超高效液相色谱-串联质谱法对饲料中15种β-肾上腺素能激动剂的测定

动物源性食品中兽药残留是近几年来国际社会开始研究的公共卫生问题之一.世界各国加强了对国际贸易动物源性食品中兽药残留的检测,而开展饲料及饲料添加剂中兽药残留的检测是从源头进行控制的有效手段[1].     

超高效液相色谱-电喷雾串联四极杆质谱法同时测定配合饲料中29种兽药

建立了同时测定配合饲料中喹诺酮类、磺胺类、大环内酯类和硝基呋喃类共计29种兽药的超高效液相色谱-电喷雾串联四极杆质谱(UPLC-ESI-MS/MS)检测方法。饲料样品用甲醇-乙腈(1:1, v/v)混合溶液提取,提取液经Oasis HLB固相萃取柱净化,采用UPLC-ESI-MS/MS检测。以甲醇和含0.1%(v/v)甲酸的水溶液作为流动相,进行梯度洗脱,用C18色谱柱分离,正离子模式扫描,多反应监测模式检测。29种兽药在0.01～5.0 mg/L范围内线性关系良好,相关系数(r)均大于0.99;在复合预混饲料、全价配合饲料、浓缩饲料中4个添加水平下29种兽药的平均回收率在61.2%～94.3%范围内,相对标准偏差(RSD)为2.2%～15.0%;方法的检出限(以信噪比大于10计)为0.01 mg/kg或0.05 mg/kg。该方法简便、快速、准确,重现性好,灵敏度高,适用于配合饲料中多种兽药的同时检测。     

用超高效液相色谱-串联质谱法同时检测浓缩饲料中的多种兽药残留

建立了浓缩饲料中6种喹诺酮类兽药、14种磺胺类兽药、3种硝基呋喃类兽药、6种大环内酯类兽药共29种兽药残留检测的超高效液相色谱-串联质谱分析方法。样品经甲醇-乙腈(体积比1∶1)混合溶液提取,提取液经Oasis HLB固相萃取柱净化,29种兽药经Waters Acquity UPLC BEH C18色谱柱分离,以乙腈和0.1%甲酸水溶液为流动相进行梯度洗脱。电喷雾正离子(ESI+)模式电离,多反应监测(MRM)模式检测,内标法定量。结果表明,29种兽药在0.01～5.0mg/mL范围内呈良好的线性关系,相关系数(r)均大于0.993;浓缩饲料中3个浓度添加水平平均回收率为61.4%～93.3%;相对标准偏差为2.62%～13.9%。     

鸡肉中兽药残留检测技术研究进展

鸡肉中兽药残留检测是食品安全检测关注的热点,作为目前普遍发展的样品前处理技术,超临界流体萃取、加速溶剂萃取、毛细管固相微萃取及其与色谱质谱联用检测兽药残留受到广泛关注。以鸡肉为主要检测基质对这些新技术的基本原理、特点及在兽药残留分析中的应用进行了综述,同时对兽药残留分析的研究方向进行了展望。     

高效液相色谱法测定水产品中喹乙醇残留量

喹乙醇又称喹酰胺醇,商品名称为快育灵、倍育诺,是一种饲料添加剂,具有良好的广谱抗菌效果,并且有提高畜禽饲料消化率、提高生长速率等作用,被广泛用于兽药及饲料添加剂中。     

超高效液相色谱-串联质谱法同时检测复合预混饲料中29种兽药

建立了复合预混饲料中29种兽药同时检测的超高效液相色谱-串联质谱分析方法。样品经甲醇-乙腈(1:1,V/V)混合溶液提取,提取液经Oasis HLB固相萃取柱净化,29种兽药经Waters Acquity UPLC BEH C18色谱柱分离,以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱。电喷雾正离子(ESI+)模式电离,多反应监测模式检测,内标法定量。结果表明:29种兽药在0.01～5.0μg/mL范围内呈良好的线性关系,相关系数(r)均大于0.993;复合预混饲料中3个浓度添加水平平均回收率为65.5%～92.2%;相对标准偏差为3.8%～15%。     

市售虾中磺胺甲噁唑的LC-MS-MS定量分析

当前各国对食品安全都极为重视,我国以及发达国家相继制定了相关兽药用量的法规和标准,尽管中国已经加入到了WTO,但我国的畜禽产品出口还是遇到了很多困难,农药、兽药、饲料添加剂等非法滥用,造成药物残留而直接影响到人类的健康。如引发癌症,婴儿畸形,代谢紊乱以及抗药性等诸多问题。其中磺胺类兽药是一类受到严格限制使用的兽药,本文研究了市售虾中磺胺甲(口恶)唑定量分析方法,检出限绝对量可达到0.5pg,完全满足发达国家的最低检出规定的苛刻标准。     

液相色谱-四极杆飞行时间质谱结合化学计量学方法筛查饲料中的未知药物

建立了液相色谱-四极杆飞行时间质谱(LC-Q-TOF/MS)对饲料中非目标污染物(兽药)的筛查方法,以及未知物的判定和确认技术。饲料样品经0.1%EDTA-乙腈-甲醇(1∶1∶1,含0.1%甲酸)混合溶液提取,Q-TOF全扫描模式采集数据,数据经过峰对齐(质量误差5 m Da以内)、噪音过滤、自动排除无用峰等处理后,进行正交偏最小二乘判别分析(OPLS-DA)处理,根据Score图、S-Plot图和VIP图判定出特征离子,结合二级信息与自建谱库比对,进行结构式确认,或通过同位素丰度比例和分子式生成功能,确认分子式,结合二级断裂信息,判定特征离子的结构式,从而判定未知物的成分。结果显示:16份饲料样品共筛查出27种兽药。采用液相色谱-三重四极杆串联质谱法对其中的12种兽药进行定性定量分析,验证结果仅出现2个假阳性,兽药含量在12.5~152.0 mg/kg之间。通过LC-Q-TOF/MS结合OPLS-DA的方法能够对饲料中的非目标物进行筛查分析,方法准确度高。     

纳米金辅助介质阻挡放电离子化质谱分析法在兽药饲料快检中的应用

介质阻挡放电离子化质谱分析法具有无需复杂样品前处理、分析速度快等优点,可实现样品原位、高效检测,成为质谱分析领域的热点技术。本研究提出了一种基于纳米金辅助纸基进样方式的介质阻挡放电离子源-质谱(DBDI-MS)联用技术,实现了氟甲喹、二甲氧苄啶、替硝唑、地美硝唑共4种兽药的快速检测。结果表明,在氦气流量为2.5 L/min、离子源温度为200℃等最优实验条件下,4种兽药检测信号强度均比无纳米金辅助时提高了8~31倍。4种兽药样品在各自的浓度范围内有较好的线性关系,相关系数(r)为0.952~0.996,且在饲料基质中的加标回收率为79.4%~96.5%。本方法能快速检测兽药饲料,具有灵敏度高、检测费用低、实用性好等优点,所得结果基本满足农业部限定标准要求,适合现场检测,在饲料质量控制方面具有较好的应用前景。     

基于色谱-质谱法的肉类食品中大环内酯类药物残留及其代谢产物分析方法研究进展

大环内酯类药物是食品安全检测关注的热点,该文论述了大环内酯类兽药分析检测的研究背景和现状,介绍了大环内酯类药物及其代谢产物在畜禽类动物体内的分布部位,并对国内外肉类食品中大环内酯类兽药残留的前处理方法及检测方法进行综述,同时对大环内酯类药物及其代谢产物残留分析的研究方向进行了展望。     

饲料中激素类药物及其检测技术研究进展

综述了饲料中常用激素类药物的现状及分析技术。对激素类药物的种类、化学特性,应用情况及危害性进行了分析,对饲料中激素类药物残留的前处理和检测方法进行了介绍。重点研究关注了激素类药物和分析方法的异同,从而了解和掌握该领域发展动态,为建立健全相关的限量标准和检测方法提供参考。     

Nontargeted screening of veterinary drugs and their metabolites in milk based on mass defect filtering using liquid chromatography–high-resolution mass spectrometry

The development of nontargeted screening strategy for veterinary drugs and their metabolites is very important for food safety. In this study, a nontargeted screening strategy was developed to find the potentially hazardous substances based on mass defect filtering (MDF) using liquid chromatography–high-resolution mass spectrometry. First, the drug metabolites of 112 veterinary drugs from seven classes of antimicrobials were predicted. Second, three MDF models were established, including the traditional rectangular MDF, the enhanced parallelogram MDF, and the polygonal MDF. Finally, the strategy was applied to nontargeted screening of veterinary drugs in 36 milk samples. The polygonal MDF model based on the distribution area of parent drugs and their metabolites showed a better filtering effect. After removing food components and performing MDF, about 10% of the substances remained, and four veterinary drugs and six drug metabolites were discovered and identified, showing the effectiveness of this strategy. The nontargeted screening strategy can rapidly remove interfering substances and find the suspected compounds. It can also be used for nontargeted screening of veterinary drugs and their metabolites in other food matrices.     

悬浮芯片法与常规酶联免疫吸附法检测3种兽药残留的比较

建立氯霉素,克伦特罗和雌二醇 3种兽药残留同时检测的悬浮芯片法.通过将3种兽药的BSA蛋白结合物偶联于悬浮芯片的固相载体--聚苯乙烯荧光微球上作为检测探针,采用间接竞争法,在液相反应体系中,3种小分子兽药抗原和微球上的兽药结合物共同竞争液相中各自特异性的生物素化单抗,再加入藻红蛋白标记的链霉亲和素,反应后检测获得荧光信号,绘制出3种兽药残留检测的标准曲线.同时进行3种兽药的常规酶联免疫吸附法标准曲线的测定.在检测技术、检出限、检测区间、特异性、盲样测定和多元分析等方面对两种方法进行比较.除了特异性外的其它指标的比较中,悬浮芯片法均具有明显优势.两种方法的特异性检测具有良好的一致性.高通量悬浮芯片技术,具有操作简单、灵敏快速和成本低廉等优点,为多种兽药残留的快速检测提供了新方法.     

QuEChERS-超高效液相色谱-串联质谱法测定鸡蛋中125种兽药残留

采用冷冻脂质过滤,结合分散固相萃取净化的QuEChERS方法,建立了超高效液相色谱-串联质谱法测定鸡蛋中125种兽药残留的检测方法。样品中的11类兽药（硝基咪唑类、苯并咪唑类、磺胺类、喹诺酮类、四环素类、大环内酯类、雄激素类、孕激素类、糖皮质醇类、雌激素类、氯霉素类）用70%（v/v）乙腈-水溶液（含0.1 mol/L EDTA）提取后,提取液在-20℃冷冻处理2 h,再经分散固相萃取法净化,净化液经稀释后,以超高效液相色谱-串联质谱法测定,外标法定量。结果显示125种兽药的线性相关系数R²≥0.99,定量限范围为2.0~60 μg/kg,回收率在60.4%~119.3%之间,相对标准偏差在0.3%~16.1%之间。该方法前处理简单、准确、成本较低,适用于鸡蛋中兽药残留的高通量快速检测分析。     

Development of fast screening methods for the analysis of veterinary drug residues in milk by liquid chromatography-triple quadrupole mass spectrometry

Two rapid multi-residue screening methods for the determination of 21 veterinary drugs in milk by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) have been developed and compared. For both methods, veterinary drugs were extracted from milk samples using a rapid extraction procedure based on the modification of the well-known QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method, and no further clean-up steps were necessary. One screening method was based on the selection of a characteristic neutral loss or product ion of the various families of compounds, whereas another one was based on the choice of a selected reaction monitoring (SRM) for each compound. These methods were compared with regards to false negatives, cut-off values and the unreliability region. The total run time for both methods was 3 min, allowing quick selection of samples that contained veterinary drugs. Non-negative samples were re-analyzed by the UHPLC-MS/MS confirmation/quantification method, which consisted in the monitoring of two SRM for each compound. The methods were validated according to international guides. The proposed analytical methods allow for the identification and confirmation of the target veterinary drugs at trace levels employing quick analysis time.     

Multiclass method for the quantification of 92 veterinary antimicrobial drugs in livestock excreta,wastewater, and surface water by liquid chromatography with tandem mass spectrometry

A simple multiresidue method was developed for detecting and quantifying 92 veterinary antimicrobial drugs from eight classes (β‐lactams, quinolones, sulfonamides, tetracyclines, lincomycins, macrolides, chloramphenicols, and pleuromutilin) in livestock excreta and water by liquid chromatography with tandem mass spectrometry. The feces samples were extracted by ultrasound‐assisted extraction with a mixture of acetonitrile/water (80:20, v/v) and edetate disodium, followed by a cleanup using solid‐phase extraction with an amino cartridge. Water samples were purified with hydrophilic–lipophilic balance solid‐phase extraction column. Urine samples were extracted with acetonitrile and edetate disodium. Detection of veterinary antimicrobial drugs was achieved by liquid chromatography with tandem mass spectrometry using both positive and negative electrospray ionization mode. The recovery values of veterinary antimicrobial drugs in feces, urine, and water samples were 75–99, 85–110, and 85–101% and associated relative standard deviations were less than 15, 10, and 8%, respectively. The limits of quantification in feces, urine, and water samples were 0.5–1, 0.5–1, and 0.01–0.05 μg/L, respectively. This method was applied to determine real samples obtained from local farms and provides reliable quantification and identification results of 92 veterinary antimicrobial drugs in livestock excreta and water.     

Simultaneous Screening for Five Veterinary Drug Residues by a Developed Multiplexed Luminex Assay

Abstract

The development of sensitive and rapid methods for simultaneous determination of multiple residual veterinary drugs in foods is increasingly important. In this study we reported a new method to simultaneously detect five veterinary drug residues, including ractopamine, clenbuterol, furazolidone, chlortetracycline and qlaquindox. The experimental results indicated that the developed multiplex assay enable to detect all the five drugs at the concentration of 0.1 ng/mL, with desirable specificity, suggesting that it could be applicable for rapid screening of five residual veterinary drugs in a faster turnaround time and higher throughput.     

QuEChERS-超高效液相色谱-串联质谱法测定动物源食品中4类29种禁限用兽药残留

采用改进的QuEChERS方法提取和净化鸡肉、鸡肝、猪肉、猪肝样品,建立了4类29种禁限用兽药残留的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。样品采用McIlvaine缓冲液-乙腈提取,提取液酸化、盐析后取乙腈相,用十八烷基硅烷(C18)和氨基(NH₂)吸附剂分散固相萃取净化,最后用电喷雾串联质谱仪在正离子多反应监测扫描(MRM)模式下进行测定,基质外标法定量。解决了四环素类兽药难与其他类兽药同时检测的难题。29种兽药在各自的浓度范围内,浓度与峰面积线性关系良好,相关系数不小于0.993。在29种兽药的定量限(LOQ)水平、20 μg/kg和50 μg/kg 3个添加水平下进行加标回收率试验,29种兽药的平均回收率在71.5%和93.2%之间,相对标准偏差在0.8%和7.7%之间。29种兽药的检出限为0.3~3.0 μg/kg,金刚烷胺的定量限为1.0 μg/kg,四环素类兽药的定量限为10.0 μg/kg,其他兽药的定量限均为5.0 μg/kg。该方法操作简便,净化效果好,灵敏度、准确度和精密度均符合多残留检测技术的要求,可为食品检验机构对动物组织中禁限用兽药残留的日常监控提供更方便、快捷的检测方法支持。