Novel advanced materials and magnetic solid phase extraction as approaches in sample preparation to enhance the analysis of ochratoxin A in peanuts.

Ochratoxin A (OTA) is a mycotoxin that can contaminate a variety of agricultural commodities, including fruit juices and wines. The capability of a magnetic solid-phase extraction (MSPE) method with a magnetic metal-organic framework (MOF) material having a three-layer core-shell structure to improve the detection of OTA in food matrices using high performance liquid chromatography is described. Analysis of the material through X-ray diffraction (XRD) indicated the successful synthesis of the magnetic nanomaterial Fe3O4@SiO2@UiO66-NH2. Scanning electron microscopy (SEM) and Zetasizer lab indicated its nano-sized morphological features. The conditions affecting the magnetic solid-phase extraction procedure, such as material dosage, pH, composition and amount of eluent, desorption solution and desorption time were investigated to obtain the optimal extraction conditions. Under optimized conditions, the recoveries of spiked analytes at three different concentrations ranged from 95.83 to 101.5%, and the relative standard deviations were below 5%. Coupling with HPLC allowed the limit of detection to be 0.3 µg kg-1. This method is simple and specific, and can effectively avoid the influence of coexisting elements and improve the sensitivity of determination through fast MSPE of OTA. It has broad development prospects in OTA detection pre-treatment.

Separation of ochratoxins by centrifugal partition chromatography.

The present research work was dedicated to developing an efficient method based on liquid-liquid chromatography (centrifugal partition chromatography, CPC) applicable to routine purifications of ochratoxins (OT) from the liquid culture of the strain A. albertensis SZMC 2107. The crude extract contained numerous components in addition to OTA (90.1 %,) and OTB (1.1 %,) according to HPLC examinations. For the separation of OTs by CPC, several tertiary systems based on acetonitrile, acetone, and short-chain alcohols were examined to find the most applicable biphasic system. The hexane/i-propanol/water 35:15:50 system supplemented with 0.1 % acetic acid was found to be the most efficient for use in CPC separation. Using liquid-liquid instrumental separation, the two OTs, namely OTA (2.23 mg) and OTB (0.031 mg), were successfully isolated with 96.3 % and-72.8 % purity, respectively, from 1 L ferment broth. The identities and purities of the purified components were confirmed and the performance parameters of each separation step and the whole procedure were determined. The developed method could be used effectively to purify OTs for analytical or toxicological applications.

Nanobody-alkaline phosphatase fusion-mediated phosphate-triggered fluorescence immunoassay for ochratoxin a detection.

Ochratoxin A (OTA) is a kind of mycotoxin that seriously harms the health of humans and animals. In this study, a nanobody-alkaline phosphatase fusion-mediated phosphate-triggered fluorescence immunoassay (Nb-AP-mediated PT-FIA) was developed for detecting OTA. Based on the constructed phosphate-triggered fluorescence sensing system for Nb-AP and the optimal working conditions, the Nb-AP-mediated PT-FIA has a half maximal inhibition concentration (IC50) of 0.46 ng/mL, a limit of detection (IC10) of 0.12 ng/mL, and a linear range (IC20-80) of 0.2-1.26 ng/mL, respectively. The recovery experiment indicated acceptable accuracy and precision of the Nb-AP-mediated PT-FIA, and the results were validated by high performance liquid chromatography with fluorescence detector. Thus this proposed method is applicable to sensitive, rapid, and low-cost detection of OTA and other toxic analytes with low molecular weight in food and environment.

Ratiometric electrochemical aptasensor for ultrasensitive detection of Ochratoxin A based on a dual signal amplification strategy: Engineering the binding of methylene blue to DNA.

A novel ratiometric electrochemical aptasensor was developed for Ochratoxin A (OTA) detection based on the binding of methylene blue (MB) to DNA with a dual signal amplification strategy. The formation of dsDNA structures between ferrocene-labeled complementary DNA (Fc-cDNA), the OTA aptamer, and complementary helper DNA (hDNA) caused Fc away from the electrode, and allowed dsDNA to bind with a certain amount of MB. Here, a small oxidation current of Fc (IFc) and a large oxidation current of MB (IMB) were obtained. In the presence of OTA, its specific recognition with the aptamer induced the release of aptamer and hDNA from the electrode and subsequently the formation of hairpin structure for cDNA, which caused Fc close to the electrode and a weaker binding ability with MB. Then, an increased IFc and a decreased IMB were obtained. Based on this principle, OTA could be accurately quantified by measuring the ratiometric signal of IFc/IMB. Herein, the dual signal amplification strategy of the introduction of hDNA and the binding with MB after the OTA recognition was exploited to amplify the response signal. The obtained aptasensor showed a linear detection range from 10 pg mL-1 to 10 ng mL-1 and a detection limit of 3.3 pg mL-1. The aptasensor was successfully applied to determine OTA in wheat, and the results were validated through HPLC-MS. Furthermore, by changing the target aptamers, this strategy could be universally used for the determination of various mycotoxins, showing promising potential applications for mycotoxins monitoring in agricultural products and foods.

Three-dimensional ordered macroporous magnetic photonic crystal microspheres for enrichment and detection of mycotoxins (II): The application in liquid chromatography with fluorescence detector for mycotoxins.

Enrichment, separation and purification are very important to accurately analyze mycotoxins in complicated samples. In the work, we developed a new enrichment, purification and high-performance liquid chromatography combined with fluorescence detector (HPLC-FLD) for aflatoxins B1 (AFB1), ochratoxin A (OTA) and Zearalenone (ZEN) assay using the macroporous magnetic 3D photonic crystal microspheres (3DPCMs). The conditions of enrichment and purification for mycotoxins have been optimized, which are as follows: pore size of 3DPCMs at 280 nm, 1:1 methanol:acetonitrile (v/v) as eluent, antibody concentrations at 60 µg/mL,60 µg/mL and 120 µg/mL for OTA, AFB1 and ZEN, respectively. The recovery rates in the rice, wheat and corn samples range from 70.01% to 100.12% and the relative standard deviation (RSD) range from 0.45% to 7.09%. The recovery rates used 3DPCMs are almost tenfold higher than that used non-macroporous PCMs in the same conditions. The developed method is simple, rapid (time including enrichment, purification and detection <2 h) and only requires small volume reagents (≤200 µL).

Tunable solvency mixtures of tetrahydrofuran:water for efficient and fast extraction/clean-up of trace contaminants.

In this study, we investigated the potential of mixtures of tetrahydrofuran (THF) and water as tunable solvents for the microextraction of contaminants in solid and in liquid matrices. These two miscible solvents have very different dielectric constant and Hildebrand solubility parameters, so that tunable mixtures spanning a wide range of dispersion and hydrogen bonding forces could be easily prepared by simply changing their composition. In this way, rapid and more efficient extraction methods can be developed. A liquid-liquid and a solid-liquid microextraction method for the determination of bisphenol A (BPA) in urine and ochratoxin A (OTA) in cereal baby food were developed as a proof of concept. Both, the chemical composition and the relative solvency of the THF-water mixtures, expressed as Teas solubility parameters, were studied in order to gain some insights into the chemical interactions governing analyte extraction. For urine, the salting-out extraction with THF:water and NaCl was evaluated, a process which is still scarcely investigated for analytical purposes. These methods featured good recoveries (above 95%), satisfactory standard deviation (5-6%) and good sensitivity (detection limits of 0.l µg L-1 for BPA and of 0.l ng g-1 for OTA) with the advantages of simplicity, rapidity and low consumption of reagents. Recoveries for other compounds and matrices (bisphenols ad phosphorus flame retardants in dust and in tap water, dyes in tap water and OTA in powder milk) were also assessed to prove the wide potential of these tunable solvent mixtures.

Silica modified with polymeric amphiphilic nanoparticles as first dimension for multidimensional separation techniques.

This paper addresses the evaluation of a new amphiphilic nanoparticle supported on silica and its application as sorbent in on-line solid phase extraction. The investigated sorbent material is a copolymer composed by [2- (Acryloyloxy) ethyl] trimethylammonium chloride (block A) and butyl acrylate (block B) prepared by reversible addition-fragmentation chain transfer. After polymerization the nanoparticles were adsorbed into silica surface by electrostatic attraction providing to the sorbent both, apolar and polar characteristics. After the fundamental studies to understand the main features of the synthesized nanoparticles supported on silica, this sorbent material was packed into an extraction column. This column was connected on line with liquid chromatography-tandem mass spectrometry and utilized for automated online determination of several analytes as Ochratoxin A, azoxystrobin, cyproconazole and difenoconazole, in wine and water samples.

Enzymatic oxydate-triggered AgNPs etching: A novel signal-on photoelectrochemical immunosensing platform based on Ag@AgCl nanocubes loaded RGO plasmonic heterostructure.

A well-defined Ag@AgCl nanocubes loaded on the reduced graphene oxide plasmonic heterostructure (Ag@AgCl/RGO) was facilely prepared by sacrificial salt-crystal-template process and ethylene glycol-assisted reduction. The Ag@AgCl/RGO heterostructure shows superior photocurrent response and stability under the visible light irradiation. The enhanced performance mainly attributes to the plasmon resonance effect of AgNPs by improving the absorbance and transfer of photogenerated electrons. Significantly, we observed that the photocurrent could be dramatically decreased with the introduction of H2O2 and experimental results demonstrated the etching effect of H2O2 to AgNPs should be responsible for this phenomenon. Inspired by this phenomenon, employing H2O2 that generated from glucose oxidase catalyzed glucose triggered AgNPs etching as a novel signal mode, an improved photoelectrochemical immunosensing platform was constructed by employing Ag@AgCl/RGO heterostructure as photoactive material. As a proof of concept application, the photoelectrochemical immunosensor employed for ochratoxin A (OTA) detection with competitive-type format and it exhibited excellent analytical performance. Under optimized conditions, the photocurrent increased with the concentration of target OTA in the dynamic range of 0.05 to 300 nM with a limit of detection (LOD) of 0.01 nM (4.0 pg mL-1). The immunosensor also showed high sensitivity, good reproducibility, and satisfactory accuracy. Although the methodology proposed here focused on OTA sensing, it could flexibly extend to monitor other targets by replacing the corresponding bio-recognition elements. Thus, this work provides a new paradigm for designing novel photoelectrochemical biosensing mode based on the plasmonic metal/semiconductor heterostructure.

Secondary metabolites produced by Sardiniella urbana, a new emerging pathogen on European hackberry.

In this study the production of secondary metabolites by a virulent strain of Sardiniella urbana, a recently described pathogen originally found on declining European hackberry trees in Italy, was investigated for the first time. Chemical analysis of the culture filtrate extracts led to the isolation of three well known compounds as R-(-)-mellein and (3R,4R)-and (3R,4S)-4-hydroxy melleins which were identified by spectroscopic methods (essentially NMR and ESIMS). The isolated compounds were tested for their phytotoxic, antifungal and zootoxic activities. Among them, only R-(-)-mellein was found to be active.

Facile combination of beta-cyclodextrin host-guest recognition with exonuclease-assistant signal amplification for sensitive electrochemical assay of ochratoxin A.

Smartly coupling exonuclease-induced target recycling signal amplifications with ß-cyclodextrin host-guest recognition, a novel "signal-on" aptamer sensor for sensitive determination of ochratoxin A (OTA) was proposed for the first time. Firstly, the formation of double-strand DNA (dsDNA) was occurred by hybridizing OTA aptamer with its complementary DNA (cDNA) and as the probe DNA the cDNA at its 3' terminal was labeled with methylene blue (MB). Next, when OTA was present, the aptamer tended to form aptamer-OTA complex with conformation of G-quadruplex instead of aptamer-cDNA duplex, leading to thus the probe DNA separating from dsDNA complex. Then the RecJf exonuclease was added, demolishing partially G-quadruplex structure and releasing a certain number of OTA. Sequentially, those released OTA would continue to react with the rest of aptamer in dsDNA, drawn into development of a new round of G-quadruplex complex, where the target cycling was realized. Meanwhile, as a signal molecule, MB modified on cDNA was liberated along with the cDNA being digested into monoucleotides by RecJf exonuclease, capable of diffusing onto the electrode surface due to host-guest recognition with ß-cyclodextrin, whereupon the signal was enriched and yielded. In this way, cycles of target with continuous output of signal indicators were undergone, in which the detection of target was in return fulfilled with signal amplification owing to the joint endeavor of exonuclease and ß-cyclodextrin. Under the optimal conditions, the raising signal maintained a linear relation with the logarithm of the target concentrations ranging from 10 pg/mL to 10.0 ng/mL and the detection limit reached as low as 3 pg/mL. This brand-new strategy was simple and low-cost but satisfactory in terms of detection limit, range and sensitivity, in all possibility to be applied extensively for diverse targets detection by easily alternating the corresponding aptamers.

In vitro selection of an XNA aptamer capable of small-molecule recognition.

Despite advances in XNA evolution, the binding capabilities of artificial genetic polymers are currently limited to protein targets. Here, we describe the expansion of in vitro evolution techniques to enable selection of threose nucleic acid (TNA) aptamers to ochratoxin A (OTA). This research establishes the first example of an XNA aptamer of any kind to be evolved having affinity to a small-molecule target. Selection experiments against OTA yielded aptamers having affinities in the mid nanomolar range; with the best binders possessing KD values comparable to or better than those of the best previously reported DNA aptamer to OTA. Importantly, the TNA can be incubated in 50% human blood serum for seven days and retain binding to OTA with only a minor change in affinity, while the DNA aptamer is completely degraded and loses all capacity to bind the target. This not only establishes the remarkable biostability of the TNA aptamer, but also its high level of selectivity, as it is capable of binding OTA in a large background of competing biomolecules. Together, this research demonstrates that refining methods for in vitro evolution of XNA can enable the selection of aptamers to a broad range of increasingly challenging target molecules.

Polymeric substances for the removal of ochratoxin A from red wine followed by computational modeling of the complexes formed.

Ochratoxin A (OTA) is a mycotoxin produced by filamentous-type fungi that contaminates a wide variety of foods and beverages such as wines. In these trials, we evaluated the capacity of the following polymers for the removal of OTA from acidic model solutions and red wine: polyvinylpolypyrrolidone (PVPP), resin of N-vinyl-2-pyrrolidinone with ethylene glycol dimethacrylate and triallyl isocyanurate (PVP-DEGMA-TAIC), and poly(acrylamide-co-ethylene glycol-dimethacrylate) (PA-EGDMA). In acidic model solution, PVP-DEGMA-TAIC and PA-EGDMA polymers removed up to 99.9% of OTA, but their trapping capacity was highly reduced by the presence of competing phenolic substances (i.e. gallic acid and 4-methylcathecol). In real red wine, PA-EGDMA polymer showed the most promising results, with more than 68.0% OTA removal and less than 14.0% reduction in total phenolic. Finally, computational chemistry analyses showed that the affinity between OTA and the polymers studied would be due to Van der Waals interactions.

A Label-Free Aptasensor for Ochratoxin a Detection Based on the Structure Switch of Aptamer.

A label-free sensing platform is developed based on switching the structure of aptamer for highly sensitive and selective fluorescence detection of ochratoxin A (OTA). OTA induces the structure of aptamer, transforms into G-quadruplex and produces strong fluorescence in the presence of zinc(II)-protoporphyrin IX probe due to the specific bind to G-quadruplex. The simple method exhibits high sensitivity towards OTA with a detection limit of 0.03 nM and excellent selectivity over other mycotoxins. In addition, the successful detection of OTA in real samples represents a promising application in food safety.

Aptamer-based polyhedral oligomeric silsesquioxane (POSS)-containing hybrid affinity monolith prepared via a "one-pot" process for selective extraction of ochratoxin A.

A novel aptamer-based polyhedral oligomeric silsesquioxane (POSS)-containing hybrid affinity monolith has been prepared with a facile "one-pot" process simultaneously via "free radical polymerization" and "thiol-ene" click reaction, and used for on-line selective extraction and practical analysis to trace ochratoxin A (OTA). By using the ternary porogenic mixture composed of water/DMF/PEG, a homogeneous polymerization mixture with POSS chemicals, acrylate-based monomers and aptamer aqueous solution was obtained, and the copolymerization of POSS chemicals, polymer monomers and aptamer aqueous solution was systematically studied. Characterizations such as the morphology, FT-IR and fluorescence spectra, mechanical stability, dynamic binding capacity, cross-reactivity and selectivity of the resultant affinity monolith were also evaluated. Attributed to the porous monolithic structure and aptamer-based affinity interaction, acceptable selective recognition and recovery yields towards trace OTA were obtained. With a 5-fold volume enrichment, the limit of detection (LOD) and limit of quantitation (LOQ) of OTA in fortified beer samples were gained at 0.025 ng/mL (S/N = 3) and 0.045 ng/mL (S/N = 10), respectively. It could be competent for the sensitive measure of actual OTA residues in real beer samples. In comparison with the previously reported strategies containing common "sol-gel" chemistry, the proposed protocol to fabricating aptamer-modified POSS-containing hybrid affinity monolith showed a simpler preparation with acceptable selectivity and higher recovery to trace OTA.

Occurrence of Toxigenic Fungi and Mycotoxins on Root Herbs from Chinese Markets.

Herbs derived from roots, leaves, flowers, or fruits of plants are unavoidably contaminated with fungi and mycotoxins during growth, harvest, and storage, thereby posing a health threat to humans. Especially, root herbs (RHs) are more easily contaminated with fungi and mycotoxins because the roots are in direct contact with the soil. Here, we investigated the occurrence of fungi, aflatoxins (AFs), and ochratoxin A (OTA) in eight RHs that are used as medicines, beverages, dietary supplements, and functional foods in China and other countries. Morphological observation and MultiGeneBlast (ß-tubulin and calmodulin) were used to identify the potentially toxigenic fungi. Of the 48 samples tested, all were contaminated by fungi, and 1,844 isolates belonging to 25 genera were detected. The genera Aspergillus and Penicillium, which contain potentially toxigenic fungal species, represented a frequency of 10 and 25%, respectively. Thirty-three isolates of Aspergillus flavus, Aspergillus parasiticus, Aspergillus niger, and Penicillium polonicum were arbitrarily selected for analysis of their toxigenic potential. Five of 13 isolates of A. flavus and 1 isolate of A. parasiticus produced AFs, whereas OTA production was not detected for any of the isolates of A. niger and P. polonicum. The occurrence of AFs and OTA in the 48 samples of eight RHs was tested by ultraperformance liquid chromatography-tandem mass spectrometry; 37.50% of samples from six RHs were contaminated with AFs and 16.67% of samples from four RHs were contaminated with OTA. Seven (14.58%) and four (8.33%) samples of ginseng, polygala, and liquorice exceeded the permissible limits of aflatoxin B1 and AFs, respectively. Because ginseng, polygala, and liquorice are widely used as herbs, dietary supplements, and functional foods, the high frequency of AF contamination of these herbs indicated by our current study warrant attention to raise public awareness.

Performance of purified grape pomace as a fining agent to reduce the levels of some contaminants from wine.

The quality of red wine depends on the absence of compounds which may affect its safety and/or stability such as ochratoxin A, biogenic amines and some metals and trace compounds. The presence of ochratoxin A in musts and wines is due to fungal contamination of the grapes and has been classified as a possible human carcinogen. Biogenic amines are formed by the microbiological decarboxylation of the corresponding amino acid precursors during the fermentation or ageing and storage, and, at high concentrations, they may induce adverse reactions in sensitive people. Trace elements may have both a nutritional and a toxic effect on health, but also can cause turbidity and stability problems. Their presence is affected mainly by natural factors such as soil mineral content and direct contact with tank surfaces and metallic tubing during winemaking. One of the best options to remove these compounds when present in excess in wine is fining. However, some fining agents commonly used may themselves present problems related with their allergenic properties or with their propensity to increase the protein content, which can cause turbidity problems. In an attempt to avoid such these problems, purified grape pomace was tested as a fining alternative since it has been seen to have a high capacity to reduce the astringency, turbidity and also the ochratoxin A content. The main aim of this work, therefore, was to study if this material can limit the presence of ochratoxin A, biogenic amines and metals and some trace elements in a Monastrell red wine, thus increasing the value and safety of this product.

Broad-specificity photoelectrochemical immunoassay for the simultaneous detection of ochratoxin A, ochratoxin B and ochratoxin C.

A broad-specific photoelectrochemical (PEC) immunosensor was developed for the simultaneous detection of ochratoxin A, ochratoxin B and ochratoxin C (OTA, OTB, OTC) by using the direct growth of CdS nanorods on FTO as the photoelectrode and Au nanoflowers-modified glass carbon electrode (GCE) as the bioelectrode. The bioelectrode was used to capture antigens and then associate corresponding antibodies, followed by using SiO2@Cu2+ nanocomposites to conjugate the secondary antibody (Ab2) and a DNA strand as the initiator. After the hybridization chain reaction (HCR) and the addition of hemin, numerous DNAzymes (G-quadruplex/hemin) were produced. Due to the similar enzymatic property with horseradish peroxidase (HRP), G-quadruplex/hemin can accelerate the oxidation of 4-chloro-1-naphthol (4-CN) with H2O2 to yield the biocatalytic precipitation (BCP) on the bioelectrode. Then, the bioelectrode was further treated with moderate acid and thus Cu2+ was released, which can decrease the photocurrent of the photoelectrode by the formation of CuS. Due to the advantages of surface effect of Au nanoflowers, DNA amplification and high photoelectrocatalytic activity, the proposed broad-specificity PEC immunosensor can detect OTA, OTB and OTC with a detection limit of 0.02, 0.04 and 0.03 pg/mL, respectively. In addition, the acceptable stability and selectivity suggest its possible application in the detection of OTA, OTB and OTC in water samples.

Aflatoxin B1, ochratoxin A and zearalenone in sorghum grains marketed in Tunisia.

A total of 64 samples of sorghum (37 Tunisian sorghum samples and 27 Egyptian sorghum samples) were collected during 2011-2012 from markets in Tunisia. Samples were analysed for contamination with aflatoxin B1, ochratoxin A and zearalenone by High-Performance Liquid Chromatography Coupled with Fluorescence Detection (HPLC-FLD). Aflatoxin B1 was found in 38 samples in the range 0.03-31.7 µg kg-1. Ochratoxin A was detected in 24 samples with concentrations ranging from 1.04 to 27.8 µg kg-1. Zearalenone was detected in 21 samples and the concentration varied between 3.7 and 64.5 µg kg-1. ANOVA analysis of the influence of the country of origin on the incidence and concentration of mycotoxins in the samples studied showed no significant difference (P > 0.05) between the two batches of samples for each of the three mycotoxins studied. The studied mycotoxins contaminate sorghum and may also co-exist because of the diversity of the mycobiota in this cereal.

Analytical, thermodynamical and kinetic characteristics of photoluminescence immunosensor for the determination of Ochratoxin A.

Ochratoxin A (OTA) is one of the most widespread and dangerous food contaminants. Therefore, rapid, label-free and precise detection of low OTA concentrations requires novel sensing elements with advanced bio-analytical properties. In the present paper we report photoluminescence (PL) based immunosensor for the detection of OTA. During the development of immunosensor photoluminescent ZnO nanorods (ZnO-NRs) were deposited on glass substrate. Then the ZnO-NRs were silanized and covalently modified by Protein-A (Glass/ZnO-NRs/Protein-A). The latest structure was modified by antibodies against OTA (Anti-OTA) in order to form OTA-selective layer (Glass/ZnO-NRs/Protein-A/Anti-OTA). In order to improve immunosensors selectivity the surface of Glass/ZnO-NRs/Protein-A/Anti-OTA was additionally blocked by BSA. Formed Glass/ZnO-NRs/Protein-A/BSA&Anti-OTA structures were integrated within portable fiber optic detection system, what is important for the development of low cost and portable immunosensors. The immunosensor has been tested in a wide range of OTA concentrations from 10-4ng/ml until 20ng/ml. Interaction isotherms were derived from analytical signals of immunosensor. Association constant and Gibbs free energy for the interaction of Glass/ZnO-NRs/Protein-A/Anti-OTA with OTA were calculated, analyzed and compared with some other related results. Sensitivity range and limit of detection were determined as 0.1-1ng/ml and 10-2ng/ml, respectively. Interaction kinetics of ZnO-NRs with OTA was evaluated. Response time of the immunosensor toward OTA was in the range of 500-800s. Some insights related to the mechanism of PL-signal generation are proposed and discussed.

Visible-light driven label-free photoelectrochemical immunosensor based on TiO2/S-BiVO4@Ag2S nanocomposites for sensitive detection OTA.

A label-free photoelectrochemical (PEC) platform with high visible-light activity for quantitative detection of the ochratoxin A (OTA) was developed by assembly of Ag2S nanoparticles (NPs) sensitized on titanium dioxide/red blood cell-like shape bismuth vanadate (TiO2/S-BiVO4) electrode via layer-by-layer (LBL) strategy. In this protocol, ascorbic acid was used as an efficient electron donor for scavenging photo-generated holes and inhibiting light driven electron-hole pair recombination. TiO2 has good photoelectric activity and large surface area. The S-BiVO4 with porous structure surfaces can contribute to the high photocurrent intensity under visible-light irradiation. Moreover, the Ag2S NPs were in-situ growth on surfaces of thioglycolic acid modified S-BiVO4, which enhanced photocurrent response and further improved the photocurrent conversion efficiency. Under optimal conditions, the PEC immunosensor exhibited a wide linear concentration range from 5pgmL-1 to 750ngmL-1, with a low detection limit of 1.7pgmL-1 (S/N = 3) for OTA. Additionally, the designed immunosensor was performed with good stability, reproducibility and selectivity, thus opening up a new promising PEC platform for some other small molecules analysis.