Study On Spectinomycin And Lincomycin Residues In Poultry Tissues,poultry Eggs And Pig Muscle By Gas Chromatography Tandam Mass Spectrometry Detection Method

The purpose of this study was to establish a method for the simultaneous determination of spectinomycin and lincomycin residues in poultry tissues(chicken muscle,chicken kidney,chicken liver,duck muscle,goose muscle),poultry eggs(eggs,duck eggs,goose eggs,pigeon eggs and quail eggs)and pig muscle by gas chromatography tandam mass spectrometry(GC-MS/MS)detection method.In this experiment,Jinghai yellow chicken,Gaoyou duck,Yangzhou goose,domestic pigeon,domestic quail and Three-way cross hybrid pig(Duroc x Landrace x Large White)were selected as the experimental animals.Accelerated solvent extraction(ASE)technology was used to extract the targets.Development and optimization of GC-MS/MS method for simultaneous determination of spectinomycin and lincomycin residues in poultry tissue,poultry eggs and pig muscle.The main results of this study are shown as follows:1.The reaction conditions of the derivatization of spectinomycin and lincomycin and bis(trimethylsilyl)trifluoroacetamide(BSTFA)were optimized,that is,100 μL 1.0 ug/mL spectinomycin and lincomycin,nitrogen is blown to dry at 40℃.Removed 200μL BSTFA and 100 μL acetonitrile were put in a 10 mL centrifuge tube,sealed and reacted at 75℃ for 60 min in the oven,then,spectinomycin-TMS and lincomycin-TMS was produced.2.A method for simultaneous extraction of the spectinomycin and lincomycin residues in poultry tissues,poultry eggs and pig muscle by accelerated solvent extraction(ASE)instrument was established and optimized.that is,chicken tissues,eggs and pig muscle were degrease with n-hexane under 60℃ and 1500 psi conditions,then,the spectinomycin and lincomycin residue from the poultry tissues,poultry eggs and pig muscle was extracted with 0.01M potassium dihydrogen phosphate buffer(pH 4.0).The static extraction time is lasted for 5 minutes and extraction 2 times.This method of extraction has advantages of less consumption of extraction solvent and matrix effect,higher extraction efficiency and recoveries.3.A GC-MS/MS method was established,optimized,and validated,to determine spectinomycin and lincomycin residue levels in poultry tissues,poultry eggs and pig muscle,respectively.Electron ionization was adopted,with SCAN mode,and AutoSRM was used for qualitative or quantitative purpose.Within the dosing level of LOQ～600.0)μg/kg for spectinomycin in chicken muscle,duck muscle,goose muscle and pig muscle,the peak area of quantificational ion showed a good linear correlation with concentration(R2≥0.9992).Within the dosing level of LOQ～2000.0 μg/kg for spectinomycin in chicken liver,chicken kidney,and poultry eggs(whole egg,albumen,yolk),the peak area of quantificational ion showed a good linear correlation with concentration(R2≥0.9991).Within the dosing level of LOQ-1000.0 μg/kg for lincomycin in chicken liver,the peak area of quantificational ion showed a good linear correlation with concentration(R2was 0.9994).Within the dosing level of LOQ-3000.0μg/kg for lincomycin in chicken kidney,the peak area of quantificational ion showed a good linear correlation with concentration(R2 was 0.9995).Within the dosing level of LOQ-200.0 μg/kg for lincomycin in chicken muscle,duck muscle,goose muscle,and poultry eggs(whole egg,albumen,yolk),the peak area of quantificational ion showed a good linear correlation with concentration(R2>0.9992).When the added concentration of spectinomycin and lincomycin were the limit of quantification(LOQ),0.5 MRL,1.0 MRL and 2.0 MRL,the recoveries were 79.72%～94.23%and 78.86%～93.64%in the poultry tissues and pig muscle,the intra-day RSD were 2.29%～5.45%and 2.24%～4.89%,the inter-day RSD were 3.46%～7.76%and 2.55%～6.17%,the LOD were 2.5～4.4 μg/kg and 3.1～6.0 μg/kg(S/N≥3)and LOQ were 5.7～8.7 μg/kg and 6.2～10.0 pg/kg(S/N≥10),respectively.The recoveries in poultry eggs(whole egg,albumen,yolk)were 80.37%～95.72%and 80.01%～95.12%,respectively;the intra-day RSD were 2.03%～5.23%and 1.93%～5.99%,respectively;the inter-day RSD were 2.23%～6.67%and 3.05%～6.71%,respectively;the LOD were 2.3～4.0 μg/kg and 2.5-4.3 μg/kg(S/N≥3),respectively;the LOQ were 5.6-8.0 μg/kg and 5.9～9.5 μg/kg(S/N≥10),respectively.This method is of simplicity,high precision,and high sensitivity and the validation parameters of this method meet the drug residue requirements of the Ministry of Agriculture and village of China,EU and the United States.