Detection Of Antibiotic Residues Research Articles

Development of a highly sensitive ampicillin sensor utilizing functionalized aptamers.

To develop a sensitive and simple ampicillin (AMP) sensor for trace antibiotic residue detection, the influencing factors of the modification effect of nanogold-functionalized nucleic acid sequences (Adenine: A, Thymine: T) were comprehensively analyzed in this study, including the modification method, base length and type. It was found that under the same base concentration, longer chains are more likely to reach saturation than shorter chains; and when the base concentration and length are both the same, A exhibits a higher saturation modification level compared to T. Based on these research findings, a highly sensitive fluorescence aptamer sensor for detecting ampicillin was constructed using the optimized functionalized sequence (ployA6-aptamer) and experimental conditions (6 hours binding time between nucleic acid aptamer and complementary strand, pH 7 working solution, 20 minutes detection time) based on the principle of fluorescence resonance energy transfer. The sensor has a detection range of 0.18 ng ml-1 to 3.11 ng ml-1 for ampicillin, with a detection limit of 0.04 ng ml-1. It exhibits significant selectivity and achieves an average recovery rate of 98.71% in tap water and 91.83% in milk. This method can be used not only for residual ampicillin detection, but also for highly sensitive detection of various antibiotics and small biological molecules by replacing the aptamer type. It provides a research basis for the design of highly sensitive fluorescence aptamer sensors and further applications of nanogold@DNA composite structures.

A highly efficient molecularly imprinted fluorescence sensor for selective and sensitive detection of tetracycline antibiotic residues in pork

Tetracyclines (TCs) residues in animal-based foods may cause consumer poisoning or allergic reactions, as well as resistance of pathogenic microorganisms. Therefore, a method to accurately monitor TCs is urgently needed. Here, a sensitive and greenificated molecularly imprinted fluorescent sensor (CDs@MIP) for detecting TCs residues using non-covalent imprinting binding method has been synthesized in a simple and safe manner. The sensor was demonstrated to contain specific adsorption sites and high adsorption capacity by static adsorption test results and Freundlich and Langmuir model analyses, and the maximum adsorption capacity was 45.8 mg/g. Moreover, the sensor exhibited intense blue-green fluorescence under UV light, and the fluorescence was gradually quenched as TCs was adsorbed to achieve the detection. The fluorescent spectrum experiment demonstrated that the prepared sensor has good recognition and detection performance for TCs with a linear range of 0.02–50 mg/L and a detection limit of 0.18 μg/L. Furthermore, This sensor is recyclable and can be used to determine TCs content in pork with a recovery rate of 98 %–108 %. Therefore, the constructed sensor provides a new convenient strategy for detecting TCs residues.

Progress in graphene-based electrochemical sensors for antibiotic residue detection

Progress in graphene-based electrochemical sensors for antibiotic residue detection

CRISPR-Cas12a-mediated colorimetric aptasensor of netilmicin based on enzymes-assisted signal amplification and nanozyme employing a rationally truncated aptamer

Netilmicin (NET) is a typical veterinary antibiotic used in aquaculture and animal husbandry, but its excessive accumulation poses a serious threat to the ecological environment and human health. Therefore, a novel and highly sensitive Crispr-Cas12a-mediated colorimetric aptasensor was developed by combining the enzyme-assisted signal amplification strategy and magnetic NMOF-Pt nanozyme composite material (NPSM) for the visual detection of veterinary antibiotic residues in real samples. A high-affinity truncated aptamer (N-20) targeting NET was evolved from a parental aptamer sequence with the help of comprehensive recognition mechanism studies. Subsequently, N-20 was elaborately designed into the isothermal enzyme-assisted strand displacement amplification reaction. The amplified products (Act-DNAs) activated the trans-cleavage activity in the Crispr Cas12a system. The NMOF-Pt nanozyme was released from NPSM after cleaving the single chain of DNA (ssDNA). The quantity of released nanozyme was positively correlated with the amount of NET to quantitatively achieve the detection with the linear range and limits of detection (LOD) of about 0.003–30 ng/mL and 1.15 pg/mL, respectively. This method was successfully applied to detect NET in Lake water, milk and pork samples with satisfactory recoveries ranging from 89.9 % to 104.3 %. The constructed aptasensor demonstrated superior analytical performance and exhibited enormous application prospects in food supervision, environment monitoring, and medical laboratory science.

Development of transcriptional factor-based whole-cell biosensors to monitor and degrade antibiotics using mutant cells obtained via adaptive laboratory evolution

With the widespread use of antibiotics and increasing environmental concerns regarding antibiotic abuse, the detection and degradation of antibiotic residues in various samples has become a pressing issue. Transcriptional factor (TF)-based whole-cell biosensors are low-cost, easy-to-use, and flexible tools for detecting chemicals and controlling bioprocesses. However, because of cytotoxicity caused by antibiotics, the application of such biosensors is limited in the presence of antibiotics. In this study, we used antibiotic-tolerant mutants obtained via adaptive laboratory evolution (ALE) to develop TF-based whole-cell biosensors for antibiotic monitoring and degradation. The biosensors had high performance and stability in detecting relatively high concentrations of tetracycline (Tc) and nisin. The ALE mutant-based Tc biosensor exhibited a 10-fold larger linear detection range than the wild-type strain-based biosensor. Then, the Tc biosensor was employed to detect residual amounts of Tc in mouse stool, serum, and urine samples and facilitate Tc biodegradation in mouse stool, demonstrating its high utility. Considering that ALE has been demonstrated to enhance cell tolerance to various toxic chemicals, our strategy might facilitate the development of whole-cell biosensors for most antibiotics and other toxic ligands.

A homogenous electrochemical biosensor for ultrasensitive ofloxacin detection based on AuNPs-enhanced synergistic amplification of enzymes

Antibiotic residue in food is still a serious health hazard to human with the various and severe adverse effects, so ultrasensitive detection of antibiotic residue becomes increasingly significant. Herein, a homogenous electrochemical biosensor is established for ultrasensitive detection of antibiotic ofloxacin (OFL) based on AuNPs-enhanced synergistic amplification of dual enzymes. Target OFL is ingeniously designed to initiate the synergistic effect of exonuclease III-catalyzed DNA cycle and terminal deoxynucleotidyl transferase-mediated polymerization, producing a great many polyA@AuNPs which subsequently adsorb onto the surface of gold electrode and finally lead to highly amplified electrochemical signal by binding with numerous electroactive indicator RuHex. Benefiting from the collaboratively multiple signal amplifications enhanced by AuNPs with good electrical conductivity and probe loading capacity, the constructed biosensor exhibits excellent quantitative detection performance for OFL with a linear range of 0.008 to 100 ng mL−1 and a limit of detection as low as 2.73 pg mL−1. Furthermore, the detection system accomplished in homogeneous solution without complex chemical modification and rebuilding of sensing architectures on the electrode endows the biosensor with the advantages of convenient operation, good reproducibility and stability. Moreover, the biosensor shows satisfactory analytical performance for OFL detection in real food samples, providing a potential approach for practical application.

Aptamer-free upconversion nanoparticle/silk biosensor system for low-cost and highly sensitive detection of antibiotic residues

The detection of antibiotics is crucial for safeguarding the environment, ensuring food safety, and promoting human health. However, developing a rapid, convenient, low-cost, and sensitive method for antibiotic detection presents significant challenges. Herein, an aptamer-free biosensor was successfully constructed using upconversion nanoparticles (UCNPs) coated with silk fibroin (SF), based on Förster resonance energy transfer (FRET) and the charge-transfer effect, for detecting roxithromycin (RXM). A synergistic FRET efficiency was achieved by utilizing alizarin red and RXM complexes as energy acceptors, with UCNP as the energy donor, and immobilizing an ultrathin SF protein corona within 10 nm. The biosensor detects RXM in deionized water with high sensitivity primarily through monolayer adsorption, with a detection range of 1.0 nM–141.6 nM and a detection limit as low as 0.68 nM. The performance of this biosensor was compared with the ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) method for detecting antibiotics in river water separately and a strong correlation between the two methods was observed. The biosensor exhibited long-term stability in aqueous solutions (up to 60 d) with no attenuation of fluorescence intensity. Furthermore, the biosensor's applicability extended to the highly sensitive detection of other antibiotics, such as azithromycin. This study introduces a low-cost, eco-friendly, and highly sensitive method for antibiotic detection, with broad potential for future applications in environmental, healthcare, and food-related fields.

Potential of a deep eutectic solvent in silver nanoparticle fabrication for antibiotic residue detection.

Deep eutectic solvents (DESs) have recently emerged as an alternative solvent for nanoparticle synthesis. There have been numerous advancements in the fabrication of silver nanoparticles (Ag NPs), but the potential of DESs in Ag NP synthesis was neither considered nor studied carefully. In this study, we present a novel strategy to fabricate Ag NPs in a DES (Ag NPs-DES). The DES composed of ᴅ-glucose, urea, and glycerol does not contain any anions to precipitate with Ag+ cations. Our Ag NPs-DES sample is used in a surface-enhanced Raman scattering (SERS) sensor. The two analytes for SERS quantitation are nitrofurantoin (NFT) and sulfadiazine (SDZ) whose residues can be traced down to 10-8 M. The highest enhancement factors (EFs) are competitive at 6.29 × 107 and 1.69 × 107 for NFT and SDZ, respectively. Besides, the linearity coefficients are extremely close to 1 in the range of 10-8 to 10-3 M of concentration, and the SERS substrate shows remarkable uniformity along with great selectivity. This powerful SERS performance indicates that DESs have tremendous potential in the synthesis of nanomaterials for biosensor substrate construction.

Foldable paper-based photoelectrochemical biosensor based on etching reaction of CoOOH nanosheets-coated laser-induced PbS/CdS/graphene for sensitive detection of ampicillin

Timely and rapid detection of antibiotic residues in the environment is conducive to safeguarding human health and promoting an ecological virtuous cycle. A foldable paper-based photoelectrochemical (PEC) sensor was successfully developed for the detection of ampicillin (AMP) based on glutathione/zirconium dioxide hollow nanorods/aptamer (GSH@ZrO2 HS@apt) modified cellulose paper as a reactive zone with laser direct-writing lead sulfide/cadmium sulfide/graphene (PbS/CdS/LIG) as photoelectrode and cobalt hydroxide (CoOOH) as a photoresist material. Initially, AMP was introduced into the paper-based reaction zone as a biogate aptamer, which specifically recognized the target and then left the ZrO2 HS surface, releasing glutathione (GSH) encapsulated inside. Subsequently, the introduction of GSH into the reaction region and etching of CoOOH nanosheets to expose the PbS/CdS/LIG photosensitive material increased photocurrent. Under optimal conditions, the paper-based PEC biosensor showed a linear response to AMP in the range of 5.0 - 2 × 104 pM with a detection limit of 1.36 pM (S/N = 3). In addition, the constructed PEC sensing platform has excellent selectivity, high stability and favorable reproducibility, and can be used to assess AMP residue levels in various real water samples (milk, tap water, river water), indicating its promising application in environmental antibiotic detection.

Aggregation-induced-emission red carbon dots for ratiometric sensing of norfloxacin and anti-counterfeiting

The detection of trace antibiotic residues holds significant importance because it’s related to food safety and human health. In this study, we developed a new high-yield red-emitting carbon dots (R-CDs) with aggregation-induced emission properties for ratiometric sensing of norfloxacin. R-CDs were prepared in 30 min using an economical and efficient microwave-assisted method with tartaric acid and o-phenylenediamine as precursors, achieving a high yield of 34.4 %. R-CDs showed concentration-dependent fluorescence and aggregation-induced-emission properties. A ratiometric fluorescent probe for detecting the norfloxacin was developed. In the range of 0–40 μM, the intensity ratio of two emission peaks (I445 nm/I395 nm) towards norfloxacin show good linear relationship with its concentrations and a low detection limit was obtained (36.78 nM). In addition, complex patterns were developed for anti-counterfeiting based on different emission phenomenon at different concentrations. In summary, this study designed a novel ratiometric fluorescent probe for detection of norfloxacin, which greatly shortens the detection time and improves efficiency compared with high-performance liquid chromatography and other methods. The study will promote the application of carbon dots in anti-counterfeiting and other related fields, laying the foundation for the preparation of low-cost photosensitive anti-counterfeiting materials.

Detection of Antibiotic Residues in Honeys from Different Regions in Türkiye by Liquid Chromatography-Tandem Mass Spectrometry Method

This study aimed to investigate 29 antibiotics and their metabolites in 27 honey samples obtained from different provinces of Turkey by Liquid Chromatography-Tandem Mass Spectrometry Method (LS-MS/MS). This study showed that the correlation coefficients of the calibration graphs were 0.999, the limit of detection (LOD) was 0.94–3.40 ng/g, and the limit of quantification (LOQ) was 3.11–11.22 ng/g. To express the accuracy of the method, intra- and inter-day recoveries were tested using three different concentrations from 0.25 to 1 μg/kg. Intra-day recoveries for antibiotics and metabolites were found to be 95.56–115.56% with relative standard deviation values between 0.43 and 6.58; inter-day recoveries were found to be 90.00–108.89% with relative standard deviation values between 0.54 and 5.31. The analysis results showed that no antibiotic residues were found in any of the honey samples. The honey did not pose any danger to food safety or public health.

Iron-driven self-assembly of dopamine into dumbbell-shaped nanozyme for visual and rapid detection of norfloxacin on a smartphone-assisted platform

Antibiotics in aquatic environments raise health concerns. Therefore, the rapid, on-site, and accurate detection of antibiotic residues is crucial for protecting the environment and human health. Herein, a dumbbell-shaped iron (Fe3+)-dopamine coordination nanozyme (Fe-DCzyme) was developed via an iron-driven self-assembly strategy. It exhibited excellent peroxidase-like activity, which can be quenched by adding l-cysteine to prevent Fe3+/Fe2+ electron transfer but restored by adding norfloxacin. Given the ‘On-Off-On’ effect of peroxidase-like activity, Fe-DCzyme was used as a colourimetric sensor for norfloxacin detection, and showed a wide linear range from 0.05 to 6.00 μM (R2 = 0.9950) and LOD of 27.0 nM. A portable smartphone-assisted detection platform using Fe-DCzyme was also designed to convert norfloxacin-induced color changes into RGB values as well as to realise the rapid, on-site and quantitative detection of norfloxacin. A good linear relation (0.10–6.00 μM) and high sensitivity (LOD = 79.3 nM) were achieved for the smartphone-assisted Fe-DCzyme detection platform. Its application was verified using norfloxacin spiking methods with satisfactory recoveries (92.66%–119.65%). Therefore, the portable smartphone-assisted Fe-DCzyme detection platform with low cost and easy operation can be used for the rapid, on-site and visual quantitative detection of antibiotic residues in water samples.

Label-free simultaneous detection of quinolone antibiotic residues using an origami paper–based electrochemical immunosensor

Label-free electrochemical sensors have gained popularity as point-of-care devices because they do not require complicated labeling procedures. However, they have difficulties detecting multiple analytes simultaneously. In this study, we developed a novel label-free electrochemical immunosensor on an origami paper–based analytical device (oPAD) for the simultaneous detection of quinolone antibiotics, in which norfloxacin (Nor) and enrofloxacin (Enr) were used as a model. The oPAD was designed with two antibody zones, each zone contains different antibodies. The oPAD has two thionine- and ferrocenecarboxylic acid-coated reagent zones, as redox species, which will generate the response current signals for the detection of quinolone antibiotics. The simultaneous current responses of thionine and ferrocenecarboxylic acid decreased proportionately when exposed to Nor and Enr concentrations, respectively. Under optimal conditions, this device provided linear range at concentrations of 0.01–10 µg/mL for the detection of Nor and Enr and limit of detection was calculated as 2.02 and 1.70 ng/mL, respectively. Moreover, this device demonstrates performance comparable to standard methods for the simultaneous detection of Nor and Enr in milk, honey, and fish. Therefore, label-free electrochemical immunosensors provide a highly potent, inexpensive, and portable method for detecting multiple quinolone antibiotics simultaneously on a single device.

Synthesis and characterization of Au@Ag/Ce-UIO-66 nanocomposite for highly sensitive detection of sulfadiazine residue

Antibiotic residue detection plays a key role in ensuring human health. However, rapid, reliable, and highly sensitive detection methods are still insufficient. To tackle this challenge, in this study, the Au@Ag/Ce-UIO-66, a nanocomposite material consisting of bimetallic gold core and silver shell (Au@Ag) introduced on the surface of Ce-UIO-66, was successfully synthesized. By meticulously regulating the exact concentration and volume of silver nitrate, we successfully generated an optimal matrix. This significantly enhanced the detection capability of the probe molecule CV to an impressive concentration of 10−9 M, with a remarkably low relative standard deviation of 7.54 %. The matrix exhibits excellent surface-enhanced Raman scattering (SERS) performance and stability, reaching adsorption equilibrium within 30 min. Moreover, employing this substrate enables the detection of sulfadiazine at concentrations as minimal as 2.5 μg/L, significantly surpassing the standards set by the European Union. Notably, the Raman spectral signal of sulfadiazine maintains stability across various pH conditions at a concentration of 10−5 M. Finally, the adsorption mechanism and enhancement mechanism between Au@Ag/Ce-UIO-66 and sulfadiazine were explored.

A Novel Approach for Single-Step Analyte Fractionation of Raw Milk Prior to Antibiotic Residue Trace Analysis as an Alternative to QuEchERS-Based Extraction.

Antibiotic residues in milk are a well-known hazard in the dairy food chain. Detection methods for these residues, such as nonspecific microbiological inhibitor tests or group-specific receptor tests, are relatively inexpensive, easy to use, and widely applied to ensure food safety. In contrast, specific detection by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-although a critical, complimentary method to confirm the results of nonspecific testing-is relatively costly, time-consuming, and laborious. Furthermore, sample processing before LC-MS/MS analysis requires unique preparation procedures for different groups of antibiotic compounds. To simplify and speed up specific antibiotic residue detection, a low-cost, passive, and single-step method to fractionate analytes in raw milk was developed. Untreated raw milk was fractionated into its water and fat/protein phases using a Fractionation of Milk for Trace Analysis of Contaminants and Residues for Antibiotics (FraMiTrACR® AB) fractionation unit. The water fraction was then analyzed by LC-MS/MS. The analyte fractionation method was evaluated against a Quick Easy Cheap Effective Rugged and Safe (QuEChERS)-based method for sample preparation. Our method allows qualitative and quantitative detection of substances from the penicillin, cephalosporin, macrolide, lincosamide, sulfonamide, tetracycline, and fluoroquinolone groups of antibiotics. Detection limits are below the legally prescribed maximum residue levels, allowing reliable, specific, and rapid validation of a positive result in nonspecific microbiological inhibitor tests. Analyte fractionation by FraMiTrACR AB is a faster alternative to QuEChERS-based sample preparation for the detection of antibiotic substances in milk. This method describes a low-cost, environmentally friendly, passive, and single-step milk analyte fractionation. As an alternative to QuEChERS-based preparation, this fractionation method simplifies and speeds up the process for specific antibiotic residue detection.

Colorimetric aptasensor utilizing MOF-235 with exceptional peroxidase-like activity for the detection of oxytetracycline residues in raw milk

In this work, a simple, convenient and cost-effective colorimetric aptasensor was successfully constructed for the detection of antibiotic residues in raw milk based on the property that aptamer (Apt) synergistically enhances the catalase-like activity of MOF-235. Under optimised conditions, the proposed colorimetric aptasensor exhibited a wide detection range (15–1500 nM) with a low detection limit (6.92 nM). Furthermore, the proposed aptasensor demonstrated high selectivity, good resistance to interference and storage stability. The proposed aptasensor was validated by spiking recovery in camel milk, cow milk and goat milk with satisfactory recoveries, which demonstrated the great potential of the aptasensor for further application in real food samples, and also suggested that MOF-235 can be used as a potential universal platform to build a sensitive detection platform for other targets.

Rapid discrimination and ratio quantification of mixed antibiotics in aqueous solution through integrative analysis of SERS spectra via CNN combined with NN-EN model

IntroductionAbusing antibiotic residues in the natural environment has become a severe public health and ecological environmental problem. The side effects of its biochemical and physiological consequences are severe. To avoid antibiotic contamination in water, implementing universal and rapid antibiotic residue detection technology is critical to maintaining antibiotic safety in aquatic environments. Surface-enhanced Raman spectroscopy (SERS) provides a powerful tool for identifying small molecular components with high sensitivity and selectivity. However, it remains a challenge to identify pure antibiotics from SERS spectra due to coexisting components in the mixture. ObjectivesIn this study, an intelligent analysis model for the SERS spectrum based on a deep learning algorithm was proposed for rapid identification of the antibiotic components in the mixture and quantitative determination of the ratios of these components. MethodsWe established a water environment system containing three antibiotic residues of ciprofloxacin, doxycycline, and levofloxacin. To facilitate qualitative and quantitative analysis of the SERS spectra antibiotic mixture datasets, we developed a computational framework integrating a convolutional neural network (CNN) and a non-negative elastic network (NN-EN) method. ResultsThe experimental results demonstrate that the CNN model has a recognition accuracy of 98.68%, and the interpretation analysis of Shapley Additive exPlanations (SHAP) shows that our model can specifically focus on the characteristic peak distribution. In contrast, the NN-EN model can accurately quantify each component's ratio in the mixture. ConclusionIntegrating the SERS technique assisted by the CNN combined with the NN-EN model exhibits great potential for rapid identification and high-precision quantification of antibiotic residues in aquatic environments.

Dual-mode detection of antibiotic drugs using ytterbium molybdate/porous carbon nanocomposite

Monitoring the levels of metronidazole (MTZ) is crucial to ensure the desired therapeutic effectiveness and to prevent potential harm to patients. However, MTZ exhibits concentration-dependent antimicrobial activity and has a narrow therapeutic range. In this study, “one stone two mango” strategy was influenced for the electrochemical and UV–vis spectroscopy detection of antibiotic residues of MTZ. The successfully synthesized ytterbium molybdate nano petals (YbMO) with porous carbon (PC) nanocomposite (NC) using ultrasonication method. The YbMO/PC NC, as well as other modified electrodes, were identified and targeted as molecules using natural pH as the probe to indicate current intensity. Also, examined the morphology and physicochemical properties of the synthesized materials. Furthermore, conducted cycling voltammetry (CV) and linear sweep voltammetry (LSV) on the modified electrodes to model MTZ analytes. Among them, the YbMO/PC NC/GCE exhibited larger reduction peak current and lower potential compared to other modified electrodes due to the synergistic effect between YbMO and PC, resulting in effective detection of MTZ. The YbMO/PC NC/GCE demonstrated exceptional sensitivity (3.28 µA µM−1 cm−2), a wide linear range (0.01–10.61 and 20.61–1630.61 µM), and a remarkably low detection limit (LOD: 0.006 µM). The proposed YbMO/PC NC/GCE exhibits the successfully analyzed MTZ in serum (101%), urine (97.9%), tablets (99.15%), lake (99.1%), and river water (99.8%). On the other hand, a UV–vis spectrometric method was employed to detect MTZ. The results showed a wide linear range (0.05–57.2 µM) and an impressive LOD (70 nM). These results are attributed to the heightened sensitivity, selectivity, repeatability, and durability of the sensor.

A double defects-dominated flexible TiO2 matrix for in-situ SERS sensing of antibiotic residues in aquatic ecosystem (fish & fishpond water) and their on-site degradation in flowing water

High-performance flexible semiconductor material can be used as an excellent multifunctional matrix for in-situ ultrasensitive surface-enhanced Raman scattering (SERS) detection and synchronous photocatalytic degradation of antibiotic residues in aquatic ecosystem. Here, a calcium-doped TiO2 flexible matrix with double defects (surface oxygen vacancy defect and Ti3+ energy level defect) was developed by its “in-situ one-step” hydrothermal synthesis on cotton fabric for the above purposes. Due to the joint contribution of double defects, a multi-channel charge transfer mode and a high-efficiency carrier separation are achieved, which endows flexible cotton fabric/Ca-doped TiO2 (Cot/Ca-TiO2) substrate with the greatly boosted SERS effect for in-situ detection of antibiotic residues on fish body surface and in fishpond water by a simple wiping or dipping sampling method, even for simultaneous identification of multi-component residues. The detection limits of three antibiotic residues (enrofloxacin, ciprofloxacin and enoxacin) are as low as 10−9 M, which are far lower than the EU standard. More meaningfully, the flexible Cot/Ca-TiO2 can be used as a multifunctional filter-membrane type photocatalyst for efficient on-site degradation of antibiotic residues in flowing fishpond water by a multi-grade photocatalysis means. Moreover, the flexible matrix exhibits good recyclability in both actual detection and photocatalysis.

Detection of antibiotic residues in groundwater with a validated multiresidue UHPLC-MS/MS quantification method

The occurrence of antibiotic residues in the environment has received considerable attention because of their potential to select for bacterial resistance. The overuse of antibiotics in human medicine and animal production results in antibiotic residues entering the aquatic environment, but concentrations are currently not well determined. This study investigates the occurrence of antibiotics in groundwater in areas strongly related to agriculture and the antibiotic treatment of animals. A multiresidue method was validated according to EU Regulation 2021/808, to allow (semi-)quantitative analysis of 78 antibiotics from 10 different classes: β-lactams, sulfonamides, tetracyclines, lincosamides, amphenicols, (fluoro)quinolones, macrolides, pleuromutilins, ansamycins and diaminopyrimidines using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). This method was used to test different storage conditions of these water samples during a stability study over a period of 2 weeks. Sulfonamides, lincosamides and pleuromutilins were the most stable. Degradation was most pronounced for β-lactam antibiotics, macrolides and ansamycins. To maintain stability, storage of samples at −18 °C is preferred. With the validated method, antibiotic residues were detected in groundwater, sampled from regions associated with intensive livestock farming in Flanders (Belgium). Out of 50 samples, 14% contained at least one residue. Concentrations were low, ranging from < LOD to 0.03 μg/L. Chloramphenicol, oxolinic acid, tetracycline and sulfonamides (sulfadiazine, sulfadoxine, sulfamethazine and sulfisoxazole) were detected. This study presents a new method for the quantification of antibiotic residues, which was applied to investigate the presence of antibiotic residues in groundwater in Flanders.