色谱2023，Vol.41Issue(12)：1106-1114,9.DOI:10.3724/SP.J.1123.2023.03008
超高效液相色谱-串联质谱测定畜肉中14种β-受体激动剂
Determination of 14 β-agonists in animal meat by ultra high performance liquid chromatography-tandem mass spectrometry
摘要
Abstract
The addition of β-agonists to animal feed can significantly improve the lean-meat rate of pigs,cattle,sheep,and other animals.However,the food residues of β-agonists are harmful to human health.When meat with β-agonist residues is consumed,poisoning symptoms such as palpitation,dizziness,and muscle tremors may develop,and damage to the cardiovas-cular system,liver,and kidney may occur.In this study,a method based on ultra high per-formance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)was established for the rapid detection of 14 β-agonists(clenbuterol,salbutamol,ractopamine,clorprenaline,terbutaline,tulobuterol,bromobuterol,bambuterol,zilpaterol,mabuterol,fenoterol,arfor-moterol,cimaterol,and cimbuterol)in animal food sources.The sample pretreatment method and chromatographic conditions were optimized.The samples were hydrolyzed with β-glucu-ronidase hydrochloride/aryl sulfate esterase in ammonium acetate buffer(pH 5.2).Enzymatic hydrolysis was performed in a constant-temperature water bath((36±2)℃)oscillator for 16 h.The samples were cooled to room temperature and extracted with 0.5%formic acid acetoni-trile.NaCl was added to separate the organic and aqueous phases,and 5 mL of the upper or-ganic layer was purified using a one-step purification solid-phase extraction column.After dr-ying with nitrogen at 50℃,the residue was dissolved in 0.4 mL of 0.2%formic acid aqueous solution.The samples were passed through a 0.22 μm filter and detected by UHPLC-MS/MS with gradient elution using acetonitrile and 0.1%formic acid aqueous solution as the mobile phases.The analytes were separated on a Phenomenex Kinetex F5 column and detected by pos-itive-ion scanning in multiple-reaction monitoring(MRM)mode.Internal and external standard methods were used for quantitative analysis.The effects of the extract pH,solid-phase extrac-tion column,purification method,and dissolved solution on the extraction efficiency were opti-mized during pretreatment.UHPLC-quadrupole time-of-flight MS was used to verify the purifica-tion effect of the one-step purification solid-phase extraction column,and the results indicated that this type of column could remove most of the phospholipids,sphingolipids,and glycerides in the sample extract.The factors influencing the different chromatographic columns and mobile phases were investigated.MS scanning was conducted in positive-ion mode with needle pump injection in mass-only mode,and the two daughter ions with the highest responses for each tar-get were selected as the quantitative and qualitative ions.The declustering potential(DP)and collision energy(CE)of each ion were separately optimized in MRM mode.The switching mode of the mass spectrum and waste liquid was used,and the mobile phase was switched to waste liquid after all the target peaks were removed.These steps ensured that impurities in the sample flowed out of the column in a timely manner and that the effects of excessive impurities on the mass spectra were avoided.The 14 β-agonists showed good linear relationships in the range of 1.0-50 μg/L,with correlation coefficients of＞0.99.The limits of detection(LODs)and quantification(LOQs)were in the range of 0.1-0.2 and 0.3-0.6 μg/kg,respectively.The average recoveries of the 14 β-agonists ranged from 70.25%to 117.48%,with relative standard deviations(RSDs)in the range of 0.63%-14.29%at low,medium,and high spiked levels.Pork,beef,and mutton samples were selected and analyzed using the developed method.The results were close to those of the national standard method,indicating that the method is accu-rate and reliable.Moreover,the proposed method has good stability and high accuracy;thus,it is suitable for the qualitative and quantitative determination of β-agonists in animal meat.关键词
超高效液相色谱-串联质谱/一步式净化固相萃取柱/β-受体激动剂/畜肉Key words
ultra high performance liquid chromatography-tandem mass spectrometry(UH-PLC-MS/MS)/one-step purification solid-phase column/β-agonist/animal meat分类
化学化工引用本文复制引用
董洁琼,肖琎,周鑫,李宁,王雪松,康俊杰..超高效液相色谱-串联质谱测定畜肉中14种β-受体激动剂[J].色谱,2023,41(12):1106-1114,9.基金项目
唐山市科技计划项目(21150220A,23150208A) （21150220A,23150208A）
唐山市科技创新领军人才(21130243A). Tangshan Science and Technology Planning Project(Nos.21150220A,23150208A) （21130243A）
Tangshan City Science and Technology Innovation Leader(No.21130243A). （No.21130243A）
