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The environmental impact of insecticides used as ectoparasitic treatments for companion animals is not well understood, since they are not subject to detailed environmental risk assessment. Many of these treatments include active ingredients such as fipronil and imidacloprid that are banned from agricultural use in the EU. These treatments are applied topically and can remain on the animal's fur for an extended period of time. Birds (adults, eggs, and nestlings) using fur as an inner layer for their nests have the potential of being exposed dermally to these chemicals. In this study, we collected 103 nests from blue and great tits, which were lined with fur. Using UHPLC-MS/MS, we detected 17 out of the 20 insecticides we screened for, with the number of insecticides detected per nest ranging from 2 to 11. Fipronil, imidacloprid, and permethrin were detected in 100 %, 89.1 %, and 89.1 % of samples, respectively. The average concentration of fipronil, imidacloprid and permethrin were respectively 115.5 ppb, 376.3 ppb, and 231.1 ppb. Dinotefuran was found at the highest concentration of 7198 ppb in a single sample. Overall, a higher number of either dead offspring or unhatched eggs was found in nests containing a higher number of insecticides, higher total concentration of insecticides or a higher concentration of fipronil, imidacloprid or permethrin, suggesting that contact exposure of eggs to insecticides in nest lining may lead to mortality and lower reproductive success. This highlights the need for a re-evaluation of the environmental risks associated with use of these potent and persistent insecticides on companion animals., 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 The Authors. Published by Elsevier B.V. All rights reserved.)

Co-extraction of multiple types of target substances is the key to achieve high throughput detection. In this work, PDA@MOF-808/PAN NFsM was prepared by co-modified polyacrylonitrile nanofiber membrane (PAN NFsM) with polydopamine (PDA) and metal-organic framework-808 (MOF-808), and its potential as a solid-phase extraction (SPE) adsorbent was investigated by using the most typical nine veterinary drug residues in eggs and chicken as model target substances. The results show that PDA@MOF-808/PAN NFsM could effectively co-extract all the target substances (adsorption efficiency ranged from 81.46 % to 96.78 %), and had good capability of sample matrix purification (matrix effect was lower than -15.26 %), so a new SPE method has been established. Combined with UPLC-MS/MS, the detection limits were 0.3 to 3.1 μg/kg, the recoveries were between 71.02 % and 106.48 %, and the relative standard deviations were lower than 12.03 %, indicating that the method has considerable good sensitivity, accuracy and precision., 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 Ltd. All rights reserved.)

Veterinary drugs are used worldwide to prevent and treat diseases and promote growth in animals, fisheries, and beekeeping. Despite their effectiveness, the illegal and improper use of these drugs can result in livestock and fishery products, potentially impacting human health by causing allergic reactions, cytotoxicity, and antimicrobial resistance. To mitigate these adverse effects, the Japanese government established a positive list system in 2006. Maximum residue levels (MRLs) have been defined for approximately 300 veterinary drugs. The regulation of certain antimicrobial drugs without defined MRLs uses in zero tolerance. Residual veterinary drugs in meat, fish, eggs, milk, and honey are inspected to ensure compliance with legal limits. Initially, samples are screened using bioassay and LC-MS/MS. If residues are detected, they are confirmed and quantified using more precise methods. We have detailed the detection of residual norfloxacin in honey. Additionally, we are currently developing new analytical methods. This study introduces an analytical method for the residue screening of sulfonamides and quinolones. After residue detection by bioassay, the same test solutions were analyzed by LC-MS/MS for accurate identification and quantification. This method has also proven to be more environmentally friendly compared to other quantitative methods. We present the ingenuity used to realize the combined method, performance evaluation results, examples of applications to actual samples, and future prospects.

Pesticides, veterinary drugs, and feed additives (hereinafter referred to as "pesticides") can remain in foods when used in agricultural and livestock products. Since consuming a variety of foods every day can result in ingesting trace amounts of these pesticides, which may be harmful to health, risk management for residual pesticides in foods is necessary to prevent adverse effects. Based on the Food Sanitation Act, the Ministry of Health, Labour and Welfare (MHLW) has established maximum residue limits (MRLs) for each pesticide and each food type. Currently, approximately 770 pesticides have MRLs set. Since May 2006, Japan has implemented a positive list system, prohibiting the distribution of food containing residual pesticides exceeding the MRLs or uniform limit of 0.01 ppm for pesticides without established MRLs. Appropriate analytical methods are required to determine whether pesticides exceed the MRLs or uniform limit. Currently, MHLW has notified ten simultaneous analytical methods and approximately 350 individual analytical methods. However, many pesticides still lack developed analytical methods. These methods should be simple, quick, and accurate, but developing them is challenging. The National Institute of Health Sciences, in cooperation with local health institutes, registered conformity assessment bodies, and universities, is working on developing these analytical methods. This lecture introduces an overview and the challenges of analytical methods for detecting residual pesticides.

A modified QuEChERS method was developed to determine multi-class pesticide and veterinary residues in aquatic products. Chitosan microspheres were conveniently synthesized and utilized as the cleanup adsorbent in the QuEChERS procedure, showcasing rapid filtration one-step pretreatment ability for the determination of drug multi-residues in aquatic products. Compared to conventional synthetic sorbents, chitosan microspheres not only have good purification performance, but also have renewable and degradable properties. This novel sorbent worked well in the simultaneous determination of 95 pesticides and veterinary drug residues in aquatic products after being combined with an improved one-step vortex oscillating cleanup method. We achieved recoveries ranging from 64.0% to 115.9% for target drugs in shrimp and fish matrix. The limits of detection and quantification were 0.5-1.0 and 1.0-2.0 μg kg -1 , respectively. Notably, hydrocortisone was detected with considerable frequency and concentration in the tested samples, underscoring the necessity for stringent monitoring of this compound in aquatic products., 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. Published by Elsevier Ltd.)

The COVID-19 pandemic caused global pandemonium, and due to an unprecedented global response, the popularity and use of (veterinary) ivermectin, amongst many other conceivable 'treatments', experienced a meteoric rise. Ivermectin is a macrocyclic lactone compound belonging to the avermectin drug class and is a registered medicine in many countries, although the most common use is as veterinary medicine. In this study, a fast HPLC method was developed and validated for the quantification of ivermectin in veterinary products that were used off-label by a substantial number of people during COVID-19. Locally used veterinary products were collected as well as commercial products acquired and were tested using the newly developed method. The ivermectin content was compared to the products' label claims and it was found that all products tested contained ivermectin within acceptable limits. However, the use of veterinary products is strongly discouraged due to high dosages and administration regimens that were tested in animals and can lead to serious adverse events in humans. The presence of untested excipients and secondary actives, such as clorsulon, which can cause unknown (long-term) health impacts in humans, further adds credence to this warning.

Livestock and poultry meat consumption play an important role in the dietary structure of Chinese residents. However, the extent of residues of veterinary drugs and heavy metal contamination in livestock and poultry meat and their by-products within Hunan province is not extensively studied. This survey aimed to fill this gap by assessing the presence of 76 veterinary drug residues in Hunan province. Additionally, heavy metals in pork and pig liver were also assessed. The obtained findings suggest that residues of veterinary drugs are still present in livestock and poultry meat, as well as their by-products, within Hunan province. However, the contamination of heavy metals remained within the food safety limits. These results underscore the significance of establishing more refined criteria for assessing human exposure, taking into account factors such as consumption patterns, product varieties and chemical compounds of interest.

Based on salting-out assisted liquid-liquid extraction (SALLE) and high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS), a simple, rapid pretreatment without complex clean-up for the determination of 22 veterinary drug residues in aquatic products was developed and validated. In order to improve the efficiency of the method, the key procedural parameters of SALLE were fabricated. Na 2 EDTA-Mcllvaine buffer/ACN was used as the extraction solvent, anhydrous MgSO 4 and NaCl as the extraction salts. The relationship between extraction efficiency and logD was initially evaluated during the optimization process. This study was well validated in various aquatic samples such as bass, large yellow croaker, carp, and shrimp, the limits of detection (LOD) and accuracy for all compounds ranged from 0.5 to 1.0 μg/kg, 71.4% to 120%. This method has the advantages of rapidity, simplicity, low cost, and high efficiency, and has broad potential for risk monitoring and evaluation of veterinary antibiotics in aquatic products., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

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 drugs play a crucial role in the treatment of various animal diseases. However, their residues, stemming from issues, such as withdrawal period lapses, overuse, or abuse, can jeopardize food safety and human health. This study addresses recent regulations in Korea concerning specific veterinary drugs (anacolin, ephedrine, menichlopholan, piperonyl butoxide, and etisazole HCl) and their ongoing discussions. This study aimed to validate two pre-developed methods for quantifying residues in livestock and fishery products using QuEChERS and liquid chromatography-tandem mass spectrometry. Both methods exhibited excellent linearity, recoveries (70.3-119%), and coefficient of variations (1.3-28%), along with low limits of detection and quantification (0.3-4 ng/g and 1-12 ng/g). This study is significant for its contribution to the detection of veterinary drugs in livestock and fishery products, given the limited research available on the methods for analyzing these substances., 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 Ltd. All rights reserved.)

Effective monitoring of veterinary drug residues in food is essential for legislation compliance and food safety, yet remains challenging due to low concentrations and complex matrices. This study introduced a miniaturized 96-well electromembrane extraction (EME) technique for pre-concentration and isolation 80 prohibited/restricted veterinary drugs from honey samples. Three liquid membranes were developed and characterized: V1 ("V" for veterinary), a mixture of 2-undecanone and 0.5 % 2-nitro-p-cymene, was optimal for extraction of quinolones, tetracyclines, and β-agonists. V2, a mixture of 2-nitrophenyl octyl ether and a deep eutectic solution (formed upon mixing 6-methylcoumarin with thymol in a 1:1 w/w ratio) in a 1:1 v/v ratio, with the addition of 0.5 % di(2-ethylhexyl) phosphate (DEHP), was suitable for macrolides and sulfonamides extraction. V3, a deep eutectic solvent formed upon 1-decanoic acid and camphor in a 1:1 w/w ratio with 1 % DEHP, was suitable for aminoglycosides and β-lactams. Operational parameters were fine-tuned to achieve optimal extraction efficiencies, with recoveries ranging from 20.1-117.5 %, 20.3-91.9 %, and 20.1-118.8 % for V1, V2, and V3, respectively. High precision was observed for compounds with recoveries over 40 % (intra-day: 0.4-27.7 %; inter-day: 2.6-33.0 %), and matrix effects were generally within 80-120 %. This work established a theoretical framework for EME in food analysis, highlighting its potential as a powerful sample pretreatment method for the detection of veterinary drug residues. The results demonstrated the feasibility and advantages of using EME for ensuring food safety and regulatory compliance., 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 Ltd. All rights reserved.)

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.)

Precise determination of veterinary pharmaceutical concentrations represents a critical foundation for delivering safe and efficacious animal healthcare interventions. Two synthetic glucocorticoids - dexamethasone sodium phosphate (DXM) and prednisolone acetate (PRD) - are extensively employed in veterinary medicine due to their potent anti-inflammatory capabilities. Our research presents a novel, cost-effective, and environmentally sustainable analytical methodology that enables simultaneous quantification of DXM and PRD within binary veterinary formulations. The method synergistically combines UV spectroscopy with dimension reduction algorithms (DRAs), representing a significant advancement in pharmaceutical analysis. A comprehensive evaluation of seventeen DRAs was conducted using four distinct performance metrics: mean squared error (MSE), mean absolute error (MAE), median absolute error (MedAE), and coefficient of determination (R 2 ). Among the assessed algorithms, mini-batch sparse principal component analysis demonstrated superior predictive accuracy for this specific analytical challenge. The developed method was validated using the accuracy profile approach, yielding results that confirm its satisfactory accuracy. An ecological impact assessment was conducted using five greenness evaluation tools: the Green Solvent Selection Tool (GSST), National Environmental Methods Index (NEMI), Green Certificate modified Eco-Scale, carbon footprint analysis, and the Modified GAPI (MoGAPI). In addition, whiteness was evaluated with Red-Green-Blue 12 (RGB 12) algorithms. The proposed method showed elevated GSST scores and a greener profile according to NEMI. The calculated carbon footprint was 0.0006 kg CO 2 equivalent per sample, with a Green Certificate modified Eco-Scale score of 84, a MoGAPI score of 81, and a whiteness assessment of 90.1 by the RGB12 algorithm. Statistical comparison between the proposed spectrophotometric method and a previously reported HPLC method for pharmaceutical dosage form analysis revealed no statistically significant differences at the 95 % confidence level. This study underscores the innovative combination of UV spectroscopy with dimension reduction algorithms, presenting substantial improvements over traditional UV techniques for drug analysis. This method enhances both the efficiency and accuracy of active ingredient determination in pharmaceutical dosage forms while also supporting environmental sustainability., 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., (Published by Elsevier B.V.)

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.)

Background: Veterinary antibiotics are chemical compounds used to kill or inhibit the growth of pathogenic bacteria associated with animal diseases. These molecules can be defined by their retention times (t R ) in liquid chromatography-mass spectrometry (LC-MS). One strategy to predict the t R of new veterinary antibiotics is the development of predictive quantitative structure-property relationships (QSPRs), which were used in this study., Results: A database of 122 antibiotics was selected in which the t R was measured using a Hypersil GOLD column. An optimal three-feature model was developed by integrating the unsupervised variable reduction, replacement method variable subset selection, and multiple linear regression. The negligible differences among the coefficient of determination and the root-mean-square error for the training set (R 2  = 0.902 and RMSEC = 0.871) and test set (Q 2  = 0.854 and RMSEP = 1.064) indicate a stable and predictive model. In a further step, a more in-depth explanation of the mechanism of action of each descriptor in predicting the t R is provided, with the construction of the theoretical chemical space for accurate predictions of new antibiotics., Conclusion: The in silico model developed in this work identified three molecular descriptors associated with aqueous solubility, octanol-water partition coefficient, and the presence of negative and lipophilic atom pairs. The QSPR developed here could be implemented by agricultural and food chemists to identify and monitor existing and new antibiotics within the framework of LC-MS. The computational model was developed in accordance with five principles outlined by the Organization for Economic Co-operation and Development. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

An HPLC-MS/MS multi-class method for quantitation of 15 different classes of veterinary drug residues (>140 analytes) in milk and poultry feed was developed and validated. Accuracy criteria for routine laboratories were met for the majority of analytes, > 83 % in milk and between 50 and 60 % in chicken feed, with an apparent recovery of 60-140 %. Extraction efficiency criteria were met for >95 % of the analytes for milk and > 80 % for chicken feed. Intermediate precision meets the SANTE criterion of RSD < 20 % for 80-90 % of the analytes in both matrices. For all analytes with an existing MRL in milk, the LOQ was below the related MRL. Twenty-nine samples of commercial milk and chicken feed were analyzed within the interlaboratory comparison. No residues of veterinary drugs were found in the milk samples. However, the feed samples exhibited high levels of nicarbazin, salinomycin, and decoquinate., 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 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 )≥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% ( 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.

Pesticides and veterinary drugs are widely employed to support food production. Assessing potential risks associated with the dietary consumption of pesticide and veterinary drug residues is, however, essential. Potential risks depend on the toxicity degree of the analyzed residue and population exposure levels. Human populations are exposed to numerous chemical substances through different pathways with varying exposure times, leading to increased health risks when compared to exposure to individual chemicals. Cumulative exposure assessments usually assess combined exposures to multiple chemicals through multiple exposure pathways. In this sense, this comprehensive review aims to provide insights into cumulative dietary pesticide and veterinary drug residue exposures. The main methodologies, strategies, and legislation employed by international agencies to this end are discussed. A review concerning articles that apply existing methodologies and approaches, as well as the challenges in this context faced by Brazil is also presented. As this is a critical issue not only for Brazilian public health but also for the global community, regulatory agencies should prioritize formulating regulations that incorporate exposure assessments regarding the simultaneous presence of residues and contaminants in foodstuffs.

Background: The presence of veterinary drug residues in food-producing animals and animal products is regulated through the enforcement of maximum residue limits (MRLs). To answer the need of the food sector to monitor these substances in a wide range of food commodities, stakeholders at AOAC INTERNATIONAL identified the need for a reliable confirmatory screening method. Such a qualitative approach is required for compliance checking and to support product release in manufacturing., Objective: Data were collected from five independent laboratories that applied the First Action Official Method 2020.04 to demonstrate adequate performance under reproducibility conditions. The probability of detection (POD) was calculated in blank test samples and test samples spiked at the screening target concentration (STC) level, with the objective to achieve PODs ≤10% and ≥90%, respectively. Additionally, the effectiveness of the screening method was evaluated by participating in 92 proficiency tests., Methods: Four streams were optimized to screen for 152 veterinary drug residues by LC-MS/MS in a wide variety of food commodities including milk-based ingredients and related products (e.g., milk fractions, infant formula, infant cereals, and baby foods), meat- and fish-based ingredients and related products (fresh, powdered, cooked, infant cereals, and baby foods), and other ingredients based on eggs, animal fat, and animal byproducts. The four streams covered 105 antibiotic residues, anti-inflammatory and antiparasitic agents (stream A), 23 beta-lactams (stream B), 14 aminoglycosides (stream C), and 10 tetracyclines (Stream D)., Results: The multilaboratory validation led to PODs at the STC ≥94% and PODs in the blank ≤9%. Further application of the multilaboratory validated method to 92 proficiency tests provided more than 99% satisfactory submitted results (n = 784)., Conclusion: The interlaboratory reproducibility determined for this method met the acceptance criteria defined in AOAC Standard Method Performance Requirement (SMPR®) 2018.010., Highlights: AOAC has approved the method for Final Action status., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Milk, as a widely consumed nutrient-rich food, is crucial for bone health, growth, and overall nutrition. The persistent application of veterinary drugs for controlling diseases and heightening milk yield has imparted substantial repercussions on human health and environmental ecosystems. Due to the high demand, fresh consumption, complex composition of milk, and the potential adverse impacts of drug residues, advanced greener analytical methods are necessitated. Among them, functional materials-based analytical methods attract wide concerns. The magnetic molecularly imprinted polymers (MMIPs), as a kind of typical functional material, possess excellent greenification characteristics and potencies, and they are easily integrated into various detection technologies, which have offered green approaches toward analytes such as veterinary drugs in milk. Despite their increasing applications and great potential, MMIPs' use in dairy matrices remains underexplored, especially regarding ecological sustainability. This work reviews recent advances in MMIPs' synthesis and application as efficient sorbents for veterinary drug extraction in milk followed by chromatographic analysis. The uniqueness and effectiveness of MMIPs in real milk samples are evaluated, current limitations are addressed, and greenification opportunities are proposed. MMIPs show promise in revolutionizing green analytical procedures for veterinary drug detection, aligning with the environmental goals of modern food production systems., (© 2024 Institute of Food Technologists®.)

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 15-channel pressure filtration purification method was presented for high throughput sample preparation of aquatic products. A cost-effective device was constructed and melamine sponge was selected as the cleanup sorbent. Upon interfacing with HPLC-MS/MS, the analytical procedure demonstrated its suitability for quantifying 160 pesticides and veterinary drug residues in aquatic products such as fish, shrimp, and crab. The method achieved sample recoveries ranging from 61.3 to 124.9 %. The detection limits were established between 0.5 and 1.0 μg/kg, while the quantitation limits were confirmed to be within the range of 1.0-2.0 μg/kg. The method was applied to quantify the pesticide and veterinary drug residues in mostly consumed aquatic products from five coastal provinces in China. The results showed significant differences between different aquatic products in the concentrations of pesticide and veterinary drug residues, implying the necessity of supervision for the accurate determination of pesticides and veterinary drugs., 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 Ltd. 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.)

An analytical method was developed for the screening of 172 veterinary drugs in traditional Chinese medicine Galli Gigerii Endothelium Corneum by high-performance liquid chromatography tandem mass spectrometry. The samples were pretreated by a modified QuEChERS method. A Zorbax Eclipse plus C18 column (1.8 μm, 3.0 × 150 mm2, Agilent) was used for the separation of analytes by gradient elution. All analytes were detected by electrospray ionization mass spectrometry with multiple reaction monitoring mode. Good linearity with R ≥ 0.99 was exhibited for all analytes within the respective range. The recoveries of all monitored analytes ranged from 55.4 to 127.6% at three spiked levels (limit of quantitation-LOQ, 2-fold LOQ, 10-fold LOQ), with relative standard deviations <17.8%. The estimated LOQ levels were 0.2-20 μg/kg. The application of this method provides a reference for the safety control of traditional Chinese medicines., (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

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.

Brazil plays an important role in ensuring its position on the international market by assuring high food safety standards for its products, and all products should meet the requirements for residues from veterinary drugs and contaminants in animal products. Statutory monitoring provides insights into the compliance of the Brazilian industry regarding these legal requirements. The objective of this study was to provide insight into the safety of Brazilian animal products by reporting the occurrence of residues from veterinary drugs and contaminants according to an analysis of an 11-year report published by the Brazilian Ministry of Agriculture, Livestock and Food Supply (MAPA). Between 2010 and 2021, 166,647 samples from animal-derived products were analyzed in Brazil, and 624 of those samples were non-compliant (0.37%) exceeding maximum residue limits (>MRLs) or showed the presence of prohibited substances. The most common types of substances found in the non-compliant samples were heavy metals, parasiticides, and antimicrobials, accounting for 82% of all documents from the MAPA. Among Brazilian products, the challenge related to occurrence of substances varied across the food supply chain, with highest incidence rates observed in the fish chain, followed by eggs, milk, equids, sheep/goat, honey, bovine, swine, and broilers chains in decreasing order. Considering the type of substance, heavy metals were found to be more prevalent in fish products, mainly arsenic in wild fish. The prevalence of contaminants and heavy metals decreased, while that of veterinary drugs increased in Brazilian products from 2010 to 2021. From these results, it can be concluded that the number of accidental incidents including those associated with environmental contaminants decreased over the last decade, opposed to those involving human adversaries and deliberate illegal actions, such as the abuse of veterinary drugs, increased. Future monitoring plans need to take this paradigm shift into account.

A rapid and highly sensitive method was established for the analysis of 37 veterinary drug residues in milk using a modified QuEChERS method based on a reduced graphene oxide-coated melamine sponge (rGO@MeS) coupled with UPLC-MS/MS. Under optimal chromatographic and mass spectrometric conditions, the effects of different dehydrated salts (MgSO 4 and Na 2 SO 4 ) and metal chelating agents (Na 2 EDTA) on extraction efficiency were first investigated. Next, the influence of a dynamic and static purification mode was evaluated in terms of drug recoveries. Calibration curves of 37 veterinary drugs were constructed in the range 0.6-500 μg kg -1 , and good linearities were obtained with all determination coefficients ( R 2 ) ≥0.992. The limits of detection (LODs) and quantitation (LOQs) were in the range 0.3-1.1 μg kg -1 and 0.6-3.5 μg kg -1 , respectively. The recoveries of all compounds were in the range 61.3-118.2% at three spiked levels (20, 100, and 200 μg kg -1 ) with RSDs ≤15.4% for both intra- and inter-day precisions. Compared to pristine melamine sponges and commercial adsorbents (C18, PSA, and GCB), rGO@MeS demonstrated an equal or even better purification performance in terms of recoveries, matrix effects, and matrix removal efficiency. This method is rapid, simple, efficient, and appropriate for the qualitative and quantitative analyses of 37 veterinary drug residues in milk, providing a new detection strategy and technical support for the routine analysis of animal-derived food.

Although filtration is one of the most common steps in sample preparation for chemical analysis, filter membrane materials can leach contaminants and/or retain some analytes in the filtered solutions. In multiclass, multiresidue analysis of veterinary drugs, it is challenging to find one type of filter membrane that does not retain at least some of the analytes before injection in ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). In this study, different filter membranes were tested for use in UHPLC-MS/MS analysis of 183 diverse drugs in bovine muscle, kidney, and liver tissues. Membranes evaluated consisted of polytetrafluoroethylene (PTFE), polyvinylidene difluoride (PVDF), polyethersulfone, nylon, and regenerated cellulose. Drug classes represented among the analytes included β-agonists, β-lactams, anthelmintics, macrolides, tetracyclines, sulfonamides, tranquilizers, (fluoro)quinolones, anti-inflammatories, nitroimidazoles, coccidiostats, phenicols, and others. Although the presence of a matrix helped reduce the binding of analytes on surface active sites, all of the filter types partially retained at least some of the drugs in the final extracts. In testing by flow-injection analysis, all of the membrane filters were also observed to leach interfering components. Ultimately, filtration was avoided altogether in the final sample preparation approach known as the quick, easy, cheap, effective, rugged, safe, efficient, and robust (QuEChERSER) mega-method, and ultracentrifugation was chosen as an alternative., (© 2024 Wiley-VCH GmbH.)

A rapid multi-residue LC-MS/MS method for the identification and determination of banned veterinary drugs in honey was developed. A total of 31 investigated veterinary drugs belonging to 4 classes including nitrofurans metabolites, nitroimidazoles, amphenicols, and quinolones were quantified by LC-MS/MS with ESI using one single injection. The sample preparation included treatment with 5-nitro-2-furaldehyde (5-NFA) in a thermostated ultrasonic bath (80 °C, 0.5М НСl, 20 min) to liberate matrix-bound residues of nitrofurans. Magnetic hypercrosslinked polystyrene (HCP/Fe 3 O 4 ) was proposed for the solid-phase extraction and clean-up of target analytes prior to LC-MS/MS analysis. To evaluate and validate the performance of method, the criteria of the Decision (EC) no 2002/657 were applied. The LOQs of the examined analytes range from 0.3 to 1 μg kg -1 , which indicates good sensitivity to quantify the target compounds in honey. The recoveries of veterinary drugs from 1 g of honey with 50 mg of the sorbent are 97-109% for nitrofuran metabolites, 84-115% for nitroimidazoles, 86-103% for amphenicols, and 97-118% for quinolones. The relative standard deviations of intra-day and inter-day precision analyses (RSD) are less than 16%. This methodology was applied to real honey samples and trace levels of some veterinary drugs were detected., 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 © 2023 Elsevier B.V. All rights reserved.)

Excessive use of veterinary drugs in livestock growth poses a threat to food safety. It is, however, challenging to quantify these multi-class veterinary drugs within animal muscles, because of their varied physicochemical properties. In this work, we presented a simple, efficient and sensitive method for the simultaneous determination of multi-class veterinary drugs with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The method involves a highly efficient extraction using a EDTA (pH 7)-ACN (30:70, v/v) solvent system, followed by a one-step solid-phase extraction cleanup approach with PRiME HLB sorbent (Reversed-phase N-vinylpyrrolidone and divinylbenzene copolymer). For all the analytes, over a wide range of polarity, satisfactory recoveries were obtained between 70% and 120%, with relative standard deviations <15%. Excellent sensitivities were achieved with the limits of quantification ranging from 0.2 μg/kg to 3.0 μg/kg. This developed method provides a new targeted strategy for the analysis of multi-class veterinary drugs in muscle matrices., 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 © 2023 Elsevier Ltd. All rights reserved.)

Veterinary drugs are of concern in terms of potential environmental pollution and their negative impacts on avian scavengers. These pharmaceuticals reach vultures through the consumption of carcasses of previously treated livestock. Here, we analysed samples from livestock carcasses (n = 159), avian scavenger tissues (n = 116) and plasma (n = 312) for 49 compounds commonly used in veterinary medicine in Aragon (NE Spain) and nearby regions. Samples were analysed using liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS/MS). We detected pharmaceuticals in 54.1% of livestock carcasses analysed (50.3% with antibiotics, 10.8% with NSAIDs). For veterinary pharmaceuticals in tissues and plasma from avian scavengers, we detected pharmaceuticals in 51.7% and 28.5% of samples, respectively. Antibiotics were detected in 50.9% and 25.3% while NSAIDs were determined in 6.0% and 5.5% of tissues and plasma from avian scavengers, respectively. Moreover, caffeine was detected in plasma in 73.7% of vultures sampled at landfill sites, indicating its usefulness as a biomarker of urban garbage ingestion. We found an association between livestock carcasses, especially pigs and chickens, and the presence of veterinary pharmaceuticals in avian scavengers. We highlight that carcass disposal for feeding avian scavengers must address the potential risks posed by veterinary pharmaceutical residues., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests Marta Herrero Villar reports administrative support was provided by Biodiversity Service of the Government of Aragon., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Analyte stability is more commonly a confounding factor in analytical chemistry than many analysts recognize. In this study, we assessed the stability of 31 common veterinary drugs in water and final extracts of bovine (milk and kidney/liver) and chicken (muscle and egg) matrices. Two different sample preparation methods were evaluated for one-month storage of the final extracts at typical room, refrigerator, and freezer temperatures. Liquid chromatography - mass spectrometry (LC-MS) by triple quadrupole and high-resolution techniques was used for analysis of the extracts spiked at different relevant concentrations for general regulatory purposes (10-1000 ng/g sample equivalent). Comparison of results between two labs demonstrated that stable drugs (≤20% loss) at all tested conditions consisted of danofloxacin, enrofloxacin, florfenicol, flubendazole, hydroxy-flubendazole, flumequine, flunixin, 5-hydroxy-flunixin, lincomycin, and meloxicam. The tested drugs found to be the most unstable (>20% loss at room temperature within a matter of days) consisted of the β-lactams (ampicillin, cefalexin, cloxacillin, and penicillin G). Curiously, the following antibiotics (mostly macrolides) were apparently more stable in sample extracts than water: emamectin, erythromycin, ivermectin, lasalocid, monensin, tilmicosin, tulathromycin, and tylosin. Those and the other drug analytes (ciprofloxacin, doxycycline, florfenicol amine, 2-amino-flubendazole, oxytetracycline, sulfadiazine, sulfadimethoxine, sulfamethazine, and trimethoprim) were mostly stable for a month in refrigerated extracts, especially at higher concentrations, but not in all cases. In practice, freezer storage of extract solutions was found to be acceptable for at least a month, with a few exceptions.

Veterinary drug residues present in foods can pose severe health threats to the population. The present study aims to develop a high-resolution mass spectral library of 158 veterinary drugs of 16 different classes for their rapid identification in food samples through liquid chromatography-high-resolution electrospray ionization-tandem mass spectrometry (LC-HR-ESI-MS/MS). Standard drugs were pooled according to their log P values and exact masses before analysis. Spectra were collected at system automated collision energy, i.e., of 25-60 eV and four predetermined collision energies (10, 20, 30, and 40 eV) for each compound using a schedule precursor list of [M + H] + , [M + Na] + , and [M + NH 4 ] + ions. The utility of the developed database was checked by analyzing food samples. A total of 17 veterinary drugs based on the reference standard retention times (RTs), HR-MS spectra, and MS/MS spectra were identified in the analyzed samples. Moreover, five veterinary drugs were selected for quantitative analysis, including doxycycline hyclate, lincomycin, sulfasalazine, moxifloxacin, and diphenoxylate, using liquid chromatography-ion trap mass-spectrometry (LC-IT-MS). Concentrations of the drug were obtained to vary from 0.0805 to 0.9731 mg/kg in food samples and were found to be exceeded in most of the cases as per the maximum residue levels described by Food and Agriculture Organization (FAO)/World Health Organization (WHO). The MS data were submitted to the MetaboLights online database (MTBLS2914). This study will help in the high-throughput screening of multiclass veterinary drugs in foodstuffs.

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.

The use of veterinary drugs (VDs) is widely administered to animals for both therapeutic and prophylactic purposes. However, their improper use may involve their occurrence in the final products intended for human consumption. In this scientific work, a method for the investigation of target (n = 30) VDs residues and retrospective suspect screening followed by confirmation using analytical standards of others 38 contaminants in ready-to-eat cooked ham by ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was developed. The extraction was performed based on the QuEChERS approach and validated in accordance with the European Regulation 2021/808. The application of the in-house validated method to ready-to-eat cooked ham showed the occurrence of fourteen VDs residues. Despite the important incidence, the concentration levels found were below the maximum residue limits set for VDs in porcine muscle, except for colchicine. Constant monitoring of animals derived food is strongly recommended to ensure the food safety of consumers., 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 Ltd. All rights reserved.)

This work aimed to develop an integrated high-throughput screening and quantification for multi-class veterinary drug residues by HPLC-Q-Orbitrap mass spectrometry. A qualitative screening mass database of 171 veterinary drugs was created using full scanning mode, which improved the screening accuracy and scope. Beef and chicken samples were chosen to validate the quantification method at three spiked concentration levels. The quantification method of 146 veterinary drug residues was developed. After enzymatic hydrolysis, beef and chicken samples were treated using optimized QuEChERS. The calibration curves showed good linearities with correlation coefficients of 0.9921-0.9994. The recovery rates were within 52.1-138.2 % with relative standard deviations 0.4-17.7 %. The limits of detection and limits of quantification were in the range of 0.15-3.03 μg/kg and 0.5-10 μg/kg, respectively. The proposed method was demonstrated to be reliable for the simultaneous analysis of multi-class veterinary drugs. It is of significance to expand the screening scope and quantitative analysis efficiency., 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 Ltd. All rights reserved.)

This study investigated the behavior of veterinary antibiotics (VAs) in a small farm ecosystem. Manure and environmental samples were collected around a large pig farm in northeast China. Thirty-four VAs in six categories were analyzed. Then, a multimedia fugacity model was used to estimate the fates of VAs in the environment. The results showed that VAs were prevalent in manure, soil, water, and sediment, but not in crops. Compared with fresh manure, VA levels were significantly lower in surface manure piles left in the open air for 3-6 months. The main VAs, tetracyclines and quinolones, decreased by 427.12 and 158.45 µg/kg, respectively. VAs from manure piles were transported to the surroundings and migrated vertically into deep soil. The concentrations of ∑VAs detected in agricultural soils were 0.03-4.60 µg/kg; > 94% of the mass inventory of the VAs was retained in soil organic matter (SOM), suggesting that SOM is the main reservoir for antibiotics in soil. Risk assessment and model analysis indicated that the negative impact of mixed antibiotics at low concentrations in farmland on crops may be mediated by indirect effects, rather than direct effects. Our findings highlight the environmental fates and risks of antibiotics from livestock farms., 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.)

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 content of veterinary drugs in manure is usually estimated by the amount of residues determined in its solid or liquid phase, individually, which previously required a separation step. As an alternative, a multiresidue method for the analysis of 48 veterinary drugs and other contaminants of emerging concern (CECs) in swine raw manure as a whole has been developed and in-house validated in this work. The impact of several experimental factors during ultrasound assisted extraction was assessed. Hence, the use of alumina seemed to especially decrease the matrix effect and improve the overall recovery of drugs, mainly those with a high octanol-water partition coefficient. CECs in the extracts were analyzed by ultra-high performance liquid chromatography coupled to mass spectrometry in tandem. A standard addition-matrix matched calibration was used for quantification. Application of the method to two related samples (raw manure and farm centrifuged raw manure) from a facility revealed that the concentrations of CECs determined in the raw manure by the comprehensive methodology were higher than those calculated by adding the concentrations measured in the solid and liquid phases, separately. This was attributed to the loss of CECs adsorbed on fine particles in the suspension during the sample preparation procedure of the liquid-phase. Furthermore, the decrease of residues in the raw manure when this is centrifuged in the farm to yield compost is shown.

Veterinary drugs are widely used to improve the health and growth of livestock. The supervision of these residues is necessary to ensure food safety. A high-throughput method based on Oasis PRiME HLB with solid phase extraction for simultaneous qualitative and quantitative analysis of 155 veterinary drugs in livestock foods was developed by the ultra-high performance liquid chromatography tandem quadrupole linear-ion-trap mass spectrometry (UHPLC-QTRAP-MS). The limits of detection and quantification ranged from 0.5 μg/kg to 5 μg/kg and 2 μg/kg to 20 μg/kg, respectively. For over 85% of the analytes, the recoveries were between 60% and 120%. The positive simulated samples perfectly matched with a purity fit value over 70% from the self-built library. The screening results of UHPLC-QTRAP-MS were almost consistent with UHPLC tandem quadrupole-exactive orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap/MS). The evaluated UHPLC-QTRAP-MS method was powerful and reliable for the screening and quantification of veterinary drugs in real samples., (Copyright © 2022 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.

Current screening capabilities for veterinary drugs (VDs) in foods are limited, requiring time-consuming and expensive trace-level analyses. For the first time, a high-throughput planar solid-phase extraction (HTpSPE) cleanup, controlled by UV/vis/FLD imaging, was developed for screening 81 VDs from 6 different groups (glucocorticoids, anthelmintics, antiparasitics, coccidiostats, nonsteroidal anti-inflammatory drugs, and antibiotics) in 4 different matrices (honey, pig muscle, cow milk, and chicken eggs). It consumed 13 times less solvent and was more eco-friendly and 5 times faster than routine methods. The VDs were automatically eluted using the autoTLC-LC-MS interface, separated online on a high-performance liquid chromatography column via a 10-min gradient, and detected by Orbitrap high-resolution tandem mass spectrometry. The screening method was validated according to the latest European Commission Implementing Regulation 2021/808. Most VDs except penicillins and cephalosporins were detected at the 5-μg/kg level in pig muscle, cow milk, and chicken eggs and 25-μg/kg level in honey.

A quantitative multi-class multi-residue analytical method was developed for the determination of veterinary drugs in milk by high-performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS). A total of 132 veterinary drugs investigated belonged to almost 15 classes including sulfonamides, β-lactams, tetracyclines, quinolones, macrolides, nitrofurans, nitroimidazoles, phenicols, lincosamides, pleuromutilins, macrocyclic lactones, quinoxaline antibiotics, benzimidazoles, anthelmintics, coccidiostats and some others. A magnetic solid-phase extraction procedure was developed using magnetic hypercrosslinked polystyrene (HCP/Fe 3 O 4 ) for the sample preparation prior to HPLC-MS/MS without deproteinization step. The results indicated recoveries of 85-107% for 14 sulfonamides, 85-120% for 13 β-lactams, 89-115% for 4 tetracyclines, 82-119% for 14 quinolones, 82-115% for 8 macrolides, 97-109% for 4 nitrofurans, 84-115% for 10 nitroimidazoles, 89-114% for 3 phenicols, 86-111% for 3 lincosamides, 97-102% for 2 pleuromutilins, 72-88% for 4 macrocyclic lactones, 87-104% for 4 quinoxaline antibiotics, 76-119% for 21 benzimidazoles, 79-115% for 12 anthelmintics, 81-118% for 12 coccidiostats and 75-119 % for 5 unclassified drugs, with relative standard deviations (RSDs) of less than 20%, and the LOQs ranged from 0.05 to 1 μg kg -1 . This methodology was then applied to field-collected real milk samples and trace levels of some veterinary drugs were detected., (Copyright © 2022 Elsevier Ltd. 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.)