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 cardiovascular system, liver, and kidney may occur. In this study, a method based on ultra high performance 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, arformoterol, cimaterol, and cimbuterol) in animal food sources. The sample pretreatment method and chromatographic conditions were optimized. The samples were hydrolyzed with β-glucuronidase 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 acetonitrile. NaCl was added to separate the organic and aqueous phases, and 5 mL of the upper organic layer was purified using a one-step purification solid-phase extraction column. After drying 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 positive-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 extraction column, purification method, and dissolved solution on the extraction efficiency were optimized during pretreatment. UHPLC-quadrupole time-of-flight MS was used to verify the purification 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 target 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 accurate 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.

利用超高效液相色谱-四极杆飞行时间质谱技术建立了生鲜牛乳中48种兽药残留的快速筛查方法.采用含0.2％甲酸的乙腈溶液作溶剂,Oasis PRIME HLB固相萃取载体作吸附剂,实现牛乳样品中48种目标物的同时提取和有效净化.目标药物经ACQUITY UPLC HSS T3 (21 mm×50 mm,1.8μm)色谱柱分离,以10 mmol/L甲酸铵-甲醇和0.1％甲酸水溶液为流动相进行梯度洗脱,使用电喷雾离子源,在正离子模式下进行全扫描,在9min内可实现48种兽药的良好分离.结果 表明:牛乳中48种兽药的定量限(LOQ,S/N=10)为5～20μg/kg在3个加标水平(10,20和50μg/kg)下进行6次重复试验,其平均回收率为68.4％～118.2％,相对标准偏差为3.1％～29.8％.该方法简便快速、灵敏度较高,可用于生鲜牛乳中48种兽药的快速筛查.

[目的]利用超高效液相色谱-四极杆飞行时间质谱技术建立虾中38种农药残留的快速筛查方法.[方法]采用50％乙腈水溶液作溶剂、QuEChERS提取盐包和Oasis PRiME HLB固相萃取载体作吸附剂,实现了虾肉样品中38种目标物的同时提取和有效净化.目标药物经ACQUITY UPLC HSS T3(1.8μm,21 mm×50 mm)色谱柱分离,以10 mmol/L乙酸铵-甲醇和0.1％甲酸水溶液为流动相进行梯度洗脱,使用电喷雾离子源,在正离子模式下进行全扫描,可在10 min内实现38种农药的良好分离.[结果]虾中38种农药的定量限(LOQ)为0.5～1.5μg/kg;3个加标水平的平均回收率为71.4％～97.9％,相对标准偏差(RSD)为4.1％～7.6％.[结论]该方法简便快速、灵敏度较高,可用于虾中38种农药的快速筛查.

文章旨在确定在不影响牛奶产量、牛奶成分、乳脂肪酸组成和牛奶感官质量的情况下,用不同水平甘油代替大麦对泌乳中后期奶牛的影响.试验选择了40头荷斯坦奶牛,采用4×4拉丁方试验设计.在部分混合日粮中,粗甘油替代大麦的含量分别为日粮干物质的0、6％、12％和18％.每天记录个体产奶量、采食量和挤奶频率,每周分析牛奶成分.对新鲜牛奶和贮藏7?d的牛奶样品进行感官分析.部分混合日粮的摄入量随甘油添加水平由0升高至12％,增加近1?kg,甘油由12％升高至18％,减少约1?kg.甘油水平由0升高至6％时,精料摄入量下降0.5?kg.随着日粮中甘油比例的增加,牛奶产量呈线性下降(P<0.05),而牛奶中的脂肪和蛋白质含量呈上升趋势(P=0.08),导致能量校正乳水平随甘油水平的升高表现为二次曲线效应,其中6％甘油组产量最高,18％甘油组产量最低.蛋白质和乳糖产量随日粮中甘油比例的增加线性下降(P<0.05).随着日粮甘油比例的增加,牛奶中棕榈酸、油酸、亚油酸、亚麻酸和共轭亚油酸的比例线性下降(P<0.05),而短链和中链脂肪酸的比例大部分呈上升趋势.综上所述,甘油作为一种能量饲料可以在不影响牛奶品质的前提下,在泌乳中后期奶牛日粮中替代大麦的水平高达18％.但日粮甘油水平超过12％时可能会降低能量校正乳的产量.