Determination Of Aminoglycoside Antibiotics And Dicyclanil In Feeds

Aminoglycosides antibiotic derived from a variety of Streptomyces and Microspora are a highly effective broad-spectrum antibiotic that can cause toxicity and nephrotoxicity.Dicyclanil is a novel insect growth regulator that has oxidative stress and carcinogenesis in rats.The quality and safety of feeds products are directly related to the safety of animal food.In order to monitor AGs and dicyclanil,high technology research is needed.A UPLC-MS/MS method for the determination of STP,DSTP,KAN,NEO,TOB,APR,GEN,HYG,AMI and SPC in the feeds was developed in this research.AGs were extracted with 10 mM potassium dihydrogen phosphate(containing 0.04 mM disodium edetate,2% trichloroacetic acid),ultrasonic extraction of the enzyme probe,the solution was centrifuged and the pH of the supernatant was adjusted at 7.5±0.1.The eluent was dreid under liquid nitrogen less than 1 mL,and add 10 ?L of heptafluorobutyric acid(HFBA).Distilled water were added and the final volume reached to 1 mL,detected by UPLC-MS/MS.The C8 column was used,gradient elution with 0.1% formic acid(containing 0.5 mM of ammonium acetate)-acetonitrile was adopted as the mobile phase.Following the methodological verification,all the AGs presented a good linear relationship within the range of 5 ?g/kg~1000 ?g/kg,the limit of detection was 5 ?g/kg~20 ?g/kg and the limit of quantification is 20 ?g/kg~80 ?g/kg.In the concentration range of 20 ?g/kg~400 ?g/kg,the recovery rate of 10 aminoglycoside antibiotics in feeds was 75.4~113.2%;the intra-day coefficient of variation was 1.1~10.1%,and the daytime coefficient of variation was 0.8~12.4%.This method was applied to the detection of actual samples,a certain amount of NEO,GEN and APR was detected in some concentrated materials and feed ingredients,and the detection rate was only 5.5%.A HPLC method for the determination of dicyclanil in the feeds was developed in this research,and confirmed by UPLC-MS/MS.Dicyclanil were extracted with acetonitrile.After centrifugation,the supernatant was purged with nitrogen to dryness and re-dissolved with 1 mL of acetonitrile for HPLC analysis,0.1% formic acid solution(98:2,v/v)was adopted as mobile phase and the wavelength used for detection was 270 nm,0.2 mg/kg and 1 mg/kg were taken as the limit of detection and quantification respectively.A good linear relationship was observed in the concentration range of 0.2 mg/L~10 mg/L.In the concentration range of 1 mg/kg~10 mg/kg,the recovery rate of the dicyclanil in the feed was 83.7~98.6%,the intra-day coefficient of variation was less than 4.6%,and the daytime coefficient of variation was less than 10.7%.Determination of dicyclanil by UPLC-MS/MS under the above sample preparation and quantification was carried out by ESI+ and MRM mode.This method was applied to the detection of actual samples,but no dicyclanil was detected.Ministry of Agriculture and the Rural Affairs reported the alarming level of the listed AGs and dicyclanil for feeds quality and safety.The method established here can provide theoretical basis for the detection of AGs and dicyclanil in the feeds.