Target Mycotoxins Research Articles

Simultaneous detection of multiple mycotoxins using MXene-based electrochemical aptasensor array and a self-developed multi-channel portable device

In response to the pressing need for highly efficient simultaneous detection of multiple mycotoxins, which are often found co-occurring in food raw materials and feed, an MXene-based electrochemical aptasensor array (MBEAA) was developed. This aptasensor array utilizes high-specificity aptamers as recognition elements, enabling the capture of electrical signal changes in the presence of target mycotoxins. Based on this platform, a multi-channel portable electrochemical device, enabling rapid, cost-effective, and simultaneous detection of aflatoxin B1 (AFB1), ochratoxin A (OTA), and zealenone (ZEN) was further developed. The developed system boasts a wide detection range of 1.0 × 10−1 to 10.0 ng mL−1, with remarkable performance characterized by ultra-low detection limits of 41.2 pg mL−1, 27.6 pg mL−1, and 33.0 pg mL−1 for AFB1, OTA, and ZEN, respectively. Successfully applied in corn samples, this method offers a portable, easy-to-operate, and cost-effective solution for simultaneous multi-mycotoxin detection. Moreover, the application of the self-developed detection system could be expanded for simultaneous detection of many different targets when their specific aptamers or antibodies were available.

Upconversion nanoparticles-modified aptasensors for highly sensitive mycotoxin detection for food quality and safety.

Mycotoxins, highly toxic and carcinogenic secondary metabolites produced by certain fungi, pose significant health risks as they contaminate food and feed products globally. Current mycotoxin detection methods have limitations in real-time detection capabilities. Aptasensors, incorporating aptamers as specific recognition elements, are crucial for mycotoxin detection due to their remarkable sensitivity and selectivity in identifying target mycotoxins. The sensitivity of aptasensors can be improved by using upconversion nanoparticles (UCNPs). UCNPs consist of lanthanide ions in ceramic host, and their ladder-like energy levels at f-orbitals have unique photophysical properties, including converting low-energy photons to high-energy emissions by a series of complex processes and offering sharp, low-noise, and sensitive near-infrared to visible detection strategy to enhance the efficacy of aptasensors for novel mycotoxin detection. This article aims to review recent reports on the scope of the potential of UCNPs in mycotoxin detection, focusing on their integration with aptasensors to give readers clear insight. We briefly describe the upconversion photoluminescence (UCPL) mechanism and relevant energy transfer processes influencing UCNP design and optimization. Furthermore, recent studies and advancements in UCNP-based aptasensors will be reviewed. We then discuss the potential impact of UCNP-modified aptasensors on food safety and present an outlook on future directions and challenges in this field. This review article comprehensively explains the current state-of-the-art UCNP-based aptasensors for mycotoxin detection. It provides insights into potential applications by addressing technical and practical challenges for practical implementation.

Development of a corn flour certified reference material for the accurate determination of zearalenone.

A certified reference material (CRM, KRISS 108-01-002) for zearalenone in corn flour was developed to assure reliable and accurate measurements in testing laboratories. Commercially available corn flour underwent freeze-drying, pulverization, sieving, and homogenization. The final product was packed in amber bottles, approximately 14 g per unit, and preserved at -70 °C. 13C18-Zearalenone was used as an internal standard (IS) for the certification of zearalenone by isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LC‒MS/MS) and for the analysis of α-zearalenol, β-zearalenol, and zearalanone by LC‒MS/MS. The prepared CRM was sufficiently homogeneous, as the among-unit relative standard deviation for each mycotoxin ranged from 2.2 to 5.7 %. Additionally, the stability of the mycotoxins in the CRM was evaluated under different temperature conditions and scheduled test periods, including storage at -70°C, -20°C, and 4°C and room temperature for up to 12 months, 6 months, and 1 month, respectively. The content of each target mycotoxin in the CRM remained stable throughout the monitoring period at each temperature. Zearalenone content (153.6 ± 8.0 µg/kg) was assigned as the certified value. Meanwhile, the contents of α-zearalenol (1.30 ± 0.17 µg/kg), β-zearalenol (4.75 ± 0.33 µg/kg), and zearalanone (2.09 ± 0.16 µg/kg) were provided as informative values.

Food Security and Foodborne Mycotoxicoses-What Should Be the Adequate Risk Assessment and Regulation?

The purpose of this review is to elucidate the actual threat of the most prevalent mycotoxins in agricultural commodities and human/animal food/feed for the induction of foodborne diseases or ailments. The underestimated hazard of combined mycotoxin uptake by animals or humans is critically discussed with regard to synergistic or additive interaction between some target mycotoxins. The real toxicity of target mycotoxin combinations as it happens in practice is evaluated and possible lower limit values or control measures are suggested in such cases. Some critical points on adequate risk assessment, hygiene control, and regulation of mycotoxins are discussed. The efficiency of current mycotoxin regulations and control measures is evaluated in regard to human/animal health hazards. The risk assessment in the case of multiple mycotoxin exposure of humans/animals via food/feed or agricultural commodities is evaluated and some suggestions are proposed in such cases. Appropriate control measures and food safety issues throughout the food supply chain are proposed in order to prevent the target foodborne diseases. Some preventive measures and possible veterinary hygiene controls or risk evaluations are proposed in some natural cases of foodborne diseases for preventing mycotoxin contamination of animal products designed for human consumption and to avoid possible public health issues.

Extracting mycotoxins from edible vegetable oils by using green, ecofriendly deep eutectic solvents.

In this work, we developed an environmentally friendly liquid-liquid microextraction method using a natural deep eutectic solvent in combination with liquid chromatography for the simultaneous determination of four mycotoxins (deoxynivalenol, alternariol, ochratoxin A and zearalenone) in edible vegetable oils. A chemometric approach assessed the effect of the operational parameters on the mycotoxin extraction efficiency. The extracts were analyzed by HPLC coupled with a diode array and fluorescence detector. The optimum NADES composition resulted in the highest extraction recoveries, and it was applied to coextract the target mycotoxins in several types of edible vegetable oils without using hazardous solvents or requiring further clean-up. The limits of detection ranged from 0.07 to 300µgkg-1, and recoveries were close to 100%, except for zearalenone (viz. 35%), with relative standard deviations below 9% in all cases. The proposed method was validated following the European Commission 2002/657/EC and 2006/401/EC.

Rapid detection of zearalenone in cereals using La3+-doped upconversion nanoparticles-based immunochromatographic assay

A novel fluorescent immunochromatographic assay (ICA) with excellent specificity and sensitivity for evaluating trace residues of zearalenone (ZEN) in cereal samples was successfully developed. La3+-doped upconversion nanoparticles (UCNPs) were assessed as antibody labels, following optimization of doping ratios. Our results indicated that 20% La3+-doped UCNPs had superior morphological structure and fluorescence characteristics, making them the optimal UCNPs. After labeling 20% La3+-doped UCNPs with ZEN monoclonal antibodies (mAbs), optimal polyacrylic acid amount, coupled mAbs amount, detection time, and fluorescent immunoprobe concentrations were determined as 2.0 mL, 20 μg, 10 min, and 8 mg/mL, respectively. Under optimized detection conditions, the limit of detection (LOD) for ZEN standard solutions was 0.25 ng/mL with coefficients of variation (CVs) ＜ 8.67% and a high linear correlation (R2 = 0.9944). Furthermore, specificity analysis demonstrated that prepared test strips were highly specific for the target mycotoxin. The LODs of ZEN in wheat and maize were both 0.5 μg/kg in spiking recovery experiments, and the recoveries were 87.65%–99.26% and 84.12%–95.12% with the CVs being 1.29%–7.14% and 4.03%–9.01%, respectively, confirming that the optimal UCNPs-ICA was efficient, straightforward, and suitable for rapid on-site screening of ZEN in cereals.

Mycotoxin Determination in Peaches and Peach Products with a Modified QuEChERS Extraction Procedure Coupled with UPLC-MS/MS Analysis.

Peaches are the most significant temperate fruit crop worldwide. However, peach fruits are susceptible to fungal and mycotoxin contamination. Consequently, monitoring the residual levels of multiple mycotoxins in peaches and related products is essential. In this study, a novel method based on QuEChERS extraction, followed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) detection, was developed for analyzing 14 mycotoxins in peaches and peach products from China. Matrix-matched calibrations were employed to accurately quantify the mycotoxins and compensate for matrix effects. Recoveries for the target analytes ranged from 84.6% to 117.6%, with intra-day and inter-day precision below 20%. The limits of quantification were 2 or 5 μg/L for the 14 mycotoxins. This method was utilized to detect the presence of target mycotoxins in 109 fresh peaches, 100 diseased peaches, and 89 peach products from China. Six mycotoxins were identified in the rotten parts of the diseased peaches, with concentrations ranging from 5.2 to 1664.3 µg/kg. In the remaining parts of the diseased peach samples, only two toxins, alternariol (AOH) and alternariol monomethyl ether (AME), were quantified at levels of 15.3 µg/kg and 15.5 µg/kg, respectively. No mycotoxins were detected in fresh peaches. For peach products, all contamination levels were below the quantitative limits and significantly lower than the maximum legal limits established for the products.

Foodborne Diseases Due to Underestimated Hazard of Joint Mycotoxin Exposure at Low Levels and Possible Risk Assessment.

The subject of this review paper is to evaluate the underestimated hazard of multiple mycotoxin exposure of animals/humans for the appearance of foodborne ailments and diseases. The significance of joint mycotoxin interaction in the development of foodborne diseases is discussed, and appropriate conclusions are made. The importance of low feed/food levels of some target mycotoxins co-contaminations in food and feedstuffs for induction of target foodborne mycotoxicoses is also studied in the available literature. The appropriate hygiene control and the necessary risk assessment in regard to possible hazards for animals and humans are also discussed, and appropriate suggestions are made. Some internationally recognized prophylactic measures, management of the risk, and the necessity of elaboration of new international regulations in regard to the maximum permitted levels are also carefully discussed and analysed in the cases of multiple mycotoxin contaminations. The necessity of harmonization of mycotoxin regulations and control measures at international levels is also discussed in order to facilitate food trade between the countries and to ensure global food safety.

Microfluidic Biosensor Integrated with Signal Transduction and Enhancement Mechanism for Ultrasensitive Noncompetitive Assay of Multiple Mycotoxins

To achieve high-throughput ultrasensitive detection of mycotoxins in food, a functional DNA-guided transition-state CRISPR/Cas12a microfluidic biosensor (named FTMB) was successfully constructed. The signal transduction CRISPR/Cas12a strategy in FTMB has utilized DNA sequences with a specific recognition function and activators to form trigger switches. Meanwhile, the transition-state CRISPR/Cas12a system was constructed by adjusting the composition ratio of crRNA and activator to achieve a high response for low concentrations of target mycotoxins. On the other hand, the signal enhancement of FTMB has efficiently integrated the signal output of quantum dots (QDs) with the fluorescence enhancement effect of photonic crystals (PCs). The construction of universal QDs for the CRISPR/Cas12a system and PC films matching the photonic bandgap produced a significant signal enhancement by a factor of 45.6. Overall, FTMB exhibited a wide analytic range (10-5-101 ng·mL-1), low detection of limit (fg·mL-1), short detection period (∼40 min), high specificity, good precision (coefficients of variation <5%), and satisfactory practical sample analysis capacity (the consistency with HPLC at 88.76%-109.99%). It would provide a new and reliable solution for the rapid detection of multiple small molecules in the fields of clinical diagnosis and food safety.

Determination of Regulated and Emerging Mycotoxins in Organic and Conventional Gluten-Free Flours by LC-MS/MS.

Gluten-free cereal products have grown in popularity in recent years as they are perceived as "healthier" alternatives and can be safely consumed by celiac patients, and people with gluten intolerance or wheat allergies. Molds that produce mycotoxins contaminate cereal crops, posing a threat to global food security. Maximum levels have been set for certain mycotoxins in cereal flours; however, little is known about the levels of emerging mycotoxins in these flours. The aim of this study was to develop an efficient, sensitive, and selective method for the detection of four emerging (beauvericin and enniatins A1, B, and B1) and three regulated (aflatoxin B1, zearalenone, and deoxynivalenol) mycotoxins in gluten-free flours. Ultrasound-assisted matrix solid-phase dispersion was used in the extraction of these mycotoxins from flour samples. The validated method was utilized for the LC-MS/MS analysis of conventional and organic wholegrain oat and rice flours. Six of the seven target mycotoxins were detected in these samples. Multi-mycotoxin contamination was found in all flour types, particularly in conventional wholegrain oat flour. Despite the low detection frequency in rice flour, one sample was found to contain zearalenone at a concentration of 83.2 μg/kg, which was higher than the level set by the European Commission for cereal flours. The emerging mycotoxins had the highest detection frequencies; enniatin B was present in 53% of the samples at a maximum concentration of 56 μg/kg, followed by enniatin B1 and beauvericin, which were detected in 46% of the samples, and at levels reaching 21 μg/kg and 10 μg/kg, respectively. These results highlight the need to improve the current knowledge and regulations on the presence of mycotoxins, particularly emerging ones, in gluten-free flours and cereal-based products.

SsDNA-C3N4 conjugates-based nanozyme sensor array for discriminating mycotoxins.

A nanozyme sensor array based on the ssDNA-distensible C3N4 nanosheet sensor elements for discriminating multiple mycotoxins commonly existing in contaminated cereals has been explored. The sensor array exploited (a) three DNA nonspecific sequences (A40, T40, C40) absorbed on the C3N4 nanosheets as sensor elements catalyzing the oxidation of TMB; (b) the presence of five mycotoxins affected the catalytic activity of three nanozymes with various degrees. The parameter (A0-A) was employed as the signal output to obtain the response patterns for different mycotoxins with the same concentration where A0 and A were the absorption peak values at 650nm of oxTMB in the absence and presence of target mycotoxins, respectively. After the raw data was subjected to principal component analysis, 3D canonical score plots were obtained. The sensor array was capable of separating five mycotoxins from each other with 100% accuracy even if the concentration of the mycotoxins was as low as 1nM. Moreover, the array performed well in discriminating the mycotoxin mixtures with different ratios. Importantly, the practicality of this sensor array was demonstrated by discriminating the five mycotoxins spiking in corn-free samples in 3D canonical score plots, validating that the sensor array can act as a flexible detection tool for food safety. A nanozyme sensor array was developed based on the ssDNA-distensible C3N4 NSs sensor elements for discriminating muitiple mycotoxins.

Comparison of two commercial methods with a UHPLC–MS/MS method for the determination of multiple mycotoxins in cereals

Immunoassay-based techniques are important on-site screening tools for the detection of mycotoxins in cereals. This study aims to evaluate the trueness, precision, repeatability and cross-reactivity of commercially available test strips, ELISA kits and UHPLC-MS/MS on analyzing zearalenone, ochratoxin A, deoxynivalenol, T-2 toxin and fumonisin B1. The results showed that false negative rate (25.7 %–37.4 %) of all tested mycotoxins by test strips was higher than the false positive rate (0 %–31.0 %). The repeatability of ELISA kits at the declared LOD dispersed from −85.7 % to +98.4 %. ELISA kits were more accurate at 50 % of the maximum residue limit (MRL) of mycotoxins than 150 % and 200 %. All the tested deoxynivalenol/zearalenone derivatives had cross-reactivity with different level, and sample matrix could reinforce this overestimation of target mycotoxin. This study emphasized that higher-quality antibody screening and more analytical performance investigations are need to address for on-site detection of mycotoxins in the future.

Reduction of Mycotoxigenic Fungi Growth and Their Mycotoxin Production by Bacillus subtilis QST 713.

The use of chemical pesticides to control the occurrence of mycotoxigenic fungi in crops has led to environmental and human health issues, driving the agriculture sector to a more sustainable system. Biocontrol agents such as Bacillus strains and their antimicrobial metabolites have been proposed as alternatives to chemical pesticides. In the present work, a broth obtained from a commercial product containing Bacillus subtilis QST 713 was tested for its ability to inhibit the growth of mycotoxigenic fungi as well as reduce their mycotoxin production. Mass spectrometry analysis of Bacillus subtilis broth allowed to detect the presence of 14 different lipopeptides, belonging to the iturin, fengycin, and surfactin families, already known for their antifungal properties. Bacillus subtilis broth demonstrated to be a useful tool to inhibit the growth of some of the most important mycotoxigenic fungi such as Aspergillus flavus, Fusarium verticillioides, Fusarium graminearum, Aspergillus carbonarius, and Alternaria alternata. In addition, cell-free Bacillus subtilis broth provided the most promising results against the growth of Fusarium graminearum and Alternaria alternata, where the radial growth was reduced up to 86% with respect to the untreated test. With regard to the mycotoxin reduction, raw Bacillus subtilis broth completely inhibited the production of aflatoxin B1, deoxynivalenol, zearalenone, and tenuazonic acid. Cell-free broth provided promising inhibitory properties toward all of the target mycotoxins, even if the results were less promising than the corresponding raw broth. In conclusion, this work showed that a commercial Bacillus subtilis, characterized by the presence of different lipopeptides, was able to reduce the growth of the main mycotoxigenic fungi and inhibit the production of related mycotoxins.

Fast extraction of aflatoxins, ochratoxins and enniatins from maize with magnetic covalent organic framework prior to HPLC-MS/MS detection

A simple, rapid and sensitive vortex-assisted magnetic solid phase extraction (MSPE) method using core–shell structured magnetic covalent organic frameworks (Fe3O4/COF-TpBD) as adsorbents was established for simultaneous determination of aflatoxins, ochratoxins and enniatins in maize before high-performance chromatography tandem spectrometry detection. Fe3O4/COF-TpBD was facilely fabricated, characterized, and applied for simultaneous extraction of ten mycotoxins from maize. Key parameters affecting MSPE efficiency were investigated. The method possessed good linearity (0.05–50 μg/kg), high sensitivity (0.02–1.67 µg/kg for limits of detection), and satisfactory recoveries (73.8–105.3 %), with relative standard deviations less than 8.5 %. The amount of adsorbent used (5 mg) and the time required (0.5 min each for adsorption and for desorption) for MSPE were greatly shortened in comparison with the previous reports. Therefore, the fabricated Fe3O4/COF-TpBD exhibited a great potential for rapid extraction of common mycotoxins in maize, which can be attributed to its multiple interactions with the target mycotoxins.

Portable chemiluminescence optical fiber aptamer-based biosensors for analysis of multiple mycotoxins

Multiple mycotoxin contamination in foodstuffs causes significant threat to human health especially considering their synergistic effects. Herein, we present a novel chemiluminescence optical fiber aptamer-based biosensors for ultrasensitive on-site assay of multiplex mycotoxins in food samples. The optical fiber aptasensor is developed based on the specific recognition of single-stranded binding protein (SSB) and mycotoxin aptamers for the selective analysis. Optical fibers are utilized not only as the carrier to immobilize SSB to prepare the sensing region, but also as the transducer of the chemiluminescent emission to detect the sensor response. Multiple-channel fiber sensors are coupled together toward the detector and a compact and portable design of the chemiluminescence optical fiber aptasensor system is proposed, allowing four mycotoxins (AFB1, FB1, OTA or ZEN) to be simultaneously determined from one run without any tedious operation procedure. The optical fiber aptasensor system can perform sensitive analysis with a wide detection range from pg/mL to 105 pg/mL and low limit-of-detection in the range of 0.015 pg/mL-0.423 pg/mL for different target mycotoxins. The method is successfully applied for simultaneous and sensitive detection of multiple mycotoxins in infant cereals, showing its practical application in on-site analysis of real food samples.

Opinion: Multi-Mycotoxin Reference Materials.

The analysis of mycotoxins in food and feed using liquid chromatography coupled with mass spectrometry is considered advantageous because the hyphenated technology enables simultaneous determination of multiple mycotoxins. Multi-mycotoxin analysis requires special consideration of quality control parameters to ensure proper evaluation of data quality for all target mycotoxins in method development and routine sample analysis. Mycotoxin matrix reference materials, especially certified reference materials, are stable and homogeneous matrices with certified traceability, concentrations, and uncertainty for mycotoxin(s) of interest. The use of these reference materials for single mycotoxin analysis has been a well-accepted practice and should be extended to multi-mycotoxin analysis. This opinion piece discusses the following essential metrological and operational components to improve data quality: (1) purposes of multi-mycotoxin reference materials; (2) comparison of reference materials, certified reference materials, and in-house quality control materials; (3) advantages of using reference materials for multi-mycotoxin analysis; (4) current trends and challenges of multi-mycotoxin reference materials. Potential applications of reference materials discussed here can improve routine mycotoxin determination and will lead to better accuracy and consistency of results. Quality control processes that incorporate reference materials in the field of mycotoxin analysis ensure successful development and implementation of liquid chromatography mass spectrometry-based multi-mycotoxin methods.

A versatile Y shaped DNA nanostructure for simple, rapid and one-step detection of mycotoxins

A versatile Y shaped DNA nanostructure has been developed for simple, rapid and one-step simultaneous detection aflatoxin B1 (AFB1) and ochratoxin A (OTA). Y shaped duplex DNA arms was formed with two DNA tweezer at the ends. The aptamer sequence at the third end can bind to its target mycotoxins with strong affinity and then release the two DNA fragments. The released DNA fragments can open the DNA tweezers at the ends of Y shaped DNA arm. The amounts of AFB1 and OTA can be quantitative detection through the recovery of the fluorescent intensities. This strategy is simple and rapid with self-powered DNA hybridization reaction to control the “open” of Y shaped DNA tweezers. Furthermore, it can be finished in 60 min with only one-step of operation. The linear range of AFB1 was from 0.5 to 200 ng/mL (R2 = 0.995) and linear relationship of OTA was obtained from 4 to 300 ng/mL (R2 = 0.990). It also has been successfully applied for mycotoxins detection in real food samples. Importantly, the target mycotoxins can be extended to others by simply replacing the corresponding aptamer sequences.

Simultaneous Removal of Mycotoxins by a New Feed Additive Containing a Tri-Octahedral Smectite Mixed with Lignocellulose.

Simultaneous removal of mycotoxins has been poorly addressed, and a limited number of studies have reported the efficacy of feed additives in sequestering a large spectrum of mycotoxins. In this study, a new mycotoxin-adsorbing agent was obtained by properly mixing a tri-octahedral smectite with a lignocellulose-based material. At a dosage of 1 mg mL−1, these materials simultaneously adsorbed frequently occurring mycotoxins and did not exert a cytotoxic effect on intestinal cells. Chyme samples obtained by a simulated GI digestion did not affect the viability of Caco-2TC7 cells as measured by the MTT test. In addition, the chyme of the lignocellulose showed a high content of polyphenols (210 mg mL−1 catechin equivalent) and good antioxidant activity. The properties of the individual constituents were maintained in the final composite, and were unaffected by their combination. When tested with a pool of seven mycotoxins at 1 µg mL−1 each and pH 5, the composite (5 mg mL−1) simultaneously sequestered AFB1 (95%), FB1 (99%), ZEA (93%), OTA (80%), T-2 (63%), and DON (22%). HT-2 adsorption did not occur. Mycotoxin adsorption increased exponentially as dosage increased, and occurred at physiological pH values. AFB1, ZEA and T-2 adsorption was not affected by pH in the range 3–9, whereas OTA and FB1 were adsorbed at pH values of 3–5. The adsorbed amount of AFB1, ZEA and T-2 was not released when pH rose from 3 to 7. FB1 and OTA desorption was less than 38%. Langmuir adsorption isotherms revealed high capacity and affinity for adsorption of the target mycotoxins. Results of this study are promising and show the potential of the new composite to remove mycotoxins in practical scenarios where several mycotoxins can co-occur.

Low background interference SERS aptasensor for highly sensitive multiplex mycotoxin detection based on polystyrene microspheres-mediated controlled release of Raman reporters

Mycotoxin contamination is a serious global issue in food safety. The accurate detection of mycotoxins in complex samples, particularly via a portable detection system that realizes multi-analyte detection, remains a great challenge. Here, a polystyrene (PS)-mediated surface-enhanced Raman scattering (SERS) aptasensor was designed for ultrasensitive and simultaneous detection of multiple mycotoxins in food samples. First, the novel Raman tags were prepared using PS as nanocontainer to encapsulate abundant Raman reporters. The PS nanocontainer with Raman reporters was then functionalized by aptamer as an effective target-recognition platform. Second, the single-strand binding protein modified magnetic nanoparticles (MNPs@SSB) were used as a magnetic capture substrate. When the target mycotoxin was present and recognized by its aptamer, the combination of PS-Apt and mycotoxins can hinder the binding between PS-Apt and MNPs@SSB. Consequently, after the magnetic separation and treatment by THF solution, fewer biological-silent reporter probes were released from PS, causing the reduction of SERS signal as a function of target mycotoxins concentration. Due to the high loading capacity of PS and the excellent signal conversion and amplification of aptamer-assisted SERS assay, the proposed strategy is reliable and ultrasensitive for the determination of multiple mycotoxins. The limit of detection was as low as 0.159 fg L−1, 2.015 fg L−1, and 1.561 fg L−1 for ZEN, OTA and AFB1, respectively. This strategy not only raises a new idea for designing novel Raman tag, but also broadens the application of SERS for multiple detection of trace target.

Simultaneous Determination of 15 Mycotoxins in Aquaculture Feed by Liquid Chromatography-Tandem Mass Spectrometry.

The use of plant-based fish feed may increase the risk of contamination by mycotoxins. The multiresidue analysis of mycotoxins in fish feed presents many difficulties due to the complexity of the matrix, the different characteristics of the compounds, and their presence in highly different concentration levels. The aim of this study was to develop a selective, sensitive, and efficient analytical method for the simultaneous determination of 15 mycotoxins (regulated and emerging mycotoxins) in aquaculture feed by LC-MS/MS. Sample extraction was performed with ultrasonic assistance, and different cleanup strategies were evaluated. The optimized method was composed by ultrasound-assisted extraction (two cycles, 55 °C, 20 min), followed by cleanup using a Captiva EMR Lipid cartridge. Then, nine commercial samples of aquaculture fish feed were analyzed. Eight of the 15 target mycotoxins were detected in the samples. Results showed that two enniatins (EENB and ENNB1), beauvericin, and fumonisin B2 were detected in all samples. These results show the multi-mycotoxin contamination of fish feed, highlighting the need to improve current knowledge on the occurrence and toxicity of mycotoxins in fish feed, mainly the emerging ones.