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A method using QuEChERS coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the determination of the residues of 19 veterinary drugs in ten animal-derived matrices, including beef, pork, sheep, horse, chicken, prawn, fish, liver, milk, and fat. This method was based on the enactment of veterinary drug compounds by Korea, Canada, the United States, and the European Union in recent years. The samples were extracted using 85% acetonitrile and separated on an ACQUITY UPLC HSS T3 column (2.1 mm × 100 mm, 1.8 μm) with a gradient elution of methanol-0.2% formic acid water as the mobile phase. The detection of the analytes was achieved through the use of positive ion electrospray ionization (ESI) and multiple reaction monitoring (MRM) modes, while the quantification was conducted via the matrix-matched external standard method. Following optimization, the linearity of the target veterinary residues in the ten matrices was observed to be satisfactory, having a range of 0.5-50.0 ng/mL (R 2 > 0.991). The limits of detection (LOD) were in the range of 0.01-1.29 μg/kg, while the limits of quantification (LOQ) were in the range of 0.02-4.31 μg/kg. The recoveries were observed to be in the range of 60.6-117.7 %, with relative standard deviations (RSDs) of ≤20.6 %. The method is straightforward and highly sensitive, and it satisfies the maximum limits set by the relevant standards of Korea, Canada, the USA, and the EU. It is well-suited for the rapid screening, qualitative, and quantitative analyses of metomidate, acetanilide, dl-methylephedrine, and other substances in foods of animal origin, providing technical assistance for cross-border food safety and testing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier B.V.)

This study presents a method utilizing solid-phase microextraction Arrow (SPME Arrow) combined with ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) for the selective detection of three veterinary drugs-thiabendazole, sulfamethazine, and clenbuterol-in milk and pork. Two-dimensional metal-organic framework nanosheets (2D-MOFs) were employed as coating materials for the SPME Arrow. Three types of 2D-MOFs (Ni, Mn, and Co based) were synthesized and characterized using Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and a physical adsorption analyzer. The 2D-MOF coatings were fabricated using the electrospinning technique, with polyacrylonitrile (PAN) serving as the binder. Comparative analysis of the three 2D-MOF coatings revealed that 2D-Ni-MOF was the optimal coating material for the SPME Arrow. Optimization of the coating preparation conditions and SPME procedures included determining the optimal mass ratio of 2D-Ni-MOF to PAN, electrospinning time, and extraction and desorption parameters. Equilibrium extraction was achieved within 60 min, and desorption was completed within 30 min. Subsequently, the 2D-Ni-MOF-SPME Arrow-UPLC-Q-TOF-MS method was established and validated under optimal conditions, demonstrating high precision with inter-day precision ranging from 3.8 % to 9.5 % and intra-day precision ranging from 5.1 % to 11.5 %. The reusability study indicated that the extraction performance of the new SPME Arrow remained consistent after 90 adsorption-desorption cycles. The method exhibited linearity in milk and pork over the ranges of 0.002-5 μg L -1 and 0.01-5 μg L -1 , respectively. The detection limits in milk and pork were 0.001-0.004 μg L -1 and 0.003-0.007 μg L -1 , respectively. This method demonstrated excellent applicability for determining residues of the three veterinary drugs in milk and pork., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Hangzhen Lan reports financial support was provided by Ningbo University. Hangzhen Lan reports financial support was provided by One Health Interdisciplinary Research Project of Ningbo University. Hangzhen Lan reports financial support was provided by J National Natural Science Foundation of China. Hangzhen Lan reports financial support was provided by Key R&D Program of Zhejiang. Hangzhen Lan reports financial support was provided by Natural Science Foundation of Zhejiang Province. Hangzhen Lan reports financial support was provided by Science and Technology Programs of Ningbo., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Antibacterial medications are receiving the most attention due to hypersensitivity reactions and the emergence of bacterial mutants resistant to antibiotics. Treating Animals with uncontrolled amounts of antibiotics will extend beyond their lives and affect humans. This study aims to determine the concentration of the residues of sulfadimidine, sulfaquinoxaline, diaveridine, and vitamin K3 in the tissues of poultry (muscles and liver) after treatment with the combined veterinary formulation. A UPLC-MS-MS method was developed using Poroshell 120 ECC 18 and a mobile phase composed of acetonitrile and distilled water, containing 0.1 % formic acid, in the ratio of (85:15 v/v) at a flow rate of 0.6 mL/min. Sample extraction solvent was optimized using response surface methodology (RSM) to be acetonitrile: methanol in the ratio (49.8: 50.2 v/v), and the method was validated according to the FDA bioanalytical method validation protocol over the range (50-1000 µg/Kg) for sulfaquinoxaline and (50-750 µg/Kg) for the other 3 drugs. The greenness of the sample preparation and analytical method was assessed by applying Analytical Eco-scale (AES) and AGREE coupled with AGREEprep. The Competence of the study was evaluated via the EVG framework known as Efficiency, validation, and greenness, to achieve a balance point represented by a radar chart. The method was applied to decide the time required for poultry products to be safe for human use after administration of the studied drugs. It was found that, after the administration of the last dose, minimally 7 days are required till the levels of the drugs drop to the maximum residue limit determined by the FDA/WHO in animal tissues., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Modified QuEChERS and triple quadrupole mass spectrometry (LC and GC-MS/MS) technology were used to sequentially analyze pesticides, veterinary drugs, and mycotoxins in feed. In order to analyze the harmful substances that may remain or occur in the feed, we performed optimization experiments for sample preparation and LC-MS/MS and GC-MS/MS conditions. Optimized sample preparation involves extracting 5 g of sample with 15 mL of 0.25 M EDTA and 10 mL of acetonitrile. And some extracts were diluted 10-fold with 100 mM ammonium formate aqueous solution and analyzed by LC-MS/MS, and some extracts were purified through 25 mg PSA and analyzed by GC-MS/MS by adding an analyte protectant. We confirmed the matrix effect of feed ingredients and compound feeds, and added a dilution process after extraction to increase on-site efficiency. Matrix-matched calibration was applied for quantification. Method validation was performed for 197 pesticides, 56 components for veterinary drugs, and 5 components for toxins. All the components showed good linearity (r2 ≥ 0.98) in the developed analytical method. For most compounds, the limit of quantitation was 0.05 mg/kg. The recovery rate experiment was repeated three times at three concentrations including LOQ in feed ingredient, compound feed for livestock, and compound feed for pets. The recovery rate was 70.09-119.76% and relative standard deviations were ≤ 18.91%. And the accuracy and precision were further verified through cross-validation between laboratories. The developed analytical method was used to monitor 414 domestically distributed and imported feeds., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The purpose of this study was to develop a modified QuEChERS method based on melamine sponge for rapid determination of multi-class veterinary drugs in milks by UPLC-MS/MS. Through simple infiltration and extrusion, fast and convenient matrix purification could be achieved within several seconds, and there was no need of extra phase separation operations. Good linearity with correlation coefficient (R 2 ) ≥0.999 was obtained for all drugs in the range of 2~500 µg·kg -1 . The obtained matrix effects were within ±20% for all monitored drugs. The recoveries of all monitored drugs ranged from 60.7% to 116.0% at three spiked levels (50, 100, and 200 µg·kg -1 ), with relative standard deviations less than 7.4%. Comparatively low LODs and LOQs were obtained in the ranges of 0.1~3.8 µg·kg -1 and 0.2~6.3 µg·kg -1 , respectively. Compared with conventional purification adsorbents, melamine sponge yielded an equal or higher purification performance with matrix removal rate as high as 52.5% and acceptable recoveries in range of 60%-120% for all monitored drugs. The satisfactory results have demonstrated the good potential of melamine sponge in matrix purification for rapid determination of multiclass residues in food safety., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier B.V.)

A rapid, simple and generic analytical method has been developed for the analysis of veterinary drugs in pork by a quadrupole time-of-flight mass spectrometry (Q-TOF MS). This method allows for the simultaneous identification, screening and quantitation of 141 veterinary drug residues and metabolites from eighteen different classes. After extraction with acetonitrile/water and clean-up with C 18 cartridges, the samples were analyzed by HPLC-Q-TOF MS. Validation of this method consisted of confirmation of identity, selectivity, linearity, limit of detection (LOD), lowest limit of quantification (LLOQ), matrix effect, recovery, precision and applicability of the method. Identification of the analytes was based on accurate mass measurements. The characteristic fragments were obtained by collisional experiments for a more reliable identification. The procedure was then applied to real pork samples. Sulfamethazine was detected in one sample and its metabolites were successfully found in one single run. This approach proved to be satisfactory for routine analysis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The first quantitative multiclass approach enabling the accurate quantification of >1200 biotoxins, pesticides and veterinary drugs in complex feed using liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed. Optimization of HPLC/UHPLC (chromatographic column, flow rate and injection volume) and MS/MS conditions (dwell time and cycle time) were carried out in order to allow the combination of five major substance classes and the high number of target analytes with different physico-chemical properties. Cycle times and retention windows were carefully optimized and ensured appropriate dwell times reducing the overall measurement error. Validation was carried out in two compound feed matrices according to the EU SANTE validation guideline. Apparent recoveries matching the acceptable range of 60-140% accounted 60% and 79% for all analytes in cattle and chicken feed, respectively. High extraction efficiencies were obtained for all analyte/matrix combinations and revealed matrix effects as the main source for deviation of the targeted performance criteria. Concerning the methods repeatability 99% of all analytes in chicken and 96% in cattle feed complied with the acceptable RSD ≤ 20% criterion. Limits of quantification were between 1-10 µg/kg for the vast majority of compounds. Finally, the methods applicability was tested in >130 real compound feed samples and provides first insights into co-exposure of agro-contaminants in animal feed., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest., (Copyright © 2020. Published by Elsevier B.V.)

Animal feeds are often reported to be contaminated with chemical residues, and when present above the maximum legal limit, these compounds can cause harmful effects to consumers of animal produce. Thus, animal feed safety is an important regulatory concern. The aim of this study was to optimise a multiresidue method for the simultaneous analysis of multi-class pesticides and a number of frequently used veterinary drugs using LC-MS/MS and GC-MS/MS. The method was validated in a range of feed matrices, including maize feed, poultry feed and mixed feed concentrate. The optimised sample preparation workflow involved extraction of feeds (5 g) with ethyl acetate (10 mL), followed by a freezing step (at -20°C) used for eliminating the matrix co-extractives. The extract was further cleaned by dispersive solid phase extraction with a combination of primary secondary amine, C18 and florisil sorbents. From the cleaned-extract, an aliquot was analysed by GC-MS/MS, while another portion of it was solvent-exchanged to acetonitrile:water (50:50) and then analysed by LC-MS/MS. This method effectively minimised the matrix interferences. A total of 192 pesticides was analysed by GC-MS/MS within a runtime of 22 min. The LC-MS/MS method was validated for 187 compounds including 17 veterinary drugs. For most of the compounds, the limit of quantification (LOQ) was 0.01 mg/kg. The recoveries at LOQ and higher levels ranged between 70% and 120%, with precision-RSDs of < 20%. The method provided a precise analysis in a wide range of market-feed samples. As shown, the method is suitable for regulatory and commercial testing purposes., Competing Interests: Declaration of Competing Interest The authors have declared no conflict of interest exists., (Copyright © 2020. Published by Elsevier B.V.)

An ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method based on selective accelerated solvent extraction and magnetic material purification was established to analyze the residues of various veterinary antibiotics and agricultural fungicides and insecticides in livestock and poultry excrement. Methanol-acetonitrile (4:1, V/V) was used as the extraction solvent and static extraction was conducted three times in 5 min. Preliminary purification was achieved by adding 0.5 g acidic alumina-florisil (1:1, W/W) to the extraction cell while the extraction was conducted. This preliminarily-purified extract was further purified using magnetic material, then analyzed using UPLC-MS/MS. Under optimal conditions, 33 types of antibiotics, including 3 amphenicols, 8 macrolides, 17 sulfonamides and 5 nitroimidazoles, as well as 37 types of pesticides, including 27 insecticides and 10 fungicides, were detected. Recoveries ranged from 60.3% to 110.0% at three spiked concentrations (10 μg/kg, 20 μg/kg and 50 μg/kg), the detection limits ranged from 0.2 to 3.5 μg/kg and the quantitative limits ranged from 0.5 to 11.5 μg/kg. This newly-established method was tested using 30 livestock and poultry excrement samples and confirmed its use for determining veterinary drugs and pesticides in practical samples., Competing Interests: Declaration of Competing Interest The authors declared that they have no conflicts of interest to this work., (Copyright © 2019. Published by Elsevier B.V.)

In this work, the use of design of experiments and posterior data modelling by artificial neural network (ANN) and least squares (LS) is presented as a suitable analytical tool for the performance optimization of a tandem mass spectrometric detector coupled to ultra-high performance liquid chromatography for the analysis of seventeen veterinary drugs. Firstly, a central composite design was built considering as factors the cone, capillary, extractor and radio frequency voltages of the mass spectrometer in order to obtain a proper combination to improve the sensitivity of the method. Secondly, a one factor design considering the collision voltage was built to define the adequate voltage for each daughter ion. The response surface methodology (RSM) was then applied, and the prediction capability of ANN and LS were compared. As conclusion, the ANN modelling provided better results than LS, both in terms of the ANOVA and predicted areas results. The accuracy of the model prediction was between 85 and 125%, confirming that the estimates of the model were correct, and endorsing the optimization procedure as a suitable way to gather excellent results. The suitability of the new approach and its implications on the simultaneous analysis of seventeen veterinary drugs by ultra-high liquid chromatography coupled to tandem mass spectrometry detection are discussed., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Cephalosporins are of particular importance in human medicine and should be reserved for second-line curative treatment in the veterinary field to avoid any emerging antimicrobial resistance. Due to misuse of ceftiofur in the poultry sector in France, it is now recommended to completely stop using cephalosporins in this sector. Methods currently used for the control of veterinary practices are mostly based on liquid chromatography coupled to mass spectrometry in a targeted mode, including parent compounds and any major metabolites. The aim of the present study was to evaluate the relevance of untargeted metabolomic approaches to highlight a possible exposure of laying hens to cephalosporins using a predictive model including selected treatment biomarkers. An experimentation carried out on living animals involved the administration of cefquinome and ceftiofur. Three biological matrices-droppings, eggs and liver-were investigated. Metabolites were extracted and analysed by liquid chromatography coupled to high resolution mass spectrometry in a full scan mode. Metabolites impacted by the treatment were selected by using univariate and multivariate statistical analyses. Predictive models built from the potential biomarkers selected in the "droppings" matrix were validated and able to classify "treated" and "control" hens. PLS-DA and logistic regression models were compared and both models gave satisfactory results in terms of prediction. Results were of less interest for other matrices in which only biomarkers of exposure to cefquinome were detected., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study presents the development and validation of a quantitation method for the analysis of multi-class, multi-residue veterinary drugs using lipid removal cleanup cartridges, enhanced matrix removal lipid (EMR-Lipid), for different meat matrices by liquid chromatography tandem mass spectrometry detection. Meat samples were extracted using a two-step solid-liquid extraction followed by pass-through sample cleanup. The method was optimized based on the buffer and solvent composition, solvent additive additions, and EMR-Lipid cartridge cleanup. The developed method was then validated in five meat matrices, porcine muscle, bovine muscle, bovine liver, bovine kidney and chicken liver to evaluate the method performance characteristics, such as absolute recoveries and precision at three spiking levels, calibration curve linearity, limit of quantitation (LOQ) and matrix effect. The results showed that >90% of veterinary drug analytes achieved satisfactory recovery results of 60-120%. Over 97% analytes achieved excellent reproducibility results (relative standard deviation (RSD) < 20%), and the LOQs were 1-5 μg/kg in the evaluated meat matrices. The matrix co-extractive removal efficiency by weight provided by EMR-lipid cartridge cleanup was 42-58% in samples. The post column infusion study showed that the matrix ion suppression was reduced for samples with the EMR-Lipid cartridge cleanup. The reduced matrix ion suppression effect was also confirmed with <15% frequency of compounds with significant quantitative ion suppression (>30%) for all tested veterinary drugs in all of meat matrices. The results showed that the two-step solid-liquid extraction provides efficient extraction for the entire spectrum of veterinary drugs, including the difficult classes such as tetracyclines, beta-lactams etc. EMR-Lipid cartridges after extraction provided efficient sample cleanup with easy streamlined protocol and minimal impacts on analytes recovery, improving method reliability and consistency., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A multiclass method was proposed for the simultaneous determination of various classes of veterinary drugs (n = 65), mycotoxins and metabolites (n = 39) in egg and milk by ultra-high performance liquid chromatography-tandem mass spectrometry. The contaminants were extracted by QuEChERS-based strategy including salt-out partitioning and dispersive solid-phase extraction for cleanup further. With the aim of maximizing throughput and extraction efficiency, Plackett-Burman design was employed initially for screening significant variables. And response surface methodology based on central composite design was conducted to achieve optimal conditions in details: 3.35% (v/v) of formic acid in acetonitrile, 1.2 g of NaCl, 0.5 g of anhydrous NaAc, 300 mg of C18 and 140 mg of primary secondary amine. Satisfactory analytical characteristics in validation, in aspects of accuracy (70%-105% for mycotoxins and quinolones, 55%-80% for sulphonamides and 40%-105% for other veterinary drugs), precision (inter-day RSDs < 14%) and sensitivity (LOQs ranged from 0.01 μg/kg to 31 μg/kg), were achieved under the optimized conditions. The matrix effects were evaluated and compensated by the use of matrix-matched calibration curves (R 2  > 0.987). In practice, 45 eggs and 30 milk samples were investigated by the established method, of which positive finding aflatoxin in milk and sterigmatocystin in eggs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This work reports the development of a multi-residue method for the identification and quantification of 82 veterinary drugs belonging to different chemical classes in swine waste lagoon. The proposed method applies a solid-phase extraction procedure with Oasis PRiME HLB cartridges that combines isolation of the compounds and sample clean-up in a single step. Analysis is performed by ultra-high performance liquid chromatography-tandem mass spectrometry, in one single injection with a chromatographic run time of only 9.5min. Linearity was studied in the range between 1 and 500μgkg -1 using standards prepared both in pure solvent and in the presence of matrix, showing coefficients of determination higher than 0.99 for all the analytes except for cefapirin in matrix. The average recoveries were in the range of 60-110% for most of the compounds tested with inter-day relative standard deviations below 17%. More than 97% of the investigated compounds had less or equal to a 5μgkg -1 quantitation limit in the studied matrix. Finally, the method was used with success to detect and quantify veterinary drugs residues in real samples with sulfonamides, quinolones, and tetracyclines being the most frequently determined compound groups., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Avermectins and milbemycin are widely used as veterinary drugs and as agricultural pesticides, and their residues have been detected in soil. This study reports a simple and high-throughput method for determining ivermectin (IVER), abamectin (ABA), doramectin (DORA), eprinomectin (EPRI), and moxidectin (MOXI) residues in soils, employing an on-line solid-phase extraction technique coupled with ultra-high performance liquid chromatography and tandem mass spectrometry (SPE-UHPLC-MS/MS). The method was validated and applied for the determination of ABA in soils from an orange plantation treated with this pesticide. The sample preparation procedure consisted of extraction of the compounds from soil using methanol (with recoveries of 73-85%), and subsequent on-line SPE cleanup and concentration using a C8 sorbent coupled to the UHPLC-MS/MS system. The optimal conditions were: water:methanol (40:60, v/v) sample solvent; water:methanol (96:4, v/v) loading solvent; 2×250μL sample volume; and elution of the analytes retained on the SPE column in back flush with 5mmolL -1 ammonium acetate:acetonitrile (10:90, v/v) chromatographic mobile phase. The method produced linear results in the ranges 0.1-10ngg -1 (IVER, ABA, DORA, and MOXI) and 0.5-10ngg -1 (EPRI), with linearity greater than 0.99. The precision of the method was better than 19% and accuracy was in the range 74-89%. The limits of quantitation were 0.2ngg -1 for EPRI and 0.1ngg -1 for the other compounds. The SPE column could be reused in more than 2000 analyses without loss of efficiency. The ABA concentration in the soil varied between 1.7 and 18ngg -1 , and no dissipation was observed during five consecutive days after application of the pesticide to the orange plantation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This paper presents a multi-residue analytical method for 210 drugs in pork using ultra-high-performance liquid chromatography-Q-Trap tandem mass spectrometry (UPLC-MS/MS) within 20min via positive ESI in scheduled multi-reaction monitoring (MRM) mode. The 210 drugs, belonging to 21 different chemical classes, included macrolides, sulfonamides, tetracyclines, β-lactams, β-agonists, aminoglycosides, antiviral drugs, glycosides, phenothiazine, protein anabolic hormones, non-steroidal anti-inflammatory drugs (NSAIDs), quinolones, antifungal drugs, corticosteroids, imidazoles, piperidines, piperazidines, insecticides, amides, alkaloids and others. A rapid and simple preparation method was applied to process the animal tissues, including solvent extraction with an acetonitrile/water mixture (80/20, v/v), defatting and clean-up processes. The recoveries ranged from 52% to 130% with relative standard deviations (RSDs)<20% for spiked concentrations of 10, 50 and 250μg/kg. More than 90% of the analytes achieved low limits of quantification (LOQs)<10μg/kg. The decision limit (CCα), detection capability (CCβ) values were in the range of 2-502μg/kg and 4-505μg/kg, respectively. This method is significant for food safety monitoring and controlling veterinary drug use., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A quick, sensitive and selective analytical reversed-phase multi-residue method using ultra-high performance liquid chromatography coupled to an ion-trap mass spectrometry detector (UHPLC-IT-MS/MS) operating in both positive and negative ion mode was developed for the simultaneous determination of 23 veterinary drug residues (β-blockers, β-agonists and Non-Steroidal Anti-inflammatory Drugs (NSAIDs)) in meat samples. The sample treatment involved a liquid-solid extraction followed by a solid-phase extraction (SPE) procedure. SBA-15 type mesoporous silica was synthetized and modified with octadecylsilane, and the resulting hybrid material (denoted as SBA-15-C18) was applied and evaluated as SPE sorbent in the purification of samples. The materials were comprehensively characterized, and they showed a high surface area, high pore volume and a homogeneous distribution of the pores. Chromatographic conditions and extraction procedure were optimized, and the method was validated according to the Commission Decision 2002/657/EC. The method detection limits (MDLs) and the method quantification limits (MQLs) were determined for all the analytes in meat samples and found to range between 0.01-18.75μg/kg and 0.02-62.50μg/kg, respectively. Recoveries for 15 of the target analytes ranged from 71 to 98%. In addition, for comparative purpose SBA-15-C18 was evaluated towards commercial C18 amorphous silica. Results revealed that SBA-15-C18 was clearly more successful in the multi-residue extraction of the 23 mentioned analytes with higher recovery values. The method was successfully tested to analyze prepacked preparations of mince bovine meat. Traces of propranolol, ketoprofen and diclofenac were detected in some samples., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A multiresidue/multiclass method for the simultaneous determination of 76 veterinary drugs and pharmaceuticals in bovine muscle tissue has been developed and validated according to the requirements of European Commission Decision 2002/657/EC. The analytes belong in 13 different classes, including aminoglycoside antibiotics, whose different physicochemical properties (extremely polar character) render their simultaneous determination with other veterinary drugs quite problematic. The method combines a two-step extraction procedure (extraction with acetonitrile followed by an acidic aqueous buffer extraction) with hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) determination, allowing confirmation and quantification in a single chromatographic run. Further cleanup with solid phase extraction was performed using polymeric SPE cartridges. A thorough ionization study of aminoglycosides was performed in order to increase their sensitivity and significant differences in the abundance of the precursor ions of the analytes were revealed, depending on the composition of the mobile phase tested. Further gradient elution optimization and injection solvent optimization were performed for all target analytes.The method was validated according to the European Commission Decision 2002/657. Quantitative analysis was performed by means of standard addition calibration. Recoveries varied from 37.4% (bromhexine) to 106% (kanamycin) in the lowest validation level and 82% of the compounds showed recovery >70%. Detection capability (CCβ) varied from 2.4 (salinomycin) to 1302 (apramycin) μgkg(-1)., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A new magnetic dispersive solid-phase extraction approach based on Eu- and Tb-coated magnetic nanocomposites, combined with ultra-high performance liquid chromatography with fluorometric detection, is reported for the extraction and simultaneous determination of veterinary antibiotics. The method is aimed at monitoring of potential residues of three tetracyclines, namely oxytetracycline, tetracycline, chlortetracycline and three acidic quinolones, such as oxolinic acid, nalidixic acid and flumequine, chosen as model analytes, in animal muscle samples. The nanocomposites were obtained by synthesizing magnetic nanoparticles by a co-precipitation method and their coating with terbium and europium ions. The limits of detection obtained using standard solutions were: 1.0, 1.5, 3.8, 0.25, 0.7 and 1.2ngmL(-1), which corresponds to 3.3, 5.0, 12.7, 0.8, 2.3 and 4.0μgkg(-1) for oxytetracycline, tetracycline, chlortetracycline, oxolinic acid, nalidixic acid and flumequine, respectively, in meat samples. The precision values, obtained in the presence of the sample matrix, were in the ranges 0.12-2.0% and 2.6-15.4% for retention times and areas, respectively. The selectivity of the method was checked by assaying different veterinary drugs, finding that most of them did not interfere at the same concentration levels as that of analytes. A recovery study was performed in the presence of chicken and pork muscle samples, which provided values in the range of 61.5-102.6%., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Subcritical water extraction was investigated as a novel and alternative technology for the separation of trace amounts of chloramphenicol, thiamphenicol, florfenicol and its major metabolite florfenicol amine from poultry tissues and its results were compared with those of conventional shaking extraction, ultrasonic extraction, and pressurized liquid extraction. Decreasing the polarity of water by successively increasing the extraction temperature from 50°C to 200°C at the moderate pressure enabled selective, highly effective extractions to be performed. Rapid quantification of the target compounds was carried out by ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). The critical parameters of subcritical water extraction such as solvent modifier, temperature, pressure, extraction time, and static cycles were varied with control. The optimized extraction procedures using subcritical water as extraction solvent, were carried out on a pressurized liquid extractor operated at 150°C and 100bar, applying two static cycles for 3min. Average recoveries of the four analytes from fortified samples ranged between 86.8% and 101.5%, with relative standard deviations (RSDs) lower than 7.7%. The limits of detection (LODs) and quantification (LOQs) for the target compounds were in the ranges of 0.03-0.5μgkg(-1) and 0.1-2.0μgkg(-1), respectively. The proposed method is fast, sensitive, water-based thus more environmental acceptable, making it a suitable replacement for conventional organic solvent extraction in veterinary drug residue analysis., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Retrospective analysis has been applied in different samples, including honey, meat, feed and nutraceutical products from ginkgo biloba, soya, royal jelly and green tea, with the aim of searching transformation products of pesticides and veterinary drugs, which were not included in an initial analysis. Generic extraction and analytical procedures based on high resolution mass spectrometry (Exactive-Orbitrap analyser was used) have been applied. All obtained data have been reprocessed and some compounds as anhydroerythromycin in honey and 3,5,6-trichloro-2-pyridinol in feed have been detected, demonstrating the applicability and the utility of the procedure. Advantages and disadvantages of retrospective approach have been highlighted., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A new analytical method was developed and validated for simultaneous analysis of 333 pesticide and veterinary drug residues in baby food. Response surface methodology was employed to optimize a generic extraction method. Ultrahigh-performance liquid chromatography and electrospray ionization quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-ESI Q-Orbitrap) was used for the separation and detection of all the analytes. The method was validated by taking into consideration the guidelines specified in Commission Decision 2002/657/EC and SANCO/12571/2013. The extraction recoveries were in a range of 79.8-110.7%, with coefficient of variation <8.3%. The 333 compounds behave dynamic in the range 0.1-1000μgkg(-1) concentration, with correlation coefficient >0.99. The limits of detection for the analytes are in the range 0.01-5.35μgkg(-1). The limits of quantification for the analytes are in the range 0.01-9.27μgkg(-1). This method has been successfully applied on screening of pesticide and veterinary drugs in ninety-three commercial baby food samples, and tilmicosin, fenbendazole, tylosin tartrate and thiabendazole were detected in some samples tested in this study. The present study is very useful for fast screening of different food contaminants., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In the European Union (EU), sulfonamides are among the most widely administrated groups of antibiotics in animal husbandry. Therefore, monitoring their residues in edible animal tissues plays an important role in the EU food safety framework. In this work, a simple and efficient method for the rapid screening of twelve prior sulfonamides frequently prescribed as veterinary drugs by high-performance thin-layer chromatography (HPTLC) was established. Sample extracts obtained with acetonitrile were tenfold concentrated and applied to HPTLC without any further cleanup. Following separation and fluram derivatization, sensitive and selective quantitation of the analytes can readily be accomplished with fluorescent densitometry. Limits of detection and quantitation were 15-40 and 35-70μg/kg, respectively. Additionally, a confirmative detection by HPTLC-electrospray ionization mass spectrometry (HPTLC-ESI/MS) was optimized, offering straightforward identification of target zones. Therefore, the risk of potential false positive findings can efficiently be reduced. The method was validated to meet the enforced commission regulation (EU) No. 37/2010, regarding different matrix complexities (bovine milk, porcine liver and kidney)., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Distillers Grain (DG) is an important by-product of ethanol production. The ethanol production process uses only the starch portion of the plant and all the remaining nutrients, protein, fat, minerals, and vitamins remain in DGs, a valuable feed material for livestock. The use of antimicrobial drugs is helpful to limit harmful bacterial growth during the early part of the fermentation process. This can lead to the possible presence of contaminants in the by-products that are used in the food and feed industries, resulting in a major concern for the development of bacterial resistance in both humans and animals. To facilitate the detection of antimicrobial and other commonly used veterinary drugs in DGs, a liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed targeting a wide range of 12 chemical classes of anti-bacterial substances and drugs, such as ionophore and non-ionophore authorized coccidiostats, banned coccidiostats, macrolides, tetracyclines, nitroimidazoles, amphenicols, quinolones, sulphonamides, tranquilizers, non-steroidal anti-inflammatory drugs and benzimidazoles. Following a simple and fast extraction step with a mixture of organic solvents, the extract was directly injected into the LC coupled to an Orbitrap mass analyzer. The identification of residues is based on accurate mass measurement. The high mass resolution of 50,000 full width at half maximum (FWHM) and corresponding narrow mass windows permitted a very selective and sensitive detection of the analytes in such a complex matrix. A single-laboratory validation procedure was carried out evaluating selectivity, sensitivity, linearity, precision and accuracy. The method showed satisfactory analytical performance for precision and trueness, and allowed the determination of the compounds at low concentration. The proposed multi-method demonstrated that liquid chromatography coupled to an Orbitrap mass spectrometer is a promising analytical technique, suitable for official residue control of a variety of veterinary drugs in DGs supporting feed safety policies., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Ionophore antibiotics (IPAs) are widely used as coccidiostats in poultry and other livestock industries to promote growth and prevent infections. Because most of the ingested IPAs are excreted in poultry litter, which is primarily applied as grassland fertilizer, a significant amount of IPAs can be released into the litter-soil-water environment. A robust analytical method has been developed to quantify IPAs (monensin (MON), salinomycin (SAL) and narasin (NAR)) in complex environmental compartments including surface runoff, soil and poultry litter, with success to minimize matrix interference. The method for water samples involves solid-phase extraction (SPE) followed by liquid-liquid extraction (LLE) post-clean up steps. The method for solid samples involves bi-solvent LLE. IPAs were detected by HPLC-MS, with optimized parameters to achieve the highest sensitivity. Nigericin (NIG), an IPA not used in livestock industry, is successfully applied and validated as a surrogate standard. The method recoveries were at 92-95% and 81-85% in runoff samples from unfertilized and litter-fertilized fields, respectively. For solids, the method recoveries were at 93-99% in soils, and 79-83% in poultry litter samples. SAL was detected at up to 22mg/kg and MON and NAR at up to 4mg/kg in broiler litter from different farms. Up to 183μg/kg of MON was detected in litter-fertilized soils. All three IPAs were detected in the rainfall runoff from litter-fertilized lands at concentrations up to 9μg/L., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A multi-residue pressurised liquid extraction (PLE) methodology has been established for the determination of the four ionophores: lasalocid, monensin, salinomycin and narasin in solid environmental matrices. The PLE methodology is combined with solid phase extraction as clean-up using liquid chromatography coupled to tandem mass spectrometry applying electrospray ionisation for detection. The samples were freeze-dried prior to extraction. The absolute recoveries for soil and sediment ranged from 71 to 123% (relative standard deviation (RSDs) below 16%) and in the range 94-133% (RSDs 9-35%) for poultry manure. The final method allowed for the detection of four ionophores down to a few hundred ngkg(-1) in natural solid matrices with limit of quantifications (LOQs) being 0.96, 0.87, 0.98, and 0.64μgkg(-1) in soil for lasalocid, monensin, salinomycin, and narasin, respectively. Corresponding LOQs in sediment were 1.28, 1.34, 1.39, and 0.78μgkg(-1) for the respective ionophores, while in manure the LOQs were 0.98, 1.01, 1.45, and 1.01μgkg(-1)., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Glucocorticoids constitute a class of molecules widely used in animal husbandry. Some of these compounds are licensed for veterinary practices while their use for growth promoting purposes is prohibited within the European Union. In order to ensure the respect of the legislation and consumers safety, several methodologies have been proposed to monitor these substances in various products, including edible matrices for which a regulatory limit has been set up (MRL). An extended range of targeted analytes together with reduced time of analysis and cost are however still current challenges regularly revisited according to the continuous technological improvements. In this context, the aim of the present study was to develop and implement a new fast and multi-residue method based on UHPLC-MS/MS for the determination of twenty glucocorticoids in bovine milk, included the screening of the three regulated MRL compounds (dexamethasone, betamethasone and prednisolone). This validated method authorises such multi-analyte measurement within a 10min runtime while the signal specificity is ensured through the SRM acquisition mode. Decision limits and detection capabilities were calculated in the range of 0.001-0.363μgL(-1), which allows a very efficient control at low trace level for a potential illegal use of these substances. The performances obtained in terms of application range, selectivity and sensitivity were found to be significantly improved in comparison to other reported approaches either for screening or confirmation purposes: regarding linearity, correlation coefficients were above 0.98 within the range of 0.01-5.0μgL(-1), repeatability and reproducibility parameters ranged from 1 to 30% with the maximum relative standard deviation (RSD) observed for cortisone (30.1%). Stability of the stock solutions and minor changes in the standard operating procedure have been included for the determination of ruggedness of the method. Identification was systematically ensured according to 4 identification points, RSD of transitions ratio (T2/T1) ranged from 3.2% and 19.3% and the RSD of the retention time was lower than 0.25%., (Copyright © 2013 Elsevier B.V. All rights reserved.)

This paper presents a sensitive and confirmatory multi-residue method for the analysis of 23 veterinary drugs and metabolites belonging to three classes (nitroimidazoles, benzimidazoles, and chloramphenicols) in porcine muscle, liver, and kidney. After extracted with ethyl acetate and basic ethyl acetate sequentially, the crude extracts were defatted with hexane and further purified using Oasis MCX solid-phase extraction cartridges. Rapid determination was carried out by ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometry. Data acquisition was performed under positive and negative mode simultaneously. Recoveries based on matrix-matched calibrations for meat, liver, and kidney ranged from 50.6 to 108.1%. The method quantification limits were in the range of 3-100ng/kg., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The performance of liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) post-run target screening for veterinary drug residue analysis (sulfonamides, tetracyclines and quinolones) in animal urine has been critically evaluated. It was found that retention time information still remains an essential information and that accurate masses together with relative isotopic abundance data alone are not sufficient for many residue applications. Post-run target screening requires the careful setting of parameters to achieve near zero false negative (above a defined threshold level) and a manageable numbers of false positive findings. HRMS offers many possibilities for the reduction of false positives (e.g. isotopic ratio, isotopic fine structure, exact mass of fragment ions). However, the successful use of such tools requires a sufficient ion intensity. This is often not available when trace level compounds are to be detected. Nevertheless, the proposed method is sufficiently sensitive to detect the veterinary drugs at the relevant concentration levels in urine. This means that the approach is well suited to significantly reduce the number of corresponding meat samples which have to be analyzed in a final step for the regulatory relevant quantification of residue levels in meat. The semi-quantitative screening of many samples for a large number of analytes within a short period of time requires the availability of software tools which provide fast and reliable answers., (Copyright © 2012 Elsevier B.V. All rights reserved.)

In the field of food contaminant analysis, the most significant development of recent years has been the integration of ultra-high pressure liquid chromatography (UHPLC), coupled to tandem quadrupole mass spectrometry (MS/MS), into analytical applications. In this review, we describe the emergence of UHPLC through technological advances. The implications of this new chromatographic technology for MS detection are discussed, as well as some of the remaining challenges in exploiting it for chemical residue applications. Finally, a comprehensive overview of published applications of UHPLC-MS in food contaminant analysis is presented, with a particular focus on veterinary drug residues., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A QuEChERS (Quick Easy Cheap Effective Rugged Safe)-like extraction method was developed for the simultaneous analysis of veterinary drugs and mycotoxins in hen eggs by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray (ESI) source. Various classes of antimicrobials (tetracyclines, ionophores, coccidiostats, penicillins, cephalosporins, fluoroquinolones, sulfonamides) and mycotoxins (enniatins, beauvericin, ochratoxins, aflatoxins) were considered for the development of this method. Particular attention was devoted to extraction optimization: different solvents (acetone, acetonitrile and methanol), different pH values and different sample to extracting volume ratios were tested and evaluated in terms of recovery, relative standard deviation (RSD) and ESI signal suppression due to matrix effect. Chromatographic and mass spectrometric conditions were optimized to obtain the best instrumental performances for most of the analytes. Quantitative analysis was performed by means of matrix-matched calibration, in a range that varied depending on the analyte and its established maximum limit, when there was one. Recoveries at 100 μg kg(-1) spiking level were >62% (3

A multi-residue analytical method was developed to quantify nine antibiotics and one hormone in soil, broiler manure and manure compost. The developed method was based on ultrasonic extraction with MeOH:ACN:EDTA:McIlvaine buffer, solid phase extraction (SPE) using HLB (3 cc/60 mg) cartridge, followed by instrumental analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with 25 min total run time. It was validated and tested on soil, broiler manure and manure compost samples and showed that the method is able to simultaneously detect and quantify the target analytes with good selectivity and sensitivity. The developed method was linear in a concentration range from its instrumental quantification limit (IQL) to 500 ng/mL, with correlation coefficients higher than 0.999. The overall method performance was good for the majority of the analytes, with recoveries range from 63% to 121% in all the sample matrices. The method quantification limit (MQL) for the 10 target analytes in the soil, broiler manure and manure compost samples were 2-10, 3-16 and 5-15 μg/kg dry weight (DW), respectively. The method has also included tilmicosin, an antibiotic known to be reported in the environment for the first time. The developed method was then applied on broiler manure samples and its relative manure amended agricultural soil samples to identify and quantify veterinary antibiotic and hormone residues in the environment. These analytes were detected in broiler manure and soil samples, with maximum concentrations reaching up to 78516.1 μg/kg DW (doxycycline) and 1331.4 μg/kg DW (flumequine), respectively. The results showed that the method can potentially be adopted for the analysis of veterinary antibiotic and hormone wastes in solid environmental matrices., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The detection of veterinary drugs in blood meal is needed since it is used as an environment-friendly agricultural material despite its origination from animal blood. A method using accelerated solvent extraction and liquid chromatographic linear ion trap quadrupole Orbitrap mass spectrometry was developed to determine sedatives and adrenergic blockers in blood meal. The determination method was established following optimizations of accelerated solvent extraction, dispersive solid-phase extraction and high resolution mass spectrometric detection. Linearity, sensitivity, accuracy, repeatability and reproducibility of the method were fully validated. The method was applied to commercial blood meal products., (Copyright © 2012 Elsevier B.V. All rights reserved.)

In this study, optimization, extension, and validation of a streamlined, qualitative and quantitative multiclass, multiresidue method was conducted to monitor >100 veterinary drug residues in meat using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Optimization centered on extensive ruggedness evaluation of the method. Various clean-up sorbents were tested and the amount of co-extractives were weighed, matrix effects were measured using post-column infusion of representative analytes, the effect of extract dilution before injecting was studied, and analyte recoveries and reproducibilities were determined. In order to extend our previous method, more drug analytes were added that possessed a wider range of chemical properties, and a re-appraisal of different types of C18 in dispersive solid-phase extraction clean-up and mobile phases in UHPLC-MS/MS was done. Ultimately, end-capped C18 and post-column infusion of ammonium formate as an ionization enhancer for the late-eluting anthelmintics were found to give improved qualitative results for greater analytical scope. A multi-day, multi-analyst validation demonstrated that the final method is suitable for screening of 113 analytes, identifying 98 and quantifying (recoveries between 70-120% and RSD<25%) 87 out of the 127 tested drugs at or below US regulatory tolerance levels in bovine muscle., (Published by Elsevier B.V.)

The present work describes the development of an analytical method, based on automated off-line solid phase extraction (SPE) followed by ultra-high-performance liquid chromatography coupled to quadrupole linear ion trap tandem mass spectrometry (UPLC-QqLIT) for the determination of 81 pharmaceutical residues, covering various therapeutic groups, and some of their main metabolites, in surface and treated waters (influent and effluent wastewaters, river, reservoir, sea and drinking water). For unequivocal identification and confirmation, two selected reaction monitoring (SRM) transitions per compound are monitored. Quantification is performed by the internal standard approach, indispensable to correct matrix effects. Moreover, to obtain an extra tool for confirmation of positive findings, an information dependent acquisition (IDA) experiment was performed, with SRM as survey scan and an enhanced product ion (EPI) scan as dependent scan. Compound identification was carried out by library search, matching the EPI spectra achieved at one fixed collision energy with those present in a library. The main advantages of the method are automation and speed-up of sample preparation by the reduction of extraction volumes for some matrices, the fast separation of a big number of pharmaceuticals, its high sensitivity (limits of detection in the low ng/L range), selectivity, due to the use of tandem mass spectrometry, reliability since a significant number of isotopically labeled compounds are used as internal standards for quantification and finally, the analysis of tap, reservoir and sea waters, since information about occurrence of pharmaceuticals in these matrices is still sparse. As part of the validation procedure, the method developed was applied to the analysis of pharmaceutical residues in waste and surface waters from different sites in Catalonia (North East of Spain)., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A database has been created for the simultaneous analysis of more than 350 pesticides and veterinary drugs (including antibiotics) using ultra-high performance liquid chromatography coupled to high resolution Orbitrap mass spectrometry (UHPLC-Orbitrap-MS). This is a comprehensive exact mass database built using the Exactive-Orbitrap analyzer. The developed database includes exact masses of the target ions and retention time data, and allows the automatic search of the included compounds. Generic chromatographic and MS conditions have been applied. The presented database is suitable for qualitative analysis, and it was also evaluated for quantitative purposes in routine analysis, after the optimization and validation of a generic extraction method in honey samples. Adequate recovery and precision values for most of the studied analytes were obtained and the limits of detection (LOD) ranged from 1 to 50 μg kg(-1). For pesticides, LODs were always lower than the MRLs established by European Union in honey, except for a few compounds. This method was applied to the analysis of 26 real honey samples and some pesticides (azoxystrobin, coumaphos, dimethoate and thiacloprid) were detected in 4 samples. Azoxystrobin and coumaphos were determined in two different samples (organic honey) at 1.5 μg kg(-1) and 5.1 μg kg(-1). Veterinary drugs were not detected in the analyzed samples., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The aim of this study was to develop an analytical method for the analysis of traces of hormonal steroids and veterinary and human drugs in soil. Thus, 31 substances were selected, including 14 veterinary products, 11 hormonal steroids and 6 other well-known human contaminant compounds. The procedure inspired by the quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction method was developed. First the acetonitrile-based extraction was optimized. This step was followed by a solid-phase extraction (SPE) clean-up using both a strong anion-exchange cartridge and a polymeric cartridge. The analysis was then performed using liquid chromatography coupled to a triple quadrupole analyser operated with tandem mass spectrometry. This analytical procedure was validated using the ICH/2005 standard by evaluating the linearity (from 0.01 ng/g to 1000 ng/g--R²>0.99), the intra-day precision (relative standard deviation (RSD)<20%), the inter-day precision (RSD<30%), recoveries (40-110% for most of the compounds) and limits of detection and quantification. This method allowed for the determination of the target analytes in the lower ng/g concentration range. The methodology was then applied to real soil samples collected in several areas of France that received different manure or sludge treatments. Some target compounds were detected at very low level (inferior to the ng/g). Veterinary antibiotics, mainly from the sulfonamide family, were found in soils treated by manure (0.02-0.12 ng/g). On the other hand, pharmaceuticals usually used by humans (carbamazepine, ibuprofen) were detected in soils treated by domestic sludge., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Veterinary drugs (VDs) can remain in milk as a consequence of their use in livestock. In order to control the levels of VD residues in milk, screening methodologies can be applied for a rapid discrimination among negative and non-negative samples. In a second stage, non-negative samples are classified as negative or positive samples by using a confirmation method. Pre-target screening methods in low resolution MS (LRMS) are normally applied, but the number of analytes is limited, whereas the information obtained by full scan acquisition in high resolution mass spectrometry (HRMS) is improved. Here, three screening methods (running time<4 min) based on Orbitrap, quadrupole-time of flight (QqTOF) and triple quadrupole (QqQ) have been compared, using in all cases ultra-high performance liquid chromatography (UHPLC). For HRMS, the identification of the VDs was based on retention time (RT) and accurate mass measurements. Confirmation was based on the monitoring of fragments generated without precursor selection. The performance characteristics of the screening method provided reliable information regarding the presence or absence of the compounds below an established value, including uncertainty region and cut-off values. Better results in terms of cut-off values (≤ 5.0 μg kg(-1), except for spiramycin with a cut-off of 13.4 μg kg(-1) for milk samples and 43.1 μg kg(-1) for powdered milk based, emamectin with a cut-off of 42.2 μg kg(-1) for milk samples and doxycycline, with a cut-off value of 15.8 μg kg(-1) in powdered milk-based infant formulae) and uncertainty region were obtained using the Orbitrap-based screening method, which was submitted to further validation and used to analyze different real milk samples. The proposed method can be used in routine analysis, providing reliable results., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A simple multi-residue method was developed for detecting and quantifying 33 analytes from 13 classes of antibiotics (tetracyclines (3), quinolones (7), penicillins (3), ionophore coccidiostats (7), macrolides (3), sulfonamides (1), quinoxalines (2), phenicols (2), lincosamides (1), diaminopyrimidines (1), polypeptides (1), streptogramins (1) and pleuromutilins (1)) in animal feeds. Extraction and clean-up procedures were optimized with spiked piglet feed. Samples were extracted by ultrasonic-assisted extraction with a mixture of methanol/acetonitrile/McIlvaine buffer at pH 4.6 (37.5/37.5/25, v/v/v) containing 0.3% of EDTA-Na(2), followed by a clean up using a dispersive solid-phase extraction (d-SPE) with PSA (primary secondary amine). Detection of antibiotics was achieved by liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) within 28 min using both positive and negative ESI mode. Average recoveries ranged from 51% (oxytetracycline) to 116% (tilmicosin) with associated relative standard deviations of 7.3% and 9.0% and an overall mean of 87%. Limits of quantification ranged from 3.8 ngg(-1) (lincomycin) to 65.0 ngg(-1) (bacitracin). Following optimization, the method was further verified for bovine and lamb feeding stuffs; negative matrix effects were evaluated and overcome by a standard addition method., (Copyright © 2010 Elsevier B.V. All rights reserved.)

A novel magnetic solid-phase extraction (MSPE) sorbent, magnetite/silica/poly (methacrylic acid-co-ethylene glycol dimethacrylate) (Fe(3)O(4)/SiO(2)/P(MAA-co-EGDMA)), was developed. This MSPE material was prepared by distillation-precipitation polymerization of MAA and EGDMA in the presence of Fe(3)O(4)/SiO(2) microspheres with the surface containing abundant reactive double bonds. The resultant sorbent material was characterized by elemental analysis, electron microscopy, X-ray diffraction and Fourier-transformed infrared spectroscopy. In this work, eleven sulfonamides (SAs) were selected as model analytes to validate the extraction performance of this new MSPE sorbent. Noticeably, the extraction can be carried out quickly, the extraction time for the SAs onto Fe(3)O(4)/SiO(2)/P(MAA-co-EGDMA) sorbent can be clearly shortened to 0.5 min. The desorption solution of SAs was analyzed by LC-MS/MS, and the results showed that the recoveries of these compounds were in the range of 87.6-115.6%, with relative standard deviations ranging between 0.9% and 10.8%; the limit of detection were in the range of 0.5-49.5 ng/L., (2010 Elsevier B.V. All rights reserved.)

LC-ion trap mass spectrometry was used to screen and confirm 38 compounds from a variety of drug classes in four species of fish: trout, salmon, catfish, and tilapia. Samples were extracted with acetonitrile and hexane. The acetonitrile phase was evaporated, redissolved in water and acetonitrile, and analyzed by gradient chromatography on a phenyl column. MS(2) or MS(3) spectra were monitored for each compound. Qualitative method performance was evaluated by the analysis over several days of replicate samples of control fish, fish fortified with a drug mixture at 1 ppm, 0.1 ppm and 0.01 ppm, and fish dosed with a representative from each drug class. Half of the 38 drugs were confirmed at 0.01 ppm, the lowest fortification level. This included all of the quinolones and fluoroquinolones, the macrolides, malachite green, and most of the imidazoles. Florfenicol amine, metronidazole, sulfonamides, tetracyclines, and most of the betalactams were confirmed at 0.1 ppm. Ivermectin and penicillin G were only detectable in the 1 ppm fortified samples. With the exception of amoxicillin, emamectin, metronidazole, and tylosin, residue presence was confirmed in all the dosed fish.

Several classes of chemical compounds, exhibiting many different chemical properties, are classified under the generic term of "veterinary drugs", among which are the antimicrobial medicines such as antibiotics or dyes, and drugs exhibiting growth promoting properties like steroids, beta-agonist compounds, thyrostats or growth hormones. For food safety purposes, the resort to these substances in animal breeding has been submitted to strict regulation within the European Union for more than 15 years. Systems of control have therefore been set up within the same period of time to ensure compliance with the regulation. The current strategy relies on targeted analytical approaches focusing on the detection of residues of the administered compounds or their metabolites in different kinds of feed, food or biological matrices. If screening methods, which provide rapid discrimination between compliant and suspect samples, may be based on several techniques such as immunoassays or mass spectrometry, confirmatory methods mainly rely on the latter, which provides adequate specificity and sensitivity for unambiguous identification of the target analytes in biological matrices at trace level. The present article reviews the main mass spectrometric strategies, from the very first, nonetheless still efficient, single MS and multidimensional and high-resolution MS through to advanced isotope ratio MS. Several applications in the field of residue analysis illustrate each of these approaches and focus on the balance between issues related to the compounds of interest (chemistry, matrix, concentration, ...) and the large offer of mass spectrometric-related technical possibilities, from the choice of the ionization conditions (EI, NCI, PCI, reagent gases, ESI+, ESI-), to the mass analyzers (single quadrupole, triple quadrupole, ion traps, time-of-flight, magnetic sectors, isotope ratio mass spectrometer) and corresponding acquisition modes (full scan, LR-SIM, HR-SIM, SRM, precursor scan, ...). All the displayed strategies, from the importance of sample preparation to MS analysis to potential derivatization steps and chromatographic separation parameters are discussed in that context. Besides the advantages of each strategy, main issues associated to such MS approaches are commented with an emphasis not only on such critical points as ion suppression and resolution, but also on the adequacy of the current regulation regarding the evolution of the technology. Finally, future trends which may lead to strong and positive impacts in the field of residue analysis are presented, including latest developments and improvements in chromatography or software dedicated to signal acquisition and data analysis.

The topic of validation is diversified. This review outlines the validation strategies which can be found in national, international and supranational regulations, compares them with one another and aims to elaborate on the main principles. European regulations and legislation, Codex alimentarius guidelines, the official methods program of the AOAC, and naturally the relevant ISO standards, particularly the ISO 5725 series, are taken into consideration. The objective of every validation is to demonstrate fitness for purpose. This varies of course in its characteristics for the diverse uses. However, all approaches have in common the objective of harmonisation of food control by using effective and reliable methods. To this end, criteria are determined and validation models developed and made compulsory. ISO 5725 is the central basis for validations for quantitative methods with its validation specifications through method collaborative studies. On the contrary, there are no valid uniform international method specifications for qualitative methods. Collaborative studies are in opposition to single-lab-validations with different sources of error. Whereas laboratory errors are predominant in collaborative studies, the single-lab-validation or in-house validation concentrates particularly on time and processing errors (intermediate precision). In new statistical models for in-house validations, the matrix mismatch error is also considered. The validation models presented here are of a general nature and can be used in principle for all analytical methods. Correct and appropriate statistical modelling is very important.

The last 2 years multi-compound methods are gaining ground as screening methods. In this study a high-resolution liquid chromatography combined with time-of-flight mass spectrometry (HRLC-ToF-MS) is tested for the screening of about 100 veterinary drugs in three matrices, meat, fish and egg. While the results are satisfactory for 70-90% of the veterinary drugs, a more efficient sample preparation or extract purification is required for quantitative analysis of all analytes in more difficult matrices like egg. The average mass measurement error of the ToF-MS for the veterinary drugs spiked at concentrations ranging from 4 to 400 microg/kg, is 3.0 ppm (median 2.5 ppm) with little difference between the three matrices, but slightly decreases with increasing concentration. The SigmaFit value, a new feature for isotope pattern matching, also decreases with increasing concentration and, in addition, shows an increase with increasing matrix complexity. While the average SigmaFit value is 0.04, the median is 0.01 indicating some high individual deviations. As with the mass measurement error, the highest deviations are found in those regions of the chromatogram where most compounds elute from the column, be it analytes or matrix compounds. The median repeatability of the method ranges from 8% to 15%, decreasing with increasing concentration, while the median reproducibility ranges from 15% to 20% with little difference between matrices and concentrations. The median accuracy is in between 70% and 100% with a few compounds showing higher values due to matrix interference. The squared regression coefficient is >0.99 for 92% of the compounds showing a good overall linearity for most compounds. The detection capability, CCbeta, is within 2 times the associated validation level for >90% of the compounds studied. By changing a few conditions in the analyses protocol and analysing a number of blank samples, it was determined that the method is robust as well as specific. Finally, an alternative validation strategy is proposed and tested for screening methods. While the results calculated for repeatability, within-lab reproducibility and CCbeta show a good comparison for the matrices meat and fish, and a reasonable comparison for the matrix egg, only 27 analyses are required to obtain these results versus 63 analysis in the traditional, 2002/657/EC, approach. This alternative is suggested as a cost-effective validation procedure for screening methods.

A simple, rapid, and sensitive method for the quantitative monitoring of five sulfonamide antibacterial residues (SAs) in milk was developed by stir bar sorptive extraction (SBSE) coupling to high performance liquid chromatography with diode array detection. The analytes were concentrated by SBSE based on poly (vinylimidazole-divinylbenzene) monolithic material as coating. The extraction procedure was very simple, milk was diluted with water then directly sorptive extraction without elimination of fats and protein in samples was required. To achieve optimum extraction performance for SAs, several parameters, including extraction and desorption time, desorption solvent, ionic strength and pH value of sample matrix were investigated. Under the optimized experimental conditions, low detection limits (S/N=3) quantification limits (S/N=10) of the proposed method for the target compounds were achieved within the range of 1.30-7.90 ng/mL and 4.29-26.3 ng/mL from spiked milk, respectively. Good linearities were obtained for SAs with the correlation coefficients (R(2)) above 0.996. Finally, the proposed method was successfully applied to the determination of SAs compounds in different milk samples and satisfied recoveries of spiked target compounds in real samples were obtained.

A comprehensive review is presented on the current trends in sample preparation for the isolation of veterinary drugs and growth promoters from foods. The objective of the review is to firstly give an overview of the sample preparation techniques that are applied in field. The review will focus on new techniques and technologies, which improve efficiency and coverage of residues. The underlying theme to the paper is the developments that have been made in multi-residue methods and particularly multi-class methods for residues of licensed animal health products, which have been developed in the last couple of years. The role of multi-class methods is discussed and how they can be accommodated in future residue surveillance.

A residue is a trace (microg kg(-1), ng kg(-1)) of a substance, present in a matrix. Residue analysis is a relatively young discipline and a very broad area, including banned (A) substances as well as registered veterinary medicinal products (B substances). The objective of this manuscript is to review future trends in the analysis of residues of veterinary drugs in meat producing animals out of historical perspectives. The analysis of residues in meat producing animals has known a tremendous evolution during the past 35-40 years. In the future, it can be foreseen that this evolution will proceed in the direction of the use of more and more sophisticated and expensive machines. These apparatus, and the necessary human resources for their use, will only be affordable for laboratories with sufficient financial resources and having guarantee for a sufficient throughput of samples.

A liquid chromatography-electrospray ionisation tandem mass spectrometry method for the simultaneous detection and quantitation of 5-nitroimidazole veterinary drugs in lyophilised pork meat, the chosen format of a candidate certified reference material, has been developed and validated. Six analytes have been included in the scope of validation, i.e. dimetridazole (DMZ), metronidazole (MNZ), ronidazole (RNZ), hydroxymetronidazole (MNZOH), hydroxyipronidazole (IPZOH), and 2-hydroxymethyl-1-methyl-5-nitroimidazole (HMMNI). The analytes were extracted from the sample with ethyl acetate, chromatographically separated on a C(18) column, and finally identified and quantified by tandem mass spectrometry in the multiple reaction monitoring mode (MRM) using matrix-matched calibration and (2)H(3)-labelled analogues of the analytes (except for MNZOH, where [(2)H(3)]MNZ was used). The method was validated in accordance with Commission Decision 2002/657/EC, by determining selectivity, linearity, matrix effect, apparent recovery, repeatability and intermediate precision, decision limits and detection capabilities, robustness of sample preparation method, and stability of extracts. Recovery at 1 microg/kg level was at 100% (estimates in the range of 101-107%) for all analytes, repeatabilities and intermediate precisions at this level were in the range of 4-12% and 2-9%, respectively. Linearity of calibration curves in the working range 0.5-10 microg/kg was confirmed, with r values typically >0.99. Decision limits (CCalpha) and detection capabilities (CCbeta) according to ISO 11843-2 (calibration curve approach) were 0.29-0.44 and 0.36-0.54 microg/kg, respectively. The method reliably identifies and quantifies the selected nitroimidazoles in the reconstituted pork meat in the low and sub-microg/kg range and will be applied in an interlaboratory comparison for determining the mass fraction of the selected nitroimidazoles in the candidate reference material currently developed at IRMM.

A simple, selective and fast multi-residue method was developed to determine 18 veterinary drugs in milk by ultra-high-pressure liquid chromatography coupled to tandem quadrupole mass spectrometry. The selected antibiotics include quinolones, sulphonamides, macrolides, anthelmintics and one tetracycline. An extraction procedure based on QuEChERS methodology (quick, easy, cheap, effective, rugged and safe), consisting of a liquid extraction of the milk samples with acetonitrile without sample clean up was performed. The extract was centrifuged and the supernatant was filtered prior to chromatographic analysis. Several chromatographic conditions were optimized in order to obtain a fast separation (less than 10 min). The antibiotics were detected by electrospray ionization in positive ion mode with multiple reaction monitoring (MRM) and mass spectrometric conditions were optimized in order to increase selectivity and sensitivity. The developed method was validated in terms of linearity, trueness, precision, limits of detection (LODs) and quantification. Mean recoveries ranged from 70% to 110% and interday precision was lower than 21%. LODs ranged from 1 to 4 microg/kg. Finally, the method was applied to real samples and only traces of tylosin and fenbendezol were detected in two samples.