Study On The Detection Method Of Veterinary Drug Residues In Sika Deer Velvet Antler

Cervi cornu pantotrichum is the young horn of Cervus nippon Temminck or Cervus elaphus Linnaeus,which is not ossified and densely hairy.It has the functions of strengthening kidney yang,benefiting essence and blood,strengthening muscles and bones,adjusting Chong and Ren,and supporting sore poison.In this paper,a rapid,sensitive,simple and reliable method for the detection of sulfonamides,hormones,cephalosporins,sedatives,tetracyclines and quinolones in Sika antler was established,which is of great significance to the health and safety of consumers and the development of deer industry.The first part:Using the pretreatment technology of QuECh ERS,an ultra high liquid chromatography-tandem mass spectrometry method was established for the simultaneous determination of 36 veterinary drugs such as sulfonamides,hormones,cephalosporins and sedatives in sika deer antler.Antler samples were extracted with acetonitrile and purified with 150 mg PSA,100 mg C₁₈8 and 70 mg neutral alumina.The UPLC analysis was carried out by a 35℃thermostatic Waters Acquity UPLC BEH C₁₈8 column（2.1 mm×100 mm,1.7μm）with a mobile phase composed of 25 mmol/L formic acid acetonitrile-12.5 mmol/L formic acid water for gradient elution at 0.3 m L/min.The detection was carried out on MS/MS with ESI⁺in positive ion in the multiple reaction monitoring mode.The content of 36 kinds of Veterinary Drugs in pilose antler was quantified by external standard method,The linear relationship of 36 veterinary drugs was good in the range of 0.1 ng/m L～50 ng/mL,the correlation coefficient R² were all greater than 0.993,the LOD was 0.08～1μg/kg,and the LOQ was 0.3～3μg/kg.36 kinds of veterinary drugs with concentrations of 2,5 and 10μg/kg were added to the blank sample,the average recovery was 77.8%～107.6%,and the relative standard deviation was less than 10%.The average recovery of 36 kinds of Veterinary Drugs was 77.8%～107.6%with RSD less than10%.The second part:a method for the simultaneous determination of tetracycline and quinolones in antler of Sika Deer by ultra-high liquid chromatography tandem mass spectrometry was established.The antler samples were purified with 0.1 mol/L Na₂EDTA mellvaine buffer and 50 mg PSA and HLB.The contents of tetracycline and quinolones in antler were determined by external standard method.15kinds of veterinary drugs have good linear relationship in the range of 0.1 ng/m L～50 ng/mL,the correlation coefficient R² is greater than 0.992,LOD is 0.5μg/kg～1μg/kg,LOQ is 1μg/kg～3μg/kg.15 kinds of veterinary drugs with concentrations of 5,10 and 20μg/kg were added to the blank sample.The average recovery rate of 15 kinds of veterinary drugs was 69.6%～82.4%,RSD was 12.8%～16.8%,and the relative standard deviation was less than 20%.4.Innovation（1）A method for the simultaneous determination of 36 kinds of Veterinary Drugs in antler velvet of Cervus by UPLC-MS/MS was developed by using the pretreatment technology of dispersed solid phase extraction.（2）A method for the simultaneous determination of 15 kinds of veterinary drugs,such as tetracyclines and quinolones,in antler of Sika Deer by ultra-high liquid chromatography tandem mass spectrometry was established by using the pretreatment technology of PSA and HLB.