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Intestinal coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses for the industry. It compromises the nutrition absorption, leading to weight loss and elevated mortality rates. Furthermore, the stress caused by the infection can compromise the immune system, making poultry more susceptible to secondary infections and reducing overall productivity. As a result, simple analytical techniques are critical for determining anticoccidial drugs. A new, sensitive, and environmentally friendly HPLC method was developed for determining amprolium (AMP), sulfaquinoxaline (SUL), diaveridine (DIV), and vitamin K3 (VIT K3) in their formulations for the first time. Stability tests were performed under diverse stress conditions to verify the safety and efficiency of the formulation throughout its designated shelf time. These investigations ascertain the influence of various environmental conditions on a drug's chemical stability and physical characteristics. A Supelcosil C18 column was used as the stationary phase, and 0.05 M KH 2 PO 4 and acetonitrile were mixed in a ratio of 80:20 (v/v) as the developing system with a flow rate of 2.0 mL min -1 . The proposed drugs were quantified at 260 nm. It was tested and found that the novel analytical method was linear for AMP and SUL between 20.0 and 60.0 µg mL -1 , 2.0-6.0 µg mL -1 for VIT K3, and 2.1-6.3 µg mL -1 for DIV. The anticipated method was validated according to ICH guidelines. Advanced evaluation tools, such as GAPI, Red Green Blue (RGB 12, whiteness), Blue Applicability Grade Index (BAGI), the Analytical Eco-Scale, and (AGREE) assessed the sustainability profile of the proposed method, illustrating its enhanced environmental friendliness and sustainability., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Declaration of generative AI and AI-assisted technologies in the writing process: During the writing of this research article, the authors relied on QuillBot.com to check for grammar errors and make the content easier to read. Upon utilizing this tool or service, the authors assume complete responsibility for the content of the publication and have reviewed and updated it as required., (© 2025. The Author(s).)

Background: Amino acid supplements are crucial for animal health and productivity. Traditional analysis methods face limitations like complexity, long testing times and toxic reagents. Therefore, a more efficient and reliable method is needed., Objectives: This study aimed to develop and validate an efficient method for the simultaneous analysis of eight amino acids commonly used in veterinary medicine: alanine, arginine, glutathione, lysine, ornithine, methionine, threonine and tryptophan., Methods: We analysed eight veterinary amino acid preparations. From 100 registered products, we selected 35. After confirming ingredients, we diluted them to 1 mg/L with 50% acetonitrile (ACN) and filtered them using a 0.2 µm RC filter for liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis., Results: All analytes showed excellent linearity (r 2  > 0.99) within 0-10 mg/L. The limits of detection (LOD) ranged from 0.04 to 0.83 mg/L, and the limits of quantification (LOQ) ranged from 0.12 to 2.52 mg/L. Average recovery ranged from 92.96% to 105.61%, with relative standard deviations (RSD) from 0.27% to 3.50%, meeting CD 2002/657/EC standards. Six out of the 35 products (17.14%) did not meet regulations., Conclusions: The method developed in this study offers an efficient and reliable approach for the simultaneous analysis of essential amino acids in veterinary medicine. Implementing this method can improve the quality control of amino acid products, enhancing animal health and productivity. This study also highlights the need for stringent domestic management and continuous monitoring. By overcoming traditional technique limitations, this validated method ensures the quality and efficacy of amino acid supplements in the veterinary industry., (© 2025 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

The endangered Pyrenean Capercaillie (Tetrao urogallus aquitanicus) inhabits perennial forests of the Pyrenees (Spain, France and Andorre). Feces of domestic animals (e.g., horses and cattle) are often found in this species' habitat as evidence of land use overlapping, especially during spring and summer. As a result, pharmaceutical residues found in feces of these domestic ungulates may be absorbed by plants and insects that are part of the diet of Pyrenean Capercaillies (e.g., blueberries [Vaccinium uliginosum, Vaccinium myrtillus], red wood ants [Formica rufa]). Based on the absence of data regarding the exposure of Pyrenean Capercaillie to residues of veterinary medicinal products (VMP), we selected 71 compounds as indicators of anthropogenically-related environmental contamination, analyzed in 90 samples collected in several subalpine forests, northwestern Spain. Residues of several VMP were detected in feces (capercaillie [ciprofloxacin, enrofloxacin, tetracycline and florfenicol], horse [ciprofloxacin, enrofloxacin, tetracycline and ivermectin], and cattle [ciprofloxacin and enrofloxacin]), and in entomofauna (ciprofloxacin and ivermectin). No VMP residues were detected in blueberry plants. Herein, we present novel data about the presence of VMP residues in the Pyrenean Capercaillie's environment, and identify potential VMP sources (i.e., livestock feces and entomofauna) and an exposure route (i.e., food chain) for Capercaillie chicks. Further studies are necessary to investigate the potential indirect or chronic effects of VMP residues in the species' breeding success and adult fitness, which must be taken into account by managers and policy makers to improve management and conservation actions., 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 The Authors. Published by Elsevier Inc. All rights reserved.)

Veterinary medications used for disease treatment and prevention may remain in animal-origin foods, such as milk, eggs, honey and meat, which could pose a risk to the public's health. These drugs come from different groups of drugs, mostly with antibiotic, anti-parasitic or anti-inflammatory actions, in a range of food matrices including milk, meat or egg. This review is intended to provide the reader with a general insight about the current status of veterinary drug residues in food products of animal origin, detection methods and their public health consequences. The discovery of antimicrobials has led to the development of antibiotics for treating and preventing cattle illnesses and encouraging growth. However, the rise of drug resistance has led to increased antibiotic consumption and resistance among microbes in the animal habitat. This resistance can be passed to humans directly or indirectly through food consumption and direct or indirect interaction. Improper and illegal use, inadequate withdrawal periods and environmental contamination from veterinary drugs are reported to be the major causes for the formation of residue in food products of animal origin. The use of veterinary products above or below the advised level may also result in short- or long-term public health issues, such as the creation of resistant strains of micro-organisms, toxicity, allergy, mutagenesis, teratogenicity and carcinogenetic effects. To ensure consumer safety, veterinary drug residues in food must be under control., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Direct application of animal waste on farmlands was banned in China recently, rendering organic fertilizer production a sound solution for disposing of animal manures and recycling their materials and nutrients. Due to the overuse of antimicrobials in livestock and poultry farms, manure-based organic fertilizers often contain elevated residues of antimicrobials and abundant antimicrobial resistance genes. Land application of such products has caused significant concerns on the environmental pollution of antimicrobials, and the transmission and development of antimicrobial resistance (AMR), which is a major global health challenge. China's recent attempt to restrict the contents of antimicrobial residues in organic fertilizers encountered strong resistance from the industry as it would hinder the utilization of animal manures as a raw material. Reducing and even eliminating the use of antimicrobials in animal farms is the ultimate solution to the challenge of manure disposal posed by the elevated levels of antimicrobial residues and AMR. Phasing out the non-therapeutic use of antimicrobials, developing substitutes of antimicrobials, enhancing animal welfare in farms, promoting diversification of animal farms, and developing antimicrobial removal and disinfection technologies for animal waste are recommended to improve the veterinary antimicrobial stewardship and manure management in China's animal agriculture. These concerted measures would enhance the sustainability of crop and animal farming systems in China and mitigate the impact of antimicrobials and AMR to agro-environmental quality and human health., 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Humans are exposed to a cocktail of food-related and environmental contaminants, potentially contributing to the etiology of chronic diseases. Better characterizing the "exposome" is a challenging task and requires broad human biomonitoring (HBM). Veterinary drugs (VDs)/antibiotics, widely used and regulated in food and animal production, however, are typically not yet included in exposomics workflows. Therefore, in this work, a previously established multianalyte liquid chromatography-tandem mass spectrometry (LC-MS/MS) method covering >80 diverse xenobiotics was expanded by >40 VDs/antibiotics and pesticides. It was investigated if the generic workflow allowed for the successful integration of a high number of new analytes in a proof-of-principle study. The expanded method was successfully in-house validated and specificity, matrix effects, linearity, intra- and inter-day precision, accuracy, limits of quantification, and detection were evaluated. The optimized method demonstrated satisfactory recovery (81-120%) for most of the added analytes with acceptable RSDs (<20%) at three spiking levels. The majority of VDs/antibiotics and pesticides (69%) showed matrix effects within a range of 50-140%. Moreover, sensitivity was excellent with median LODs and LOQs of 0.10 ng/mL and 0.31 ng/mL, respectively. In total, the expanded method can be used to detect and quantify more than 120 highly diverse analytes in a single analytical run. To the best of the authors' knowledge, this work represents the first targeted biomonitoring method integrating VDs with various other classes of pollutants including plasticizers, PFAS, bisphenols, mycotoxins, and personal care products. It demonstrates the potential to expand targeted multianalyte methods towards additional groups of potentially toxic chemicals., (© 2024. The Author(s).)

Milk is an important consumer product with high nutritional value. The presence of veterinary drug residues in milk owing to the indiscriminate use of veterinary drugs may affect consumer health. In the mass spectrometric analysis of trace compounds, chromatographic co-eluting components easily interfere with the mass spectral signals obtained, affecting the accuracy of qualitative and quantitative analyses. Matrix purification is a promising method to reduce the matrix effect. Chitosan is a natural biopolymer with numerous active functional groups such as amino, acetyl, and hydroxyl groups; these groups can adsorb lipids through hydrophobic and electrostatic interactions. Chitosan also has the advantages of low production cost, stable chemical properties, and convenient modification. Novel chitosan-based materials are promising candidates for lipid purification. In this study, a chitosan membrane was modified with trimethoxyoctadecylsilane (C 18 -CSM). C 18 -CSM was prepared through one-step hydrolysis and used as a dispersive solid phase extraction (DSPE) adsorbent to purify the matrix during milk pretreatment. We combined C 18 -CSM with ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry (UHPLC-Q/Exactive Orbitrap MS) to develop an effective method for the extraction and determination of ofloxacin, enrofloxacin, ciprofloxacin, diazepam, and metronidazole in milk. C 18 -CSM was characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, and water contact angle testing. The results indicated that the material has a rough surface and uniformly dense cross-section. The water contact angle of C 18 -CSM was 104°, indicating its good hydrophobicity. The pretreatment conditions (extraction solvent, dosage of NaCl, extraction frequency, and dosage of C 18 -CSM) that influenced the recoveries of the five veterinary drugs were investigated in detail. The optimal conditions were established as follows: 5% formic acid in acetonitrile, 1 g NaCl, extraction 1 time, 20 mg C 18 -CSM. Separation was performed on a Hypersil GOLD VANQUISH column (100 mm×2.1 mm, 1.9 μm). The mobile phase consisted of 0.1% formic acid aqueous solution and 0.1% formic acid in acetonitrile, and was flowed at a rate of 0.3 mL/min. The sample injection volume was 1 μL, and the column temperature was maintained at 25 ℃. Mass spectrometric analysis was performed in positive electrospray ionization mode. To verify the necessity of the purification material, the matrix effect was investigated using the matrix-matched standard curve method. The use of C 18 -CSM reduced the matrix effects of the five necessity drugs from the range of -22%-8.8% to the range of -13%-3.6%, indicating that C 18 -CSM is a highly efficient DSPE material. Under optimal conditions, the developed method showed good linearities within the range of 0.5-100 μg/L, with correlation coefficients ( r 2 )≥0.9970. The limits of detection(LODs) and quantification (LOQs) were 0.2 μg/L and 0.5 μg/L, respectively. To assess the accuracy and precision of the method, we prepared milk samples with three spiked levels (low, medium, and high). The recoveries of the five veterinary drugs were ranged from 79.5% to 115%, and the intra-day and inter-day relative standard deviations were 7.0%-13% ( n =6) and 1.3%-11% ( n =3), respectively. This study provides a simple, accurate, and reliable method for the rapid and simultaneous determination of the five veterinary drug residues in milk.

The contamination risks of plant-derived foods due to the co-existence of pesticides and veterinary drugs (P&VDs) have not been fully understood. With an increasing number of unexpected P&VDs illegally added to foods, it is essential to develop a non-targeted screening method for P&VDs for their comprehensive risk assessment. In this study, a modified support vector machine (SVM)-assisted metabolomics approach by screening eligible variables to represent marker compounds of 124 multi-class P&VDs in maize was developed based on the results of high-performance liquid chromatography-tandem mass spectrometry. Principal component analysis and orthogonal partial least squares discriminant analysis indicate the existence of variables with obvious inter-group differences, which were further investigated by S-plot plots, permutation tests, and variable importance in projection to obtain eligible variables. Meanwhile, SVM recursive feature elimination under the radial basis function was employed to obtain the weight-squared values of all the variables ranging from large to small for the screening of eligible variables as well. Pairwise t -tests and fold changes of concentration were further employed to confirm these eligible variables to represent marker compounds. The results indicate that 120 out of 124 P&VDs can be identified by the SVM-assisted metabolomics method, while only 109 P&VDs can be found by the metabolomics method alone, implying that SVM can promote the screening accuracy of the metabolomics method. In addition, the method's practicability was validated by the real contaminated maize samples, which provide a bright application prospect in non-targeted screening of contaminants. The limits of detection for 120 P&VDs in maize samples were calculated to be 0.3~1.5 µg/kg.

This work compares the electroanalytical performance of two electroanalytical systems based on (1) the glassy carbon electrode (GCE), and (2) the electrified liquid-liquid interface (eLLI), for the detection of fluoroquinolone antibiotic-danofloxacin (DANO). Our aim was to define the optimal conditions to detect the chosen analyte with two employed systems, extract a number of electroanalytical parameters, study the mechanism of the charge transfer reactions (oxidation at GCE and ion transfer across the eLLI), and to provide physicochemical constants for DANO. Detection of the chosen analyte was also performed in the spiked milk samples. To the best of our knowledge, this is the first work that directly compares the electroanalytical parameters obtained with solid electrode (in this case GCE) and eLLI. We have found that for DANO the latter provides better electroanalytical parameters (lower LOD and LOQ) as well as good selectivity when the milk was analyzed., (© 2024. The Author(s).)

A rapid and online microvolume flow-through dialysis probe designed for sample preparation in the analysis of veterinary drug residues is introduced. This study addresses the need for efficient and green sample preparation methods that reduce chemical waste and reagent use. The dialysis probe integrates with liquid chromatography and mass spectrometry (LC-MS) systems, facilitating automated, high-throughput analysis. The dialysis method utilizes minimal reagent volumes per sample, significantly reducing the generation of solvent waste compared to traditional sample preparation techniques. Several veterinary drugs were spiked into tissue homogenates and analyzed to validate the probe's efficacy. A diagnostic sensitivity of >97% and specificity of >95% were obtained for this performance evaluation. The results demonstrated the effective removal of cellular debris and particulates, ensuring sample integrity and preventing instrument clogging. The automated dialysis probe yielded recovery rates between 27 and 77% for multiple analytes, confirming its potential to streamline veterinary drug residue analysis, while adhering to green chemistry principles. The approach highlights substantial improvements in both environmental impact and operational efficiency, presenting a viable alternative to conventional sample preparation methods in regulatory and research applications.

A method was established for the simultaneous detection of 12 prohibited veterinary drugs, including β 2 -receptor agonists, nitrofuran metabolites, nitroimidazoles, chlorpromazine, and chloramphenicol, in pig urine. The sample was pretreated by enzymolysis, acid hydrolysis/derivatization, and liquid-liquid extraction combined with solid-phase extraction. Detection was performed using ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Ammonium acetate solution (0.2 mol/L, 4.5 mL) and β -glucuronidase/aryl sulfatase (40 μL) were added to the sample, which was subsequently enzymolized at 37 ℃ for 2 h. Then, 1.5 mL of 1.0 mol/L hydrochloric acid solution and 100 μL of 0.1 mol/L o -nitrobenzaldehyde solution were added to the sample. The mixture was incubated at 37 ℃ for 16 h, and the analytes were extracted with 8 mL of ethyl acetate by liquid-liquid extraction. The lower aqueous phase obtained after extraction was extracted and purified using a mixed cation-exchange solid-phase extraction column. The extracts were combined, the extraction solution was blow-dried with nitrogen, and the residue was redissolved for determination. The samples were analyzed under multiple-reaction monitoring mode with both positive and negative electrospray ionization, and quantified using an isotope internal standard method. The correlation coefficients ( r ) of the 12 compounds were >0.99. The limits of detection (LODs) and quantification (LOQs) of chloramphenicol were 0.05 and 0.1 μg/L, respectively, and the LODs and LOQs of the other compounds were 0.25 and 0.5 μg/L, respectively. The mean recoveries and RSDs at 1, 2, and 10 times the LOQ were 83.6%-115.3% and 2.20%-12.34%, respectively. The proposed method has the advantages of high sensitivity, good stability, and accurate quantification; thus, it is suitable for the simultaneous determination of the 12 prohibited veterinary drug residues in pig urine.

The consumption of poultry eggs has increased in recent years owing to the abundance of production and improvements in living standards. Thus, the safety requirements of poultry eggs have gradually increased. At present, few reports on analytical methods to determine banned veterinary drugs during egg-laying period in poultry eggs have been published. Therefore, establishing high-throughput and efficient screening methods to monitor banned veterinary drugs during egg-laying period is imperative. In this study, an analytical method based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with QuEChERS-based techniques was developed for the simultaneous determination of 31 banned veterinary drugs encompassing nine drug classes (macrolides, antipyretic and analgesic drugs, sulfonamides, antibacterial synergists, anticoccidials, antinematodes, quinolones, tetracyclines, amphenicols) in different types of poultry eggs. The main factors affecting the response, recovery, and sensitivity of the method, such as the extraction solvent, purification adsorbent, LC separation conditions, and MS/MS parameters, were optimized during sample pretreatment and instrumental analysis. The 31 veterinary drug residues in 2.00 g eggs were extracted with 2 mL of 0.1 mol/L ethylene diamine tetraacetic acid disodium solution and 8 mL 3% acetic acid acetonitrile solution, and salted out with 2 g of sodium chloride. After centrifugation, 5 mL of the supernatant was cleaned-up using the QuEChERS method with 100 mg of octadecylsilane-bonded silica gel (C 18 ), 50 mg of N -propylethylenediamine (PSA), and 50 mg of NH 2 -based sorbents. After nitrogen blowing and redissolution, the 31 target analytes were separated on a Waters CORTECS UPLC C 18 analytical chromatographic column (150 mm×2.1 mm, 1.8 μm) at a flow rate, column temperature, and injection volume of 0.4 mL/min, 30 ℃, and 5 μL, respectively. Among these analytes, 26 analytes were acquired in dynamic multiple reaction monitoring (MRM) mode under positive electrospray ionization (ESI + ) conditions using (A) 5 mmol/L ammonium acetate (pH 4.5) and (B) acetonitrile as mobile phases. The gradient elution program was as follows: 0-2.0 min, 12%B-30%B; 2.0-7.5 min, 30%B-50%B; 7.5-10.0 min, 50%B; 10.0-10.1 min, 50%B-100%B; 10.1-12.0 min, 100%B; 12.0-12.1 min, 100%B-12%B; The five other target analytes were acquired in MRM mode under negative electrospray ionization (ESI - ) conditions using (A) H 2 O and (B) acetonitrile as mobile phases. The gradient elution program was as follows: 0-2.0 min, 12%B-40%B; 2.0-6.0 min, 40%B-80%B; 6.0-6.1 min, 80%B-100%B; 6.1-8.0 min, 100%B; 8.0-8.1 min, 100%B-12%B. Matrix-matched external standard calibration was used for quantification. The results showed that all the compounds had good linear relationships within their respective ranges, with correlation coefficients of >0.99. The limits of detection (LODs) and quantitation (LOQs) were 0.3-3.0 μg/kg and 1.0-10.0 μg/kg, respectively. The average recoveries of the 31 banned veterinary drugs spiked at three levels (LOQ, maximum residue limit (MRL), and 2MRL) in poultry eggs ranged from 61.2% to 105.7%, and the relative standard deviations (RSDs) ranged from 1.8% to 17.6%. The developed method was used to detect and analyze banned veterinary drugs in 30 commercial poultry egg samples, including 20 eggs, 5 duck eggs, and 5 goose eggs. Enrofloxacin was detected in one egg with a content of 12.3 μg/kg. The proposed method is simple, economical, practical, and capable of the simultaneous determination of multiple classes of banned veterinary drugs in poultry eggs.

Food safety is recognized as a main requirement for consumers, food industries, and official laboratories. Here, we present the optimization and screening qualitative validation of two multianalyte methods in bovine muscle tissues by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry with an Orbitrap-type analyzer, operated with a heated ionization source in positive and negative mode. This aims for not only the simultaneous detection of veterinary drugs regulated in Brazil but also the prospection of antimicrobials not yet monitored. Two different sample preparation procedures were applied: method A-generic solid-liquid extraction with 0.1% formic acid ( v / v ) in an aqueous solution of EDTA 0.1% ( w / v )-acetonitrile-methanol (1:1:1, v / v / v ), followed by an additional ultrasound-assisted extraction and method B-QuEChERS. In both procedures, selectivity showed satisfactory conformity. From a detection capability (CCβ) equivalent to ½ the maximum residue limit, >34% of the analyte resulted in a false positive rate of <5%, preponderant by the QuEChERS method, which exhibited a higher yield of the sample. The results showed the potential application of both procedures in the routine analysis of foods by official laboratories, enabling the expansion of this methodological portfolio as well as its analytical scopes, thus optimizing the control of residues of veterinary drugs in the country.

Background: Otitis externa is a commonly diagnosed dermatological disorder in canines. The pathogens primarily involved in canine otitis externa (COE) include Staphylococcus pseudintermedius, Pseudomonas aeruginosa, Proteus mirabilis, and Malassezia pachydermatis. As COE tends to be superficial, medications delivered topically are often effective and practical in managing the condition. As such, there is a wide variety of approved topical products currently available in the market. The efficacy of topical dosage forms can be dependent on various factors such as the pharmacology of active constituents and the physicochemical properties of the formulation, including pH, viscosity, spreadability, and bio-adhesion. Currently, there is a lack of published literature available on the optimal properties of topical COE products. In this study, we compared the physicochemical properties of nine commercially available otic veterinarian products in Australia used clinically to manage COE., Results: Based on our comparative analysis, the pH (6.26 ± 0.04) of an aqueous-based product was similar to a healthy dog's external auditory canal. Products containing polymers exhibited higher viscosity and bio-adhesion. Spreadability was inversely related to viscosity and Osurnia ® a product with high viscosity demonstrated the lowest spreadability. Aqueous-based otic products showed better syringebility whereas oil-based systems required higher force to expel the products. Variability in droplet size was noted. Derm Otic, Baytril Otic, and Aurizon Ear Drops had the lower standard deviation which indicates they would give a more consistent dose., Conclusions: Findings from this work provide considerations for industry researchers or formulation scientists working in the area of otic dosage formulations., (© 2023. The Author(s).)

Modern agriculture depends highly on pesticides and pharmaceutical preparations, so controlling exposure to these substances in the feed and food chain is essential. This article presents the first study on residues of a broad spectrum of pesticides and veterinary drugs in the diets of dairy cattle. One hundred and two representative samples of the complete diets, including basal feed rations and additional fed concentrate, were collected in three Austrian provinces (Styria, Lower and Upper Austria) in 2019 and 2020. The samples were tested for >700 pesticides, veterinary drugs and related metabolites using a validated method based on liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS). In total, 16 residues (13 pesticides and three veterinary drug residues) were detected. > 90% of the diets contained pesticide residues and <10% veterinary drug residues, whereas banned pesticides were not found. The most frequent pesticide residues were fluopyram (62%), piperonyl butoxide (39%) and diethyltoluamide (35%). The following pesticides exceed the default EU maximum residue level (MRL) (10 μg kg -1 ) for products exclusively used for animal feed production: Benzovindiflupyr (proportion of samples > MRLs: 1%), bixafen (2%), fluopyram (6%), ipconazole (1%) and tebuconazole (3%). Three residues (dinitrocarbanilide, monensin and nicarbazin) of veterinary drugs were identified, all below the MRLs. Over 60% of the evaluated samples contained mixtures of two to six residues/sample. Only one pesticide (diethyltoluamide) presented a significant difference among regions, with higher concentrations in Upper Austria. Brewery's spent grains were the dietary ingredient that showed the strongest correlation to pesticide residues. These findings evidence the realistic scenario of highly occurrent low doses of pesticides cocktails in the feed/food chain, which may affect the animal, human and environmental health. Since the risk assessments are based on single pesticides, the potential synergistic effect of co-occurring chemicals ("cocktail effect") requires further investigations., 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 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The presence of unauthorized substances, such as residues of veterinary medicines or chemical contaminants, in food can represent a possible health concern. For this reason, a complete legislative framework has been established in the European Union (EU), which defines the maximum limits allowed in food and carries out surveillance programs to control the presence of these substances. Official food control laboratories, in order to ensure a high level of consumer protection, must respond to the challenge of improving and harmonizing the performance of the analytical methods used for the analysis of residues of authorized, unauthorized, or prohibited pharmacologically active substances. Laboratories must also consider the state of the art of the analytical methodologies and the performance requirements of current legislation. The aim of this work was to develop a multiresidue method for the determination of antibiotics in milk, compliant with the criteria and procedures established by Commission Implementing Regulation (EU) 2021/808. The method uses an LC-Orbitrap-HRMS for the determination of 57 molecules of antibiotic and active antibacterial substances belonging to different chemical classes (beta-lactams, tetracyclines, sulfonamides, quinolones, pleuromutilins, macrolides, and lincosamides) in bovine, ovine, and goat milk samples. It provides a simple and quick sample pretreatment and a subsequent identification phase of analytes, at concentrations equal to or lower than the maximum residual limit (MRL), in compliance with Commission Regulation (EU) 2010/37. The validation parameters: selectivity, stability, applicability, and detection capability (ccβ), are in agreement with the requirements of Commission Implementing Regulation (EU) 2021/808 and demonstrated the effectiveness of the method in detecting veterinary drug residues at the target screening concentration (at the MRL level or below), with a false positive rate of less than 5%. This method represents an effective solution for detecting antibiotics in milk, which can be successfully applied in routine analyses for official food control plans.

A method was developed for the rapid and quantitative analysis of 30 veterinary drugs belonging to 17 classes (amphenicols (1), anthelmintics (1), cephalosporins (4), coccidiostats (1), lincosamides (1), macrolide (1), nitroimidazole (1), penicillins (3), phenylhydrazines (1), polypeptides (1), pyrethrins (1), quinolones (5), sulfonamides (3), tetracycline (3), neuroleptic agents (1), triazene trypanocidal agents (1), other. (1)) in feeds. The proposed method with a modified Quick Polar Pesticides (QuPPe) sample preparation was validated for the determination of 30 veterinary drugs in feed samples by liquid chromatography triple-quadrupole mass spectrometry (LC-MS/MS). The sample was extracted with methanol containing 1% acetic acid and purified by dispersive solid-phase extraction (d-SPE) with C18. Good linearity ( r 2 ≥ 0.98) was observed, and the LOQ values ranged from 10 to 200 µg/kg. Average recoveries ranged from 70.8 to 118.4%, and the relative standard deviation was ≤ 18.7%. This validated method was used in the determination of 30 veterinary drugs in 142 feed samples obtained from South Korea. The results show that lincomycin was present in only one of the tested feed samples, although it was detected at a value lower than the LOQ. In conclusion, this multi-residue method can be used for screening through the detection and quantitation of residual multiclass veterinary drugs in feed samples.

Manure is a major source of soil and plant contamination with veterinary drugs residues. The aim of this study was to evaluate the uptake of 14 veterinary pharmaceuticals by parsley from soil fertilized with manure. Pharmaceutical content was determined in roots and leaves. Liquid chromatography coupled with tandem mass spectrometry was used for targeted analysis. Screening analysis was performed to identify transformation products in the parsley tissues. A solid-liquid extraction procedure was developed combined with solid-phase extraction, providing recoveries of 61.9-97.1% for leaves and 51.7-95.6% for roots. Four analytes were detected in parsley: enrofloxacin, tylosin, sulfamethoxazole, and doxycycline. Enrofloxacin was detected at the highest concentrations (13.4-26.3 ng g -1 ). Doxycycline accumulated mainly in the roots, tylosin in the leaves, and sulfamethoxazole was found in both tissues. 14 transformation products were identified and their distribution were determined. This study provides important data on the uptake and transformation of pharmaceuticals in plant tissues.

Objectives: Antibiotics are important for improving animal health and production. However, the deposition of its residues in food of animal origin intended for human consumption at non-permissible levels has generated global health concern and the need to tackle this using the "One Health Approach". This study assessed the knowledge and practice of 286 cattle handlers in Kwara State, Nigeria., Methods: A web-based cross sectional online survey using a semi-structured questionnaire was conducted from November to December, 2019. Univariate, bivariate and multivariate analyses were performed at 95% confidence interval to determine predictors of good knowledge and practices towards Antibiotic Residues in Meat and Milk among cattle handlers., Results: This study revealed that majority (52.7% n = 165/286) of the cattle handlers were not aware of antibiotic residues. Knowledge and practices regarding antibiotic residues were generally poor among the study population; 36.7% and 35.5% had satisfactory knowledge and practice respectively. The age (p = 0.026), gender (p = 0.006) and business duration (p = 0.001) of participants were significantly associated with their knowledge of antimicrobial residues. The effect of education on knowledge was modified by age. The odds of having poor knowledge on antibiotic residues increased 4 times among participants who were ≤40 years old than those above 40 years (Stratum Specific OR = 3.65; CI = 1.2, 11.1; p = 0.026). Knowledge levels of participants were statistically associated with their practice levels p<0.05 (OR = 2.43; CI = 1.45. 4.06; p = 0.0006)., Conclusion: This implies that poor knowledge is a risk factor to having poor practice among cattle handlers. Deliberate efforts towards educating cattle farmers on best farm practices in antibiotic use would prevent antibiotic residues in meat and milk. Also, an effective surveillance system for monitoring the use of veterinary drugs in Kwara State, Nigeria is crucial., Competing Interests: The authors have declared that no competing interests exist.

An innovative, rapid and stable method for simultaneous determination of three tetracycline (oxytetracycline, tetracycline and doxycycline) and two fluoroquinolone (ciprofloxacin and enrofloxacin) residues in poultry eggs by ultra-high performance liquid chromatography-fluorescence detection (UPLC-FLD) was established and optimized. The samples were homogenized and extracted with acetonitrile/ultrapure water (90:10, v/v ) and then purified by solid-phase extraction (SPE). LC separation was achieved on an ACQUITY UPLC BEH C18 column (1.7 µm, 2.1 mm × 100 mm), and the mobile phase was composed of acetonitrile and a 0.1 mol/L malonic acid solution containing 50 mmol/L magnesium chloride (the pH was adjusted to 5.5 with ammonia). When the five target drugs were spiked at the limit of quantification, 0.5 times the maximum residue limit (MRL), 1.0 MRL and 2.0 MRL, the recoveries were above 83.5% and the precision ranged from 1.99% to 6.24%. These figures of merit complied with the parameter validation regulations of the EU and U.S. FDA. The limits of detection and quantifications of the targets were 0.1-13.4 µg/kg and 0.3-40.1 µg/kg, respectively. The proposed method was easily extended to quantitative analyses of target drug residues in 85 egg samples, thus demonstrating its reliability and applicability.

Detection of relevant contaminants using screening approaches is a key issue to ensure food safety and respect for the regulatory limits established. Electrochemical sensors present several advantages such as rapidity; ease of use; possibility of on-site analysis and low cost. The lack of selectivity for electrochemical sensors working in complex samples as food may be overcome by coupling them with molecularly imprinted polymers (MIPs). MIPs are synthetic materials that mimic biological receptors and are produced by the polymerization of functional monomers in presence of a target analyte. This paper critically reviews and discusses the recent progress in MIP-based electrochemical sensors for food safety. A brief introduction on MIPs and electrochemical sensors is given; followed by a discussion of the recent achievements for various MIPs-based electrochemical sensors for food contaminants analysis. Both electropolymerization and chemical synthesis of MIP-based electrochemical sensing are discussed as well as the relevant applications of MIPs used in sample preparation and then coupled to electrochemical analysis. Future perspectives and challenges have been eventually given.

In 2018, AOAC International issued Standard Method Performance Requirements (SPMR) 2018.010 - Screening and Identification Method for Regulated Veterinary Drug Residues in Food. In response, we compared 4 different multiresidue methods of sample preparation using the same analytical method entailing ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Tilapia was chosen for testing, and the analytes and monitoring levels were from SPMR 2018.010. The methods consist of efficient procedures with published validation results from the US Department of Agriculture (USDA), Food and Drug Administration (FDA), and Canadian Food Inspection Agency (CFIA), and an enhanced-matrix removal (EMR)-Lipid protocol from China. Each method was used to prepare 102 final extracts of tilapia spiked or not at different levels with the 78 targeted analytes plus metabolites. The same FDA/USDA rules of mass spectral identification were employed in all analyses to assess rates of false positives and negatives. Quantitative accuracy of the methods was also compared in terms of recoveries and reproducibility of spiked tilapia, incurred catfish, and spiked and certified reference material of bovine muscle. Each method yielded generally acceptable results for the targeted veterinary drugs, but the USDA "extract & inject" method was the fastest, simplest, and cheapest to achieve equally or more acceptable results for the widest scope of analytes for the tested food matrices.

Chloramphenicols, nitroimidazoles, lincosamides, and macrolides are common antibiotics used in veterinary medicine. Overdoses of these drugs will lead to residual substances in animal-derived foods and accumulate in the body through the food chain, thereby exerting adverse effects on human health. Therefore, regulation of veterinary drug levels is imperative to ensure the quality of animal-derived foods and safeguard the health of consumers. In this study, a method based on ultra performance liquid chromatography-tandem mass spectrometry coupled with solid phase extraction (SPE-UPLC-MS/MS) was developed for the simultaneous determination of eight prohibited and restricted veterinary drugs and three metabolite residues across four categories (chloramphenicols, nitroimidazoles, lincosamides, and macrolides) in eggs, liquid milk, chicken, and freshwater fish. The main factors affecting the response, recovery, and sensitivity of the method, such as the type and pH values of the extraction solvent, dilution solution for the analytes, type of chromatographic column, and type and proportion of the mobile phase, were optimized during sample pretreatment and instrument analysis. The samples were hydrolyzed and dispersed in 0.1 mol/L phosphate buffer solutions (pH 9.0) and extracted with acetonitrile. The extract was further extracted using ethyl acetate. After centrifugation, the supernatant ethyl acetate was concentrated to near dryness in nitrogen below 40 ℃. The residue was dissolved in 0.3 mL methanol, followed by the addition of 5.7 mL phosphate buffer solution. After shaking, the solutions were purified and enriched on an Oasis HLB SPE column. The target analytes were separated on an ACQUITY UPLC BEH C18 chromatographic column (100 mm×2.1 mm, 1.7 μm) at a column temperature of 40 ℃ with a flow rate of 0.4 mL/min. The injection volume was 10 μL. Gradient elution was carried out with methanol and 0.1% formic acid aqueous solution as the mobile phases. Multiple reaction monitoring (MRM) was conducted in the positive and negative electrospray ionization modes. The isotope internal standard method was used for quantitative analysis. Under optimal conditions, each analyte showed a good linear relationship in each range, and the correlation coefficient ( R 2 ) was greater than 0.99. The limits of detection (LODs) ranged from 0.050 to 0.50 μg/kg, and the limits of quantification (LOQs) ranged from 0.20 to 1.5 μg/kg. With eggs, freshwater fish, chicken, and liquid milk as the matrix samples, the recoveries in spiked blank samples were determined at different addition levels in compliance with the current legislation. The average recoveries of the 11 analytes were 65.3% to 108%. The relative standard deviations (RSDs) were between 0.40% and 21%. The matrix effects of the analytes were between 0.0124% and 46.80% in four different samples after purification on the Oasis HLB column. The practicality of the proposed approach for routine analyses of the eight prohibited and restricted veterinary drugs, and three metabolite residuals was evaluated by applying it to the determination of these compounds in animal-derived food samples. The samples, including 80 eggs, 80 chicken, 40 liquid milk, and 32 freshwater fish, were procured from a supermarket and a farm product market. The results of the positive samples were consistent with those observed with the standard methods. The method described herein is easy to operate, sensitive, and accurate. It is suitable for the simultaneous and rapid determination of various prohibited and restricted veterinary drug residues and metabolites in animal-derived foods.

Access to quality veterinary antimicrobial products contributes to efficient treatment of diseases in Vietnamese livestock and to reducing antimicrobial resistance (AMR). Poor quality antimicrobial drugs can lead to treatment failure, potentially influencing the inappropriate use of antimicrobials products, including increasing the dose, combining drugs, or changing to a broader spectrum antimicrobial. The objective of the study was to determine the actual concentration of antimicrobial active ingredient (AAI) in commercially available veterinary antimicrobial products as an indicator of their quality. A total of 144 veterinary antimicrobial products were purchased from randomly selected veterinary drug stores in 34 districts in eight provinces. For the qualitative analysis, we observed criteria linked to form, colour, and labelling information according to the Department of Animal Health regulations. For the quantitative analysis, high-performance liquid chromatography was used to determine the actual concentration of AAI in each sample. Of the 144 samples, 131 (91%) met the national standard of quality of being within ±10% of the labelled concentration. Ten antimicrobials (6.9%) contained less than half of the labelled content concentrations. Veterinary antimicrobial product quality control is an important part of addressing AMR. To support the national action plan to lower AMR, a veterinary drug quality control program should be implemented at all stages of the supply chain to assure high quality drugs and effective treatment of sick animals., Competing Interests: The authors have declared that no competing interests exist.

Monitoring of food contaminants and residues has undergone a significant improvement in recent years and is now performed in an intensive manner. Achievements in the area of chromatography-mass spectrometry coupling techniques enabled the development of quantitative multi-target approaches covering several hundred analytes. Although the majority of methods are focusing on the analysis of one specific group of substances, such as pesticides, mycotoxins, or veterinary drugs, current trends are going towards the simultaneous determination of multiclass compounds from several families of contaminants and residues. This work provides an overview of relevant multiclass concepts based on LC-MS/MS and LC-HRMS instruments. Merits and shortcomings will be critically discussed based on current performance characteristics of the EU legislation system. In addition, the discussion of a recently developed multiclass approach covering >1000 substances is presented as a case study to illustrate the current developments in this area.

The excessive use or abuse of pesticides and veterinary drugs leads to residues in food, which can threaten human health. Therefore, there is an extremely urgent need for multi-analyte analysis techniques for the detection of pesticide and veterinary drug residues, which can be applied as screening techniques for food safety monitoring and detection. Recent developments related to rapid multi-residue detection methods for pesticide and veterinary drug residues are reviewed herein. Methods based on different recognition elements or the inherent characteristics of pesticides and veterinary drugs are described in detail. The preparation and application of three broadly specific recognition elements-antibodies, aptamers, and molecular imprinted polymers-are summarized. Furthermore, enzymatic inhibition-based sensors, near-infrared spectroscopy, and SERS spectroscopy based on the inherent characteristics are also discussed. The aim of this review is to provide a useful reference for the further development of rapid multi-analyte analysis of pesticide and veterinary drug residues.

Honey is a natural food widely consumed due to its high content in nutrients and bioactive substances. In order to prevent hive infections, xenobiotics such as pesticides and antibiotics are commonly used. Chloramphenicol (CAP) is a broad-spectrum antibiotic used to treat honeybee larvae diseases. However, CAP has toxic and nondose-dependent effects in sensitive subjects; for this reason, its use has been prohibited in food-producing animals, such as the honeybee. In this study, we proposed a rapid, simple, and cheap analytical method, based on salting-out assisted liquid-liquid extraction coupled with UHPLC MS/MS detection for the accurate determination of CAP in honey to be used in routine analyses. The parameters that influence the extraction efficiency have been optimized using an experimental design in order to maximize the recovery of the analyte by reducing the matrix effects. Therefore, the developed method was internally validated according to the 2002/657/EC Decision guidelines and applied to the analysis of 96 honey samples.

Hazardous chemicals in food are an important cause of food safety problems. Mass spectrometry is an effective tool for the qualitative and quantitative analysis of these substances. In this paper, the fragmentation mechanisms for several chemical hazardous substances, including pesticides, veterinary drugs, mycotoxins, and other chemical pollutants classified by structural analogs, are reviewed. For each class of compounds, we summarize the characteristic fragments and neutral loss generated by cleavage in the mass spectrometry analysis. We also summarize the mechanisms applied to screen and discover new structural analogs in food. This review can help researchers analyze and confirm the structure of compounds and provide a theoretical basis for the discovery of new structural analogs in food.

In vivo, drug molecules interact with their biological targets (e.g., enzymes, receptors, ion channels, transporters), thereby eliciting therapeutic effects. Assays that measure the interaction between drugs and bio-targets may be used as drug biosensors, which are capable of broadly detecting entire drug classes without prior knowledge of their chemical structure. This Trends article covers recent developments in bio-target-based screening assays for detecting drugs associated with the following areas: illicit products marketed as dietary supplements, food-producing animals, and bodily fluids. General challenges and considerations associated with using bio-target assays are also presented. Finally, future applications of these assays for drug detection are suggested based upon current needs.

A method was established for the rapid determination of 28 prohibited and restricted veterinary drugs of five classes (sulfonamides, quinolones, chloramphenicols, nitrofurans metabolites and triphenylmethane) in fish and shrimp by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) coupled with simultaneous extraction and purification. After optimizing the sample pretreatment methods and chromatographic conditions, the samples were hydrolyzed with hydrochloric acid. The five kinds of residual drugs were simultaneously extracted with acetonitrile after nitrofuran metabolites derivatized with 2-nitrobenzaldehyde. The extraction solutions were dehydrated with sodium chloride and salted out. The acetonitrile layer was purified with n -hexane and concentrated to near dryness. The residues were dissolved in 1.0 mL methanol. Then, the solutions were purified by dispersive solid phase extraction with C18, N -( n -propyl)ethylenediamine (PSA) and amino bonded silica gel (NH2) sorbents. The residues were separated on a ZORBAX C18 chromatographic column (50 mm×2.1 mm, 1.8 μm). The MS data were acquired in positive and negative multiple reaction monitoring (MRM) modes, and quantitated by the isotope internal standard method. With this approach, the linear relationships of the 28 veterinary drugs were good. The linear correlation coefficients were greater than 0.99. The limits of detection and limits of quantification were in the range of 0.1-0.8 μg/kg and 0.3-2.7 μg/kg, respectively. The average recoveries at three spiked levels (1, 5, and 20 fold of the LOQs) were 70.0%-120% with relative standard deviations of 0.9%-10.0%. The method is simple, rapid, accurate and sensitive, and it is suitable for the rapid determination of various prohibited and restricted drugs in fish and shrimp.

This study was performed to optimize the analytical method for multi-residues of 60 compounds in flatfish samples. Three sample preparation methods were tested to identify the optimal recovery conditions for target analytes. As a result, 10 mL of water/acetonitrile (1:4, v / v ) was used to extract analytes from fish samples. For purification, C 18 and 10 mL of acetonitrile saturated hexane were used to treat the samples. After evaporation and reconstitution, the fish samples were analyzed by ultra-performance liquid chromatography-tandem mass spectrometry. The proposed method was validated according to the CODEX guidelines (CAC/GL-71). Our results showed the recoveries of 73.2%-115% and coefficients of variation of 1.6%-22.1%. The limit of quantification was 0.0005-0.005 mg/kg in the fishery products. In analysis of real samples, no samples exceeded the limit of quantification. This analytical method can be used for multi-residue screening and confirmation of the residues of veterinary drugs in fishery products., Competing Interests: The authors declare no conflict of interest.

Recent years have seen the increased utilization of ionic liquids (ILs) in the development and optimization of analytical methods. Their unique and eco-friendly properties and the ability to modify their structure allows them to be useful both at the sample preparation stage and at the separation stage of the analytes. The use of ILs for the analysis of pharmaceuticals seems particularly interesting because of their systematic delivery to the environment. Nowadays, they are commonly detected in many countries at very low concentration levels. However, due to their specific physiological activity, pharmaceuticals are responsible for bioaccumulation and toxic effects in aquatic and terrestrial ecosystems as well as possibly upsetting the body's equilibrium, leading to the dangerous phenomenon of drug resistance. This review will provide a comprehensive summary of the use of ILs in various sample preparation procedures and separation methods for the determination of pharmaceuticals in environmental and biological matrices based on liquid-based chromatography (LC, SFC, TLC), gas chromatography (GC) and electromigration techniques (e.g., capillary electrophoresis (CE)). Moreover, the advantages and disadvantages of ILs, which can appear during extraction and separation, will be presented and attention will be given to the criteria to be followed during the selection of ILs for specific applications.

In the last years, the European Commission has adopted restrictive directives on food quality and safety in order to protect animal and human health. Veterinary drugs represent an important risk and the need to have sensitive and fast analytical techniques to detect and quantify them has become mandatory. Over the years, the availability of different modes, interfaces, and formats has improved the versatility, sensitivity, and speed of capillary electrophoresis (CE) techniques. Thus, CE represents a powerful tool for the analysis of a large variety of food matrices and food-related molecules with important applications in food quality and safety. This review focuses the attention of CE applications over the last decade on the detection of different classes of drugs (used as additives in animal food or present as contaminants in food products) with a potential risk for animal and human health. In addition, considering that the different sample preparation procedures have strongly contributed to CE sensitivity and versatility, the most advanced sample pre-concentration techniques are discussed here.

Although the poultry production sector plays a key role in sustaining the majority of animal protein demand in Egypt, the deleterious effects of widespread antibiotic resistance on health and environment are currently not well recognized. Litter and dropping samples from broiler and layer poultry farms as well as, tilapia samples from the Nile River and aquaculture farms were collected from Upper Egypt. Samples were extracted and examined for tetracycline residues [tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC) and doxycycline (DC)] using HPLC. In addition, tetracycline resistance genes [tet (M), tet (W), tet (Q) and tet (G)] were screened from pooled intestinal contents collected from twelve broiler farms in Upper Egypt. The antibiotic resistance genes results revealed that tet (W) was confirmed to be expressed in all intestinal samples. In contrast, tet (Q) and tet (M) were detected only in 42% and 17% of the samples, respectively. CTC and OTC were the antimicrobial compounds with the highest concentrations in poultry litter and droppings, with concentrations of 6.05 and 2.47 μg g -1 (CTC) and 5.9 and 1.33 μg g -1 (OTC), respectively. However, the concentrations of DC were significantly higher than those of the other compounds in both aquaculture and Nile River tilapia. The tetracycline residue levels in aquaculture tilapia were significantly higher than those in Nile River tilapia. The hazard quotients (HQs) exceeded 1 for OTC, CTC and DC, which highlights the great risk of using broiler litter to fertilize agricultural land. Moreover, the presence of DC and CTC indicates that consumption of aquaculture tilapia poses a considerable health risk. Therefore, poultry litter or droppings containing tetracycline residues and tet resistance determinants used for aquaculture or as farmland fertilizers could be major sources of antibiotic resistance in fish, humans and environment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Maduramicin (MD) is a type of monoglycoside polyether ionophore antibiotic that can effectively treat coccidiosis and facilitate animal growth. However, its extensive and excessive use brings potential risk to human health. Herein, an electrochemical immunosensor based on indirect competitive format was fabricated for analysis of MD residue in eggs by a multiple signal amplification system. Initially, Au nanoparticles were deposited onto glassy carbon electrode surface to load the coating antigen MD-BSA and to improve conductivity. Then the signal amplification platform was constructed by encapsulating hemin into Fe-MIL-88 NH 2 metal-organic frameworks (hemin@MOFs), and then the obtained composites were decorated with AuPt nanoparticles. The synthesized hemin@MOFs/AuPt was not only used as a signal amplification mediator, but also utilized as a carrier for immobilization of horseradish peroxidase-conjugated affinipure goat anti-mouse antibody (Ab 2 -HRP) and horseradish peroxidase (HRP). The constructed hemin@MOFs/AuPt-Ab 2 -HRP bioconjugates could effectively amplify the current signal since hemin@MOFs, AuPt and HRP all exhibited high catalytic activity towards the hydrogen peroxide. Moreover, the established immunosensor showed high sensitivity and stability during the detection procedure. With the synergistic catalytic effect of hemin@MOFs, AuPt and HRP, a wide detection range of 0.1-50 ng mL -1 and a low detection limit of 0.045 ng mL -1 were achieved (S/N = 3), respectively. Ultimately, the developed method displayed excellent performance in practical applications, providing a promising probability to detect other veterinary drug residues to guarantee food safety., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A method was established to rapidly determine 20 kinds of veterinary drug residues including three catagories of antibiotics (sulfonamides, quinolones, and chloramphenicols) and two kinds of triphenylmethanes (malachite green (MG) and leucomalachite green (LMG)) in fish and shrimp, based on dispersive solid phase extraction purification-ultra high performance liquid chromatography-tandem mass spectrometry. The samples were first hydrolyzed using a dipotassium hydrogen phosphate solution, and then extracted using acetonitrile. Afterward, the extraction solution was dehydrated and salted out with sodium chloride and condensed to nearly dry using a rotating evaporator. This residue was dissolved in 1.0 mL methanol. The resulting solution was purified by dispersive solid phase extraction method with C18 and PSA sorbents, and filtered through a filter. The target compounds were separated employing a ZORBAX C18 column. The mass spectrometer datas were acquired by multiple reaction monitoring (MRM) of positive and negative modes and quantitated applying the isotope internal standard method. The 20 veterinary drugs showed a good linear relationship in the range of 0.2-300 μg/L. The limits of detection and the limits of quantification were 0.1-0.6 and 0.3-1.8 μg/kg, respectively, while the correlation coefficients were greater than 0.99. The average recoveries at the three spiked levels (1, 5, and 20 times of quantitative limits) ranged between 72.5%-118%, with the relative standard deviations of 1.9%-9.8%. The advantages of method include a simple pretreatment, a high detection efficiency, and a low cost. Moreover, it is suitable for the simultaneous determination of multiple veterinary drug residues in fish and shrimp.

We developed a QuEChERS (quick, easy, cheap, effective, rugged and safe) method for the high through-put determination of 28 common veterinary antibiotics in swine wastewater using one-step dispersive solid-phase extraction (d-SPE) for sample cleanup and ultra-performance liquid chromatography-tandem mass spectrometry for detection. The orthogonal test method was used to systematically investigate the parameters that might influence d-SPE efficiency. The optimal d-SPE procedure utilized 40 mg primary secondary amine sorbent and 3 g L -1 Na 2 EDTA. The recoveries ranged from 50 to 100% with relative standard deviations <20% for all target analytes except for enrofloxacin and chlortetracycline. The limits of detection and limits of quantification for all the analytes ranged from 0.002 to 0.200 ng mL -1 and 0.005-0.500 ng mL -1 , respectively. The developed method was successfully applied to the analysis of 28 antibiotic residues in swine wastewater from 10 pig farms located in central China. Fourteen antibiotics including 4 sulfonamides (sulfadiazine, sulfamerazine, sulfamonomethoxine and trimethoprim), 5 fluoroquinolones (norfloxacin, ciprofloxacin, pefloxacin, enrofloxacin, and ofloxacin), 1 lincosamide (lincomycin) and 4 tetracyclines (doxycycline, tetracycline, oxytetracycline, and chlortetracycline) were detected at levels ranging from 0.0560 to 1793 ng mL -1 . Our results demonstrated that the optimized method is a simple but reliable analytical technique for the routine monitoring of veterinary antibiotics in swine wastewater. Swine wastewater samples that we analyzed from 10 pig farms in Jiangxi Province, China were highly contaminated and pose a serious threat to ecosystems and to public health., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Through agricultural soil fertilization using organic manure, antibiotic residues can accumulate in the environment. In order to assess the risks of environmental pollution by veterinary drugs, monitoring of manure for antibiotic residues is necessary. As manure is a complex matrix, extraction of antibiotics proved to be challenging. In this study, 24 extraction solvents were assessed for the extraction of residues from manure representing ten antibiotics from the antibiotic classes tetracyclines, quinolones, macrolides, lincosamides and sulfonamides. Especially for the tetracyclines and quinolones the extraction solvent selection is critical, due to high fractions of non-extractable residues especially when using aqueous solvents (62-77% and 90-95% respectively when using milli-Q water). In contrast, sulfonamides can effectively be extracted with aqueous solvents. Overall, 0.125% trifluoroacetic acid in acetonitrile in combination with McIlvain-EDTA buffer proved to be the most effective extraction solvent. A longitudinal study pointed out that most antibiotics bind to solid manure particles instantaneously after addition. Trimethoprim is an exception, but because by using the optimal extraction solvent, the optimum fraction of bound residues is desorbed, this does not hamper quantitative analysis when using spiked manure quality control samples. Based on these new insights, the current in-house multi-residue LC-MS/MS method for manure analysis, containing 48 antibiotics, was revised, additionally validated and applied to 34 incurred manure samples., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The analysis of veterinary drugs in organic fertilizers is crucial for an assessment of their potential risks to microbial communities in soils and to human health. A method has been developed for the simultaneous determination of ten fluoroquinolones in organic fertilizers by using the QuEChERS method coupled with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The sample was extracted with disodium ethylenediaminetetraacetic acid (Na 2 EDTA)-McIlvaine buffer (pH=4.0) and acetonitrile. The target compounds were separated on an Atlantis T3 C18 column (250 mm×4.6 mm, 5 μm) with a mobile phase of 0.2% (v/v) formic acid in acetonitrile and analyzed in the multiple reaction monitoring (MRM) mode via electrospray ionization (ESI). Ten fluoroquinolones were quantified by an internal standard method. The calibration curves of the ten fluoroquinolones exhibited good linearity over the range of 10-500 μg/kg, and the correlation coefficients were above 0.9930. The limits of detection were 0.5-2.5 μg/kg; the limits of quantification were 1.7-8.3 μg/kg. The analytical method was successfully applied in a survey of veterinary drug contamination in nine compost samples. The average recoveries were 82.5%-117.1%, with relative standard deviations of 3.4%-10.2%. The method is accurate, reliable, and sensitive and supports the simultaneous detection of various veterinary drug residues. Therefore, this method can provide a basis for the risk assessment of veterinary drugs in organic fertilizers.

The objectives of the present study was to investigated the ractopamine (RAC) distribution and depletion process in various tissues of goat including liver, kidney, spleen, lung, heart, fat, bile, brain and the eyes. The experiment was carried out on 21 goats (18 treated and 3 controls). Treated goats were orally administered RAC in a dose of 1 mg/kg body mass per day for last 28 days and randomly sacrificed on withdrawal days of 0.25, 1, 3, 7, 14 and 21. RAC in all matrices were determined by ultra-high performance liquid chromatography-quadrupole orbitrap high resolution mass spectrometry. After 21 days treatment discontinuation, the levels of RAC in bile reached at 13.48 ± 3.36 mg/L, which was significantly higher than that in the other tissues. The concentrations of RAC were followed by kidney, the excretory organ and liver, the major metabolic organ (4.49 ± 0.16 mg/kg for kidney and 1.81 ± 0.11 mg/kg for liver, respectively). The residual concentration of the drug in the eyes of goat was less than that in bile, kidney, liver, lung and spleen on withdrawal days 0.25. RAC residues was higher than the limits of detection = 0.15 μg/mL in liver on Day 21. These findings demonstrated that liver can serve as an alimentary matrix and as a matrix for the control of RAC abuse hypothetically except for urine., (© The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Veterinary medicines, especially antibiotics, are among the most important components related to animal feed production. Generally, the main use of antibiotics in animals is for the treatment and prevention of diseases and growth promotion. Antibiotic usage in animals may result antibiotic residues in foodstuffs such as milk, egg and meat. These residues may cause various side effects such as transfer of antibiotic resistant bacteria to humans, immunopathological effects, allergy, mutagenicity, nephropathy (gentamicin), hepatotoxicity, reproductive disorders, bone marrow toxicity (chloramphenicol) and even carcinogenicity (sulphamethazine, oxytetracycline, furazolidone). The most important adverse effect of antibiotic residues is the transfer or antibiotic resistant bacteria to the humans due to the mobile properties of resistance. Because of these undesirable effects, it is important to regulate the use of antibiotics in food animals. The individuals and the local procedures should be aware of the problem through education by authorities. In this review, antibiotic use in the foodstuffs and their effects on the human health will be discussed., (Copyright © 2019. Published by Elsevier Ltd.)

In this work, two sample treatment procedures have been evaluated for the determination of veterinary drug residues in milk. In order to cover a wide range of polarities, a total of 66 veterinary drugs with log K ow ranging from - 1 to 5 were selected. Two sample cleanup steps, (i) dispersive solid-phase extraction (dSPE) using enhanced matrix removal lipid as sorbent and (ii) solid-phase extraction (in pass-through mode) using Oasis HLB PRiME cartridges, were critically assessed in terms of sample throughput, recovery, matrix effect, cleanliness of extracts, limit of quantification, and repeatability. The veterinary drugs tested (viz. benzimidazoles, cephalosporins, imidazothiazoles, macrolides, NSAIDs, penicillins, quinolones, steroids, sulfonamides, and β-agonists) were analyzed by ultra-high-performance liquid chromatography tandem mass spectrometry. According to the results, both methods exhibited similar recovery rates between 70 and 120% for most of compounds tested. Matrix effects were satisfactory for both methodologies, although the tolerance to matrix effects was slightly higher with HLB PRiME with nearly negligible matrix effects in most cases. Limits of quantitation were also well below the current maximum residue levels established by the European Union. Notably, sample throughput was higher in the case of HLB PRiME, since this pass-through SPE cleanup approach involved fewer steps than the EMR-Lipid dSPE approach. The results in terms of analysis time, sensitivity, precision, cleanliness of extracts, and matrix effect showed the suitability of both procedures for the monitoring of veterinary drugs residues in milk samples in a single run. Graphical abstract.

We examined a total of 369 bovine liver and muscle samples for the detection of oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), and doxycycline (DOX) residues by implementation and validation of a LC-MS/MS method. The method showed good recovery values between 86% and 92% at three levels of concentrations. The linearity tests revealed r² > 0.996 for all the tetracyclines examined. Furthermore, the Youden test revealed that the method was robust. Only 14.4% of the samples showed OTC and TC residues in a concentration range of 10.4⁻40.2 µg kg -1 . No CTC and DOX residues were found in all the samples analyzed. Liver samples showed the highest average values (31.5 ± 20.6 and 21.8 ± 18.9 for OTC and TC, respectively). The results showed a low incidence of TCs in all the samples examined, in comparison with other studies reported in the literature. A significant decrease in TC residues frequency was found from 2013 ( p < 0.05). This work reports for the first time epidemiological data on the presence of TC residues in liver and muscle samples of cattle farmed in Sicily (Southern Italy). The very low incidence of TC residues indicates a continuous improvement in farming techniques in Southern Italy, which is essential to ensure consumers' protection.

A comprehensive pretreatment system was developed to simultaneously extract and purify 120 veterinary antibiotics, possessing various physicochemical properties in eight categories. This system was coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the detection of the antibiotics. The samples were dissolved and dispersed by a Na 2 EDTA-Mcllvaine buffer, extracted with acetonitrile containing 1.5% (v/v) formic acid, and cleaned using an Oasis PRiME HLB SPE system followed by n -hexane liquid purification. The separation of residue targets was performed on an Atlantis ® T3 column (100 mm×2.1 mm, 3 μm) with a gradient elution of 0.1% (v/v) formic acid aqueous solution and acetonitrile as mobile phases and detected by UPLC-MS/MS with the multiple reaction monitoring (MRM) mode via ESI + ionization. The pH of the extraction solvent and the purification method were optimized to promote the target recoveries. All targets showed good linear ranges from 1.0 to 50.0 μg/L, while all their correlation coefficients ( r 2 ) were higher than 0.9953, including r 2 values of 89 targets being over 0.9990. The limits of quantification (LOQs) of seven targets were 10.0 μg/kg; 21 targets, 5.0 μg/kg; and the remaining 92 targets, not higher than 2.0 μg/kg. The average recoveries on three spiked levels (low, medium, and high) for all targets ranged from 71.5%-109.2%, with RSD ranging from 0.6%-15.3%. The combined system, exhibiting satisfying recovery and stable repeatability, can be suitably employed for the simultaneous determination of multiple veterinary antibiotics in animal-derived meat products.

Soil bacterial communities have complex regulatory networks, which are mainly associated with soil fertility and ecological functions, and are likely to be disturbed due to antibiotics applications. The impact of antibiotics, particularly in mixtures form, on bacterial communities in different paddy soils is poorly understood. Using pyrosequencing techniques of 16 S rRNA genes, this study investigated the synergistic effects of veterinary antibiotics (sulfadiazine, sulfamethoxazole, trimethoprim, florfenicol, and clarithromycin) on bacterial communities in a soil-bacteria-plant system. Rice was grown under controlled greenhouse conditions where unplanted and planted treatments were doped with 200 µg kg -1 of combined antibiotics over a period of 3 months. Bacterial richness remained unaltered, while a significant decline was observed in bacterial diversity due to antibiotics in the four paddy soils. Bacteroidetes and Acidobacteria were increased, while Actinobacteria and Firmicutes decreased under antibiotics exposure. Despite antibiotics perturbation, compositional variations were mainly attributed to the different paddy soils which harbor distinct bacterial communities. Haliangium and Gaiella were among the sensitive genera that were negatively correlated to antibiotics perturbation. Additionally, electrical conductivity, total organic carbon, and total nitrogen of soil solution were the key physiochemical indices which significantly influenced the structure of bacterial communities in the paddy soils. These findings expanded our knowledge of effects from synergistic antibiotics application and variations in bacterial communities among different paddy soils., (Copyright © 2018 Elsevier Inc. All rights reserved.)

A method was established for the determination of 50 kinds of antibiotic residues (macrolides, quinolones, sulfonamides, tetracyclines, nitroimidazoles, lincomycin and chloramphenicol) in drone pupa powder by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted with perchloric acid and lead acetate solution, and the protein was precipitated. The extraction solution was adjusted to pH 8 using dipotassium hydrogen phosphate, then the solution was purified by solid phase extraction (SPE) and analyzed by LC-MS/MS. The target compounds in the drone pupa powder were determined quantitatively and quantitatively by using multiple reaction monitoring and positive ion or negative ion modes. The recoveries of the 50 antibiotics were in the range of 70.2%-118.3%, and the relative standard deviations (RSDs) were 1.8%-13.6%. The method is simple, selective, and can be suitable for the analysis and confirmation of veterinary drug residues in drone pupa powder.

Background: With increasing media coverage of food safety incidents, such as that of clenbuterol residues in pork, food safety has become a major public health concern in China. Rapidly developing online markets attract increasing numbers of Chinese consumers to purchase food on the Internet. However, the quality and safety of food sold online are uncertain and are less reported on., Objective: This research aimed to systematically study the quality and safety of chilled pork from wet markets, supermarkets, and online markets in China., Results: The chilled pork samples from online markets were fresher than those from wet markets and supermarkets based on the surface redness (a* value). Chilled pork contained high levels of nutritional elements, especially the magnesium and phosphorus levels in samples from online markets. The levels of heavy metal element residues and veterinary drug residues in all chilled pork samples were within the standards limits. In addition, huge differences existed in the quality and freshness of the chilled pork samples from online markets according to principal component analysis (PCA)., Conclusions: Most chilled pork sold in Chinese markets was qualified and safe. It is necessary to establish an effective online market supervision system for chilled pork.

Residue of veterinary antibiotics (VAs) in the animal breeding industry has become a problematic environmental issue. However, the residual levels of VAs as well as their variation tendency, degradation mechanisms and relationships with other parameters during animal wastewater disposal processes are still obscure. This study measured different samples during wastewater disposal processes from three farms, and systematically analyzed the residue, migration and removal of 58 kinds of typical VAs (6 classes) in Tianjin, China. The results showed that about 44 kinds of VAs were quantitatively detected. Tetracycline antibiotics (TCs) usually had higher residual concentrations than other classes of VAs in the raw wastewater; the highest residual concentration was 130.67 ± 5.90 μg/L which occurred for chlortetracycline (CTC). Pig farms generally had more VAs species and higher residual concentrations than dairy farms, and the proportion of different VAs was similar for dairy farms. The final removal rates of different VAs classes varied largely (negative to > 99.87%), and the highest removal rates usually occurred in biological processes for adsorption and biodegradation effects, and occasionally occurred in the final effluents. The correlation coefficients between VAs removal rates and chemical oxygen demand (COD) removal rates were much higher than those of total nitrogen (TN), total phosphorus (TP) and ammonia nitrogen (NH 4 -N) in pig farms, while opposite conclusion was obtained in dairy farms. Among different classes, TCs presented the highest daily mass loading of ND ~ 10,453.8 ± 471.7 mg/d in the influent and ND ~ 1141.6 ± 58.9 mg/d in the effluent in farm 1., (Copyright © 2018 Elsevier Inc. All rights reserved.)

An ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) method was developed for determination of 39 veterinary drugs in Procambarusclarkii. Samples were extracted with acidified acetonitrile and purified with enhanced matrix removal-lipid (EMR-Lipid) adsorbent and graphitized multi-walled carbon nanotubes (GMWNTs). Extracts were analyzed by UHPLC-Q-TOF-MS. Target compounds were screened according to accurate mass, isotopic ratio, and retention time, and confirmed by library search. Target analytes were quantified using the chromatographic peak area of each compound. For each of the drugs of interest, calibration curves were linear in their respective ranges with correlation coefficients greater than 0.99. The limits of quantification (LOQs) were 3-15 μg/kg. The recoveries for target compounds spiked in Procambarusclarkii ranged from 62.4% to 105.8%, with relative standard deviations (RSDs) ranging from 2.5% to 13.5%. This method is simple, rapid, accurate, and suitable for simultaneous determination of 39 veterinary drugs, including sulfonamides, quinolones, and triphenylmethane, extracted from Procambarusclarkii.