HPLC-MS/MS快速测定动物肌肉中6种聚醚类抗生素

建立动物肌肉组织中6种聚醚类抗生素(莫能菌素、马杜拉霉素、盐霉素、和拉沙洛菌素、甲基盐霉素、尼日利亚茵素)的高效液相色谱—电喷雾串联质谱快速测定法.样品经乙腈提取,并以乙腈饱和的正已烷净化,采用甲酸水溶液—甲醇体系为流动相,梯度洗脱,电喷雾离子源正离子多反应监测模式进行质谱分析.结果表明,在优化的条件下对样品进行分析检测,各抗生素的回收率为88.0％～108.7％,RSD为2.3％～6.1％(C=10μg/kg,n=5),6种待测物在1.0～150ng/mL范围内均呈线性,线性回归系数R2均大于0.99,检出限为0.05～0.2μg/kg.该方法操作简单,灵敏度高,适用于动物肌肉组织中6种聚醚类药物的批量检测分析.     

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.     

改进的QuEChERS结合超高效液相色谱—串联质谱法同时测定水产品中19种兽药残留

目的:实现水产品中19种磺胺、氟喹诺酮类兽药同步检测。方法:利用1.0%甲酸乙腈溶液作为提取液,超声1 min对样品中目标分析物进行提取,采用QuEChERS多功能针式过滤器净化样品,样品净化与过滤除杂同步进行,采用超高效液相色谱结合三重四极杆质谱进行检测。结果:19种磺胺、喹诺酮类兽药的检出限为0.5～1.0μg/kg,与国家标准方法相比,检测灵敏度有所提高。添加回收率为81.0%～113.0%,相对标准偏差(RSD)为0.2%～11.7%。结论:该方法通过超声辅助萃取、QuEChERS多功能针式过滤器提高样品前处理效率,具有方法检测灵敏度高、准确度高、重现性良好等优点,适用于大批量水产品样品中磺胺、喹诺酮类兽药日常监测。     

UPLC-MS/MS法同时测定动物源性食品中8种多肽类抗生素

建立了一种超高效液相色谱-串联质谱（UPLC-MS/MS）同时测定动物源性食品中8种多肽类抗生素残留量分析方法。试样经甲醇-水-甲酸（40∶60∶0.1,v/v/v）提取,固相萃取法净化后,以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相,梯度洗脱,经C18色谱柱分离后,在正离子扫描模式下,采用选择反应监测（SRM）,外标法定量。结果表明,8种多肽抗生素方法检出限为0.1¹0μg/kg,去甲万古霉素和万古霉素在20¹000μg/L范围内,多粘菌素B和E在40¹000μg/L范围内,杆菌肽A在10¹000μg/L范围内,杆菌肽B在6⁶14μg/L范围内,维吉尼霉素M1和S1在1¹00μg/L范围内,线性关系良好,相关系数r>0.999,加标回收率为61.0%⁹9.6%,RSD为1.2%⁹.8%。该方法简便快速、灵敏度高、重现性好,可满足动物源性食品中多肽类抗生素残留的同时测定。      

Multi-residue and multi-class method for the determination of antibiotics in bovine muscle by ultra-high-performance liquid chromatography tandem mass spectrometry

A multi-residue quantitative screening method covering 41 antibiotics from 7 different families, by ultra-high-performance–liquid-chromatography tandem mass spectrometry (UHPLC–MS/MS), is described. Sulfonamides, trimethoprim, tetracyclines, macrolides, quinolones, penicillins and chloramphenicol are simultaneously detected after a simple sample preparation of bovine muscle optimized to achieve the best recovery for all compounds. A simple sample treatment was developed consisting in an extraction with a mixture of acetonitrile and ethylenediaminetetraacetic acid (EDTA), followed by a defatting step with n-hexane. The methodology was validated, in accordance with Decision 2002/657/EC by evaluating the required parameters: decision limit (CCα), detection capability (CCβ), specificity, repeatability and reproducibility. Precision in terms of relative standard deviation was under 20% for all compounds and the recoveries between 91% and 119%. CCα and CCβ were determined according the maximum residue limit (MRL) or the minimum required performance limit (MRPL), when required.     

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.     

Determination of tetracyclines in multi-specie animal tissues by pressurized liquid extraction and liquid chromatography–tandem mass spectrometry

A specific, sensitive and robust pressurized liquid extraction (PLE) and liquid chromatography tandem mass spectrometry (LC–MS/MS) method for determining tetracycline, chlortetracycline, oxytetracycline and doxycycline in bovine, swine, poultry and lamb muscle tissues is presented. PLE was performed using an ASE^® 200 from Dionex and water as extractant, followed by solid-phase extraction (SPE) using an Oasis HLB cartridge. The method was validated for beef, chicken, pork and lamb meat in compliance with the requirements set by Commission Decision, 2002/657/EC [Commission Decision 2002/657/EC (2002). Implementing Council Directive 96/23/EC concerning the performance of analytical methods and interpretation of results. Official Journal of European Communities, L239, 66–98. (Available at: <http://europe.eu.int>)]. The average recoveries of the different meat samples, spiked with the four tetracyclines at three levels (1, 100 and 200 μg kg⁻¹ of each tetracycline), were always higher than 89% with intraday and interday precision lower than 15% and 17%, respectively. A good linearity was established for the four tetracyclines in the range from 5 to 10,000 μg kg⁻¹ with r > 0.995. The limits of quantification (LOQs) were between 0.5 and 1 μg kg⁻¹, which are well below the tolerance levels set by the European Union. The decision limit (CCα) and the decision capability (CCβ) were in the range 101–116 and 112–130 μg kg⁻¹, respectively. Compared with previous methods, sample preparation time required for the analysis and LOQs, are reduced. The method demonstrated its successful application for the analysis of 100 meat samples. Two samples of beef and one sample of chicken out of 25 of each type tested positive while none of 25 samples of either, lamb or pork, tested positive.     

高效液相色谱-串联质谱法测定猪肉中氯霉素和五种磺胺类药物的残留量

目的:建立超高效液相色谱-串联质谱（UPLC-MS/MS）同时快速测定猪肉中氯霉素、磺胺甲嘧啶、磺胺二甲嘧啶、磺胺噻唑、甲氧苄啶和磺胺嘧啶残留量的分析方法。方法:猪肉加入乙酸乙酯后匀浆,转移有机层N₂吹干,残渣用10%乙腈水溶液溶解,正己烷除脂,采用BEH C₁8（2.1 mm×50 mm,1.7μm）色谱柱分离,以乙腈和水为流动相进行梯度洗脱,采用电喷雾离子源（ESI）,以多反应监测（MRM）方式检测。结果:在该实验条件下,氯霉素和5种磺胺类药物分别在9.6³84 ng/kg和0.51²0.8μg/kg浓度范围呈良好的线性关系,R²均大于0.99;氯霉素检出限为3.2 ng/kg,定量限为9.6 ng/kg,5种磺胺类药物检出限为0.158⁰.173μg/kg,定量限为0.475⁰.52μg/kg;方法的平均回收率范围为70.84%¹03.99%,相对标准偏差为1.98%⁸.01%。结论:此方法具有简便快捷、灵敏度高、定量准确等特点。      

Development and validation of rapid multiresidue and multi-class analysis for antibiotics and anthelmintics in feed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry

A new multi-residue method for the analysis of veterinary drugs, namely amoxicillin, chlortetracycline, colistins A and B, doxycycline, fenbendazole, flubendazole, ivermectin, lincomycin, oxytetracycline, sulfadiazine, tiamulin, tilmicosin and trimethoprim, was developed and validated for feed. After acidic extraction, the samples were centrifuged, purified by SPE and analysed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry. Quantitative validation was done in accordance with the guidelines laid down in European Commission Decision 2002/657/CE. Matrix-matched calibration with internal standards was used to reduce matrix effects. The target level was set at the authorised carryover level (1%) and validation levels were set at 0.5%, 1% and 1.5%. Method performances were evaluated by the following parameters: linearity (0.986 < R² < 0.999), precision (repeatability < 12.4% and reproducibility < 14.0%), accuracy (89% < recovery < 107%), sensitivity, decision limit (CCα), detection capability (CCβ), selectivity and expanded measurement uncertainty (k = 2).This method has been used successfully for three years for routine monitoring of antibiotic residues in feeds during which period 20% of samples were found to exceed the 1% authorised carryover limit and were deemed non-compliant.     

同位素稀释-超高效液相色谱-串联质谱法检测蜂蜜中泛酸含量

建立了同位素稀释-超高效液相色谱-串联质谱法（UPLC-MS/MS）检测蜂蜜中泛酸（维生素B5）的快速定量和确证方法。样品经0.1%氨水（V/V）提取,MAX固相萃取柱净化后,以0.1%甲酸乙腈-0.1%甲酸水为流动相梯度洗脱,使用HSS T3色谱柱进行分离,电喷雾正离子多反应监测（MRM）模式检测,同位素内标法定量。在优化条件下,泛酸在0.500～500 μg/L范围内线性良好,相关系数（R2）为0.999 6;方法检出限为1.0 μg/kg（S/N=3）,定量限为3.0 μg/kg（S/N=10）;回收率在98.3%～98.7%之间,相对标准偏差（RSD）为2.4%～3.1%。通过市售蜂蜜样品测试表明,该方法操作简单、检测结果准确,可用于蜂蜜中泛酸的快速检测。     

Liquid chromatography tandem mass spectrometry determination of maduramycin residues in the tissues of broiler chickens

Maduramycin is a polyether ionophoric coccidiostat used to prevent coccidiosis in poultry at a prescribed concentration over a certain time interval. Due to public health concerns about the presence of coccidiostat residues in poultry, the aim of the present study was to determine the level of maduramycin residues in the tissues of broiler chickens fed commercially produced feed containing 5 mg kg^?1 of maduramycin in complete feed throughout the 5-day withdrawal period (WP). The residues were investigated by liquid chromatography (LC) coupled with electrospray ionisation (ESI) tandem mass spectrometry (MS/MS). The limit of detection (LOD) and limit of quantification (LOQ) of the method were 0.3 and 0.8 µg kg^?1, respectively. The average recovery based on matrix-fortified calibrations for chicken tissues was 90%. Maduramycin was found to be rapidly distributed in all tissues. The highest concentrations of maduramycin residues were found in the heart followed by the skin, liver, gizzard, kidneys and, finally, muscle (thigh and breast). On day 5 of the WP, residue concentrations of maduramycin did not decline below the LOQ of the method. Our results emphasize the need to establish a maximum residue limit (MRL) for maduramycin to control its residue levels in edible tissues from chickens before slaughter.     

超高效液相色谱串联质谱法检测玛咖产品中非法添加西地那非

建立了检测玛咖中西地那非的超高效液相色谱-串联质谱（UPLC-MS/MS）法。色谱条件为Agilent SB-C18柱;柱温25℃;流动相为甲醇（10 mmol/L乙酸铵）:水（10 mmol/L乙酸铵,m/V）（90∶10,v/v）等度洗脱;流速:0.3 m L/min;进样量:5μL。质谱条件为正离子模式（ESI+）,多反应监测模式（MRM）;三重四级杆质量分析器;分子离子峰[M+H]+m/z 475.3,子离子为m/z 282.9,碰撞能50 e V;m/z 58.1碰撞能为60 e V。西地那非在0.2²0μg/L范围内线性关系良好,相关系数R2为0.9994。建立的方法准确可靠、灵敏度高,可应用于玛咖产品中西地那非的含量测定。      

Multiclass mycotoxin analysis in edible oils using a simple solvent extraction method and liquid chromatography with tandem mass spectrometry

ABSTRACT

A simple and rapid method for the simultaneous determination of 11 mycotoxins – aflatoxins B₁, B₂, G₁ and G₂; fumonisins B₁, B₂ and B₃; ochratoxin A; zearalenone; deoxynivalenol; and T-2 toxin – in edible oils was established using liquid chromatography tandem mass spectrometry (LC-MS/MS). In this study, QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe), QuEChERS with dispersive liquid–liquid microextraction, and solvent extraction were examined for sample preparation. Among these methods, solvent extraction with a mixture of formic acid/acetonitrile (5/95, v/v) successfully extracted all target mycotoxins. Subsequently, a defatting process using n-hexane was employed to remove the fats present in the edible oil samples. Mass spectrometry was carried out using electrospray ionisation in polarity switching mode with multiple reaction monitoring. The developed LC-MS/MS method was validated by assessing the specificity, linearity, recovery, limit of quantification (LOQ), accuracy and precision with reference to Commission Regulation (EC) 401/2006. Mycotoxin recoveries of 51.6–82.8% were achieved in addition to LOQs ranging from 0.025 ng/g to 1 ng/g. The edible oils proved to be relatively uncomplicated matrices and the developed method was applied to 9 edible oil samples, including soybean oil, corn oil and rice bran oil, to evaluate potential mycotoxin contamination. The levels of detection were significantly lower than the international regulatory standards. Therefore, we expect that our developed method, based on simple, two-step sample preparation process, will be suitable for the large-scale screening of mycotoxin contamination in edible oils.     

Multiresidue method for simultaneous analysis of aflatoxin M1, avermectins,organophosphate pesticides and milbemycin in milk by ultra-performance liquid chromatography coupled to tandem mass spectrometry

A method developed for the simultaneous analysis of aflatoxin M₁, abamectin, doramectin, eprinomectin, ivermectin, moxidectin, acephate, azinphos-ethyl, azinphos-methyl, diazinon, methamidophos, methidathion, mevinphos, pirimiphos-ethyl and pirimiphos-methyl in whole raw milk, based on the QuEChERS method for extraction and clean-up, with detection and quantification by ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) is described. The method was validated according to parameters of the Analytical Quality Assurance Manual from the Brazilian Ministry of Agriculture and Commission Decision 2002/657/EC, and proved suitable for analysis of these analytes within the proposed working range, with recovery values between 77% and 110%, a standard deviation lower than 20%, limits of detection between 0.05 and 0.99 µg l^?1, and limits of quantification between 0.15 and 1.98 µg l^?1. Samples from animals treated with abamectin, doramectin, ivermectin and diazinon were analysed by the validated method. Residues of aflatoxin M₁ were also found in field samples at levels below the established maximum residue limit.     

超高效液相色谱-串联质谱法测定谷物中的总烟酸

建立一种快速、准确的测定谷物中总烟酸含量的超高效液相色谱-串联质谱方法。样品加入烟酸-D4同位素内标校正,经氢氧化钠水解,采用HLB固相萃取柱净化后,以10 mmol/L乙酸铵溶液（含0.1%甲酸）和乙腈作为流动相进行梯度洗脱,采用HSS T3液相色谱柱分离,正离子MRM模式进行定性定量分析。实验结果表明,烟酸在0.1²00 ng/m L浓度范围内线性关系良好,相关系数r>0.999。线性范围内,平均加标回收率为99.3%¹02.6%,相对标准偏差在4.24%⁴.74%。仪器的检出限为0.04 ng/m L,定量限为0.1 ng/m L。该方法具有灵敏度高、重现性好、分析时间短等优点,可以为谷物中总烟酸含量的测定提供技术支持。      

Assessment of liquid chromatography–tandem mass spectrometry approaches for the analysis of ceftiofur metabolites in poultry muscle

The use of cephalosporin antibiotics in veterinary practice is likely to play an important role in the development of β-lactam-resistant bacteria. To detect off-label cephalosporin antibiotic usage, an analytical method is needed that, besides the native compound, also detects their active metabolites. In this paper, the applicability of three approaches for the quantitative analysis of ceftiofur using LC–MS/MS is assessed, viz. (A) analysis of ceftiofur, desfuroylceftiofur and/or desfuroylceftiofur cystein disulfide, (B) derivatisation of ceftiofur metabolites to desfuroylceftiofur acetamide and (C) chemical hydrolysis using ammonia, to produce a marker compound for ceftiofur. We found that approach A was not suited for quantitative analysis of total ceftiofur concentration or for effectively detecting off-label use of ceftiofur. Approach B resulted in adequate quantitative results, but was considered a single compound method because it depends on cleavage of a thioester group, which is present in only a limited number of cephalosporin antibiotics. Approach C showed adequate quantitative results but, in contrast to approach B, it is applicable to a range of cephalosporin antibiotics. Therefore, it is applicable as a broad quantitative screening of cephalosporin compounds in poultry tissue samples to indicate off-label use of cephalosporins in poultry breeding. Based on this study, it was concluded that approach C is the most suitable to detect off-label use of a range of cephalosporin antibiotics.     

Liquid chromatography–electrospray ionization–tandem mass spectrometry method for determination of twenty multi-class pesticide residues in cashew

This paper describes an analytical methodology employing the extraction QuEChERS (Quick, Easy, Cheap, Effective, Robust and Safe) method and liquid chromatography coupled to mass spectrometry using electrospray ionization source (LC–ESI–MS/MS). The methodology was validated for the determination of twenty multiclass pesticides in cashew. It was evaluated for selectivity, linearity, limits of detection (LOD) and quantification (LOQ), matrix effect, as well as the precision and accuracy in terms of percentage of recovery. Analyses were performed by selected reaction monitoring (SRM) using two transitions (precursor ion → ion product). All pesticides studied showed good linearity with correlation coefficient greater than 0.99. LODs and LOQs ranged from 0.10 to 0.25 ng g⁻¹ and 0.30 to 0.75 ng g⁻¹, respectively. Due to the complexity of the sample, study the effect matrix was performed. Cashew apples samples spiked (5, 50 and 100 ng g⁻¹) showed recovery results ranging from 66 to 119 g/100 g, and RSD ≤ 12%, which is in accordance with standard recommended by Brazilian rules (ABNT NBR 14029/2005), except simazine. The validated method was applied to commercial samples obtained in Fortaleza-CE, Brazil, but no contamination of pesticides residues was observed.     

超高效液相色谱-三重四极杆串联质谱法同时测定劲酒中8种活性成分含量

目的 建立超高效液相色谱-串联质谱( UPLC-MS/MS) 法同时测定中国劲酒中8种成分(甜菜碱、腺苷、香草酸、芦丁、阿魏酸、香草醛、黄芩苷、黄芪甲苷)的含量。方法 采用Agilent EC-C18色谱柱(4.6 mm×50 mm, 1.8 μm),以流动相为水0.1%甲酸水-0.1%甲酸甲醇为流动相,梯度洗脱,流速0.2mL.min^-1。采用电喷雾离子源进行正离子模式监测,多反应监测模式(MRM)用于定量分析。结果 中国劲酒中的8个成分在考察的浓度范围内具有良好的线性关系(r>0.998 1);回收率(n=6)在 96.7%～103.3%内,RSD均<4.38%;3批次间8个成分的含量差异较大。结论 该方法简便快速、专属性强、灵敏度高,可用于中国劲酒的质量控制。     

Rapid multi-class multi-residue method for the confirmation of chloramphenicol and eleven nitroimidazoles in milk and honey by liquid chromatography-tandem mass spectrometry (LC-MS)

A confirmatory method was developed to allow for the analysis of eleven nitroimidazoles and also chloramphenicol in milk and honey samples. These compounds are classified as A6 compounds in Annex IV of Council Regulation 2377/90 (European Commission 1990) and therefore prohibited for use in animal husbandry. Milk samples were extracted by acetonitrile with the addition of NaCl; honey samples were first dissolved in water before a similar extraction. Honey extracts underwent a hexane wash to remove impurities. Both milk and honey extracts were evaporated to dryness and reconstituted in initial mobile phase. These were then injected onto a liquid chromatography-tandem mass spectrometry (LC-MS/MS) system and analysed in less than 9 min. The MS/MS was operated in multiple reaction monitoring (MRM) mode with positive and negative electrospray ionization. The method was validated in accordance with Commission Decision 2002/657/EC and is capable of analysing metronidazole, dimetridazole, ronidazole, ipronidazole and there hydroxy metabolites hydroxymetronidazole, 2-hydroxymethyl-1-methyl-5-nitroimidazole, and hydroxyipronidazole. The method can also analyse for carnidazole, ornidazole, ternidazole, tinidazole, and chloramphenicol. A recommended level of 3 µg l^?1/µg kg^?1 for methods for metronidazole, dimetridazole, and ronidazole has been recommended by the Community Reference Laboratory (CRL) responsible for this substance group, and this method can easily detect all nitroimidazoles at this level. A minimum required performance level of 0.3 µg l^?1/µg kg^?1 is in place for chloramphenicol which the method can also easily detect. For nitroimidazoles, the decision limits (CCα) and detection capabilities (CCβ) ranged from 0.41 to 1.55 µg l^?1 and from 0.70 to 2.64 µg l^?1, respectively, in milk; and from 0.38 to 1.16 µg kg^?1 and from 0.66 to 1.98 µg kg^?1, respectively, in honey. For chloramphenicol, the values are 0.07 and 0.11 µg l^?1 in milk and 0.08 and 0.13 µg kg^?1 in honey. Validation criteria of accuracy, precision, repeatability, and reproducibility along with measurement uncertainty were calculated for all analytes in both matrices.     

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。