Dual-Modal Immunochromatographic Test for Sensitive Detection of Zearalenone in Food Samples Based On Biosynthetic Staphylococcus aureus-Mediated Polymer Dot Nanocomposites.

The rapid detection of toxins is of great significance to food security and human health. In this work, a dual-modality immunochromatographic test (DICT) mediated by Staphylococcus aureus (SA)-biosynthesized polymer dots (SABPDs) was constructed for sensitive monitoring of zearalenone (ZEN) in agro products. The SABPDs as potent microorganism nanoscaffolds with excellent solubility, brightness, and stability were ingeniously fabricated employing hydroquinone and SA as precursors in the Schiff base reaction and a self-assembly technique. Thanks to the fact that they not only preserved an intact microsphere for loading Fc regions of monoclonal antibodies (mAbs) and the affinity of their labeled mAbs to antigen but also generated superb colorimetric-fluorescent dual signals, the versatile SABPDs manifested unique possibilities as the new carriers for dual-readout ICT with remarkable enhancement in sensitivity in ZEN screening (limit of detection = 0.036 ng/mL, which was 31-fold lower than that of traditional gold nanoparticle-based ICT). Ultimately, the proposed immunosensor performed well in millet and corn samples with satisfactory recoveries, demonstrating its potential for point-of-care testing. This work offers a bio-friendly strategy for biosynthesizing cell-based PD vehicles with bimodal signals for food safety analysis.

Single-crystal ordered macroporous metal-organic framework as support for molecularly imprinted polymers and their integration in membrane formant for the specific recognition of zearalenone.

Zearalenone is a fungal contaminant that is widely present in grains. Here, a novel molecularly imprinted membrane based on SOM-ZIF-8 was developed for the rapid and highly selective identification of zearalenone in grain samples. The molecularly imprinted membrane was prepared using polyvinylidene fluoride, cyclododecyl 2,4-dihydroxybenzoate as a template and SOM-ZIF-8 as a carrier. The factors influencing the extraction of zearalenone using this membrane, including the solution pH, extraction time, elution solvent, elution time, and elution volume, were studied in detail. The optimized conditions were 5 mL of sample solution at pH 6, extraction time of 45 min, 4 mL of acetonitrile:methanol = 9:1 as elution solvent, and elution time of 20 min. This method displayed a good linear range of 12-120 ng/g (R2  = 0.998) with the limits of detection and quantification of this method are 1.7 and 5.5 ng/g, respectively. In addition, the membrane was used to selectively identify zearalenone in grain samples with percent recoveries ranging from 87.9 to 101.0% and relative standard deviation of less than 6.6%. Overall, this study presents a simple and effective chromatographic pretreatment method for detecting zearalenone in food samples.

Preparation of magnetic molecularly imprinted polymers for the identification of zearalenone in grains.

This study was based on the specific binding ability of magnetic molecularly imprinted polymers (MMIPs) combined with a high-performance liquid chromatography-fluorescence detector (HPLC-FLD) for the rapid determination of zearalenone (ZEN) in cereals. A novel magnetic molecularly imprinted polymer was prepared by surface imprinting technology. Warfarin was used as a virtual template, 3-aminopropyl triethoxysilane (APTES) was used as the functional monomer, and tetraethyl orthosilicate (TEOS) was used as the cross-linking agent. Analysis by a vibrating sample magnetometer (VSM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) showed that MMIPs were prepared with a particle size about 450 nm, the imprinted molecular layer accounting for 10.7% of the total mass, and saturation magnetization of about 34.54 emu/g. The maximum adsorption capacity (Qmax) of the thermodynamic and kinetic adsorption experiments were 13.90 mg/g and 8.71 mg/g, respectively. The Langmuir model showed that the binding sites were uniformly distributed on the surface of the MMIPs. The Scatchard analysis showed that MMIPs had two types of binding sites with Qmax of 8.22 mg/g and 15.37 mg/g, respectively. In actual sample detection, the limit of detection (LOD) and limit of quantification (LOQ) were 0.4 ng/kg and 0.9 ng/kg, respectively. The sample recovery rate was 90.56-99.96%, the daytime stability was 1.35-2.87%. These results showed that MMIPs had good performance in selectively identifying ZEN and were suitable for determining ZEN in cereals.

Three kinds of lateral flow immunochromatographic assays based on the use of nanoparticle labels for fluorometric determination of zearalenone.

Colloidal gold, quantum dots and polystyrene microspheres were used as labels in three kinds of lateral flow immunochromatographic assays (ICAs) for the detection of zearalenone (ZEN) in cereal samples. The assays allow ZEN to be quantified within 20 min. The LODs are 10 µg·L-1 of ZEN for the colloidal gold-based ICA, and 1 µg·L-1 for both the quantum dot and polystyrene microsphere based ICAs. The respective data are 60 µg·kg-1, 6 µg·kg-1 and 6 µg·kg-1, respectively, for spiked samples and cereals. Only minor cross-sensitivity occurred between ZEN and fusarium toxins, and no cross-sensitivity if found for aflatoxin B1, T-2 mycotoxin, ochratoxin A, deoxynivalenol, and fumonisin B1. LODs of the three assays are lower than the maximum limits of ZEN set by most standardization agencies. Graphical abstract Schematic presentation of three lateral flow immunochromatographic assays (ICAs) based on the use of (a) colloidal gold (CG), (b) fluorescent quantum dots (QD), and

Development of a microfluidic device to enrich and detect zearalenone in food using quantum dot-embedded molecularly imprinted polymers.

Mycotoxins are secondary metabolites of certain moulds, prevalent in 60-80% of food crops and many processed products but challenging to eliminate. Consuming mycotoxin-contaminated food and feed can lead to various adverse effects on humans and livestock. Therefore, testing mycotoxin residue levels is critical to ensure food safety. Gold standard analytical methods rely on liquid chromatography coupled with optical detectors or mass spectrometers, which are high-cost with limited capacity. This study reported the successful development of a microfluidic "lab-on-a-chip" device to enrich and detect zearalenone in food samples based on the fluorescence quenching effect of quantum dots and selective affinity of molecularly imprinted polymers (MIPs). The dummy template and functional polymer were synthesized and characterized, and the detailed microfluidic chip design and optimization of the flow conditions in the enrichment module were discussed. The device achieved an enrichment factor of 9.6 (±0.5) in 10 min to quantify zearalenone spiked in food with high recoveries (91-105%) at 1-10 mg kg-1, covering the concerned residue levels in the regulations. Each sample-to-answer test took only 20 min, involving 3 min of manual operation and no advanced equipment. This microfluidic device was mostly reusable, with a replaceable detection module compatible with fluorescence measurement using a handheld fluorometer. To our best knowledge, the reported device was the first application of an MIP-based microfluidic sensor for detecting mycotoxin in real food samples, providing a novel, rapid, portable, and cost-effective tool for monitoring mycotoxin contamination for food safety and security.

Preparation of immunomagnetic composite nanostructures with bifunctional four-arm PEG derivatives as linkers for the ultrafast enrichment of zearalenone and its metabolites.

It is challenging to prepare sample pretreatment materials with simple use, strong selectivity and satisfactory enrichment performance. In this study, the antibody (3D4) that can specifically recognize zearalenone (ZEN) and its metabolites was immobilized on the surface of gold-coated magnetic Fe3O4 nanoparticles (GMN) by streptavidin (SA)-biotin interaction using GMN as the substrate and our designed four-arm PEG derivative (HS-4ARMPEG10K-(CM)3) as the linker. The immunomagnetic nanoparticles (GMN-4ARMPEG10K-SA-3D4) prepared by this strategy can achieve rapid enrichment (only 5 min) of analytes directly in the matrix, and higher enrichment capacity compared with the previous immunomagnetic particles. The sensitive and accurate analysis of ZEN and its metabolites can be achieved coupled with HPLC-MS/MS. The LODs and LOQs were 0.02-0.05 µg/kg and 0.05-0.10 µg/kg, respectively. The recoveries were 84.13%-112.67%, and the RSDs were 1.09%-9.39%. The method can provide a powerful tool for highly sensitive and rapid monitoring of mycotoxins in complex matrices due to its' strong selectivity and resistance to matrix interference.

Quantum dot loaded liposomes as fluorescent labels for immunoassay.

Liposomes loaded with water-soluble and water-insoluble quantum dots (QD) were for the first time applied as labels in different heterogeneous immunoassays for the determination of food contaminants, using mycotoxin zearalenone (ZEN) as a model. A great deal of work was devoted to the optimal choice of phospholipids for the liposomes preparation and to the factors which are important for the stability and size of obtained liposomes. Thin-film hydration and reverse-phase evaporation techniques were evaluated in terms of stability of the obtained liposomes and their efficiency for QD loading. Conjugation of liposomes with proteins and the influence of cross-linkers to the nonspecific interaction of the obtained liposomes with the surface of microtiter plates and cartridges were investigated and 3-(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester was found as the optimal cross-linker. The limits of detection (LOD) for ZEN of fluorescence-labeled immunosorbent assays were 0.6 µg kg(-1), 0.08 µg kg(-1), and 0.02 µg kg(-1), using QD, liposomes loaded with water-soluble QD, and water-insoluble QD, respectively. Similarly, the developed qualitative on-site tests using the different QD labels and taking into account the EU maximum residues level for ZEN in unprocessed cereals showed cutoff levels of 100, 50, and 20 µg kg(-1).

Simultaneous determination of zearalenone and alternariol mycotoxins in oil samples using mixed molecularly imprinted polymer beads.

This work reports the optimization of a method using Molecularly Imprinted Polymers (MIPs) for the simultaneous determination of zearalenone and alternariol mycotoxins. The method was optimized using a chemometric approach where in the optimized conditions, the cartridges with a mixture (50:50, w/w) of both MIPs, were loaded with 30 mL of sample, washed with 2 mL of ACN/water (20/80, v/v) and eluted with 2.5 mL of trifluoroacetic acid/MeOH (3/97, v/v). The extracts were analyzed by HPLC coupled to a fluorescence detector (FLD). The optimized method has been applied and validated to the analysis of the mycotoxins in maize, sunflower and olive oils samples with a limit of detection of 5 and 2 µg kg-1, respectively. Recoveries were in the range of 94 % to 108 % (RSD < 6 %) for zearalenone and 92 % to 113 % (RSD < 5 %) for alternariol. The results were confirmed by HPLC-MS/MS.

How different microfilters affect the recovery of eleven EU-regulated mycotoxins.

Microfiltration is a common step in liquid chromatography - tandem mass spectrometry (LC-MS), a method of choice in determining several mycotoxins in a solution at once. However, microfiltration may entail filter-analyte interactions that can affect the accuracy of the procedure, and underestimate exposure. The aim of our study was to assess how five different membrane materials for syringe filters (nylon, polytetrafluoroethylene, polyethersulphone, mixed cellulose ester, and cellulose acetate) affect microfiltration and recovery of EU-regulated mycotoxins, including aflatoxins B1, B2, G1, and G2, deoxynivalenol, fumonisins B1 and B2, zearalenone, T-2 and HT-2 toxins, and ochratoxin A. Polytetrafluoroethylene filters turned out to least affect microfiltration through mycotoxin loss, followed by more commonly used nylon filters, whereas the remaining three filter membrane materials had such a negative effect on recoveries that we found them incompatible with the procedure. Our findings clearly suggest that it is important to select a proper filter type that suits analyte properties and solution composition and to discard the first few filtrate drops to ensure the accuracy of the analytical procedure.

How does multiannual plastic mulching in strawberry cultivation influence soil fungi and mycotoxin occurrence in soil?

The production of mycotoxins is often interpreted as fungal response to cope with unfavorable growth conditions induced by toxic substances, environmental and biological factors. Soil covers influence soil environment, which consequently can change the abundance and composition of microbial communities. We investigated how plastic coverage (PC) influence soil fungi and mycotoxin occurrence (deoxynivalenol, nivalenol and zearalenone) compared to the traditional straw coverage (SC) in dependence of soil depth and time in a 3-year field experiment in strawberry cultivation. In total, 300 soil samples, resulting from two treatments, three soil layers, and ten sampling dates (n = 5), were analyzed for mycotoxins and ergosterol (proxy for soil fungal biomass) with liquid chromatography high resolution mass spectrometry and high-performance liquid chromatography with UV-detection, respectively. The modified microclimate under PC had no significant influence on fungal biomass, whereas SC promoted fungal biomass in the topsoil due to C-input. Mycotoxins were detected under both cover types in concentrations between 0.3 and 21.8 µg kg-1, mainly during strawberry establishment period and after fungicide application. Deoxynivalenol had the highest detection frequency with 26.3% (nivalenol: 8.3%, zearalenone: 8.7%). This study confirmed the in situ production of mycotoxins in soil, which seems mainly triggered by field treatment (fungicide application) and plant growth stage (establishment period) rather than on mulching type. Further investigations are necessary to better understand the influence of different agricultural practices and soil types on the production and fate of mycotoxins.

Application of magnetic hydroxyapatite surface-imprinted polymers in pretreatment for detection of zearalenone in cereal samples.

This study is about the combination of magnetic solid phase extraction (MSPE) and high performance liquid chromatography-fluorescence detector (HPLC-FLD) for the pre-concentration and determination of Zearalenone (ZEA) in grain sample extracts. The novel sorbent (Fe3O4-HAP@MIPs), for selective and intelligent extraction of ZEA, was synthesized by doping Fe3O4 into the fibrous structure of hydroxyapatite nanoparticle (Fe3O4-HAP) and subsequently wrapping with molecularly imprinted polymers. The characteristic and morphology of magnetic particles were studied by infrared spectroscopy, vibrating sample magnetometer and scanning electron microscopy. The maximum adsorption capacity was 2.89 µg/mg. It could reach the adsorption equilibrium within 5 min. The adsorption isotherm of ZEA by the Fe3O4-HAP@MIPs were simulated. The results showed that the extraction process of ZEA with the sorbent accorded with Langmuir isotherm. The important factors affecting the extraction efficiency include elution solvent, washing solvent and the volume of them. After a serious of experiments, the optimum conditions were as follows: the volume of elution solvent was 4 mL of methanol and the washing solvent was acetonitrile-water 2:8（v/v). The calibration curve for ZEA was linear in the range of 10.00-300.00 µg/kg. The limit of detection and limit of quantitative was 2.00 µg/kg and 6.65 µg/kg, respectively. This method could provide a good reusability of 8 times and enough recoveries at three spiked levels (3, 5 and 8 ng/mL) ranging between 61.97% and 95.15% with the relative standard deviations of 1.94%∼7.44%. These results demonstrated that Fe3O4-HAP@MIPs could be used for separation, concentration and detection of ZEA from real samples.

Rainbow latex microspheres lateral flow immunoassay with smartphone-based device for simultaneous detection of three mycotoxins in cereals.

The coexistent pollution of multiple mycotoxins displays a synergistic toxicity effect that significantly threatens human health. Therefore, it is essential to establish a rapid detection method for multi-mycotoxins in food. In this study, red, green, and blue latex microspheres (LMs) were applied as the aflatoxin B1 (AFB1), T-2 toxins (T-2), and zearalenone (ZEN) antibodies labeled tracer, respectively. Based on the principle of spatial resolution, a rainbow "traffic light" pattern latex microspheres lateral flow immunoassay (LMs-LFIA) integrated with a portable and user-friendly smartphone-based device was first developed to detect three kinds of mycotoxins simultaneously. The cut-off values of the method for AFB1/T-2/ZEN in cereals were 1/15/40 µg kg-1, the limits of detection were 0.04/0.40/1.21 µg kg-1, respectively. The recoveries ranged from 82.1% to 107.5%, with the coefficient of variation from 3.0% to 8.1%. A parallel analysis in 26 naturally contaminated cereal samples was confirmed by commercial ELISA kits; the results showed a good correlation (R2＞0.99), indicating the practical reliability of the rainbow LMs-LFIA. This method provided a visually enjoyable, portable, and sensitive detection mode for multi-target detection of mycotoxins or other small molecule hazard factors in food.

Determination of zearalenone and its metabolites in endometrial cancer by coupled separation techniques.

This study presents a selective method of isolation of zearalenone (ZON) and its metabolite, α-zearalenol (α-ZOL), in neoplastically changed human tissue by accelerated solvent and ultrasonic extractions using a mixture of acetonitrile/water (84/16% v/v) as the extraction solvent. Extraction effectiveness was determined through the selection of parameters (composition of the solvent mixture, temperature, pressure, number of cycles) with tissue contamination at the level of nanograms per gram. The produced acetonitrile/water extracts were purified, and analytes were enriched in columns packed with homemade molecularly imprinted polymers. Purified extracts were determined by liquid chromatography (LC) coupled with different detection systems (diode array detection--DAD and mass spectrometry--MS) involving the Ascentis RP-Amide as a stationary phase and gradient elution. The combination of UE-MISPE-LC (ultrasonic extraction--molecularly imprinted solid-phase extraction--liquid chromatography) produced high (R≈95-98%) and repeatable (RSD<3%) recovery values for ZON and α-ZOL.

Magnetic hyperbranched molecularly imprinted polymers for selective enrichment and determination of zearalenone in wheat proceeded by HPLC-DAD analysis.

A novel magnetic surface molecular imprinted polymers with 2, 4, 6-trisacrylamido-3, 5-triazine (TAT) as a functional monomer was successfully synthesized and used for the enrichment and determination of zearalenone. The molecular imprinting is reported herein at first time for application of zearalenone in wheat. The magnetic imprinted materials possessed excellent magnetism and uniform appearance, which were characterized by fourier transform infared spectroscopy and transmission electron microscope. The results proved the magnetic molecular imprinted polymers was successfully prepared. The magnetic molecular imprinted polymers exhibited satisfactory sensitivity, stability and potential reusability. The binding affinity was investigated by selectivity experiment, which possessed high selectivity. To obtain the optimal application conditions, the amount of adsorption, extraction time, elution solvent and time were optimized. The limited detection of zearalenone was 0.55 ng g-1 and the recoveries of zearalenone were 92.1-96.0%. The relative standard deviation was lower than 5.4%. This indicated that a simple, efficient and low-cost method was established and successfully applied in spiked wheat sample.

Lateral Flow Immunoassay Based on Polydopamine-Coated Gold Nanoparticles for the Sensitive Detection of Zearalenone in Maize.

In this work, polydopamine-coated gold nanoparticles (Au@PDAs) were synthesized by the oxidative self-polymerization of dopamine (DA) on the surface of AuNPs and applied for the first time as a signal-amplification label in lateral flow immunoassays (LFIAs) for the sensitive detection of zearalenone (ZEN) in maize. The PDA layer functioned as a linker between AuNPs and anti-ZEN monoclonal antibody (mAb) to form a probe (Au@PDA-mAb). Compared with AuNPs, Au@PDA had excellent color intensity, colloidal stability, and mAb coupling efficiency. The limit of detection of the Au@PDA-based LFIA (Au@PDA-LFIA) was 7.4 pg/mL, which was 10-fold lower than that of the traditional AuNP-based LFIA (AuNP-LFIA) (76.1 pg/mL). The recoveries of Au@PDA-LFIA were 93.80-111.82%, with the coefficient of variation of 1.08-9.04%. In addition, the reliability of Au@PDA-LFIA was further confirmed by the high-performance liquid chromatography method. Overall, our study showed that PDA coating can chemically modify the surface of AuNPs through a simple method and can thus significantly improve the detection sensitivity of LFIA.

Preparative isolation and purification of zearalenone from rice culture by combined use of macroporous resin column and high-speed counter-current chromatography.

Zearalenone is one of the most harmful mycotoxins found in grains and there is a large demand for zearalenone substrate for research purposes. A new separation method was developed for the preparative purification of zearalenone from rice culture of Fusarium graminearum by utilizing macroporous resin column combined with high-speed counter-current chromatography. Zearalenone was adsorpted on XAD-2 resin at 25 °C, neutral pH and a feed flow of 4 BV/h, followed by dynamic desorption by 60% ethanol solution. Further purification was achieved by high-speed counter-current chromatography using an optimized biphasic solvent system. A total of 267 mg of zearalenone crystal was obtained in one single run from 4.2 g of crude extract. The purity of the final product was 98.9% and the total recovery yield of zearalenone in this study was 73.9%. This dual-step purification procedure provided an effective way to obtain the costly mycotoxin for both toxicological and detoxification studies on zearalenone.

A Novel Magnetic Molecular Imprinted Polymer for Selective Extraction of Zearalenone from Cereal Flours before Liquid Chromatography-Tandem Mass Spectrometry Determination.

Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by various Fusarium species and commonly occurring in corn and other cereals. Even though its acute toxicity is low, still the estrogenic activity of ZEN and metabolites is a matter of concern. In this work, a new magnetic molecularly imprinted polymer (mMIP) for the selective extraction of ZEN from cereal flours is presented. The mMIP was synthesized previously using quercetin as dummy template, and here we wanted to test its applicability to complex food samples. Analyte determination was carried out by high-performance liquid chromatography coupled to tandem mass spectrometry. The selectivity of the mMIP and the main validation method parameters were assessed. In particular, even in samples as complex as cereals, matrix effect was negligible. Although the mMIP showed cross-selectivity towards both ZEN-related and quercetin-related compounds, nonetheless ZEN recovery was > 95% for the two lower spiking levels, and the quantification limit was 0.14 ng g-1, i.e., ca. 500 times lower than the maximum limit fixed for most cereals by European law. Therefore, the material, also in comparison with a commercial sorbent, appears suitable for the application in food analysis, also to isolate ZEN at trace levels.

Fabrication of pioneering 3D sakura-shaped metal-organic coordination polymers Cu@L-Glu phenomenal for signal amplification in highly sensitive detection of zearalenone.

Low molecular weight pollutants from foods have aroused global attention due to their toxicity after long-time exposure. There is an increased demand for appropriate methods to detect these pollutants in foods. In this study, a brand-new type of nano metal-organic coordination polymers (MOCPs) nanocarriers (3D sakura-shaped copper (II) ions@L-glutamic acid (L-Glu)) has been first synthesized. We herein demonstrate a facile chelated method that allows the combination of copper (II) ions and L-Glu. A series of controlled experiments have revealed that the reaction time and the ratio of reactants played the crucial roles in affecting the morphology of the final product. 3D sakura-shaped Cu@L-Glu combined with palladium-platinum nanoparticle (Pd-PtNPs) to obtain Cu@L-Glu/Pd-PtNPs acting as the signal tag, which applied in electrochemical aptasensor for ultrasensitive detection of zearalenone (ZEN). A glassy carbon electrode was first modified with spherical Au-PANI-Au nanohybrids to enhance the conductivity and immobilize more amino modified ZEN aptamer. Cu@L-Glu/Pd-PtNPs were labeled with Complementary DNA (partial matching with ZEN aptamer) to form bioconjugates for signal amplification. After the hybridization reaction of ZEN aptamer and the bioconjugates, a significant electrochemical signal from the catalysis of H2O2 by Cu@L-Glu/Pd-PtNPs can be observed. ZEN competed with bioconjugates for binding to ZEN aptamer, resulting in decreased the electrochemical signal. Chronoamperometry was applied to record the final electrochemical signals. Under optimal conditions, the electrochemical aptasensor exhibited desirable sensitive detection of ZEN with a wide linearity ranging from 1 fg/mL to 100 ng/mL and a relatively low detection limit of 0.45 fg/mL (S/N = 3). Furthermore, the proposed electrochemical aptasensor shows excellent selectivity to the ZEN in the presence of possible interfering substances, and has potential application for ZEN detection in food samples.

Preparation of dummy molecularly imprinted polymers for extraction of Zearalenone in grain samples.

Surface molecularly imprinted polymers (MIL-101@MIPs) were prepared using MIL-101 as supporting core, imprinted polymers as selective shell synthesized with coumarin-3-carboxylic acid as dummy template of Zearalenone (ZEN), methacrylic acid as functional monomer, and ethylene glycol dimethacrylate hydroxyethyl methacrylate as cross-linker. The polymers were characterized by scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction, and particle-size distribution analyses. MIL-101@MIPs were used as the sorbent to compose the self-made cartridge. The cartridge was used to purify and enrich ZEN from real samples. Under optimized SPE conditions, a self-made cartridge can be reused for at least seven cycles. Elution was monitored with a high-performance liquid chromatography-fluorescence detection system. The linearity of the method ranged within 6.25-250 ng kg-1. The limits of detection ranged within 2.09 - 4.16 ng kg-1, and the limits of quantification ranged within 6.25 -12.50 ng kg-1, respectively. The matrix effects of four real samples were discussed. The spiking recoveries of ZEN ranged within 81.70%-90.10% with relative standard deviations lower than 5.56%. The performance of the self-made cartridge and immunoaffinity column was compared by chromatography.

Effect of the template and functional monomer on the textural properties of molecularly imprinted polymers.

Molecularly imprinted polymers (MIPs) for zearalenone analysis have been synthesized using the template mimics cyclododecyl 2,4-dihydroxybenzoate (CDHB), resorcinol and resorcylic acid. The MIPs are photochemically prepared from 2-(diethylamino)ethyl methacrylate (2-DAEM), 4-vinylpyridine (VIPY), 2-hydroxyethyl methacrylate (HEMA) or 1-allylpiperazine (1-ALPP) as the functional monomers, trimethylolpropane trimethacrylate (TRIM) as cross-linker, azobis(isobutyronitrile) as initiator and acetonitrile as porogen. Non-imprinted polymers have been also synthesized for reference purposes. The textural properties of the novel polymers (BET areas, pore volumes and pore size distributions) have been determined from nitrogen adsorption-desorption isotherms. These parameters have shown to be strongly dependent on the presence of the template and the monomer nature. Scanning electron microscopy and solvent uptake experiments support these findings. Microporosity contributes less than 7% to the total pore volume for all the polymers prepared. Interestingly, a 3.5 nm pore opening is observed for all the polymers and additional pore apertures in the 20-40 nm region for VIPY-, HEMA- and 2-DAEM-based MIPs whereas a much wider opening size distribution has been measured for the 1-ALPP-based MIP. Molecular modeling and, particularly, (1)H NMR experiments demonstrate the strong (2:1) complex formed between 1-ALPP and the diphenolic CDHB (K(11)=4.7 x 10(4)M(-1) and K(12) = 2.6 x 10(2)M(-1) in acetonitrile) that make the corresponding MIP the most suitable for zearalenone recognition in real samples.