Drug Residues In Food Research Articles

Evaluation of certain veterinary drug residues in food

Evaluation of certain veterinary drug residues in food

An in vitro study to assess the impact of tetracycline on the human intestinal microbiome

The human intestinal microbiome, a generally stable ecosystem, could be potentially altered by the ingestion of antimicrobial drug residues in foods derived from animals. Data and the scientific published literature on the effects of antimicrobial residues on the human intestinal microbiome are reviewed by national regulatory authorities as part of the human food safety evaluation of veterinary antimicrobial agents used in food-producing animals. In this study, we determined if tetracycline, at low residue concentrations, could impact the human intestinal microbiome structure and the resistance-gene profile, following acute and subchronic exposure. The effects of 0.15, 1.5, 15, and 150 μg/ml of tetracycline, after 24 h and 40 days of exposure, in 3% human fecal suspensions, collected from three individuals (A, B, and C) were investigated using in vitro batch cultures. Results were variable, with either no change or minor changes in total bacterial 16S rRNA gene copies after exposure of fecal samples to tetracycline, because of the inter-individual variation of human gastrointestinal tract microbiota. Bacterial community analysis using rRNA-based pyrosequencing revealed that Firmicutes and Bacteroidetes were the predominant phyla in the three fecal samples; the ratio of phylotypes varied among individuals. The evaluation of bacterial community changes at the genus level, from control to tetracycline-treated fecal samples, suggested that tetracycline under the conditions of this study could lead to slight differences in the composition of intestinal microbiota. The genus Bacteroides (of the Bacteroidetes) was consistently altered from 1.68 to 5.70% and 4.82–8.22% at tetracycline concentrations of 0.15 μg/ml or above at both time points for individual A, respectively, and increased 5.13–13.50% and 10.92–22.18% for individual B, respectively. Clostridium family XI increased 3.50–25.34% in the presence of tetracycline at 40 days for individual C. Principal Component Analysis (PCA) confirmed the pyrosequencing findings of inter-individual variability of the ratio of phylotypes and the effect of tetracycline. Among the 23 tetracycline resistance genes (TRGs) screened, four tet genes (tetO, Q, W, and X) were major TRGs in control and tetracycline-dosed fecal samples. A variable to slight increase of copy number of TRGs appeared to be related to tetracycline treatment, interindividual variability and duration of exposure. Despite, the inherent variability of the intestinal microbiota observed among or within individuals, this pilot study contributes to the knowledge base of the impact of low residue concentrations of tetracycline on the human intestinal microbiome on the potential for antimicrobial resistance.

Rapid method for the quantification of 13 sulphonamides in milk by conventional high-performance liquid chromatography with diode array ultraviolet detection using a column packed with core-shell particles

In the present study, a column packed with core-shell particles was used for the separation and the quantification of 13 sulphonamides in milk by conventional high-performance liquid chromatography coupled with diode array ultraviolet detection (HPLC/UV-DAD). Preliminary experiments were carried out to investigate selectivity of different stationary phases. Best results were achieved using a C18 column packed with 2.6μm core-shell particles (diameter 4.6mm, length 75mm). A binary gradient elution based on acetate buffer solution at pH 4.50 and a mixture of methanol acetonitrile 50:50 (v/v) was employed at the flow rate of 1.2mLmin−1 with an injection volume of 6μL. These chromatographic conditions allowed the efficient separation of 13 sulphonamides in about 8min. To evaluate the suitability of the method for official control analysis, the most important validation parameters were investigated according to the European Decision 657/2002/EC as established for analysis of drug residues in food. Sulphonamides were recovered from milk samples by a simple and quick preparation procedure consisting of an extraction step with chloroform/acetone and a purification step with n-hexane. Mean recoveries from raw milk ranged between 55% and 86% at the Maximum Residual Limit of 100μgkg−1, and RSDs% resulted lower than Thompson and Horwitz RSD% reference values for all sulphonamides. The LOQ value (2.7–15μgkg−1) was low enough to satisfy legal limits suggested by European Regulation 37/2010/EC.

Determination of multiresidue analysis of β-agonists in muscle and viscera using liquid chromatograph/tandem mass spectrometry with Quick, Easy, Cheap, Effective, Rugged, and Safe methodologies

The official analytical method of the Taiwan Food and Drug Administration, Ministry of Health and Welfare for testing for veterinary drug residues in foods is the multiresidue analysis of β-agonists. Samples are pretreated through liquid–liquid extraction and solid-phase extraction. This method is time consuming and requires the intensive use of solvents. To improve analytical efficiency and reduce costs, our study incorporated QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) techniques to establish a new method of multiresidue analysis of β-agonists in animal muscle and viscera. The pretreatment time was shortened and solvent usage was minimized. The modified analysis was conducted using liquid chromatography/tandem mass spectrometry (LC–MS/MS) and quantification was performed using multiple reaction monitoring. The results demonstrated that the correlation coefficients of the tissue calibration curve were higher than 0.99 and the limit of quantification (LOQ) was 1 ppb. The average recoveries in spiked samples varied from 70% to 120%, and the relative difference between duplicated analysis results was lower than 10%. On the basis of the results, the proposed method was concluded to be an appropriate procedure for determining the presence of β-agonists, and demonstrated the advantages of high recovery rates in spiked samples, high precision, reduced analysis time and solvent usage, and lower costs.

An update discussion on the current assessment of the safety of veterinary antimicrobial drug residues in food with regard to their impact on the human intestinal microbiome.

The human gastrointestinal tract ecosystem consists of complex and diverse microbial communities that have now been collectively termed the intestinal microbiome. Recent scientific breakthroughs and research endeavours have increased our understanding of the important role the intestinal microbiome plays in human health and disease. The use of antimicrobial new animal drugs in food-producing animals may result in the presence of low levels of drug residues in edible foodstuffs. There is concern that antimicrobial new animal drugs in or on animal-derived food products at residue-level concentrations could disrupt the colonization barrier and/or modify the antimicrobial resistance profile of human intestinal bacteria. Therapeutic doses of antimicrobial drugs have been shown to promote shifts in the intestinal microbiome, and these disruptions promote the emergence of antimicrobial-resistant bacteria. To assess the effects of antimicrobial new animal drug residues in food on human intestinal bacteria, many national regulatory agencies and international committees follow a harmonized process, VICH GL36(R), which was issued by a trilateral organization of the European Union, the USA, and Japan called the International Cooperation on Harmonization of Technical Requirements for Veterinary Medicinal Products (VICH). The guidance describes a general approach currently used by national regulatory agencies and international committees to assess the effects of antimicrobial new animal drug residues in animal-derived food on human intestinal bacteria. The purpose of this review is to provide an overview of this current approach as part of the antimicrobial new animal drug approval process in participating countries, give insights on the microbiological endpoints used in this safety evaluation, and discuss the availability of new information. Copyright © 2016 John Wiley & Sons, Ltd.

Recent analysis of veterinary drug residues in food

Recent analysis of veterinary drug residues in food

Multiclass and multi-residue determination of antibiotics in bovine milk by liquid chromatography–tandem mass spectrometry: Combining efficiency of milk control and simplicity of routine analysis

Antibiotics are widely used in veterinary medicine and incorrect practices may lead to drug residues in food, including milk. Fast and simple sample preparation methods have been developed for determination of quinolones, fluoroquinolones, tetracyclines, sulphonamides, trimethoprim and bromexine using liquid chromatography–tandem mass spectrometry. Small volumes of sample (0.5 mL) were extracted using organic solvent followed by centrifugation and evaporation and/or dilution. Validation took into account the maximum residue limits (MRLs). Recoveries of 62–108% were obtained. Linearity (r2) above 0.96 was achieved for all compounds using concentrations in the range 0.25–2.0 × MRL. Intraday precision with coefficients of variation (n = 6) lower than 15.0% and inter-day precision lower than 17.3% were obtained for three concentration levels in the range 0.5–1.5 × MRL. Accuracy was 87–108%. Limits of detection and quantitation, as well as decision limit, detection capability, robustness and applicability, were evaluated using >600 real milk samples.

Simultaneous Determination of Residues of Non-Steroidal Anti-Inflammatory Drugs and Glucocorticosteroids in Animal Muscle by Liquid Chromatography-Tandem Mass Spectrometry

A method for the determination of a wide range residues of anti-inflammatory drugs (16 acidic non-steroidal anti-inflammatory drugs and four metamizole metabolites and five corticosteroids) has been was developed. In the first step of sample preparation, acetate buffer was added to minced muscle samples and 15-min ultrasound-assisted enzymatic hydrolysis was performed. Next, the samples were extracted twice with acetonitrile, freezed and analysed. The analytes were separated on a C18 column with a 25-min gradient of methanol/acetonitrile (8:2) and 0.05 M ammonium formate at pH 5.0 and determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method was validated according to the requirements described in the Commission Decision 2002/657/EC: linearity, precision (repeatability and within-laboratory reproducibility), accuracy, decision limit CCα and detection capability CCβ were calculated. The method developed fulfilled the performance criteria and can be used in the official control of veterinary drug residues in food of animal origin. The method was positively verified in the proficiency test and in the analysis of incurred material.

EFSA's assistance for the 2015 Codex Committee on Residues of Veterinary Drugs in Food (CCRVDF) in relation to rBST

EFSA's assistance for the 2015 Codex Committee on Residues of Veterinary Drugs in Food (CCRVDF) in relation to rBST

Structural characterization of product ions by electrospray ionization and quadrupole time-of-flight mass spectrometry to support regulatory analysis of veterinary drug residues in foods. Part 2: Benzimidazoles, nitromidazoles, phenothiazines, and mectins.

Analysis for identification and quantification of regulated veterinary drug residues in foods is usually achieved by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). The instrumental method requires the selection of characteristic ions, but structural elucidation is seldom performed to help ensure accuracy. This study is a continuation of previous work to characterize selected product ions in support of regulatory monitoring programs. The tandem mass spectra of 28 veterinary drugs from a previously published LC/MS/MS method were acquired with a high-resolution quadrupole time-of-flight (Q-TOF) mass spectrometer using electrospray ionization (ESI) in positive mode. The TOF analyzer was calibrated to achieve a mass accuracy error <5 ppm for the MS and MS/MS modes, and samples were infused for data acquisition. The high mass accuracy achieved in Q-TOF allowed elucidation of the formulae of the product ions previously selected for qualitative identification. Rational interpretation of results was made and compared with the published literature, and the structure for the MS/MS product ions of four classes of regulated drugs (mectins, benzimidazoles, nitroimidazoles, and phenothiazines), totaling 28 compounds, were examined leading to the report of new structures or confirmation of published structures using low-resolution MS. Structural characterization of the product ions selected for identification and quantification of veterinary drug residues is important information for regulatory monitoring programs in defense of regulatory enforcement actions. This study has allowed structural elucidation of 84 MS/MS product ions previously selected for the LC/MS/MS analysis of 28 drug analytes.

Human Health, Environmental Sustainability and Food Production: Interfaces with Nanomedicine

In the case of animal production, use of high-density animals in the farm will be accompanied by challenges such as optimizing the use of inputs (e.g. antibiotics, antiparasitics, and nutrients), greater sanitary control, minimizing the risk of transmission of zoonoses, greater control of drug residues in food, and ensuring low environmental impact. These challenges are a window of opportunity for the use of nanotechnology. Coping with the loss of efficiency of pharmaceutical drugs against common pathogens is a great challenge to public health. Resistant pathogens have emerged due to indiscriminate use of drugs, and the massive use of drugs in food animals may be contributing to this situation. Worldwide, most antimicrobials and antiparasitics that are being used in animal production have to be used in a better way. Drug delivery systems may be important tools to optimize veterinary drug use and then mitigate its use. Nanoencapsulation of drugs can lead to therapeutic gains and reduce the amount of drugs used for animal treatment with goal of control and prevention infectious diseases. This can occur because of the changes in the pharmacokinetic profile of the drugs, reduced toxicity and appropriate delivery.

Veterinary Drug Residues in Food-animal Products: Its Risk Factors and Potential Effects on Public Health

The use of veterinary drugs in food-producing animals has the potential to generate residues in animal derived products (meat, milk, eggs and honey) and poses a health hazard to the consumer. There are many factors influencing the occurrence of residues in animal products such as drug’s properties and their pharmacokinetic characteristics, physicochemical or biological processes of animals and their products. The most likely reason for drug residues might be due to improper drug usage and failure to keep the withdrawal period. The major public health significances of drug residue are development of antimicrobial drug resistance, hypersensitivity reaction, carcinogenicity, mutagenicity, teratogenicity, and disruption of intestinal normal flora. The residual amount ingested is in small amounts and not necessarily toxic. However, there is limited information on the magnitude of veterinary drug residue worldwide. Hence, an extensive work has to be carried out to determine the magnitude of the problem, to prevent the occurrence of veterinary drug residues, and to familiarize all animal health professionals with the knowledge of pharmacokinetics, pharmacodynamics and toxicological effects of veterinary drugs to minimize the potential public health hazards due to drug residues in food of animal origin.

Résidus d’antibiotiques et denrées d’origine animale en Afrique : risques de santé publique

The authors report on the current status of work on residues of veterinary medicinal products and, in particular, antimicrobial residues in foods of animal origin. This review focuses on residues of veterinary antimicrobials, antimicrobials used in livestock production, the concept of residues, and antimicrobial residues in foods of animal origin. Only one antimicrobial substance has been approved in the West African Economic and Monetary Union, compared with 16 substances in Benin and 56 in the European Union. The issue of antimicrobial residues in foods of animal origin has rarely been a serious concern in developing countries, in contrast to the situation in Europe. However, while the prevalence of veterinary drug residues in foods of animal origin is less than 1% in Europe, in some African countries it can be as high as 94%. Antimicrobial residues in foods of animal origin can cause allergies, cancer, alterations in the intestinal flora, bacterial resistance and the inhibition of fermentation in the dairy industry. The harmonisation of regulations in Africa could reduce the circulation of prohibited antimicrobials and lead to the implementation of a plan for the control and surveillance of residues from veterinary medicinal products in foods of animal origin.

Structural characterization of product ions by electrospray ionization and quadrupole time-of-flight mass spectrometry to support regulatory analysis of veterinary drug residues in foods

Monitoring of veterinary drug residues in foods is often conducted using liquid chromatography/tandem mass spectrometry (LC/MS/MS). Results have high economic stakes for producers, but the ions monitored are usually selected due to signal intensities without structural interpretation. In this study, the ion transitions were characterized by high-resolution mass spectrometry. The 62 veterinary drugs from the LC/MS/MS method consisted of sulfonamides, β-lactams, phenicols, macrolides, tetracyclines, fluoroquinolones, non-steroidal anti-inflammatory drugs (NSAIDs), and corticosteroids. They were individually infused into a quadrupole time-of-flight (Q-TOF) mass spectrometer using electrospray ionization (ESI) operated in positive mode. The MS and collision-induced dissociation (CID) MS/MS spectra for each analyte were obtained for structural elucidation. The Q-TOF instrument was calibrated to obtain a mass accuracy error <5 ppm for the MS and MS/MS spectra. The use of high-resolution ESI-Q-TOF-MS for the generation of the MS/MS product ions allowed for the determination of chemical formulae for the analytes, some of which led to new findings. Assigned structures were based on rational interpretation of the most stable possible products with comparison with the scientific literature. In difficult cases, isotopically labeled drugs or hydrogen/deuterium (H/D) exchange experiments were used to help confirm the structures of the product ions. The use of ESI-Q-TOF-MS in this study has allowed structure elucidation of 186 MS/MS product ions previously selected for the LC/MS/MS analysis of 62 veterinary drugs. This serves to reduce the chances of false positives and negatives in the monitoring program, and provides justification and defense in regulatory enforcement actions.

O estado da arte na determinação de resíduos de medicamentos veterinários em alimentos de origem animal empregando técnicas cromatográficas acopladas à espectrometria de massas

The determination of veterinary drug residues in foods of animal origin is an important issue because of the risk these compounds pose to human health in addition to their persistence and tendency to bioaccumulate. In recent years, significant progress has been made in the area and this review presents the state of the art in sample preparation procedures associated with chromatographic techniques coupled to mass spectrometry for multiresidue determination of veterinary drugs in food of animal origin at concentration levels suitable for the control of residues and contaminants in food.

Development of an analytical method to detect metabolites of nitrofurans: Application to the study of furazolidone elimination in Vietnamese black tiger shrimp (Penaeus monodon)

Nitrofurans, banned antibiotics in the European Union (EU), have often been found in imported aquaculture products in the past and are still found nowadays according to the Rapid Alert System for Feed and Food (RASFF) of the European Commission. A quantitative method based on liquid chromatography coupled to isotopic dilution tandem mass spectrometry (LC–IDMS/MS) was developed for the determination of the residues of four nitrofuran antibiotic residues in shrimps. The experimental protocol consisted of an acid-catalysed release of protein-bound metabolites, followed by derivatisation with 2-nitrobenzaldehyde (NBA). Then, a double liquid–liquid extraction with ethyl acetate was performed before LC–IDMS/MS analysis by positive electrospray ionisation (ES+) with multiple reaction monitoring (MRM) of two transitions per compound. An “in-house” validation of the method for shrimp analysis was conducted according to the EU criteria for the analysis of veterinary drug residues in foods. The decision limits (CCalpha) were 0.08–0.36μgkg−1 and the detection capabilities (CCbeta) were 0.12–0.61μgkg−1, which are both below the minimum required performance limit (MRPL) set at 1μgkg−1 by the EU. The developed method was applied to evaluate the elimination of furazolidone residues in shrimp muscles after a contamination experiment. After 28days of decontamination, a concentration of 115μgkg−1 of furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) was still measured in the shrimp muscle.

C18/C8-Functionalized Magnetic Silica Nanospheres (Fe3O4@Si-C8/C18) as Capture Probes for Highly Efficient and Rapid Purification of Veterinary Drug Residues

The purpose of this study is to develop a rapid and efficient purification method for determination of veterinary drug based on magnetic silica nanospheres. We have described a scheme for preparing C18/C8-functionalized magnetic silica nanospheres (Fe3O4@Si-C8/C18) for potential use as capture probes in sample pretreatment. The magnetic Fe3O4 particles were firstly coated by tetraethyl orthosilicate to form magnetic silica nanospheres, which were then modified by both trimethoxy(octadecyl)silane (C18) and trimethoxy(octyl)silane (C8). The Fe3O4@Si-C8/C18 thus prepared could exhibit strong superparamagnetism, highly efficient capture ability, easy-to-use for isolation, and enrichment of veterinary drug residues in food.

The quest for a universal vaccine against ticks: Cross-immunity insights

As blood-sucking parasites, ticks inflict great damage to animals and humans in many parts of the world. The continued use of chemical acaricides is not sustainable due to increasing tick resistance, growing public concern over drug residues in food and in the environment, and the high cost of developing new acaricides. Therefore, an alternative control strategy is urgently needed. Vaccines against ticks have been shown to be functionally feasible, as highlighted by the success of Bm86 vaccines against Rhipicephalus (Boophilus) microplus and closely related tick species. However, a limited number of tick antigens with cross-protective epitopes have been characterized so far, limiting widespread deployment of the available vaccines, including those derived from Bm86. Therefore, identifying tick antigens with potential broad-spectrum protection against multiple tick species is subject of vigorous research at present. In this paper, progress towards effective anti-tick vaccines is reviewed in the light of emerging data from studies including heterologous tick challenge. Taken together, these studies indicate that the decades-long search for a universal tick vaccine is making progress, with such a vaccine likely to be based on multiple cross-reactive antigens.

Anthelmintic drug residues in beef: UPLC-MS/MS method validation, European retail beef survey, and associated exposure and risk assessments

Anthelmintic drugs are widely used to control parasitic infections in cattle. The ProSafeBeef project addressed the need for data on the exposure of European consumers of beef to potentially harmful drug residues. A novel analytical method based on matrix solid-phase dispersive extraction and ultra-performance liquid chromatography-tandem mass spectrometry was validated for 37 anthelmintic drugs and metabolites in muscle (assay decision limits, CCα, = 0.15–10.2 µg kg−1). Seven European countries (France, Spain, Slovenia, Ireland, Italy, Belgium and Portugal) participated in a survey of retail beef purchased in local shops. Of 1061 beef samples analysed, 26 (2.45%) contained detectable residues of anthelmintic drugs (0.2–171 µg kg−1), none above its European Union maximum residue limit (MRL) or action level. Residues detected included closantel, levamisole, doramectin, eprinomectin, moxidectin, ivermectin, albendazole and rafoxanide. In a risk assessment applied to mean residue concentrations across all samples, observed residues accounted for less than 0.1% of the MRL for each compound. An exposure assessment based on the consumption of meat at the 99th percentile of consumption of adults in 14 European countries demonstrated that beef accounted for less than 0.02% of the acceptable daily intake for each compound in each country. This study is the first of its kind to apply such a risk-based approach to an extensive multi-residue survey of veterinary drug residues in food. It has demonstrated that the risk of exposure of the European consumer to anthelmintic drug residues in beef is negligible, indicating that regulation and monitoring is having the desired effect of limiting residues to non-hazardous concentrations.

Nanofiltration and sensing of picomolar chemical residues in aqueous solution using an optical porous resonator in a microelectrofluidic channel

For the first time the use of a porous microresonator placed in a microelectrofluidic system for integrated functions of nanofiltration and sensing of small biomolecules and chemical analytes in extremely dilute solution was proposed and investigated. As an example, aminoglycosides in drug residues in food and livestock products were considered as the trace chemical analyte. The filtration process of the charged analyte in aqueous solution driven by an applied electrical field and the accompanying optical whispering-gallery modes in the resonator are modeled. The dynamic process of adsorption and desorption of the analyte onto the porous matrix is studied. Deposition of the analyte inside the porous structure will alter the material refractive index of the resonator, and thus induce an optical resonance frequency shift. By measuring the optical frequency shift, the analyte concentration as well as the absorption/desorption process can be analyzed. Through an intensive numerical study, a correlation between the frequency shift and the analyte concentration and the applied electrical voltage gradient was obtained. This reveals a linear relationship between the resonance frequency shift and the analyte concentration. The applied electrical voltage substantially enhances the filtration capability and the magnitude of the optical frequency shift, pushing the porous resonator-based sensor to function at the extremely dilute picomolar concentration level for small bio/chemical molecules down to the sub-nanometer scale. Moreover, use of the second-order whispering-gallery mode is found to provide better sensitivity compared with the first-order mode.