.1,2,20μg/mL concentration in plasma,the CV%within day were 0.10%,0.19%,0.08%,0.05%.the average was 0.10±0.06%.The between day CV% were 0.13%,0.20%,0.13%,0.10%.The average was 0.14±0.04%.At 0.1,0.5,10,100μg/mL concentration in tissue,the CV%within day were 0.08%,0.06%,0.03%,0.02%.The average was 0.04±0.03%.The between day CV%was 0.12%,0.13%,0.13%,0.06%.The average was 0.11±0.03%.At in liver,the CV%within day were 0.06%,0.08%,0.04%,0.03%,The average was 0.05±0.02%;The between day CV%were0.22%,0.15%,0.09%,0.11%,The average was 0.14±0.04%。The experiment demonstrated that HPLC was a sensitive and reliable method in analysis Ampicillin residue in swine.Ampicillin was injected into pigs of treated with 20mg/kg.i.m,The plasma,mules,and liver at 2,4,6,12,24,48,72hours were collected.With calibration linear,the Ampicillin was not found at 24hours in plasma.The Ampicillin was 0.06μg/g at 12hours in plasma;The Ampicillin was 0.02μg/g at 48hours in mules,but not found at 72hours;The Ampicillin was still 0.016μg/g at 72hours in liver. |
