Determination Of Aflatoxin B1 Research Articles

Preparation of CeO2QDs-g-C3N4/C composites and electrochemical determination of aflatoxin B1

The presence of Aflatoxin B1 (AFB1) in food represents a significant threat to human health, leading to the development of cancer. The key factor for effective trace detection technology lies in the utilization of sensor materials that exhibit excellent selectivity and high sensitivity properties. In this study, a successfully synthesis of g-C3N4/C with a high specific surface area uses melamine and houttuynia cordata stem as starting materials. A CeO2QDs-g-C3N4/C composite was prepared by hydrothermally anchoring cerium oxide quantum dots (CeO2QDs) onto the surface of g-C3N4/C, the high redox efficiency of CeO2QDs and the small size limitation effect significantly improve the sensing performance. The composite was characterized by XRD, XPS, SEM, TEM, and N2 adsorption-desorption. The results revealed that, the CeO2QDs-g-C3N4/C composite sensor material exhibited significant advantages with a large specific surface area, a well-defined micropore structure, and abundant reactive sites. In addition, electrochemical activity tests are conducted using EIS, CV, and DPV for the purpose of electrochemical investigations. The CeO2QDs-g-C3N4/C/GCE exhibits exceptional electrocatalytic activity against AFB1, with a wide linear response range of 100-1300 pg ml-1, an impressively low detection limit (LOD) of 6.61 fg ml-1 (S/N=3), and a sensitivity of 0.985 pg μM ml-1μA-1. Furthermore, the stability, repeatability, and interference experimental response errors all remain below 5%. It is also noteworthy that, the sensor material demonstrates excellent practicality in AFB1 assays conducted on real samples, which serves to illustrate its immense potential for applications in food safety evaluation.

Development of a reusable polymeric fluorescence sensor based on acryloyl β-cyclodextrin for the determination of aflatoxin B1 in grain products

AFB1 is a harmful substance that can be found in agricultural products and can seriously affect human health, even in trace amounts. Therefore, monitoring AFB1 levels to ensure food safety and protect public health is crucial. New, highly reliable, selective, and rapid detection methods are needed to achieve this goal. Our work involves the development of a polymeric membrane sensor using radical polymerization that can accurately detect AFB1. Various spectroscopic techniques (Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM)) were used to obtain information about the structural and morphological properties of the prepared sensor. The sensor displayed fluorescence selectively responsive to AFB1 at the excitation wavelength of 376 nm and emission wavelength of 423 nm. The polymeric fluorescence sensor showed good sensitivity and a wide linear range from 9.61 × 10−10 and 9.61 × 10−9 mol/L for AFB1quantification. The limit of detection (LOD) is as low as 3.84 × 10−10 mol/L for AFB1. Other mycotoxins, such as aflatoxin B2 and aflatoxin G1, did not interfere with the sensor’s high selectivity towards AFB1. To test the sensor’s effectiveness in detecting AFB1 in real samples, three different grain samples – peanuts, hazelnut butter, and peanuts with a sauce known to contain AFB1 – were utilized. The results were satisfactory and demonstrated that the sensor can be successfully employed in real samples, with an error range of 0.43 % to 12.10 %.

Development of a Voltammetric Methodology Based on a Methacrylic Molecularly Imprinted Polymer-Modified Carbon-Paste Electrode for the Determination of Aflatoxin B1

Aflatoxin B1 (AFB1) is one of the most dangerous mycotoxins found in food, necessitating the development of precise and reliable methodologies for its detection. In this study, a novel electrochemical sensor based on a molecularly imprinted polymer (MIP) integrated with a carbon-paste electrode was developed for the voltammetric determination of AFB1. The innovative aspect of this work lies in the use of methacrylic acid (MAA) as the functional monomer, which enhances the sensor’s selectivity and binding affinity. The developed electrochemical sensor exhibited a linear response range from 20.8 to 80 ng/L, with a limit of detection (LOD) of 2.31 ng/L and a sensitivity of 19.83 µA (ng/L)−1 cm−2. The sensor demonstrated outstanding analytical performance, with reproducibility and repeatability yielding relative standard deviations (RSDs) of 3.24% and 1.41%, respectively. To validate the sensor’s practical applicability, its performance was tested in real samples of corn and wheat using the standard addition method. Samples were prepared following official Mexican standard methods. Detected AFB1 concentrations were 0.0147 μg/L and 0.0138 μg/L for corn and wheat, respectively. A statistical comparison using the Student’s t-test confirmed no significant matrix effects, underscoring the high selectivity and accuracy of the MIP-modified sensor. This work introduces a highly selective, sensitive, and reproducible methodology for AFB1 detection, which could significantly advance food safety monitoring.

Fluorescence/visual aptasensor based on Au/MOF nanocomposite for accurate and convenient aflatoxin B1 detection.

Adual-mode fluorescence/visual aptasensor was developed for straightforward and accurate determination of aflatoxin B1 (AFB1) based on an Au/metal-organic framework (Au/MOF) composite. Aptamer-modified Au/Fe3O4 (Apt/Au/Fe3O4) served as the recognition element, and Au/MOF modified with complementary chains and 3,3',5,5'-tetramethylbenzidine (cDNA/TMB/Au/MOF) acted as the fluorescence and visual probes. These components are integrated to form conjugates (Apt/Au/Fe3O4-cDNA/TMB/Au/MOF). Upon the introduction of AFB1, some cDNA/TMB/Au/MOF dissociated from Apt/Au/Fe3O4, enabling the use of detached probes for visual detection. The undecomposed conjugates were isolated magnetically for use in fluorescence detection. As the AFB1 concentration increases, the visual signal intensifies and fluorescence intensity diminishes. Thus, the proposed aptasensor achieves the simultaneous fluorescence and visual determination of AFB1, obviating the need for material and reagent substitutions. The detection limits were established at 0.07ngmL-1 for the fluorescence mode and 0.08ngmL-1 for the visual mode. The effectiveness of the aptasensor was further validated by quantifying AFB1 in real samples.

Immunochromatographic Strip Based on Tetrahedral DNA Immunoprobe for the Detection of Aflatoxin B1 in Rice Bran Oil.

Aflatoxin B1 (AFB1), a widespread contaminant in food and feeds, poses a threat to the health of animals and humans. Consequently, it is significant to develop a rapid, precise and highly sensitive analytical method for the detection of AFB1. Herein, we developed an immunochromatographic strip (ICS) based on a tetrahedral DNA (TDN) immunoprobe for AFB1 determination in rice bran oil. Three sizes of TDN immunoprobes (AuNP-TDN13bp-mAb, AuNP-TDN17bp-mAb, AuNP-TDN26bp-mAb) were constructed, and the performance of these three immunoprobes, including the effective antibody labeling density and immunoaffinity, was measured and compared with that of the immunoprobe (AuNP-mAb) developed using the physical adsorption method. Subsequently, the optimal TDN immunoprobe, namely AuNP-TDN13bp-mAb, was selected to prepare the immunochromatographic strip (ICS) for the qualitative and quantitative detection of AFB1 in rice bran oil. The visual limits of detection (vLODs) of the ICS based on AuNP-TDN13bp-mAb and AuNP-mAb were 0.2 ng/mL and 2 ng/mL, with scanning quantitative limits (sLOQs) of 0.13 ng/mL and 1.4 ng/mL, respectively. The ICS demonstrated a wide linear range from 0.02 ng/mL to 0.5 ng/mL, with good specificity, accuracy, precision, repeatability, and stability. Moreover, a high consistency was observed between the constructed ICS and ultra-high-performance liquid chromatography (UPLC) in the quantification of AFB1. The results indicated that the introduction of TDN was beneficial for promoting efficient antibody labeling, protecting the bioactivity of immunoprobes, and increasing the sensitivity of detection, which would provide new perspectives for the achievement of the highly sensitive detection of mycotoxins.

Colorimetric and ECL dual-mode aptasensor for smartphone-based onsite sensitive detection of aflatoxin B1 in combination with ZnO@MWCNTs/g-C3N4 nanosheets and CuO@CuPt nanocomposites

The development of dual-mode strategies with superior sensitivity and accuracy have garnered increasing attention for researchers in Aflatoxin B1 (AFB1) analysis. Herein, a colorimetric-electrochemiluminescence (ECL) dual-mode biosensor was constructed for onsite and ultrasensitive determination of AFB1. The multi-wall carbon nanotubes (MWCNTs) were integrated with the ZnO metal organic frameworks (MOFs) to accelerate the electron transfer and boost the ECL intensity of g-C3N4 nanoemitters. Through the aptamer-based DNA sandwich assay, the CuO@CuPt nanocomposites were introduced onto the electrode and acted as the dual functional signal nanoprobes. Due to the good spectrum overlap between the CuO@CuPt nanoprobes and g-C3N4 nanosheets, ECL signal could be efficiently quenched. Additionally, the CuO@CuPt nanoprobes show superior catalytic properties towards the TMB and H2O2 colorimetric reactions, and an obvious color alteration from colorless to blue can be observed using the smartphone. Under optimized conditions, a sensitive and accurate dual-mode analysis of the AFB1 was accomplished with the colorimetric detection limit of 3.26 fg/mL and ECL detection limit of 0.971 fg/mL (S/N = 3). This study combines innovative nanomaterial properties of ZnO@MWCNTs, g-C3N4 and CuO@CuPt for ultrasensitive dual-mode detection, which offers new opportunities for the innovative engineering of the dual-mode sensors and demonstrates significant potential in food safety analysis.

Simultaneous Determination of Aflatoxin B1 and Ochratoxin A in Cereals by a Novel Electrochemical Aptasensor Using Metal-Organic Framework as Signal Carrier.

A novel electrochemical aptasensor was prepared for the simultaneous determination of aflatoxin B1 (AFB1) and ochratoxin A (OTA). Composites of Au nanoparticles and polyethyleneimine-reduced graphene oxide (AuNPs/PEI-RGO) with good electrical conductivity and high specific surface area were employed as the supporting substrate, demonstrating the ability to provide more binding sites for aptamers and accelerate the electron transfer. Aptamers were immobilized on a AuNPs/PEI-RGO surface to specifically recognize AFB1 and OTA. A metal-organic framework of UiO-66-NH2 served as the signal carrier to load metal ions of Cu2+ and Pb2+, which facilitated the generation of independent current peaks and effectively improved the electrochemical signals. The prepared aptasensor exhibited sensitive current responses for AFB1 and OTA with a linear range of 0.01 to 1000 ng/mL, with detection limits of 6.2 ng/L for AFB1 and 3.7 ng/L for OTA, respectively. The aptasensor was applied to detect AFB1 and OTA in cereal samples, achieving results comparable with HPLC-MS, with recovery results from 92.5% to 104.1%. With these merits of high sensitivity and good selectivity and stability, the prepared aptasensor proved to be a powerful tool for evaluating contaminated cereals.

Development of tri-mode lateral flow immunoassay based on tailored porous gold nanoflower for sensitive detection of aflatoxin B1

Aflatoxin B1 (AFB1) is regarded as the most toxic mycotoxin and a proven human carcinogen. The development of detection assays for AFB1 is important for human health and environmental protection. In this study, the hollow porous gold nanoflower (HPGN) was synthesized to achieve colorimetric and photothermal performance. Especially, the HPGN exhibited excellent surface-enhanced Raman scattering (SERS) performance by loading with 4-mercaptobenzoic acid (4-MBA), thus SERS determination of AFB1 could be achieved. Compared with gold nanoparticles (AuNPs), the HPGN possessed unique morphology and excellent properties, providing the basis for sensitive and selective AFB1 detection. Under the optimized experimental conditions, the limits of detection (LODs) of lateral flow immunoassay (LFIA) for AFB1 through colorimetric, photothermal and SERS modes were 0.09 ng/mL, 0.09 ng/mL and 0.05 ng/mL, respectively. The tri-mode detection strategy provided a novel concept for immunosensing of mycotoxins or other small molecules.

Portable dual-mode paper chips for highly sensitive and rapid determination of aflatoxin B1 via an aptamer-gated MOFs

Paper chip as a representative microfluidic device has been mushroomed for rapid identification of contaminants in agro-food. However, the sensitivity and accuracy have still been challenged by inevitable background noise or interference in food matrix. Herein, we designed and fabricated a dual-mode paper chip (DPC) by assembling a patterned paper electrode with a platinum nanoparticles-treated colorimetric region through a flow channel. Dual-mode outputs were guided by an aptamer-gated UiO-66-NH2 metal-organic frameworks (MOFs). UiO-66-NH2 loaded with 3, 3′, 5, 5′-tetramethylbenzidine (TMB) was controlled by a switch comprised of CdS quantum dots-aptamer. Aflatoxin B1 (AFB1, a kind of carcinogenic mycotoxin) target came and induced TMB release, triggering colorimetric and ECL signals on DPC, ultra-high sensitivity with a detection limit of 7.8 fg/mL was realized. The practicability of the DPC was also confirmed by spiking AFB1 in real corn samples. This portable paper-based device provides an ideal rapid detection platform tailored for diverse food contaminants analysis.

A highly sensitive fluorescence biosensor for aflatoxins B1 detection based on polydiacetylene liposomes combined with exonuclease III-assisted recycling amplification.

Afluorescence biosensor for determination of aflatoxin B1 (AFB1)based on polydiacetylene (PDA) liposomes and exonuclease III (EXO III)-assisted recycling amplification was developed. The AFB1 aptamer partially hybridizes with complementary DNA (cDNA), which is released upon recognition of AFB1 by the aptamer. Subsequently, the cDNA hybridizes with hairpin H to form double-stranded DNA that undergoes digestion by EXO III, resulting in the cyclic release of cDNA and generation of capture DNA for further reaction. The capture DNA then hybridizes with probe modified on PDA liposomes, leading to aggregation of liposomes and subsequent fluorescence production. This strategy exhibited a limit of detection of 0.18 ng/mL within the linear range 1-100 ng/mL with adetermination coefficient > 0.99. The recovery ranged from 92.81 to 106.45%, with relative standard deviations (RSD) between 1.73 and 4.26%, for corn, brown rice, peanut butter, and wheatsamples. The stability, accuracy, and specificity of the method demonstrated the applicability for realsampleanalysis.

Phycocyanin-based rapid fluorometric immunoassay for the determination of aflatoxin B1, deoxynivalenol, and zearalenone in food and feed matrices

This study presents a new multiplex lateral flow immunoassay (LFIA) for the detection of aflatoxin B1 (AFB1), deoxynivalenol (DON), and zearalenone (ZEN). The multi-LFIA utilized phycocyanin (PC), a protein with excellent photophysical properties, as a fluorescent indicator for quantifying the three mycotoxins on a lateral flow strip platform. PC and monoclonal antibodies (mAbs) against AFB1, DON, and ZEN were immobilized onto carboxylated latex nanospheres (LNS) through a one-step coupling process. The resulting conjugates with strong red fluorescence were then used for multiplex sensing of AFB1, DON, and ZEN in wheat, corn, and animal feed. The visual results of the assay were obtained using LED UV lights coupled with a smartphone in less than 25 min. Under the optimal conditions, the new multi-LFIA achieved minimum detection limits of 1.04, 1.6, and 2.08 ng/mL for AFB1, 2.2, 6.45, and 2.9 ng/mL for DON, and 1.74, 1.67, and 2.11 ng/mL for ZEN in wheat, corn, and feed, respectively. The proposed method demonstrated high selectivity for AFB1, DON, and ZEN and showed strong correlation with LC-MS/MS results in spiked samples. In summary, the proposed method is feasible, rapid, and could serve as a highly sensitive platform for detecting AFB1, DON, and ZEN and other analytes in a single format.

Enhanced sensitivity for Aflatoxin B1 detection in food through a h-Zn1Cd5S-based photoelectrochemical aptasensor

Designing photoelectrochemical sensors for detecting ultra-trace amounts of Aflatoxin B1 (AFB1) in complex samples poses a significant challenge. In pursuit of this objective, we engineered hollow-structured ZnCdS (denoted as h-Zn1Cd5S) material and applied an electrodeposition method to introduce Au nanoparticles (AuNPs) on the surface. This approach resulted in a significant enhancement of the photocurrent response of the photoelectrode, attributed to the synergistic effects of the hollow structure and surface plasmon resonance, thereby underscoring its potential for advanced applications. Utilizing density functional theory, we computed the transfer route and energy band formation of photogenerated electrons. Subsequently, we developed an AFB1 photoelectrochemical (PEC) aptasensor based on the competition mechanism, coupled with aptamer probe technology. This aptasensor allows for the ultra-trace determination of AFB1, exhibiting a remarkable detection limit as low as 1.32 fg/mL and a linear range from 1.0 × 10−2 to 100.0 pg/mL. The PEC aptasensor for AFB1 demonstrates excellent sensitivity and selectivity. When applied to the detection of AFB1 in Tsaoko, wheat, and corn samples, its performance aligns closely with high-performance liquid chromatography (HPLC) results, validating its applicability for analyzing real-world samples.

Determination and risk assessment of aflatoxin B1 in the kernel of imported raw hazelnuts from Eastern Azerbaijan Province of Iran

Aflatoxin B1 (AFB1) is widespread and seriously threatens public health worldwide. This study aimed to investigate AFB1 in imported hazelnut samples in northwest of Iran (Eastern Azerbaijan Province) using High-Performance Liquid Chromatography with a Fluorescent Detector (HPLC-FLD). In all tested samples AFB1 was detected. The mean concentration of AFB1 was 4.20 μg/kg and ranged from 3.145 to 8.13 μg/kg. All samples contained AFB1 levels within the maximum acceptable limit except for one sample. Furthermore, the human health risk assessment of AFB1 from consuming imported hazelnuts by Iranian children and adults was evaluated based on the margin of exposure (MoE) and quantitative liver cancer risk approaches. The MoE mean for children was 2529.76, while for adults, it was 8854.16, indicating a public health concern. The present study found that the risk of developing liver cancer among Iranian children was 0.11100736 per 100,000 people, and in the Iranian adult population was 0.0314496 cancers per 100,000 people. Since environmental conditions potentially affect aflatoxin levels in nuts, countries are advised to monitor aflatoxin contents in imported nuts, especially from countries with a conducive climate for mold growth.

Determination of Aflatoxin B1 in Grains by Aptamer Affinity Column Enrichment and Purification Coupled with High Performance Liquid Chromatography Detection.

Aflatoxin B1 (AFB1) is a highly teratogenic and carcinogenic secondary metabolite produced by Aspergillus. It is commonly detected in agricultural products such as cereals, peanuts, corn, and feed. Grains have a complex composition. These complex components severely interfere with the effective extraction and separation of AFB1, and also cause problems such as matrix interference and instrument damage, thus posing a great challenge in the accurate analysis of AFB1. In this study, an aptamer affinity column for AFB1 analysis (AFB1-AAC) was prepared for the enrichment and purification of AFB1 from grain samples. AFB1-AAC with an AFB1-specific aptamer as the recognition element exhibited high affinity and specificity for AFB1. Grain samples were enriched and purified by AFB1-AAC, and subsequently analyzed by high performance liquid chromatography with post-column photochemical derivatization-fluorescence detection (HPLC-PCD-FLD). The average recoveries of AFB1 ranged from 88.7% to 99.1%, with relative standard deviations (RSDs) of 1.4-5.6% (n = 3) at the spiked levels of 5.0-20.0 μg kg-1. The limit of detection (LOD) for AFB1 (0.02 μg kg-1) was much below the maximum residue limits (MRLs) for AFB1. This novel method can be applied to the determination of AFB1 residues in peanut, corn, and rice.

Polycarboxyl ionic liquid functionalized Yb-MOFs nanoballs based dual-wavelength responsive photoelectrochemical aptasensor for the simultaneous determination of AFB1 and OTA

Developing an accurate and precise approach for the simultaneous detection of ochratoxin A (OTA) and aflatoxin B1 (AFB1) is significant for food safety surveillance. Herein, a photoelectrochemical sensing platform was constructed based on polycarboxylic ionic liquid functionalized metal-organic framework integrated with gold nanoparticles (Yb-MOFs@AuNPs). Sulfhydryl functionalized hairpin DNA (hDNA) was immobilized on a Yb-MOFs@AuNPs modified glassy carbon electrode (GCE) surface through Au-S bond. After blocking residual active binding sites with BSA, gold nanoparticles-labeled AFB1 aptamer (AuNPs-Apt 1) and gold nanorods-labeled OTA aptamer (AuNRs-Apt 2) were introduced to construct a photoelectrochemical aptasensor for the simultaneous determination of AFB1 and OTA. Due to the surface plasmon resonance effect and the nanometer size effect of gold nanomaterials, the photoelectrochemical aptasensor can output photocurrent responses as being excited with different wavelengths at 520 nm and 808 nm, respectively. When the AFB1 and OTA concentration in the range of 0.001–50.0 ng mL−1, a good linear relationship between the photocurrent difference (ΔI) before and after recognizing targets and the logarithm of AFB1 or OTA concentration was obtained. The detection limits for AFB1 and OTA were 0.40 pg mL−1 and 0.19 pg mL−1, respectively. AFB1 and OTA in corn samples were detected simultaneously by the photoelectrochemical aptasensor.

A dual-signal aptasensor based on cascade amplification for ultrasensitive detection of aflatoxin B1

Aflatoxin B1 (AFB1) is considered as a serious carcinogenic mycotoxin that was widely detected in grains and foods, and its sensitive analysis is of key importance to avoid the health threats for consumers. In this study, a dual-signal aptasensor based on cascade of entropy-driven strand displacement reaction (ESDR) and linear rolling circle amplification (LRCA) was fabricated for ultrasensitive determination of AFB1. At the sensing system, the complementary strand would be released after the aptamer combined with AFB1, which will bring about the functional domains exposed, triggering the subsequent ESDR. Meanwhile, the two strands that were outputted by ESDR would incur the downstream LRCA reaction to produce a pair of long strands to assist in the generation of fluorescence and absorbance signals. Under the optimized conditions, the proposed aptasensor could achieve excellent sensitivity (limit of detection, 0.427 pg/mL) with satisfactory accuracy (recoveries, 92.8–107.9 %; RSD, 2.4–5.0 %), mainly ascribed to the cascade amplification. Importantly, owing to the flexibility design of nucleic acid primer, this analytical method can be applied in monitoring various hazardous substances according to the specific requirements. Our strategy provides some novel insights at signal amplification for rapid detection of AFB1 and other targets.

Quantitative Determination of Aflatoxin B1 in Maize and Feed by ELISA and Time-Resolved Fluorescent Immunoassay Based on Monoclonal Antibodies.

In this study, a highly sensitive monoclonal antibody (mAb) was developed for the detection of aflatoxin B1 (AFB1) in maize and feed. Additionally, indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and time-resolved fluorescence immunoassay assay (TRFICA) were established. Firstly, the hapten AFB1-CMO was synthesized and conjugated with carrier proteins to prepare the immunogen for mouse immunization. Subsequently, mAb was generated using the classical hybridoma technique. The lowest half-maximal inhibitory concentration (IC50) of ic-ELISA was 38.6 ng/kg with a linear range of 6.25-100 ng/kg. The limits of detections (LODs) were 6.58 ng/kg and 5.54 ng/kg in maize and feed, respectively, with the recoveries ranging from 72% to 94%. The TRFICA was developed with a significantly reduced detection time of only 21 min, from sample processing to reading. Additionally, the limits of detection (LODs) for maize and feed were determined to be 62.7 ng/kg and 121 ng/kg, respectively. The linear ranges were 100-4000 ng/kg, with the recoveries ranging from 90% to 98%. In conclusion, the development of AFB1 mAb and the establishment of ic-ELISA for high-throughput sample detection, as well as TRFICA for rapid detection presented robust tools for versatile AFB1 detection in different scenarios.

Development of a label-free, sensitive gold nanoparticles-poly(adenine) aptasensing platform for colorimetric determination of aflatoxin B1 in corn.

In this work, a sensitive colorimetric bioassay method based on a poly(adenine) aptamer (polyA apt) and gold nanoparticles (AuNPs) was developed for the determination of aflatoxin B1 (AFB1). The polyA apt, adsorbed on the AuNPs, especially can bind to the analyte while deterring non-specific interactions. This nano aptasensor uses cationic polymer poly(diallyl dimethyl ammonium chloride) (PDDA), as an aggregating agent, to aggregate gold nanoparticles. PolyA apt-decorated gold nanoparticles (AuNPs/polyA apt) show resistance to PDDA-induced aggregation and maintains their dispersed state (red color) with the optical absorbance signal at λ = 520 nm. However, in the presence of AFB1 in the assay solution, the specific aptamer reacts with high affinity and folds into its three-dimensional form. Aggregation of AuNPs induced by PDDA caused their optical signal shift to λ = 620 nm (blue color). AFB1 concentration in the bioassay solution determines the amount of optical signal shift. Therefore, optical density ratio in two wavelengths (A620/520) can be used as a sturdy colorimetric signal to detect the concentration of aflatoxin B1. AFB1 was linearly detected between 0.5 and 20 ng mL-1, with a detection limit of 0.09 ng mL-1 (S/N = 3). The fabricated aptasensor was applied to the detection of AFB1 in real corn samples.

Europium nanospheres based ultrasensitive fluorescence immunosensor for aflatoxin B1 determination in feed

In this work, a new competitive immunosensor for aflatoxin B1 (AFB1) detection was developed using europium (Eu) fluorescent nanospheres and magnetic beads. Firstly, Eu nanospheres were synthesized through two steps including carboxylated polystyrene nanospheres and Eu-doped polystyrene nanospheres preparation. Then Eu nanospheres were covalently tagged to anti-AFB1 monoclonal antibody (anti-AFB1 mAb) through an EDC coupling method. Carboxylated Fe3O4 magnetic beads were conjugated to AFB1-BSA through EDC/NHS crosslinking to obtain AFB1-BSA-Fe3O4. In the absence of AFB1, Eu-anti-AFB1 mAb were incubated with AFB1-BSA-Fe3O4 to form Eu-anti-AFB1 mAb-AFB1-BSA-Fe3O4 in PBS buffer. However, in the presence of AFB1, the competitive interaction of AFB1 and AFB1-BSA-Fe3O4 to bind with Eu-anti-AFB1 mAb occurred. With the increasing concentration of AFB1, less Eu-anti-AFB1 mAb-AFB1-BSA-Fe3O4 formed. So the fluorescence intensity of Eu-anti-AFB1 mAb-AFB1-BSA-Fe3O4 was gradually decreased after magnetic separation. The degree of fluorescence decrease was linear with respect to the logarithm of AFB1 concentration in the range of 0.01–2 ng/mL in both buffer solution and feed samples and the detection limit was 0.003 ng/mL. What's more, the immunosensor showed excellent specificity for AFB1 without being interfered by other mycotoxins. In consideration of the excellent performance of this immunosensor, we can speculate that the proposed method could be widely used in detecting food contaminants.

A versatile CuCo@PDA nanozyme-based aptamer-mediated lateral flow assay for highly sensitive, on-site and dual-readout detection of Aflatoxin B1

Mycotoxin contaminations in food and environment seriously harms human health. Constructing sensitive and point-of-test early-warning tools for mycotoxin determination is in high demand. In this study, a CuCo@PDA nanozyme-based aptamer-mediated lateral flow assay (Apt-LFA) has been elaborately designed for on-site and sensitive determination of mycotoxin Aflatoxin B1 (AFB1). Benefiting from the rich functional groups and excellent peroxidase-like activity, the CuCo@PDA with original dark color can be conjugated with the specific recognition probe (i.e., aptamer), generating colorimetric signal on the test lines of Apt-LFA via a competitive sensing strategy. The signal can further be amplified in-situ by catalytic chromogenic reaction. Therefore, a visual and dual-readout detection of AFB1 has been realized. The developed Apt-LFA provides a flexible detection mode for qualitative and quantitative analysis of AFB1 by naked-eyes observation or smartphone readout. The smartphone-based LFA platform shows a reliable and ultrasensitive determination of AFB1 with the limit of detection (LOD) of 2.2 pg/mL. The recoveries in the real samples are in the range of 95.11–113.77% with coefficients of variations less than 9.84%. This study provides a new approach to realize point-of-test and sensitive detection of mycotoxins in food and environment using nanozyme-based Apt-LFAs.