Residues In Animal-derived Food Research Articles

A physiologically based pharmacokinetic model to optimize the dosage regimen and withdrawal time of cefquinome in pigs.

Cefquinome is widely used to treat respiratory tract diseases of swine. While extra-label dosages of cefquinome could improve clinical efficacy, they might lead to excessively high residues in animal-derived food. In this study, a physiologically based pharmacokinetic (PBPK) model was calibrated based on the published data and a microdialysis experiment to assess the dosage efficiency and food safety. For the microdialysis experiment, in vitro/in vivo relative recovery and concentration-time curves of cefquinome in the lung interstitium were investigated. This PBPK model is available to predict the drug concentrations in the muscle, kidney, liver, plasma, and lung interstitial fluid. Concentration-time curves of 1000 virtual animals in different tissues were simulated by applying sensitivity and Monte Carlo analyses. By integrating pharmacokinetic/pharmacodynamic target parameters, cefquinome delivered at 3-5 mg/kg twice daily is advised for the effective control of respiratory tract infections of nursery pig, which the bodyweight is around 25 kg. Based on the predicted cefquinome concentrations in edible tissues, the withdrawal interval is 2 and 3 days for label and the extra-label doses, respectively. This study provides a useful tool to optimize the dosage regimen of cefquinome against respiratory tract infections and predicts the concentration of cefquinome residues in edible tissues. This information would be helpful to improve the food safety and guide rational drug usage.

A new semiconductor heterojunction SERS substrate for ultra-sensitive detection of antibiotic residues in egg

Antibiotic residues in animal-derived food (egg) are threatening human health. Semiconductor heterojunction surface-enhanced Raman scattering (SERS) substrates can be used for ultra-sensitive detection of antibiotic residues in egg. Here, a TiO2/ZnO heterojunction was developed as a new SERS substrate based on an interface engineering strategy. Due to strong interfacial coupling and efficient carrier separating in heterostructure, utilization rate of photo-induced electrons in substrate was improved greatly, which realized the efficient charge transfer in substrate-molecule system, resulting in a prominent SERS enhancement. Taking the detection of enrofloxacin residue in egg as an example, the limit of detection (LOD) is only 13.1 μg/kg, which is far below the European Union standard, and lower than LODs of other conventional analytical methods and existing noble metal-based SERS methods. More importantly, benefiting from high sensitivity and selectivity of heterojunction and fingerprint characteristics of SERS, multiple antibiotic residues in egg can be identified simultaneously.

Few Layer Ti3C2 MXene-Based Label-Free Aptasensor for Ultrasensitive Determination of Chloramphenicol in Milk.

Quantitative detection of veterinary drug residues in animal-derived food is of great significance. In this work, a simple and label-free electrochemical aptasensor for the highly sensitive detection of chloramphenicol (CAP) in milk was successfully developed based on a new biosensing method, where the single- or few-layer Ti3C2 MXene nanosheets functionalized via the specific aptamer by self-assembly were used as electrode modifiers for a glassy carbon electrode (aptamer/Ti3C2 MXene/GCE). Differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), scanning electron microscopy (SEM), atomic force microscope (AFM), and so on were utilized for electrochemical and morphological characterization. Under the optimized conditions, the constructed aptasensor exhibited excellent performance with a wider linearity to CAP in the range from 10 fM to 1 μM and a low detection limit of 1 fM. Aptamer/Ti3C2 MXene/GCE demonstrated remarkable selectivity over other potentially interfering antibiotics, as well as exceptional reproducibility and stability. In addition, the aptasensor was successfully applied to determine CAP in milk with acceptable recovery values of 96.13% to 108.15% and relative standard deviations below 9%. Therefore, the proposed electrochemical aptasensor is an excellent alternative for determining CAP in food samples.

High-throughput screening of multi-pesticide residues in animal-derived foods by QuEChERS-online gel permeation chromatography-gas chromatography-tandem mass spectrometry

Improvements in living standards have led to an increase in the consumption of animal-derived foods. Pesticides may be used illegally during animal breeding as well as meat production and processing for pest control and preservation. Pesticides applied to crops may also be enriched in animal tissues through the food chain, thereby increasing the risk of pesticide residue accumulation in muscles and visceral tissues and endangering human health. China has stipulated maximum residue limits for pesticide residues in livestock and poultry meat and their viscera. Many other major developed countries and organizations, including the European Union, Codex Alimentarius Commission, and Japan, have also set maximum residue limits for these residues (0.005-10, 0.004-10, and 0.001-10 mg/kg, respectively). Research on pretreatment technologies for pesticide residue detection in plant-derived foods is widely available, but insufficient attention has been paid to animal-derived foods. Thus, high-throughput detection technologies for pesticide residues in animal-derived foods are limited. The impurities that can interfere with the detection process for plant-derived foods mainly include organic acids, polar pigments, and other small molecular compounds; by contrast, the matrix of animal-derived foods is much more complex. Macromolecular proteins, fats, small molecular amino acids, organic acids, and phospholipids can interfere with the detection of pesticide residues in animal-derived foods. Thus, selecting the appropriate pretreatment and purification technology is of great importance. In this study, the QuEChERS technique was combined with online gel permeation chromatography-gas chromatography-tandem mass spectrometry (GPC-GC-MS/MS) to determine 196 pesticide residues in animal-derived foods. The samples were extracted with acetonitrile, purified using the QuEChERS technique coupled with online GPC, detected by GC-MS/MS, determined in multiple reaction monitoring mode (MRM), and quantified using the external standard method. The effects of the extraction solvent and purification agent type on the extraction efficiency and matrix removal of the method were optimized. The purification effect of online GPC on the sample solution was investigated. The optimal distillate receiving time was obtained by studying the recoveries of the target substances and matrix effects over different distillate receiving periods to achieve the effective introduction of target substances and efficient matrix removal. Further, the advantages of the QuEChERS technique combined with online GPC were evaluated. The matrix effects of 196 pesticides were assessed; ten pesticide residues showed moderate matrix effects, while four pesticide residues showed strong matrix effects. A matrix-matched standard solution was used for quantification. The 196 pesticides showed good linearity in the range of 0.005-0.2 mg/L, with correlation coefficients greater than 0.996. The limits of detection and quantification were 0.002 and 0.005 mg/kg, respectively. The recoveries of 196 pesticides at spiked levels of 0.01, 0.05, and 0.20 mg/kg were 65.3%-126.2%, with relative standard deviations (RSDs) of 0.7%-5.7%. The proposed method is rapid, accurate, and sensitive; thus, it is suitable for the high-throughput screening and detection of multiple pesticide residues in animal-derived foods.

A Review of Veterinary Drug Residue Detection: Recent Advancements, Challenges, and Future Directions

Veterinary drug residues of common food (milk, meat) have posed serious threats to the environment and human health, making the quality and safety of agricultural, livestock, and aquatic products increasingly prominent. With the widespread use of veterinary drugs and the requirements for food safety, it has become urgent to detect veterinary drug residues in animal-derived foods. So far, few studies have systematically reviewed the progresses, challenges, and future directions in veterinary drug residue detection. A thorough review on the current advancements, challenges, and potential future directions of veterinary drug residue detection will be extremely beneficial and timely. This study reviewed recent developments of detection technology of veterinary drug residues. The current issues and challenges in veterinary drug residue detection were examined and highlighted. Finally, future proposals on directions and prospects for veterinary drug residue detection were suggested. High-throughput and high-sensitivity veterinary drug detection technology, sample pretreatment technology for rapid processing, and the fusion of multiple detection methods were recommended as the main directions for the future development of veterinary drug residue detection. It was suggested to develop the analysis and detection technologies of veterinary drug residue towards high automation, high sensitivity, and high throughput in the future. This review provides new ideas and strategies for the rapid development of animal husbandry industry and protecting consumers’ physical health and food safety.

Evaluation of Antibiotics Residues in Milk and Meat Using Different Analytical Methods.

Veterinary drugs are pharmacologically and biologically active chemical agents. At present, veterinary drugs are extensively used to prevent and treat animal diseases, to promote animal growth, and to improve the conversion rate of feed. However, the use of veterinary drugs in food-producing animals may leave residues of the parent compounds and/or their metabolites in food products resulting in harmful effects on humans. To ensure food safety, sensitive and effective analytical methods have been developing rapidly. This review describes sample extraction and cleanup methods, and different analytical techniques are used for the determination of veterinary drug residues in milk and meat. Sample extraction methods, such as solvent extraction, liquid-liquid extraction, and cleanup methods such as dispersive solid-phase extraction and immunoaffinity chromatography, were summarized. Different types of analytical methods such as microbial, immunological, biosensor, thin layer chromatography, high-performance liquid chromatography, and liquid chromatography-tandem mass spectrometry were discussed for the analysis of veterinary drug residues in animal-derived foods. Liquid chromatography-tandem mass spectrometry is the most widely used analytical technique for the determination of antibiotic drug residues. This is due to the powerful separation of LC and accurate identification of MS, and LC-MS/MS is more popular in the analysis of veterinary drug residues.

Strong antibacterial effects in animal-derived food detected via non-target planar bioassays

Screening for antibiotic residues in animal-derived food is largely done by in vitro assays. However, a non-negligible number of positive screening results (sum values) remains with no confirmation by high-performance liquid chromatography coupled to high-sophisticated mass spectrometers. To investigate this inconsistency, a hyphenated strategy was sought that combines on the same surface planar chromatography−multi-imaging with antibacterial planar bioassays. Strong antibacterial activity against Gram-positive and Gram-negative bacteria was revealed in four animal-derived food extracts (pig muscle, cow milk, chicken eggs, and honey), which, however, were previously proven to be veterinary drug residue-free. This antibacterial effect was strong, extremely exceeding the effect from a mixture of 81 veterinary drugs spiked at 300 µg/kg, which is about 3 times the maximum residue limit of most of these veterinary drugs. The unknown antibacterial zones were online-eluted to high-resolution mass spectrometry to obtain molecular formulas. Endogenous fatty acids and lipids were identified to be responsible for the strong antibacterial effects, confirmed by co-analyzed reference compounds. These results resolved the methodological inconsistency but also question the prevailing understanding: Current food safety only based on target analysis of regulated antibiotic residues neglects consumer protection when the much stronger antibacterial impact of food on the human microbiome is overlooked.

Affinity-Based Analysis Methods for the Detection of Aminoglycoside Antibiotic Residues in Animal-Derived Foods: A Review.

With the increasingly serious problem of aminoglycoside antibiotic residues, it is imperative to develop rapid, sensitive and efficient detection methods. This article reviews the detection methods of aminoglycoside antibiotics in animal-derived foods, including enzyme-linked immunosorbent assay, fluorescent immunoassay, chemical immunoassay, affinity sensing assay, lateral flow immunochromatography and molecular imprinted immunoassay. After evaluating the performance of these methods, the advantages and disadvantages were analyzed and compared. Furthermore, development prospects and research trends were proposed and summarized. This review can serve as a basis for further research and provide helpful references and new insights for the analysis of aminoglycoside residues. Accordingly, the in-depth investigation and analysis will certainly make great contributions to food safety, public hygiene and human health.

A Robust Homogeneous Fluorescence Polarization Immunoassay for Rapid Determination of Erythromycin in Milk.

Erythromycin (ERY) is one of the most common macrolides applied in veterinary medicine to treat diseases or as a feed additive for animal growth promotion. Long-term irrational use of ERY could lead to residues in animal-derived food and the emergence of drug-resistant strains, posing potential threats to human health. In this study, a highly sensitive, specific, robust, and rapid fluorescence polarization immunoassay (FPIA) for the determination of ERY in milk has been described. Herein, to achieve high sensitivity, five tracers of ERY with different fluorescein structures were synthesized and paired with three monoclonal antibodies (mAbs). Under the optimized conditions, the combination of mAb 5B2 and tracer ERM-FITC achieved the lowest IC50 value in the FPIA with 7.39 μg/L for ERM. The established FPIA was used to detect ERY in milk, revealing a limit of detection (LOD) of 14.08 μg/L with recoveries of 96.08-107.77% and coefficients of variations (CVs) of 3.41-10.97%. The total detection time of the developed FPIA was less than 5 min from the addition of samples to the result readout. All the above results showed that the proposed FPIA in this study was a rapid, accurate, and simple method for the screening of ERY in milk samples.

Rapid and Visual Detection of Oxytetracycline Using Mesoporous Silica Nanochannels Modified Interdigital Electrode

Oxytetracycline (OTC) is one of the widely used antibiotics in veterinary practices because of its broad-spectrum antibacterial activity and low cost. However, drug abuse has triggered adverse effects on human health, which brings a growing demand for on-site diagnosis of OTC residues in animal-derived foods. In this study, we demonstrated the combined use of interdigital electrode microarray and electrochemiluminescence (ECL) imaging for parallel multiplex measurement of OTC. Well-ordered and vertically aligned mesoporous silica nanochannels modified on microarray substrate could exert a strong electrostatic attraction to the ECL luminophores and accelerate their mass transport to generate enhanced ECL signal. The performance of the integrated ECL microarray sensor was fully validated with respect to linearity (0.5 μM to 50 μM), sensitivity (limit of detection 0.26 μM), accuracy (recovery rate between 96.78% and 106.1%), low operating sample volume (480 nL), short assay time (1.5 min), and antifouling ability toward complex media. The multiplex microarray platform can serve as a simple, fast, and low-cost tool for the detection of a wide spectrum of antibiotics in the field of food safety.

Construction of a self-sufficient DNA circuit for amplified detection of kanamycin

Improper use of kanamycin can lead to trace kanamycin residues in animal-derived foods, which can pose a potential threat to public health. Isothermal enzyme-free DNA circuits have provided a versatile toolbox for detecting kanamycin residues in complicated food samples, yet they are always limited by low amplification efficiency and intricate design. Herein, we present a simple-yet-robust nonenzymatic self-driven hybridization chain reaction (SHCR) amplifier for kanamycin determination with 5800-fold sensitivity over that of the conventional HCR circuit. The analyte kanamycin-activated SHCR circuitry can generate numerous new initiators to promote the reaction and improve the amplification efficiency, thus achieving an exponential signal gain. With precise target recognition and multilayer amplification capability, our self-sustainable SHCR aptasensor facilitated the highly sensitive and reliable analysis of kanamycin in buffer, milk, and honey samples, thus holding great potential for the amplified detection of trace contaminants in liquid food matrices.

Construction of a Molecularly Imprinted Sensor Modified with Tea Branch Biochar and Its Rapid Detection of Norfloxacin Residues in Animal-Derived Foods.

Norfloxacin (NOR) is a common antibiotic used in humans and animals, and its high levels can cause intolerance or poisoning. Therefore, NOR levels in animal-derived foods must be monitored due to potential side effects and illegal use phenomena. This research centered on the development of an environmentally friendly electrochemical sensor for NOR detection. Potassium carbonate activated tea branch biochar (K-TBC) as an efficient use of waste was coated on the surface of glassy carbon electrode (GCE), and a molecular-imprinted polymer (MIP) layer was subsequently electropolymerized onto the modified electrode. NOR was used as template molecule and o-phenylenediamine (o-PD) and o-aminophenol (o-AP) were used as bifunctional monomers. The electrochemical sensor was built and its electrochemical behavior on NOR was investigated. The sensor demonstrated an excellent linear current response to NOR concentrations in the ranges of 0.1-0.5 nM and 0.5-100 nM under ideal experimental circumstances, with a detection limit of 0.028 nM (S/N = 3). With recoveries ranging from 85.90% to 101.71%, the designed sensor was effectively used to detect NOR in actual samples of milk, honey, and pork. Besides, the fabricated sensor had low price, short detection time, good selectivity and stability, which can provide a theoretical and practical basis for the actual monitoring of NOR residues.

An aptamer-based colorimetric/SERS dual-mode sensing strategy for the detection of sulfadimethoxine residues in animal-derived foods.

Residues of sulfadimethoxine (SDM) in animal-derived foods have attracted widespread public concern. Herein, we propose an aptamer-based colorimetric/SERS dual-mode sensing strategy for the determination of SDM based on hexadecyl trimethyl ammonium bromide (CTAB) induced aggregation of nanoparticles. In the absence of SDM, the SDM aptamer formed a supramolecular composite with CTAB, and the 4-mercaptopyrimidine functionalized gold nanoparticles (AuNPs@4-MPY) remained dispersed due to the lack of CTAB. Upon the addition of SDM, the SDM aptamer preferentially combined with SDM, resulting in the release of CTAB and subsequent aggregation of AuNPs@4-MPY, and the solution color changed from red to blue and presented a dynamic UV-absorbance curve based on different aggregation states. On the other hand, when, gold-silver core-shell nanoparticles (Au@AgNPs) were added additionally, the released CTAB narrowed the nanogap between AuNPs@4-MPY and Au@AgNPs, thus exhibiting enhanced SERS intensity of 4-MPY. This strategy achieved colorimetric detection of SDM with a linear range of 4.00-200.00 ng mL-1 and a detection limit of 2.41 ng mL-1, while SERS had a detection range of 1.20-120.00 ng mL-1 with a detection limit of 0.89 ng mL-1. This strategy is simple and cost-effective for the rapid detection of SDM within 20 minutes. It was further applied for the detection of SDM in spiked milk and honey samples with satisfactory recoveries. Therefore, it exhibits great potential for fast and on-site SDM detection.

Quick and high-throughput quantification of 22 β-agonists residues in animal-derived foods using enzymatic probe sonication

A high demand exists in veterinary drug residual analysis for rapid, automatic and high-throughput analytical techniques that produce data simultaneous and faster. Here, we describe a combined automated solid-phase extraction (SPE) and enzymatic probe sonication (EPS), subsequently ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the quick extraction, automated clean-up as well as simultaneous quantification of 22 β-agonists residues in animal-derived foods. Enhanced by the ultrasonic probe, only 2 min was needed for exhaustively extraction of β-agonists from foods of animal-origin; whereas traditional enzymatic digestion methods require hours or even days, which making it more appropriate for high-throughput biomonitoring. Moreover, the clean-up and pre-concentration procedures were conducted on the automatic SPE, which allowed 36 samples being performed simultaneously within 30 min. The method was successfully applied for analyzing 56 animal-derived food samples, 53.6 % of which contained detectable levels of at least one kind of β-agonists. Interestingly, both the detection rate and residual level of β-agonists in the ruminants (bovine and sheep) were higher as compared with the swine. Concerning the real food samples analyzed, the findings of this study suggest that stricter measures should be adopted to control the illegal usage of β-agonists on the farm animals, particularly for the ruminants.

A Novel Metabolite as a Hapten to Prepare Monoclonal Antibodies for Rapid Screening of Quinoxaline Drug Residues

Quinoxalines (Qx) are chemically synthesized antibacterial drugs with strong antibacterial and growth-promoting effects. Qx is heavily abused by farmers, resulting in large residues in animal-derived foods, which pose a serious threat to human health. Desoxyquinoxalines (DQx), which have the highest residue levels, have been identified as the major toxicant and have become a new generation of residue markers. In this study, we prepared monoclonal antibodies (mAb) based on a new generation metabolite (desoxymequindox, DMEQ) and establish an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for the rapid determination of Qx residues in food. The mAb exhibited high sensitivity with half maximal inhibitory concentration (IC50) and a linear range of 2.84 µg/L and 0.8–12.8 µg/L, respectively. Additionally, the cross-reactivity (CR) of the mAb showed that it recognized multiple DQx to varying levels. The limits of detection (LOD), limits of quantification (LOQ), and recoveries for the ic-ELISA assay of pork, swine liver, swine kidney, chicken, and chicken liver were 0.48–0.58 µg/kg, 0.61–0.90 µg/kg, and 73.7–107.8%, respectively, and the coefficients of variation (CV) were less than 11%. The results of the ic-ELISA showed a good correlation with LC–MS/MS in animal-derived foods. This suggests that this analytical method can be used for the rapid screening of QX residues.

Fluorescent Sensing of Ciprofloxacin and Chloramphenicol in Milk Samples via Inner Filter Effect and Photoinduced Electron Transfer Based on Nanosized Rod-Shaped Eu-MOF.

Rapid, facile, and accurate detection of antibiotic residues is vital for practical applications. Herein, we designed a sensitive, visual, and rapid analytical method for sensitive detection of ciprofloxacin and chloramphenicol based on a nanosized rod-shaped Europium metal organic framework (Eu-MOF). The fluorescent Eu-MOF was firstly synthesized by a simple synthetic route at room temperature, which displays a red emission. The mechanisms of detecting ciprofloxacin and chloramphenicol were confirmed to be the inner filter effect (IFE) and photoinduced electron transfer (PET). Under the optimized experimental conditions, the detection limits of the developed method for ciprofloxacin and chloramphenicol detection were 0.0136 and 3.16 μM, respectively. Moreover, the sensor was effectively applied for quantitative determination of ciprofloxacin and chloramphenicol milk samples with satisfactory recoveries of 94.5–102% and 97–110%, respectively. This work developed a new method for rapid detection of ciprofloxacin and chloramphenicol residues. In addition, the established method has potential practical application value for on-site safety regulation on antibiotic residues in animal-derived food.

Broad-Specificity Aptamer of Sulfonamides: Isolation and Its Application in Simultaneous Detection of Multiple Sulfonamides in Fish Sample.

Sulfonamide antibiotics (SAs) are widely used in animal husbandry and aquaculture, and the excess residues of SAs in animal-derived foods will harm the health of consumers. In reality, various SAs were alternately used in animal husbandry and aquaculture, and thus, it is urgent need to develop simple and high-throughput methods for simultaneously detecting multiple SAs or groups of SAs in order to realize rapid screening of total SAs residues in animal-derived foods. We herein isolated a broad-specificity aptamer for SAs by using a multi-SAs systematic evolution of ligands by exponential enrichment (SELEX) strategy. The isolated broad-specificity aptamer has a higher binding affinity to five different SAs including sulfaquinoxaline (SQ), sulfamethoxypyridazine (SMPZ), sulfametoxydiazine (SMD), sulfachloropyridazine (SCP), and sulfapyridine (SPD) and, thus, can be used as a bioreceptor for developing various high-throughput methods for the simultaneous detection or rapid screening of above five SAs. Based on the isolated broad-specificity aptamer and Cy7 (diethylthiatricarbocyanine) displacement strategy, a colorimetric aptasensor was developed for the simultaneous detection of SQ, SMPZ, SMD, SCP, and SPD with a visual detection limit of 2.0-5.0 μM and a spectrometry detection limit of 0.2-0.5 μM. The colorimetric aptasensor was successfully used to detect SQ, SMPZ, SMD, SCP, and SPD in fish muscle with a recovery of 82%-92% and a RSD (n = 5) < 7%. The success of this study provided a promising bioreceptor for developing various high-throughput methods for on-site rapid screening of multiple SAs residues, as well as a simple method for the rapid and cost-effective screening of total SQ, SMPZ, SMD, SCP, and SPD in seafood.

The Simultaneous Detection of Multiple Antibiotics in Milk and Pork Based on an Antibody Chip Biosensor.

In the modern farming industry, the irrational or illegal use of veterinary drugs leads to residues in animal-derived food, which can seriously threaten human health. Efficient detection of low concentrations of drug residues in animal products in a short time is a key challenge for analytical methods. This study proposes to use an antibody chip biosensor for rapid and automated analysis of cephalosporins, aminoglycosides, and sulfonamide antibiotics in pork and milk. 3D polymer slides were applied for the preparation of antibody chips. Ovalbumin (OVA) or bovine serum albumin (BSA) conjugates of the haptens were immobilized as spots on disposable chips. Monoclonal antibodies (mAbs) against cefalexin, ceftiofur, gentamicin, neomycin, and sulfonamides allowed the simultaneous detection of the respective analytes. Antibody binding was detected by a second antibody labeled with Cy3-generating fluorescence, which was scanned a with chip scanner. The limits of detection (LOD) for all the analytes were far below the respective maximum residue limits (MRLs) and ranged from 0.51 to 4.3 µg/kg. The average recoveries of all the analytes in each sample were in the range of 81.6–113.6%. The intra- and inter-assay CV was less than 12.9% and showed good accuracy and precision for all the antibiotics at the MRL level. The sample pretreatment method is simple, and the results are confirmed to be accurate by LC–MS/MS; therefore, this method is valuable for the quality control of animal-derived food.

“Three-in-one” nanohybrids as synergistic nanozymes assisted with exonuclease I amplification to enhance colorimetric aptasensor for ultrasensitive detection of kanamycin

Kanamycin (KAN) residues in animal-derived food can cause serious threats to human health. Herein, a colorimetric aptasensor was described using “three-in-one” nanohybrids (hemin@Fe-MIL-88NH2/PtNP) as synergistic nanozymes assisted with the exonuclease I (Exo I) signal amplification for the ultrasensitive detection of KAN. In the presence of KAN and Exo I, the KAN aptamer (APT) was specifically bound to KAN, and the remaining APT complementary strand (cDNA1) captured hemin@Fe-MIL-88NH2/PtNP labeled with the cDNA1 complementary strand (cDNA2). Due to the synergistic effect of hemin, Fe-MIL-88NH2 and platinum nanoparticles (PtNPs), hemin@Fe-MIL-88NH2/PtNP exhibited superior catalytic performance and could efficiently catalyze the 3,3′,5,5′-tetramethylbenzidine (TMB)-H2O2 system for signal development. Moreover, Exo I could digest APT and release KAN into a new cycle for signal amplification. Based on multiple signal amplification effects, our proposed aptasensor achieved a wide detection range from 0.01 to 100 ng mL−1 with a low detection limit of 2 pg mL−1. This assay also successfully detected KAN-added milk and shrimp samples with added recovery ranges of 93.58%–106.08% and relative standard deviations (RSDs) of 2.20%–5.50%. The aptasensor enabled ultrasensitive, specific, and fast detection of KAN, and exhibited promising applications in the efficient detection of food pollutants.

Rapid Determination of Cephalexin in Animal-Derived Food by an Indirect Competitive Enzyme-Linked Immunosorbent Assay

Cephalexin (CFE) residues in animal-derived food and its deleterious effects on human health has attracted extensive attention. In this study, a specific anti-cephalexin monoclonal antibody (mAb) was used to develop an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) with a half-maximal inhibitory concentration (IC50) of 1.83 µg/L that was suitable for the determination of cephalexin in animal-derived foods. The limit of detection (LOD) and limit of quantitation (LOQ) for cephalexin were from 0.44 to 0.58 µg/kg and 0.57 to 0.67 µg/kg, respectively. The recovery was from 90.1 to 103.6% with coefficients of variation (CV) less than 10.0%. In addition, the established ELISA method and high-performance liquid chromatography – mass spectrometry (HPLC-MS/MS) were used to analyze the samples with good correlation between the results. The results show that the established ic-ELISA is suitable for the determination of cephalexin in animal-derived food.