Highly Sensitive Microarray Immunoassay for Multiple Mycotoxins on Engineered 3D Porous Silicon SERS Substrate with Silver Nanoparticle Magnetron Sputtering.

A high-throughput, rapid, and highly sensitive surface-enhanced Raman spectroscopy (SERS) microarray for screening multiple mycotoxins has been developed on a three-dimensional silver nanoparticle porous silicon (3D AgNP-Psi) SERS substrate, which was easy to be engineered by electrochemical etching and magnetron sputtering technology. The etching current density, etching waveform, and target material for magnetron sputtering have been investigated to obtain an optimal 3D SERS substrate. The optimized 3D AgNP-Psi SERS substrate showed an enhancement factor of 2.3 × 107 at 400 mA/cm2 constant current density etching for 20 s and Ag target magnetron sputtering for 200 nm thickness on the surface of Psi. The simulation electric field distribution showed the near-field enhancement can reach 3× higher than that of AuNPs. A protein microarray has been designed to screen multiple mycotoxins by AuNP Raman tags and a competitive immunoassay protocol on the surface of the 3D SERS substrate. The SERS protein microarray displayed wide linear detection ranges of 0.001-100 ng/mL for ochratoxin A, 0.01-100 ng/mL for aflatoxin B1, 0.001-10 ng/mL for deoxynivalenol, along with pg/mL low limit of detection, good recovery rates, repeatability, and reproducibility. The 3D SERS protein microarray is easily engineered and has a great potential application in medicine, environment, and food industry fields.

Tailoring the physicochemical properties of starch: impact of integrated ultrasonic and ethanol pretreatment on the oil uptake of infrared fried ginkgo seeds.

BACKGROUND: The structural changes of starch would have a more crucial impact on oil absorption and quality changes in starch-rich fruits and vegetables during frying process with enhanced heat transfer (such as infrared frying). In the present study, the influence of integrated ultrasonic and ethanol (US + ethanol) pretreatment on oil uptake in infrared fried (IF) ginkgo seeds was evaluated regarding modifications in the physicochemical properties of starch. The pretreatment was performed with ultrasonic (40 kHz, 300 W) and ethanol osmotic (95%, v/v) treatment individually or integrated for 40 min. RESULTS: The mass transfer in the pretreatment was facilitated by combined ultrasound and ethanol. The swelling power, solubility, and gelatinization degree of starch was significantly increased. Low-frequency-NMR curves and images revealed that the bound water fraction in ginkgo seeds was increased and the water distribution was homogenized. The results of Fourier transform-infrared spectrum and differential scanning calorimeter revealed that the crystalline regions of starch were reduced and the thermal enthalpy was decreased after US + ethanol pretreatment. The total, surface and structural oil content in IF ginkgo seeds with US + ethanol pretreatment was reduced by 29.10%, 34.52% and 29.73%, respectively. The US + ethanol pretreatment led to a thinner crust layer with increased porosity and smaller-sized pores in the IF ginkgo seeds as observed by stereo microscopy and scanning electron microscopy. CONCLUSION: The changes in structural and physicochemical properties of starch by combined ultrasound and ethanol affect the crust ratio and pore characteristics in fried high-starch fruits and vegetables, thereby reducing oil absorption. © 2024 Society of Chemical Industry.

Recent advances in physical fields-based frying techniques for enhanced efficiency and quality attributes.

Frying is one of the most common units in food processing and catering worldwide, which involves simultaneous physicochemical and structural changes. However, the problems of traditional frying technology, such as low thermal utilization and poor processing efficiency, have been gradually exposed to industrial production. In this paper, strategies of applying physical fields, such as pressure field, electromagnetic field, and acoustic field in frying technology separately or synergistically with improved efficiency and quality attributes are reviewed. The role of physical fields in the frying process was discussed with modifications in heat and mass transfer and porous structures. The effects of physical fields and their processing parameters on moisture loss kinetics, oil uptake, texture, color, and nutrients retention of fried food are introduced, respectively. Recent advances in multi-physical field-based frying techniques were recommended with synergistic benefits. Furthermore, the trends and challenges that could further develop the multi-physical field-based frying techniques are proposed, showing further commercial prospects for the purpose. The application of physical fields has brought new inspiration to the exploitation of efficient and high-qualified frying technologies, while higher technical levels and economic costs need to be taken into consideration.HighlightsThe role of physical fields in pretreatments and frying process were reviewed.The mechanism of physics fields on frying efficiency and quality was summarized.The physicochemical and microstructure changes by physics fields were discussed.The synergy of physical fields in frying technology were outlined.The trends for further multi-physical field-based frying techniques were proposed.

Synthesis and characterization of lotus seed protein-based curcumin microcapsules with enhanced solubility, stability, and sustained release.

BACKGROUND: Lotus seed protein (LSP) was extracted from lotus seed and used to encapsulate curcumin with or without complexing with pectin. The physicochemical properties of LSP-based microcapsules, including solubility, stability, and in vitro sustained release, were determined. The mechanism of interaction between curcumin, LSP, and pectin was revealed. RESULTS: The encapsulation efficiency of curcumin was found to depend on LSP concentration and was highest (86.32%, w/w) at 50 mg mL-1 . The curcumin in curcumin-LSP and curcumin-LSP-pectin powder particles achieved a solubility of 75.15% and 81.39%, respectively, which was a remarkable enhancement. The microencapsulation with LSP and LSP-pectin matrix showed a significant improvement in the antioxidant activity, photostability, thermostability, and storage stability of free curcumin. The microencapsulated curcumin showed sustained control release at the gastric stage and burst-type release in the subsequent intestinal stage, presenting cumulative release rates of 64.3% and 72.4% from curcumin-LSP and curcumin-LSP-pectin particles after gastrointestinal digestion. The LSP-pectin complex produced microcapsules with higher solubility, smaller particle size, enhanced physicochemical stability, and increased bioaccessibility. Fourier transform infrared, circular dichroism spectra, and differential scanning calorimetry data indicated that the encapsulated curcumin interacted with LSP and pectin mainly through hydrogen bonding, hydrophobic, and electrostatic interactions. CONCLUSION: This work shows that LSP can be an alternative encapsulant for the delivery of hydrophobic nutraceuticals with enhanced solubility, stability, and sustained release. The results may contribute to the design of novel food-grade delivery systems based on LSP vehicles, thereby broadening the applications of LSP in the fields of functional food. © 2021 Society of Chemical Industry.

Comparison of functional and structural properties of ginkgo seed protein dried by spray and freeze process.

The influences of spray-drying and freeze-drying processes on functional properties of ginkgo seed proteins (GSP) were systematically investigated. It was revealed that GSP dried by spray (SGSP) displays an significantly improved water holding capacity and superior emulsifying properties than the freezing-drying GSP (FGSP), whereas, the oil binding capacity is higher in FGSP. The difference in properties of SGSP and FGSP can be attributed to their different structural characteristics. Comparing with FGSP, SGSP was demonstrated having more disulfide bonds, more amorphous and less ordered structure, accounted for big differences in functional properties. With the outstanding functional characteristics, GSP could be potentially applied in oil-in-water type food system, such as milk and mayonnaise. Finally, it is important to choose the suitable drying method according to the requirements of the specific food system.

Antioxidant Blueberry Anthocyanins Induce Vasodilation via PI3K/Akt Signaling Pathway in High-Glucose-Induced Human Umbilical Vein Endothelial Cells.

Blueberries are rich in antioxidant anthocyanins. The hypotensive effects of blueberry anthocyanins in endothelial cells was investigated here. Pretreatment with blueberry anthocyanin extract, malvidin, malvidin-3-glucoside, and malvidin-3-galactoside significantly ameliorated high-glucose-induced damage by enhancing endogenous antioxidant superoxide dismutase (SOD) and heme oxygenase-1 (HO-1), lowering reactive oxygen species (ROS) generation and NADPH oxidase isoform 4 (NOX4) expression, and increasing the cell vitalities. They also effectively induced a vasodilatory effect by increasing the vasodilator nitric oxide (NO) and its promoters endothelial NO synthase (eNOS) and peroxisome proliferator-activated receptor-γ (PPARγ) levels as well as by decreasing the vasoconstrictor angiotensin-converting enzyme (ACE), xanthine oxidase-1 (XO-1), and low-density lipoprotein (LDL) levels. The activation of phosphoinositide 3-kinase (PI3K)/Akt signaling pathway and the breakdown of protein kinase C zeta (PKCζ) pathway were involved in the bioactivities. The results indicated blueberry anthocyanins protected endothelial function against high-glucose (HG) injury via antioxidant and vasodilatory mechanisms, which could be promising molecules as a hypotensive nutraceutical for diabetes patients.

A competitive aptamer chemiluminescence assay for ochratoxin A using a single silica photonic crystal microsphere.

We designed a competitive aptamer chemiluminescence assay for ochratoxin A (OTA) on the surface of a single silica photonic crystal microsphere (SPCM) in cereal samples. The structural color of SPCMs is used to recognize and trace the microspheres during process of detection. Anti-aptamer was immobilized on the surface of SPCM. OTA and anti-aptamer competed to bind to aptamer when OTA and its aptamer (labeled by biotin at 5'end) were added in the system. The chemiluminescence signal was developed by the horseradish peroxidase (HRP), luminol and H2O2. The molecules on the single SPCM can produce enough chemiluminescence signal intensity for quantitative detection for OTA. The linear detection range for OTA was from 1 pg/mL to 1 ng/mL and recovery rates were 89%-95%, 81%-92% and 94%-105% in rice, wheat and corn, respectively. The results showed that the developed method for OTA using a single SPCM has a great application potential in cereal samples.

High-Throughput Low-Background G-Quadruplex Aptamer Chemiluminescence Assay for Ochratoxin A Using a Single Photonic Crystal Microsphere.

We reported a novel hemin-G-quadruplex aptamer chemiluminescence assay platform for ochratoxin A (OTA) using the single silica photonic crystal microsphere (SPCM). The oligonucleotide A sequence containing aptamer sequences of hemin and OTA is immobilized on the surface of SPCM. The other oligonucleotide B sequence containing a partially complementary sequence with one part OTA aptamer and one part hemin aptamer is used as a blocking chain. The hybridization between chain A and chain B will be influenced by the presence or absence of OTA in the system, which will affect the bioactivity of DNAzyme. Thus, the chemiluminescence signal depends on the concentration of OTA in the samples. In the single particle assay platform, the signal/noise is remarkably enhanced, and the background signal can be ignored by separating hemin from the surface of SPCM. The limit of detection of the new method reaches to the pg/mL scale, and the linear detection range is 4 orders of magnitude for OTA. The new assay platform can provide a sensitive, cost-efficient, simple, and high-throughput screening for OTA.

Levels and formation of α-dicarbonyl compounds in beverages and the preventive effects of flavonoids.

ABSTRACT: Methylglyoxal (MGO) and glyoxal (GO), α-dicarbonyl compounds found in the Maillard reaction, progressively and irreversibly modify proteins. Beverages are an exogenous source of α-dicarbonyl compounds and may potentially increase MGO and GO levels in vivo. Using GC-FID method, we detected the MGO and GO contents of 86 beverages in Chinese supermarkets. The highest MGO and GO 587.5 µg/100 mL and 716.7 µg/100 mL respectively found in soyamilk and coffee. Herbal beverages, which contained bioactive components, had lower average levels of MGO (48.1 µg/100 mL) and GO (25.9 µg/100 mL). A box-and-whisker plot was used to display variation of the same group drinks, and comparing distributions between six different groups. It was further discovered that fat, protein and flavonoids, in addition to sweeteners, had notable effects on the formation of MGO and GO in soybean milk. The result of LC/MS indicated that quercetin could prevent the formation of MGO by trapping MGO to form the mono-MGO and di-MGO adducts during soybean milk manufacturing.

Multiplex chemiluminescent immunoassay for screening of mycotoxins using photonic crystal microsphere suspension array.

A novel multiplex chemiluminescent mycotoxin immunoassay suspension array system was developed by combining the silica photonic crystal microspheres (SPCMs) encoding technique and a chemiluminescent immunoassay (CLIA) method. The SPCMs were used as a carrier of the suspension array and encoded by their reflectance peak positions, which overcome fluorescence photobleaching, and the potential interference between the encoding fluorescence and detection fluorescence. Aflatoxin B1 (AFB1), fumonisin B1 (FB1) and ochratoxin A (OTA) artificial antigens were immobilized on the surfaces of SPCMs by using 3-glycidoxypropyltrimethoxysilane as a linker. Horseradish peroxidase (HRP) was used as a labeling enzyme for the secondary antibody in the enzyme-catalyze H2O2-luminol chemiluminescence system. The CLIA detection system was easily integrated with a multifunctional microplate reader and displayed a two to three orders of magnitude dynamic linear detection range from 0.001 to 1, 0.001 to 1, and 0.01 to 1 ng mL(-1) for AFB1, FB1 and OTA with 50% inhibitory concentrations (IC50) of 0.01, 0.036, and 0.04 ng mL(-1), respectively. The recovery rates are in the range of 63.5 to 121.6% for the three mycotoxins in three kinds of spiked cereal samples. The results of detection in 12 naturally contaminated cereal samples were consistent with that of the classic enzyme-linked immunosorbent assay (ELISA) method. This proposed system is simple, rapid, low cost and high throughput for multiplex mycotoxin assay.

High sensitive immunoassay for multiplex mycotoxin detection with photonic crystal microsphere suspension array.

A novel, sensitive, and high throughput competitive immunoassay for multiplex mycotoxins was established by immobilizing the artificial antigens (Ags) of mycotoxins on the surfaces of three kinds of silica photonic crystal microsphere (SPCM) suspension arrays. The SPCMs were encoded by their reflectance peak positions. Aflatoxin B1 (AFB1), fumonisin B1 (FB1), and citrinin (CIT) spiked in the cereals were extracted, and the fluorescein isothiocyanate (FITC) labeled antibodies (Abs) of these mycotoxins were added into the centrifuge tube which contained the SPCMs of the modified artificial antigens (Ags). The fluorescence signal was collected by an array fluorescent scanner. The limit of detection (LOD) was as low as 0.5, 1, and 0.8 pg/mL for AFB1, FB1, and CIT, respectively. The new method provided a wide linear detection range from 0.001 to 10, 0.001 to 10, and 0.001 to 1 ng/mL for AFB1, FB1, and CIT, respectively. The mean recovery rates are in range of 74.7 ± 4.0% to 127.9 ± 4.4% for the three mycotoxins in corn, peanuts, and wheat. The developed method for mycotoxins was used to assay the AFB1, FB1, and CIT level in 10 naturally contaminated cereal samples, and the results of detection were in agreement with that of a classic enzyme-linked immunosorbent assay (ELISA) method. This method saves a large amount of reagents (10 µL volume) and detection time (<3 h) for multiplex mycotoxin assay.

Preparation and characterization of octenyl succinic anhydride-modified ginkgo seed starch with enhanced physicochemical and emulsifying properties.

A quick preparation of octenylsuccinylated (OS)-ginkgo seed starch was proposed by lipase-coupling esterification within 30 min, and the physicochemical and emulsifying properties of OS-ginkgo seed starch were evaluated. High-performance liquid chromatography results revealed that ginkgolic acid in ginkgo seed starch was too low to be detected, which improved the biosafety and application range of OS-ginkgo seed starch. The degree of substitution (DS) of OS-ginkgo starch varied from 0.006 to 0.0169 depending on the lipase concentration increased from 0% to 1% (w/w, based on the volume of starch solution), and the reaction efficiency obtained the highest value of 68.5% at the lipase concentration of 1%. Fourier transform infrared spectra of OS-ginkgo seed starch confirmed ester carbonyl splicing in the starch molecular with the characteristic peaks at 1722 and 1567 cm-1 . Scanning electron microscopy observations revealed that the esterification occurred mainly in the amorphous regions with slight morphological modification. X-ray diffractions suggested that no crystal change occurred on the starch granule. The thermal analysis revealed that OS-ginkgo seed starch showed a lower temperature and endothermic enthalpy for gelatinization, and presented enhanced and DS-dependent emulsifying properties and in vitro antidigestion properties. PRACTICAL APPLICATION: Results indicated that OS-ginkgo seed starch prepared by lipase-coupling esterification would be an alternative emulsion stabilizer for encapsulation and delivery of hydrophobic components. This study would provide an alternative method for the efficient and economical production of OS-ginkgo seed starch, thereby broadening its application in commercial exploitation.

Evaluation of the infrared frying on the physicochemical properties of fried apple slices and the deterioration of oil.

The influence of infrared frying (IF) on the physicochemical properties of fried apple slices and the oil deterioration was investigated, considering conventional frying (CF) as a reference. IF had a more favorable impact on the heating rate and thermal efficiency, which subsequently resulted an accelerated moisture removal rate. The oil uptake in infrared-fried apple slices were reduced by 12.9%-17.3%, when compared to the CF, as attributed to the denser and smoother morphological microstructure. The color of apple slices was better preserved in IF and the total phenolic and flavonoid contents had a higher retention rate with the optimal infrared power (2000 W in this study). Additionally, infrared frying was proved to be a promising technology to slow down the oil deterioration rate as was observed from lower values of acid value, and carbonyl value, which was also supported by the results of gas chromatography, FT-IR, and LF-NMR analysis.

Surface modified porous silicon with chitosan coating as a pH-responsive controlled delivery system for lutein.

Surface modified pH-responsive porous silicon (PSi) carriers were developed for efficient delivery of lutein. PSi particles were prepared by the electrochemical etching method and modified with two chemical groups: hydroxyl and octadecyl silane, respectively. Chitosan (CS) was used for coating of PSi to ensure pH-responsive release. The loading conditions, release properties, cytotoxicity and toxicity were investigated. The highest loading percentage of lutein could be obtained with oxidized PSi and the structure of the microparticles was characterized by Fourier transform-infrared spectroscopy. The surface area and pore size of the microparticles were obtained from the N2 adsorption-desorption isotherm. The CS-PSi-Lut microparticles showed the minimum surface area of 220.30 m2 g-1 and a relatively larger average pore width of 179.00 Å. In vitro release experiments showed a pH-responsive and controlled release of lutein, with the fastest release rate and highest cumulative release rate of 97% under acidic conditions (pH 5.0) within 7 h. PSi, chitosan and lutein showed synergistic toxic effects, and the CS-PSi-Lut microparticles could effectively inhibit the proliferation of HT-29 cells in a dose-dependent manner, with an inhibition rate of 77% when the lutein concentration reached 40 µg mL-1. The in vivo toxicological evaluation of CS-PSi-Lut microparticles indicated good biocompatibility in the range of experimental doses. The chitosan-coated oxidized PSi capable of delivering bioactive compounds in a targeted and controlled manner provides a novel platform for the development and application of lutein.

Highly Sensitive Simultaneous Detection of Multiple Mycotoxins Using a Protein Microarray on a TiO2-Modified Porous Silicon Surface.

A new protein microarray method for multiplex mycotoxin detection in parallel has been established on a stable TiO2-modified porous silicon (PSi) surface. A typical competitive immunoassay microarray protocol has been developed for simultaneous detection of multiplex mycotoxins including aflatoxin B1 (AFB1), ochratoxin A (OTA), and fumonisin B1 (FB1) on the TiO2-PSi surface. The epoxy groups were selected to modify the surface of a TiO2-PSi wafer for the immobilization of artificial antigens of mycotoxins because of their high signal-to-noise ratios. Under optimal conditions, the developed method showed wide linear detection ranges of 0.01-1 ng/mL for OTA, 0.001-1 ng/mL for AFB1, and 0.01-1 ng/mL for FB1 and low limit of detections (LODs) of 0.433 ng/mL for OTA, 0.243 ng/mL for AFB1, and 0.093 ng/mL for FB1. The microarray method can specifically identify the three mycotoxins and their analogues. The recovery rates in real samples were within 75-120%, which were in agreement with that of the classical ELISA method. The new method has great application potential for rapid, sensitive, and high-throughput screening of multiplex mycotoxins and other target molecules.

Three-dimensional ordered macroporous magnetic photonic crystal microspheres for enrichment and detection of mycotoxins (I): Droplet-based microfluidic self-assembly synthesis.

Ordered porous materials are attracting enormous attention due to their uniform pore structures, particularly the magnetic photonic crystal microspheres (PCMs) which not only possess unique photonic crystal structure but also can achieve separation easily based on magnet. Here, a two-phase microfluidic self-assembly synthetic system was established simply and employed for the preparation of three dimensional PCMs (3DPCMs) by using the emulsion droplet approach. One phase (dispersed phase) was an aqueous emulsion containing Fe3O4, silica (SiO2) and polystyrene (PS) nanoparticles; another phase (continuous phase) was pure silicone oil. The droplets were formed by introducing the dispersed phase into the continuous phase through a tee valve. By heating the droplets, the water would evaporate and the nanoparticles would finally assemble into solid microspheres, which could be changed into macroporous 3DPCMs after removal of the PS nanoparticles by calcination. The contents and particle sizes of Fe3O4, SiO2 and PS nanoparticles in the dispersed phase were investigated in detail and optimized to prepare macroporous magnetic 3DPCMs with high quality. The morphologies, surface crystal structure, magnetic property, particle size distribution, specific surface area and pore size of the macroporous magnetic 3DPCMs were characterized. The expected 3DPCM displayed regular and uniform photonic crystal structure, narrow particle size distribution and strong magnetic property. The macroporous magnetic 3DPCMs grafted with vomitoxin (DON)-antibodies could be applied for selective enrichment of DON in real samples.

Scavenging of Acrolein by Food-Grade Antioxidant Propyl Gallate in a Model Reaction System and Cakes.

Reactive carbonyl species (RCS), such as acrolein (ACR), glyoxal (GO), and methylglyoxal (MGO), have received extensive attention recently as a result of their high activity and toxicity in vitro and in vivo. In the present study, propyl gallate (PG), a common food antioxidant, was found to effectively trap more ACR than butylated hydroxytoluene and butylated hydroxyanisole through the formation of mono-ACR adducts (PG-ACR) and di-ACR adducts (PG-2ACR). The two adducts were successfully purified, and their structures were elucidated on the basis of their high-resolution mass spectrometry and 1H, 13C, and two-dimensional nuclear magnetic resonance data. We further identified that PG-ACR had the ability to continue to trap GO and MGO to form PG-ACR-GO and PG-ACR-MGO, respectively, by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Furthermore, we verified that PG could inhibit the production of ACR, GO, and MGO via trapping these RCS simultaneously to form the corresponding adducts in pound cakes using LC-MS/MS.

Oral Curcumin via Hydrophobic Porous Silicon Carrier: Preparation, Characterization, and Toxicological Evaluation In Vivo.

Curcumin has antioxidant, anti-inflammatory, antimicrobial, and anticarcinogenic activities. However, the clinical application of curcumin has been restricted by the poor water solubility and low bioavailability of this molecule. In this work, hydrophobic porous silicon (pSi) particles were prepared by electrochemical etching method and grafted with the different hydrophobic groups on their surfaces. The loading efficiency of curcumin in pSi has been investigated. The properties of pSi particles have been characterized by scanning electron microscopy (SEM) and Fourier transform-infrared spectroscopy (FTIR). The highest loading efficiency of curcumin can be obtained with pSi surface modified with the octadecyl silane group. The release properties of curcumin in hydrophobic pSi have been researched in vitro and in vivo. The curcumin in the hydrophobic pSi surface keeps a high antioxidant bioactivity. The toxicological evaluation of the hydrophobic pSi particles indicates they have a high in vivo biocompatibility within the observed dose ranges. The hydrophobic pSi particles could provide an effective and controlled release delivery carrier for curcumin, which may provide a new tool platform for the further development of curcumin.

Porous Silicon Carrier Delivery System for Curcumin: Preparation, Characterization, and Cytotoxicity in Vitro.

A novel carrier delivery system for curcumin based on porous silicon (pSi) has been developed. The pSi film was prepared by electrochemical etching method and the microparticles of pSi were obtained by ultrasonication. The pSi film and particles of pSi were characterized by scanning electron microscopy (SEM). Sodium nitrite can induce curcumin into pSi and improve the drug loading (DL) and encapsulation efficiency (EE) of curcumin in double-distilled water loading buffer solution. Curcumin on the pSi surface was confirmed by Fourier transform infrared spectroscopy (FTIR) and UV-spectroscopy. The optimal loading conditions of curcumin in pSi are investigated. The curcumin in pSi keeps more than 95% bioactivity for 3 h and good repeatability. The cumulative release ratio of curcumin from PSi can reach 35% after 10 h. The in vitro cytotoxicity of curcumin loaded pSi was evaluated with HT-29 and NCM460 cell lines. The pSi delivery carrier can provide a controlled release and efficacy system for curcumin.

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).