肉中的兽药和瘦肉精残留及其检测技术研究进展

为提高猪肉中多兽药残留检测的效率,建立了QuEChERS结合超高效液相色谱-串联质谱（UPLC-MS/MS）同时测定猪肉中5类76种兽药及其代谢物残留量的方法,包括喹诺酮类、磺胺类、硝基咪唑类、大环内酯类、苯并咪唑类。样品经酸化乙腈提取,无水硫酸钠盐析,提取液经C₁₈和无水硫酸镁净化后分为两部分,一部分用于22种弱极性化合物的直接测定,另一部分氮吹浓缩后复溶,用于54种强极性化合物的测定。76种化合物均采用电喷雾电离正离子模式多反应监测（MRM）,基质匹配外标法定量分析。结果表明,76种兽药及其代谢物线性关系良好,相关系数均大于0.995,方法定量限均低于10 μg/kg,加标回收率为61.3%~96.2%,相对标准偏差（RSD）为2.0%~14.6%。方法应用于170批市售猪肉,其中23批次样品检出2类5种兽药残留。该方法简便快速、准确可靠,适用于猪肉中多类兽药残留的快速筛查和精准检测。     

超高效液相色谱-串联质谱法测定动物源性食品中多种肾上腺素类药物残留

目的 建立一种超高效液相色谱-串联质谱法同时测定动物源性食品中肾上腺素类、拟肾上腺素类、肾上腺素代谢物等10种肾上腺素类药的方法。方法 乙腈提取,分散固相萃取法（QuEChERS）净化,Shim-pack GIST C18色谱柱,甲醇和含0.1%甲酸的5 mmol/L乙酸铵水溶液作为流动相,UPLC-MS/MS确证检测,外标法定量。结果 10种肾上腺素类物质在0.1~200 ng/mL线性范围内关系良好,相关系数均高于0.9946, 方法定量限为0.2~2.0 μg/kg,回收率75.6%~107.5%,相对标准偏差1.1%~8.7%。结论 该方法具有检测药物种类多、最低检测限低、稳定性好、简便快捷、节省试剂等优点,为更好的控制市场上动物源性食品的品质提供的技术支持支撑。     

Multi-residue method for the analysis of pesticides in Arabica coffee using liquid chromatography/tandem mass spectrometry

Coffee is a major tropical agricultural commodity and represents a significant fraction of the economy of many countries. However, certain plant and animal species can damage coffee crops, affecting trade. A solution to this issue is the use of pesticides, some of which are harmful to human health and the environment. This work consisted of the development of a multi-residue method for the analysis of pesticides in coffee by using LC-MS/MS. The QuEChERS extraction procedure was used. The following analytical parameters were optimised: selectivity, analytical range, linearity, LOD, LOQ, precision (RSD%) and recovery of the method. The results showed that the method is selective, as they were linear in the range of 10.0–100.0 µg kg^?1. The sensitivity, recovery and precision were adequate for the multi-residue analysis of pesticides in coffee. The method was applied to the analyses of 15 Brazilian coffee samples.     

通过式净化-液相色谱-串联质谱快速筛查水产品中15类107种兽药多残留的方法研究

目的 建立水产品基质中15类包含107种药物多残留的快速筛查方法。方法 通过多溶剂分步提取-通过式净化, 结合液相色谱-串联质谱仪, 在MRM模式下正、负离子同时扫描监测水产品, 实现水产品中15类107种药物多残留的快速筛查, 并用于实际水产样品中药物残留含量的测定。结果 所有药物在35 min内分离良好, 方法线性范围为0.1~200 ng/mL, 相关系数均大于0.995; 检出限为0.1~5.0 μg/kg (S/N>3), 定量限为0.3~10 μg/kg (S/N>10); 回收率为40.0%~120%, 相对标准偏差均小于15%。结论 方法的准确性和灵敏度较高, 重现性好, 适合水产基质中药物多残留的高通量、快速筛查, 能够满足国家限量法规的要求。     

超高效液相色谱-串联质谱法检测动物组织中10种抗胆碱类药物残留方法建立

采用超高效液相色谱-串联质谱建立动物组织中丙哌维林、贝那替嗪、哌仑西平、美卡拉明等10种抗胆碱类药物残留检测方法。样品经过1%甲酸乙腈提取、Oasis PRiME HLB柱净化后,经Acquity UPLC BEH C₁₈(50 mm×2.1 mm,1.7 μm)分离,以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱,正离子MRM信号采集数据,10种抗胆碱类药物能在10.5 min内完好分离,在1.0~50.0 μg/L浓度范围内,10种抗胆碱类药物线性良好,相关系数均在0.9951以上;最低定量限均低于1.0 μg/kg,通过1.0、2.0、10 μg/kg三个水平的添加回收实验表明,猪肉中平均回收率在64.7%~108.2%,批内变异系数为1.09%~10.5%,批间变异系数为2.31%~11.4%,猪肝中平均回收率在62.6%~108.0%,批内变异系数为0.38%~10.7%,批间变异系数为2.24%~11.2%。该方法较好的满足了动物组织中该药物多残留检测要求,对有效防范药物滥用和非法添加行为和打击注水肉起到了一定的技术支撑作用。     

微量QuEChERS-超高效液相色谱-串联质谱法检测凉茶中59种非法添加化学药物

目的：建立一种微量QuEChERS（micro-QuEChERS）结合超高效液相色谱-串联质谱（UPLC-MS /MS）同时检测凉茶中59种非法添加药物的分析方法。方法：样品用乙腈超声提取5 min,提取液用NaCl及无水Na₂SO₄进行盐析分层,上清液用C₁₈吸附剂进行净化后,经ACQUITY HSS T3（100 mm×2.1 mm,1.8 μm）色谱柱分离,以0.01%甲酸水溶液（A）-乙腈:甲醇（8:2）（B）为流动相进行梯度洗脱,并采用电喷雾正负离子多反应监测模式检测,空白基质外标法定量。结果：59种化合物在其线性浓度范围内,线性关系良好,决定系数均大于0.999,检出限为5.0～10.0 μg/L,定量限为10.0~25.0 μg/L。在25.0、50.0、100.0 μg/L 3个添加水平下,平均回收率为60.3%～128.8%（n=6）,相对标准偏差（RSD）1.0%～13.7%。结论：该方法操作简单,净化效果好,灵敏度高,可用于凉茶中59种非法添加化学药物的快速分析。     

Multi-residue and multi-class determination of antibiotics in gilthead sea bream (Sparus aurata) by ultra high-performance liquid chromatography-tandem mass spectrometry

This paper describes a method for the determination of 41 antibiotics from seven different classes in gilthead sea bream (Sparus aurata) by ultra-high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). Sulfonamides, trimethoprim, tetracyclines, macrolides, quinolones, penicillins and chloramphenicol were simultaneously determined. Fourteen procedures for sample treatment were tested and an extraction with acetonitrile and ethylenediaminetetra acetic acid was found to be the best option. The methodology was validated in accordance with Decision 2002/657/EC. Precision in terms of relative standard deviation (RSD) was under 17% for all compounds, and the recoveries ranged from 92% to 111%. CCα and CCβ were determined according to the maximum residue limit or the minimum required performance limit, when necessary. The validation provided evidence that the method was suitable for application in routine analysis for the detection and confirmation of antibiotics in muscle of gilthead sea bream, an important and intensively produced fish in aquaculture.     

超高效液相色谱-串联质谱法测定保健食品中78种非法添加化学药物

建立了超高效液相色谱-串联质谱法快速筛查减肥、抗疲劳和增强免疫力类保健食品中非法添加78种化学药物的方法.保健食品样品经曱醇溶剂提取、QuEChERS净化,试样以C18柱分离,多反应监测(MRM)模式进行定量与定性分析,采用外标法定量.采用ACQUITYUPLCHSST3柱(2.1×100mm,1.7μm)进行分离,以...     

超高效液相色谱-串联质谱法检测猪肉中多兽药残留

建立一种超高效液相色谱-串联质谱(ultra performance liquid chromatography/tandem mass spectrometry,UPLC-MS/MS)法同时检测猪肉中β-受体激动剂类、磺胺类、砜类抑制剂类、喹诺酮类、糖皮质激素类以及大环内酯类等20种兽药残留。待测药物使用80%体积分数的乙腈水溶液提取,经固相萃取小柱净化和氮气吹干后使用体积分数10%甲醇水进行复溶,复溶液经微孔滤膜过滤后进行UPLC-MS/MS测定。使用BEH C_(18)色谱柱梯度洗脱分离,选用电喷雾电离源,正离子扫描及多反应监测模式测定。20种药物检出限(limit of detection,LOD)在0.5～5μg/kg之间,定量限(limit of quantitation,LOQ)在1.7～16.7μg/kg之间,当加标水平为1、5、10μg/kg时,平均回收率在62.1%～118.3%范围内,相对标准偏差(relative standard deviation,RSD)在0.4%～16.7%范围内。     

Multi-residue method for detecting coccidiostats at carry-over level in feed by HPLC–MS/MS

A multi-residue HPLC–ESI–MS/MS method has been developed for the simultaneous extraction, detection and confirmation of the 11 coccidiostats referenced by Regulation 2009/8/EC (lasalocid sodium, narasin, salinomycin sodium, monensin sodium, semduramicin sodium, maduramicin ammonium alpha, robenidine hydrochloride, decoquinate, halofuginone hydrobromide, nicarbazin, and diclazuril) in feedstuffs at carry-over level. The sensitivity of the method allows quantification and confirmation for all coccidiostats below target concentration. The method was in-house validated and meets all criteria of European legislation (2002/657/EC). The precision of the method was determined under repeatability and within-laboratory reproducibility conditions; RSD_r and RSD_R were below the maximum permitted values for every tested concentration. The specificity was checked by analysing representative blank samples and blank samples fortified with potentially interfering substances (benzimidazoles, corticosteroides, triphenylmethane dyes, quinolones, nitrofurans, nitroimidazoles, phenicols); no interference were found. Concerning quantification, a quadratic regression model was fitted to every calibration curve with a regression coefficient r ² above 0.99 on each data set. Finally, the expanded uncertainty U was calculated with data obtained within the laboratory while applying the method during validation and in routine tests.     

QuEChERS-UPLC-MS/MS法检测粮食中的黄曲霉毒素 （网络首发、推荐阅读）

构建QuEChERS-超高压液相色谱串联质谱(UPLC-MS/MS)检测粮食中6种黄曲霉毒素（B1、B2、G1、G2、M1、M2）的方法。样品经过乙腈-0.1%甲酸溶液（84∶16）提取后,经N-丙基乙二胺（PSA）净化,以0.1%甲酸-乙腈作为流动相,采用UPLC-MS/MS在多反应检测模式下进行测定,外标法定量。结果显示：6种毒素标准曲线相关系数均大于0.99,线性良好,加标回收率为80.83%～117.5%,相对标准偏差（RSD）为1.53%～15.59%。在不同粮食基质中,6种黄曲霉毒素的基质效应为0.81～1.12。该方法的准确度和精确度符合相关标准要求,具有前处理技术简单、净化效果好、精确度高等优势,适合小麦、大米、玉米、绿豆和红豆样品中6种黄曲霉毒素的定量分析。     

Deposition and depletion of maduramicin residues in eggs after oral administration to laying hens determined by LC-MS

The coccidiostat maduramicin has been approved as a feed additive for chickens and turkeys, although it is prohibited for use in laying hens. In the present study, laying hens were divided into three groups and fed for 14 days with medicated feed containing maduramicin, at three different concentrations: 50, 100 and 500 µg kg^?1. Eggs were collected during treatment and for 26 days after the end of feeding with medicated feed. Maduramicin residues were found exclusively in egg yolk, with the highest concentration in egg yolk of 459 µg kg^?1 for the highest dose. The maximum concentration of maduramicin in whole egg was 16.6 µg kg^?1 for the group receiving feed containing the maximum permitted level of maduramicin in feed (50 µg kg^?1). The half-life of elimination of maduramicin, calculated for post-treatment days 1–10, was 6.5 days. Twelve days after drug administration, the concentration of the maduramicin in egg yolk for Group 3 (fed with 500 µg kg^?1 maduramicin) still exceeded 20 µg kg^?1, while the concentrations for Groups 1 and 2 were 1.2 and 2.7 µg kg^?1, respectively.     

QuEChERS结合超高效液相色谱-串联质谱法同时测定梅花鹿鹿茸中18种磺胺

该文采用分散固相萃取的前处理技术,建立超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)法同时测定梅花鹿鹿茸中18种磺胺的检测方法。以乙腈为提取剂,70 mg PSA (乙二胺基-N-丙基)、100 mg C_(18)(十八烷基硅烷)与50 mg的中性氧化铝为净化剂,液相色谱条件:Acquity UPLC BEH柱(2.1 mm×100 mm,1.7μm),流动相为25 mmol/L甲酸乙腈-水,柱温35℃,流速0.3 mL/min;质谱条件:正离子模式(ESI+),多反应监测(multiple reaction monitoring,MRM)模式。鹿茸中磺胺含量采用外标法定量,结果18种磺胺在0.025～50 ng/mL内线性关系良好,相关系数R~2均大于0.999 1,7种磺胺的LOD(limits of detection)为0.025μg/kg,LOQ(limits of quantitation)为0.075μg/kg,11种磺胺的LOD为0.075μg/kg,LOQ为0.25μg/kg。18种磺胺的平均加样回收率为80.4%～107.2%,相对标准偏差(relative standard deviation, RSD)均小于5%。该方法具有操作简便,净化效果显著,灵敏度高、准确性好、检出限低等优点,可用于梅花鹿鹿茸中18种磺胺的检测。     

咖啡中3种赭曲霉毒素QuEChERS-UPLC-MS/MS检测方法

目的:建立同时测定咖啡中3种赭曲霉毒素的QuEChERS-超高效液相色谱—串联质谱检测方法。方法:样品经乙腈—水—甲酸(V_乙腈∶V_水∶V_甲酸为55∶40∶5)超声提取,利用QuEChERS盐包进行脱水盐析,过ZanChERS-Myco17净化小柱净化。样品采用0.2%甲酸水—乙腈作为流动相经Waters BEH C₁₈(1.7μm, 2.1 mm×100 mm)色谱柱梯度洗脱分离。电喷雾正离子模式,多反应监测扫描,内标法定量。结果:3种目标物在0.1～20.0 ng/mL的质量浓度范围内线性关系良好,相关系数≥0.999 46,方法检出限和定量限分别为0.1～0.2,0.2～0.7μg/kg。咖啡基质中3个添加水平目标物平均回收率为79.0%～103.3%,相对标准偏差为1.5%～7.6%。结论:该方法前处理简单,重现性好,分析时间短,能够适用于不同咖啡基质样品中3种赭曲霉毒素残留的高通量检测。     

Detection and quantification of 47 antibiotic residues in farmed European sea bass (Dicentrarchus labrax) using a multi-class and multi-residue UHPLC-MS/MS method

In the present study, a multi-class multi-residue method for the simultaneous detection and determination of antibiotics in European sea bass (Dicentrarchus labrax) was developed and validated. The method based on UHPLC-MS/MS proved to be a rapid, highly selective and sensitive tool, requiring minimum sample preparation, for screening and detection of 47 compounds from eight different classes. The validation was performed according to EU regulation 2002/657/EC, proving the method’s suitability for application in routine analysis. The method was applied to the analysis of 30 samples of farmed European sea bass purchased in different supermarkets in Portugal. Antibacterial residues were detected in 6 of the 30 analysed samples, namely enrofloxacin and oxytetracycline, in concentrations ranging from 0.1 to 12 µg kg^?1.     

QuEChERS/UPLC-MS/MS法同时测定梅花鹿鹿茸中7种镇静剂

采用分散固相萃取的前处理技术,建立了一种UPLC-MS/MS(ultra performance liquid chromatography-tandem mass spectrometry)法同时测定梅花鹿鹿茸中7种镇静剂的检测方法。鹿茸样品以2%氨水乙酸乙酯为提取剂,200 mg C₁₈为净化剂。液相色谱条件:ACQUITY BEH C₁₈柱(2.1 mm×100 mm,1.7 μm),流动相A为0.1%甲酸乙腈,B为水,柱温35 ℃;质谱条件:正离子模式(ESI⁺),多反应监测(multiple reaction monitoring,MRM)模式,鹿茸中镇静剂含量采用外标法定量。结果表明,7种镇静剂在0.3~100 ng/mL内线性关系良好,决定系数R²均大于0.992,7种镇静剂的LOD(limits of detection)为0.3 μg/kg,LOQ(limit of quantitation)为1 μg/kg,平均加样回收率为82.6%~107.3%,相对标准偏差RSD(relative standard deviation)均小于5%。该方法具有操作简便,净化效果显著,灵敏度高、准确性好、检出限低等优点,可用于梅花鹿鹿茸中7种镇静剂的检测。     

QuEChERS-超高效液相色谱-串联质谱法测定葡萄中388种农药残留

目的:建立了同时测定葡萄中388种农药残留量的方法。方法:样品经1%冰醋酸乙腈振荡提取后,加入无水醋酸钠、无水硫酸镁,提取液经300 mg硅胶、300 mg C₁₈、600 mg N-丙基乙二胺(PSA)和900 mg无水硫酸镁分散固相萃取吸附剂净化,上清液经氮吹浓缩,以乙腈-0.1 mol/L乙酸铵溶液(1:1)溶解进样,采用超高效液相色谱-串联质谱仪(UPLC-MS/MS)测定,分析物均采用电喷雾离子源,正离子扫描,多反应监测(MRM)模式,基质匹配内标法定量。结果:388种农药在0.005~0.4 mg/L浓度范围内线性关系良好,相关系数(r)在0.99以上,方法检出限均为0.005 mg/kg,3个加标水平的回收率为60%~120%,相对标准偏差均小于20%(n=7)。结论:该方法可同时测定388种农药残留,耗时较短,灵敏度高,分析物出峰均匀稳定,适用于葡萄中多农药残留的日常检测。     

改进 QuEChERS-超高效液相色谱-串联质谱法测定绿茶中 16 种真菌毒素含量

目的 建立一种改进QuEChERS 技术结合超高效液相色谱-串联质谱法（UPLC-MS/MS）用于检测绿茶中16种真菌毒素的方法。方法 本方法以1%甲酸乙腈作为提取溶剂,经涡旋或震荡提取后,使用 N-丙基乙二胺 （PSA）和十八烷基键合硅胶吸附剂（C18）作为净化材料净化后,经 ACQUITY BEH C18色谱柱（100 mm × 2.1 mm,1.7 ?m）实现分离,采用电喷雾正负离子电离方式（ESI±）、多反应监测（MRM）模式进行检测,基质匹配曲线外标法定量。结果 16种真菌毒素在相应浓度范围内呈良好的线性关系,相关系数（r）均大于 0.998,此方法的检出限（LOD）为 0.2-100.0 μg/kg,定量限（LOQ） 为 0.5-300.0 μg/kg。在 1、2、10 倍 定量限的加标水平下,16种真菌毒素的回收率范围为 74.4%-110.9%,相对标准偏差（RSD）值均小于5%。应用新建立的方法对 10 批次绿茶样品进行分析检测,结果在 2 批次样品中检出了黄曲霉毒素B2,1 批次样品中检出玉米赤霉烯酮。结论 该方法灵敏度好,准确度高,可作为绿茶中真菌毒素的日常监测的有效检测方法。     

QuEChERS净化结合超高效液相色谱串联质谱法测定奶粉中的季铵盐

在乳制品的生产过程中,常用季铵盐等消毒剂对生产设备等进行消毒,因此乳制品中存在季铵盐等消毒剂的污染。本文基于QuEChERS净化技术和超高效液相色谱串联质谱(UPLC-MS/MS),建立了一种测定奶粉中5种季铵盐消毒剂残留的新方法。奶粉样品先按比例加水复原成液态乳,然后经乙腈沉淀蛋白,提取液以PSA分散净化后,采用亲水色谱柱(HILIC)分离,在质谱的多反应监测(MRM)模式下测定。结果表明,5种季铵盐在0.2~50μg/L范围内线性关系良好,其相关系数(R)均在大于0.999,奶粉样品的方法检出限(S/N=3)在4.0~14.4μg/kg之间,样品基质的3个添加水平的平均回收率为96.5%~115%,日内精密度(n=6)在3.1%~7.5%之间,日间精密度(n=5)在4.3%~6.7%之间。实际样品检测结果表明奶粉中存在季铵盐消毒剂的残留污染情况。本方法准确灵敏,快速简便,精密度良好,适用于奶粉中以及其他类型乳制品中5种季铵盐消毒剂的分析检检测。     

Multiclass method for fast determination of veterinary drug residues in baby food by ultra-high-performance liquid chromatography–tandem mass spectrometry

A simple and rapid multiresidue method for the determination of different veterinary drug residues in meat-based baby food (MBF) and powdered milk-based infant formulae (PMIF) has been developed. The method involves an extraction procedure based on buffered QuEChERS (quick, easy, cheap, effective, rugged and safe) methodology, without any further clean-up step, followed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The method has been validated in two baby food matrices (MBF and PMIF) at three different concentration levels, obtaining suitable recoveries and precision (inter and intra-day precision) values. Quantification was carried out using matrix-matched standard calibration. Furthermore, the decision limit (CCα) and the decision capability (CCβ) were evaluated, ranging from 0.5 to 16.2 μg/kg and from 1.2 to 22.4 μg/kg, respectively. Finally, the method was applied to the analysis of several kinds of baby food samples and traces of some veterinary drugs were detected.