Determination Of Fumonisin B1 Research Articles

Design of a new electrochemical aptasensor based on screen printed carbon electrode modified with gold nanoparticles for the detection of fumonisin B1 in maize flour

A new aptasensor for detecting fumonisin B1 (FB1) in the maize samples was developed based on DNA- aptamer recognition and electrochemical technique. A thiol-modified single-stranded DNA (ss-HSDNA) was immobilized on a screen printed carbon electrode (SPCE) electrodeposited by gold nanoparticles (AuNPs). The morphology and structure of SPCE and AuNPs/SPCE were evaluated via scanning electron microscopy (SEM) equipped with energy dispersive spectroscopy (EDS). The SEM results demonstrated that the SPCE had a flat sheet-like structure, and the AuNPs were homogeneously electrodeposited on the SPCE. Cyclic voltammetry (CV) experiments in the [Fe(CN)6]− 3/− 4 solution were conducted to investigate each step of electrode modification as well as aptasensor performance. Aptamer-FB1 interaction prevented the electron transfer permitting the determination of FB1 in the range of 0.5–500 ng/mL with a low detection limit (0.14 ng/mL). The designed aptasensor was also shown high selectivity, acceptable repeatability and reproducibility, good long-term stability, and excellent recovery. Furthermore, there was a strong correlation between the findings achieved via the designed aptasensor and high performance liquid chromatography (HPLC). Therefore, a simple construction process and satisfactory electrochemical performance of the proposed aptasensor have a great potential for the detection of FB1 in maize samples.

Determination of Fumonisin B1 by Aptamer-Based Fluorescence Resonance Energy Transfer.

Fumonisin FB is produced by Fusarium moniliforme Sheld, of which FB1 is the most common and the most toxic. The establishment of a rapid detection method is an important means to prevent and control FB1 pollution. A highly sensitive fluorescent sensor based on an aptamer for the rapid detection of fumonisin B1 (FB1) in corn was established. In this study, 5-carboxyfluorescein (FAM) was labeled on the aptamer of FB1 (F10). F10 was adsorbed on the surface of graphene oxide (GO) by π-π stacking. The FAM fluorescence signal could be quenched by fluorescence resonance energy transfer between fluorescent molecules and graphene oxide (GO). In the presence of FB1, the binding efficiency of the aptamer to GO was reduced. Therefore, the content of FB1 in corn samples was determined by fluorescence measurements of mixed FAM-labeled F10, GO and corn samples. This method had a good linear relationship in an FB1 concentration range of 0-3000 ng/mL. The equation was y = 0.2576x + 10.98, R2 = 0.9936. The limit of detection was 14.42 ng/mL, and the limit of quantification was 43.70 ng/mL. The recovery of a spiked standard in the corn sample was 89.13-102.08%, and the time of detection was 30 min.

A stimuli-responsive colorimetric aptasensor based on the DNA hydrogel-coated MOF for fumonisin B1 determination in food samples

Fumonisin B1 (FB1), the most occurring fumonisin, is capable of disrupting sphingolipid metabolism and poses a serious threat to human and animal health. The high incidence and toxicity of FB1 have created an urgent need for alternative quantitative determination strategies. Herein, a colorimetric aptasensor was developed by combining the catalytical metal–organic frameworks with the polyacrylamide-DNA hydrogel. The porphyrin-based MOF provided the aptasensor with robust peroxidase-like activity while maintaining good stability. Concurrently, the polyacrylamide-DNA hydrogel constructed via strand-induced hybridization chain reaction (HCR) endowed the aptasensor with outstanding stimuli-responsiveness towards FB1 molecules. The developed aptasensor demonstrated remarkable selectivity and ease of use for assays conducted in solution and food samples. It also exhibited superior analytical performance with a linear range of 0.05–100 ng mL−1 and a limit of detection (LOD) of 0.024 ng mL−1. The strategy could serve as a model to develop diverse stimuli-responsive aptasensors targeting hazards in food safety.

Nuclease Triggered "Signal-On" and Amplified Fluorescent Sensing of Fumonisin B1 Incorporating Graphene Oxide and Specific Aptamer.

Remarkable advancements have been achieved in the development of rapid analytic techniques toward fumonisin B1 (FB1) monitoring and even trace levels for food safety in recent years. However, the point-of-care testing for quantitative and accurate FB1 determination is still challenging. Herein, an innovative aptasensor was established to monitor FB1 by utilizing graphene oxide (GO) and nuclease-triggered signal enhancement. GO can be utilized as a fluorescence quenching agent toward a fluorophore-modified aptamer, and even as a protectant of the aptamer from nuclease cleavage for subsequent target cycling and signal amplification detection. This proposed sensing strategy exhibited a good linearity for FB1 determination in the dynamic range from 0.5 to 20 ng mL−1 with a good correlation of R2 = 0.995. Its limit of detection was established at 0.15 ng mL−1 (S/N = 3), which was significantly lower than the legal requirements by three orders of magnitude. The interferent study demonstrated that the introduced aptasensor possessed high selectivity for FB1. Moreover, the aptasensor was successfully applied to the detection of wheat flour samples, and the results were consistent with the classical ELISA method. The rapid response, sensitive and selective analysis, and reliable results of this sensing platform offer a promising opportunity for food mycotoxin control in point-of-care testing.

Comparative Study of the Performance of Two Different Luciferases for the Analysis of Fumonisin B1 in Wheat Samples

AbstractThe development of two different immunoassays for the determination of fumonisin B1 in wheat samples is reported. A previously described mimopeptide for fumonisin B1 (FB1) was used to produce fusion proteins in combination with two different luciferases: Gaussia luciferase (GLuc) and NanoLuc luciferase (NLuc). The production, expression and the development of two immunoassays based on these fusion proteins (A2‐GLuc and A2‐NLuc) is detailed. The assay showing the best performance, A2‐NLuc, with a limit of detection of 0.61 ng mL−1 and a dynamic range from 1.9 to 95 ng mL−1, was employed for the analysis of spiked wheat samples, a reference matrix material, as well as naturally contaminated wheat samples. The recoveries obtained in the spiked samples were acceptable, between 81.5 and 109 %, with relative standard deviations lower than 14 %. The analysis of naturally contaminated wheat was validated by a liquid chromatography coupled to tandem mass detection method.

Tetrahedral DNA nanostructure-enabled electrochemical aptasensor for ultrasensitive detection of fumonisin B1 with extended dynamic range

Fumonisin B1 (FB1) is common contaminant in agricultural and food products, and sensors with high sensitivity and broad dynamic range are extremely required for FB1 determination. Herein, we reported the tetrahedral DNA nanostructure (TDN)-enabled electrochemical aptasensor for FB1 detection at sub-fg mL−1 level with a dynamic range of 7-folds magnitude. TDN with side length of 5.67 nm was designed to fabricate the substrate for FB1 aptamer anchoring, aiming to achieve the controllable and stable assembly of sensing interface by tuning the assembly density of TDN; TDN at the electrode acts as the anchor for aptamer, absorption of methylene blue as signal generator. Assistant aptamer was added into the sample solution to deplete FB1, reducing the amount of free FB1, thereby extending dynamic range of sensing system. Simultaneously, the negatively-charged TDN can efficiently depress absorption of free aptamer in sample solution. In this way, the utilization of TDN helps to enable the sensor with higher sensitivity and extended dynamic range for FB1. Consequently, the developed sensor allowed a linear dynamic range of 0.500 fg mL−1~1.00 ng mL−1, while the limit of detection was offered to be 0.306 fg mL−1, which was the lowest among the available methods. The applicability of aptasensor was validated by rice sample analysis, which exhibited the comparable reliability of liquid-chromatography tandem mass spectrometry/mass spectrometry. Our work has provided an efficient sensing platform for FB1, and the strategy can be also extended to fabricate sensors for other mycotoxins.

ИЗУЧЕНИЕ СОРБЦИОННОЙ АКТИВНОСТИ БИОСОРБЕНТОВ ПО ОТНОШЕНИЮ К ФУМОНИЗИНУ В1

The aim of this study was to evaluate the effectiveness of four biosorbents in comparison with commercial adsorbents Vetohit and Zaslon in reducing the toxic effects of fumonisin B1. Quantitative determination of fumonisin B1 in supernatants from simulated digestion in different parts of the gastrointestinal tract was carried out by enzyme-linked immunosorbent assay using the RIDASCREEN® FUMONISIN kit (R-Biopharm). In vitro studies (pH 2.0, 8.0) showed that sample № 1, obtained on the basis of extracellular polysaccharides synthesized by strain 574 of the bacterium P. mucilaginosus and calcined bentonite, adsorbed more fumonisin B1 than other biosorbents and commercial preparations (9.43 μg versus 8.60, 8.83, 8.90, 9.09 and 8.13 μg, respectively). This suggests that the developed biosorbents are effective in reducing the toxic effects of fumonisin B1.

A direct competitive nanozyme-linked immunosorbent assay based on MnO2 nanosheets as a catalytic label for the determination of fumonisin B1.

A direct competitive nanozyme-linked immunosorbent assay (dcNLISA) based on MnO2 nanosheets (MnO2 NSs) as a nanozyme label was developed for the highly sensitive determination of fumonisin B1 (FB1). MnO2 NS-labeled fumonisin B1-bovine serum albumin was easily synthesized as a competing antigen for the dcNLISA. And color changes derived from the MnO2-3,3',5,5'-tetramethylbenzidine (TMB) system were exploited as the output signals of the dcNLISA. Several experimental parameters including the concentrations of the coating antibody, pH values, ionic strength and methanol concentration were optimized. Under the optimal conditions, the proposed method demonstrated a linear range (1.17-20.74 ng mL-1) with a reliable correlation coefficient (R2 = 0.9989), a satisfactory limit of detection (0.63 ng mL-1) and high selectivity for the detection of FB1. The recoveries of FB1 in spiked corn and wheat samples were in the range of 85.31-108.16% with coefficients of variation (CVs) ranging from 6.14% to 9.23%. Meanwhile, the testing results showed good consistency (R2 = 0.9892) between the developed dcNLISA and the reference method, liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS) method. The proposed method was proven to be simple, sensitive, cost-effective and reliable for the screening of FB1 in cereals.

Determination of fumonisin B1 and B2 in corn products

An analytical method using immunoaffinty column and HPLC was developed for determination fumonisins in corn and corn products. The detection limits for fumonisin B1 (FB1) and fumonisin B2 (FB2) were 0.03 ppm and 0.07 ppm, respectively. Spiked FB1 in dried corn (kernel and flour), fresh corn, corn snack and corn flake are at 0.2-3.0 ppm level, with recovery ranged from 79.2-108.8% and relative standard deviation (RSD) of 2.9-21.9%. FB2 spiked under the same condition as FB1 had recovery ranges of 70.0-106.0% and RSD of 4.7-20.0%. A total of 76 samples were collected, including 20 dried corn (kernel or flour), 5 fresh corn, 15 corn snacks, 10 corn flakes, 5 corn starch, 5 canned corn from commercial markets and 16 raw corn materials were purchased from corn snack manufacturers. From the total 76 samples, 11 samples (14.5%) were detected with FB1 and/or FB2. Seven samples of corn were at 0.05-0.13 ppm level, 1 fresh corn with 0.15 ppm, 2 corn snacks with 0.5 and 0.16 ppm, respectively, where as 1 corn raw material was at the 0.09 ppm level. The highest contamination was only 0.16 ppm. Nevertheless, no fumonisin was detected in corn flake, corn starch and canned corn samples.

Electrochemical determination of fumonisin B1 using a chemosensor with a recognition unit comprising molecularly imprinted polymer nanoparticles

Fumonisin B1 (FB1) is a carcinogenic mycotoxin contaminating food and animal feed. Although the sensitivity of chromatography and immunoassays is high for FB1 determination, these techniques are laborious, costly, and challenging in making them portable. Herein, a highly sensitive and selective electrochemical sensor using molecularly imprinted polymer nanoparticles (nanoMIPs) for FB1 recognition was devised and fabricated. NanoMIPs were prepared by free-radical polymerisation using the solid-phase synthesis and characterised by DLS, SPR, SEM, AFM, and FTIR spectroscopy. The chemosensor fabrication was prepared in two steps. First, a film of the conducting polypyrrole-(zinc porphyrin) composite was deposited on a Pt electrode by electropolymerisation. Then, nanoMIPs were covalently attached to this film. Both electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) served for analytical signal transduction. The respective chemosensor sensitivity was 0.442 kΩ/pM (R2 = 0.98) and 0.281 μA/pM (R2 = 0.96) in the concentration range of 1 fM to 10 pM FB1. The limit of detection was impressively low, equalling to 0.03 and 0.7 fM, respectively, while the DPV determined imprinting factor was appreciably high 6.28. This electrochemical sensor revealed no cross-reactivity to other mycotoxins. The FB1 recovery in the FB1 spiked maize was high (96–102 %). Thus, the chemosensor proved useful as an effective alternative tool for FB1 determination in maize.

Aptamer and gold nanorod-based fumonisin B1 assay using both fluorometry and SERS.

An aptamer-based assay is presented for the determination of fumonisin B1 (FB1). It is bimodal in that both surface-enhanced Raman spectroscopy (SERS) and fluorometry are applied for quantitation. It makes use of platinum-coated gold nanorod (AuNR) and DNA sequences. The complementary DNA of aptamer (cDNA) against FB1 is immobilized on the surface of AuNR. The aptamer of FB1 modified with Cy5.5 are complementarily hybridized with cDNA. In the absence of FB1, the aptamer and its cDNA associate. In this situation, strong SERS and weak fluorescence signals are obtained. In the presence of FB1, the aptamer disassociates with its cDNA and binds the target. As the concentration of FB1 increases, the SERS and fluorescence signal intensities of the mixture are gradually decreased and increased, respectively. Under optimized conditions, the SERS signal at 1366cm-1 decreases linearly in the 10-500pgmL-1 concentration range with the calibration equation of y = 1997lgx-594 (the coefficient of determination is 0.998). The fluorescence signal at 670nm increases linearly in the 10-250pgmL-1 concentration range with the calibration equation of y = 500lgx-383 (the coefficient of determination is 0.991). The assay was applied to the determination of FB1 contents in spiked corn samples. The average recoveries ranged from 92 to 107%, confirming the practicality of this method. The results obtained by this assay are in good agreement with that of LC-MS/MS method. Graphical abstractSchematic illustration of a bimodal aptasensor based on surface enhanced Raman scattering (SERS) and fluorescence change for the detection of fumonisin B1 (FB1).

Europium nanosphere-based fluorescence strip sensor for ultrasensitive and quantitative determination of fumonisin B1.

Contamination of grains and related products by fumonisins (FBs) is increasingly becoming a serious food security issue. The aim of this work was to develop a europium fluorescent microsphere-based time-resolved fluorescence immunochromatographic assay (TRFICA) for FB1 detection in different grains, including corn, corn flour, wheat, rice and brown rice. Standard curves for the five types of grain matrix were established, and showed good linearity (R2 > 0.975), LOD of 8.26 μg kg-1, and a wide working range of 13.81-1000 μg kg-1. The recoveries of TRFICA for FB1 detection ranged from 82.85-103.62% with variation coefficients of 1.92-15.33%. Two corn reference materials and other natural samples were tested using TRFICA. The same samples analyzed by liquid chromatography tandem mass spectrometry further confirmed the TRFICA results. The entire detection time of TRFICA was within 30 min. Thus, this developed TRFICA can be used for onsite detection and quantitation of FB1 in grains.

A novel electrochemical aptasensor for fumonisin B1 determination using DNA and exonuclease-I as signal amplification strategy

In this work, using DNA and exonuclease-I (Exo-I) as signal amplification strategy, a novel and facile electrochemical aptasensor was constructed for fumonisin B1 (FB1) detection. The G-rich complementary DNA (cDNA) was immobilized onto the electrode surface. Then, aptamer of FB1 was hybridized with cDNA to form double-stranded DNA. In the absence of FB1, double-stranded DNA and G-rich cDNA on the electrode surface promoted effectively methylene blue (MB) enrichment and amplified the initial electrochemical response. In the presence of FB1, the combination of aptamer and FB1 led to the release of aptamer from the electrode surface and the expose of 3′ end of single-stranded cDNA. When Exo-I was added onto the electrode surface, the single-stranded cDNA was degraded in the 3′–5′ direction. The decrease of double-stranded DNA and G-rich cDNA resulted in the less access of MB to the electrode surface, which decreased the electrochemical signal. The experimental conditions including incubation time of FB1, the amount of Exo-I and incubation time of Exo-I were optimized. Under the optimal conditions, the linear relationship between the change of peak current and the logarithmic concentration of FB1 was observed in the range of 1.0 × 10−3–1000 ng mL−1 with a low limit of detection of 0.15 pg mL−1. The experimental results showed that the prepared aptasensor had acceptable specificity, reproducibility, repeatability and stability. Therefore, this proposed aptasensor has a potential application in the food safety detection.

Colorimetric determination of fumonisin B1 based on the aggregation of cysteamine-functionalized gold nanoparticles induced by a product of its hydrolysis.

A colorimetric method was developed for the determination of the mold toxin fumonisin B1 (FB1). It is based on the aggregation of cysteamine-capped gold nanoparticles (Cys-AuNPs). The assay involves alkaline hydrolysis of FB1 to obtain hydrolyzed fumonisin B1 (HFB1). The latter induces the aggregation of Cys-AuNPs which results in a color change from wine-red to blue-gray, best at a pH value of 9.0. A plot of absorbance ratio at 645/520nm versus FB1 concentration is linear in the 2-8μgkg-1 FB1 concentration range, and the detection limit is 0.90μgkg-1. Inter-day and intra-day precisions are <6.2%, and recoveries from spiked samples ranged from 93 to 99%. The assay was successfully applied to the determination of FB1 in corn samples. It has a high selectivity over other competitive mycotoxins including aflatoxin, zearalenone, citrinin and patulin. The method is more selective than the detection of FB1 directly which may lead to false-positive errors. Graphical abstract Schematic representation of colorimetric assay of fumonisin B1 (FB1). FB1 was alkali-hydrolyzed and its product (hydrolyzed fumonisin B1) induces cysteamine-capped gold nanoparticles (Cys-AuNPs) via hydrogen bondings. The aggregation of Cys-AuNPs causes changes in color from wine-red to blue-gray.

Determination of Fumonisin B1 in maize using molecularly imprinted polymer nanoparticles-based assay

Fumonisin B1 (FB1) is a carcinogenic mycotoxin produced by Fusarium species contaminating maize. At present, fumonisin determination is performed using costly and demanding chromatography techniques or immunoassays. Recently, a molecularly imprinted polymer nanoparticles (nanoMIPs) – based assay (MINA) has been developed for FB1 detection. Herein, we have applied MINA for the determination of FB1 in naturally contaminated maize samples and results were compared with those obtained with ELISA and a reference HPLC method (AOAC No. 2001.04). The nanoMIPs as a recognition element mimicking antibodies used in ELISA were produced by solid phase synthesis and used in MINA for FB1 determination in 53 maize samples. As a result, 18 maize samples were contaminated with FB1 at levels higher than 0.25 mg/kg. Fumonisin concentrations from samples measured by MINA were well correlated with those using ELISA and HPLC. Therefore, MINA could be used as an alternative technique for FB1 determination in maize.

Development of a noncompetitive idiometric nanobodies phage immumoassay for the determination of fumonisin B1

ABSTRACT Here we demonstrate a noncompetitive idiometric nanobodies phage immunoassay for the determination of FB1, utilizing two types of anti-idiotypic nanobody, that is specific for different epitopes within the hypervariable region of the primary mAb. Three alphatype anti-idiotypic nanobodies were obtained from a naive nanobodies phage-display library. A noncompetitive idiometric immunoassay was established with a combination of betatype anti-idiotypic nanobody and phage-displayed alphatype anti-idiotypic nanobody. The half-maximal saturation of the signal value (SC50) was 0.68 ng/mL, with the limit of detection (LOD) was 0.19 ng/mL. When the noncompetitive assay was compared to the competitive ELISA (LOD = 3.41 ng/mL), an over 17-fold improvement in sensitivity was observed. A correlation (R 2) was 0.988 between the data of the noncompetitive assay and LC–MS/MS for the determination of FB1 in cereal samples. The noncompetitive idiometric immunoassay would have a broad utility for monitoring small molecules in food and environment.

Magnetic Reduced Graphene Oxide/Nickel/Platinum Nanoparticles Micromotors for Mycotoxin Analysis.

Magnetic reduced graphene oxide/nickel/platinum nanoparticles (rGO/Ni/PtNPs) micromotors for mycotoxin analysis in food samples were developed for food-safety diagnosis. While the utilization of self-propelled micromotors in bioassays has led to a fundamentally new approach, mainly due to the greatly enhanced target-receptor contacts owing to their continuous movement around the sample and the associated mixing effect, herein the magnetic properties of rGO/Ni/PtNPs micromotors for mycotoxin analysis are additionally explored. The micromotor-based strategy for targeted mycotoxin biosensing focused on the accurate control of micromotor-based operations: 1) on-the-move capture of free aptamers by exploiting the adsorption (outer rGO layer) and catalytic (inner PtNPs layer) properties and 2) micromotor stopped flow in just 2 min by exploiting the magnetic properties (intermediate Ni layer). This strategy allowed fumonisin B1 determination with high sensitivity (limit of detection: 0.70 ng mL-1 ) and excellent accuracy (error: 0.05 % in certified reference material and quantitative recoveries of 104±4 % in beer) even in the presence of concurrent ochratoxin A (105-108±8 % in wines). These results confirm the developed approach as an innovative and reliable analytical tool for food-safety monitoring, and confirm the role of micromotors as a new paradigm in analytical chemistry.

An ultrasensitive chemiluminescence immunoassay for fumonisin B1 detection in cereals based on gold-coated magnetic nanoparticles.

In the present study, a novel highly sensitive magnetic enzyme chemiluminescence immunoassay (MECLIA) was developed to detect fumonisin B1 (FB1 ) in cereal samples. The gold-coated magnetic nanoparticles (Fe3 O4 @Au, GoldMag) were used as solid phase carrier to develop a competitive CLIA for detecting FB1 , in which FB1 in samples would compete with FB1 -ovalbumin coated on the surface of Fe3 O4 @Au nanoparticles for binding with FB1 antibodies. Successively, horseradish peroxidase labeled goat anti-rabbit IgG (HRP-IgG) was conjugated with FB1 antibodies on the microplate. In substrate solution containing luminol and H2 O2 , HRP-IgG catalyzed luminol oxidation by H2 O2 , generating a high chemiluminescence signal. The FB1 immune GoldMag particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscope and zeta potential analysis, etc. RESULTS: The concentrations and the reaction times of these immunoreagents were optimized to improve the performances of this method. The established method could detect as low as 0.027 ng mL-1 FB1 from 0.05 ng mL-1 to 25 ng mL-1 , demonstrating little cross-reaction (less than 2.4%) with other structurally related compounds. The average intrassay relative SD (RSD) (n = 6) was 3.4% and the average interassay RSD (n = 6) was 5.4%. This method was successfully applied for the determination of FB1 in corn and wheat and gave recoveries of between 98-110% and 91-105%, respectively. The results of the present study suggest that the MECLIA approach has potential application for high-throughput fumonisin screening in cereals. © 2018 Society of Chemical Industry.

Development and Validation of a UPLC-MS/MS and UPLC-HR-MS Method for the Determination of Fumonisin B1 and Its Hydrolysed Metabolites and Fumonisin B2 in Broiler Chicken Plasma.

A sensitive and specific method for the quantitative determination of Fumonisin B1 (FB1), its partially hydrolysed metabolites pHFB1a+b and hydrolysed metabolite HFB1, and Fumonisin B2 (FB2) in broiler chicken plasma using ultra-performance liquid chromatography combined with tandem mass spectrometry (UPLC-MS/MS) was developed. The sample preparation was rapid, straightforward and consisted of a deproteinization and phospholipid removal step using an Oasis® OstroTM 96-well plate. Chromatography was performed on an Acquity HSS-T3 column, using 0.3% formic acid and 10 mM ammonium formate in water, and acetonitrile as mobile phases. The MS/MS instrument was operated in the positive electrospray ionization mode and the two multiple reaction monitoring transitions were monitored for each component for quantification and identification, respectively. The method was validated in-house: matrix-matched calibration graphs were prepared and good linearity (r ≥ 0.99) was achieved over the concentration ranges tested (1–500 ng/mL for FB1 and FB2; 0.86–860 ng/mL for pHFB1a; 0.72–1430 ng/mL for pHFB1b and 2.5–2500 ng/mL for HFB1). Limits of quantification (LOQ) and detection (LOD) in plasma ranged between 0.72 to 2.5 ng/mL and 0.03 to 0.17 ng/mL, respectively. The results for the within-day and between-day precision and accuracy fell within the specified ranges. Moreover, the method was transferred to an UPLC high-resolution mass spectrometry (HR-MS) instrument in order to determine potential metabolites of HFB1, such as N-acyl-HFB1s and phase II metabolites. The method has been successfully applied to investigate the toxicokinetics and biotransformation of HFB1 in broiler chickens.

Molecularly imprinted polymer nanoparticle-based assay (MINA): application for fumonisin B1 determination.

The enzyme-linked immunosorbent assay (ELISA) has been used as a standard tool for monitoring food and animal feed contamination from the carcinogenic fumonisin B1 (FB1). Unfortunately, ELISA is not always efficient due to the instability of the antibody and enzyme components in the immunoassay, the presence of natural enzyme inhibitors in the samples and the high levels of non-specific protein binding. Additionally, the production of antibodies for ELISA can be time-consuming and costly, due to the involvement of animals in the manufacturing process. To overcome these limiting factors, a molecularly imprinted nanoparticle based assay (MINA) has been developed, where the molecularly imprinted nanoparticles (nanoMIPs) replace the primary antibody used in a competitive ELISA. Herein, computational modelling was used to design the nanoMIPs by selecting monomers that specifically interact with FB1. The affinity of the monomers to FB1 was verified by measuring their binding in affinity chromatography experiments. The nanoMIPs were produced by solid phase synthesis and the results showed that nanoMIPs had a hydrodynamic diameter of around 249 ± 29 nm. The assay tested in model samples is highly selective and does not show cross-reactivity with other mycotoxins such as fumonisin B2 (FB2), aflatoxin B1 (AFB1), citrinin (CTT), zearalenone (ZEA), and deoxynivalenol (DON). The MINA allows the detection of FB1 in the concentration range of 10 pM-10 nM with a detection limit of 1.9 pM and a recovery of 108.13-113.76%.