One-Pot Preparation of Mixed-Mode Reversed-Phase Anion-Exchange Silica Sorbent and its Application in the Detection of Cyclopiazonic Acid in Feeds and Agricultural Products.

A novel co-bonded octyl and pyridine silica (OPS) sorbent was prepared and applied for the solid phase extraction (SPE) of cyclopiazonic acid (CPA, a type of mycotoxin) in feed and agricultural products for the first time. A simple mixed-ligand one-pot reaction strategy was employed for OPS sorbent preparation. Nitrogen adsorption-desorption measurements, elemental analysis (EI), thermal gravimetric analysis (TGA), and Fourier transform infrared spectroscopy (FT-IR) analysis demonstrated the successful immobilization of octyl and quaternary ammonium groups onto the surface of silica gel. The large specific surface area, high-density functional groups, and mixed-mode anion-exchange characteristics of these silica particles made them the ideal material for the efficient extraction of CPA. Additionally, the OPS sorbents displayed excellent batch-to-batch reproducibility, satisfactory reusability, and low cost. The SPE parameters were optimized to explore the ionic and hydrophobic interactions between CPA and the functional groups, and the ultra-high performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) parameters were optimized to obtain a desirable extraction efficiency and high sensitivity to CPA. Meanwhile, the OPS sorbent presented a satisfactory extraction selectivity and low matrix effect. Under the optimized conditions, our developed CPA detection method was used to determine CPA level in rice, wheat flour, corn flour, peanut, and feed samples, exhibiting a lower detection limit, better linearity, higher sensitivity, and satisfactory extraction recovery rate than previously reported methods. Therefore, our method can be preferentially used as a method for the detection of CPA in agricultural products and feeds.

[Modified QuEChERS method combined with ultra performance liquid chromatography-tandem mass spectrometry for detection of cyclopiazonic acid in feeds].

Mycotoxins are toxic secondary metabolites produced by fungal species that can cause acute, subacute, and chronic toxicity in humans and animals. Thus, these toxins pose a significant threat to health and safety. Owing to the lack of effective antimold measures in the agricultural industry, feed ingredients such as corn, peanuts, wheat, barley, millet, nuts, oily feed, forage, and their byproducts are prone to mold and mycotoxin contamination, which can affect animal production, product quality, and safety. Cyclopiazonic acid (CPA), which is mainly biosynthesized from mevalonate, tryptophan, and diacetate units, is a myotoxic secondary metabolite produced by Penicillium and Aspergillus fungi. CPA is widely present as a copollutant with aflatoxins in various crops. Compared with some common mycotoxins such as aflatoxins, fumonisins, ochratoxins, zearalenones, and their metabolites, CPA has not been well investigated. In the United States, a survey showed that 51% of corn and 90% of peanut samples contained CPA, with a maximum level of 2.9 mg/kg. In Europe, CPA was found in Penicillium-contaminated cheeses as high as 4.0 mg/kg. Some studies have shown that CPA can cause irreversible damage to organs such as the liver and spleen in mice. Therefore, the establishment of a rapid and efficient analytical method for CPA is of great significance for the risk assessment of CPA in feeds, the development of standard limits, and the protection of feed product quality and safety. The QuEChERS method, a sample pretreatment method that is fast, simple, cheap, effective, and safe, is widely used in the analysis of pesticide residues in food. In this study, a modified QuEChERS method combined with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to determine CPA levels in feeds. The chromatographic separation and MS detection of CPA as well as the key factors affecting the extraction efficiency of CPA, including the type of extraction solvent, type of inorganic salt, and type and dosage of adsorbent, were optimized in detail. During the optimization of the chromatographic-separation step, the acid and salt concentrations of the mobile phase affected the separation and detection of CPA. During the optimization of the QuEChERS method, the addition of a certain amount of acetic acid improved the extraction efficiency of CPA because of its acidic nature; in addition, GCB and PSA significantly adsorbed CPA from the feed extract. Under optimal conditions, the CPA in the feed sample (1.0 g) was extracted with 2 mL of water and 4 mL of acetonitrile (ACN) containing 0.5% acetic acid. After salting out with 0.4 g of NaCl and 1.6 g of MgSO4, 1 mL of the ACN supernatant was purified by dispersive solid-phase extraction using 150 mg of MgSO4 and 50 mg of C18 and analyzed by UPLC-MS/MS. The sample was separated on a Waters HSS T3 column (100 mm×2.1 mm, 1.8 µm) using 2 mmol/L ammonium acetate aqueous solution with 0.5% formic acid and ACN as the mobile phases and then analyzed by positive electrospray ionization in multiple reaction monitoring mode. CPA exhibited good linearity in the range of 2-200 ng/mL, with a high correlation coefficient (r=0.9995). The limits of detection and quantification of CPA, which were calculated as 3 and 10 times the signal-to-noise ratio, respectively, were 0.6 and 2.0 µg/kg, respectively. The average recoveries in feed samples spiked with 10, 100, and 500 µg/kg CPA ranged from 70.1% to 78.5%, with an intra-day precision of less than 5.8% and an inter-day precision of less than 7.2%, indicating the good accuracy and precision of the proposed method. Finally, the modified QuEChERS-UPLC-MS/MS method was applied to the analysis of CPA in 10 feed samples obtained from Wuhan market. The analysis results indicated that the developed method has good applicability for CPA analysis in feed samples. In summary, an improved QuEChERS method was applied to the extraction and purification of CPA from feeds for the first time; this method provides a suitable analytical method for the risk monitoring, assessment, and standard-limit setting of CPA in feed samples.

Molecularly imprinted fluorescence assay based on lead halide perovskite quantum dots for determination of benzo(a)pyrene.

Molecularly imprinted polymers with methylammonium lead halide perovskite quantum dots (MIP@MAPbBr3 PQDs) have been prepared and applied to the determination of benzo(a)pyrene (BaP) for the first time. The photoluminescence (PL) of MIP@MAPbBr3 PQDs was enhanced due to the surface passivation of defects by BaP. PL excitation and emission spectra, X-ray diffraction, Fourier transform infrared, and time-resolved PL studies suggest that the interaction between MIP@MAPbBr3 PQDs and BaP is a dynamic process. After MIP@MAPbBr3 PQDs were incubated with BaP, the benzene ring in the molecular structure of BaP can interact with MIP@MAPbBr3 PQDs through π electrons, which reduces non-radiative recombination of MIP@MAPbBr3 PQDs and lengthens excited state lifetime. The PL intensity of the MIP@MAPbBr3 PQDs-BaP system was monitored at 520 nm with 375 nm excitation. Under optimized conditions, the PL intensity of MIP@MAPbBr3 PQDs is linear with the concentration of BaP in the 10 to 100 ng·mL-1 range, with a detection limit of 1.6 ng·mL-1. The imprinting factor was 3.9, indicating excellent specificity of MIP@MAPbBr3 PQDs for BaP. The MIP@MAPbBr3 PQDs were subsequently applied to the PL analysis of BaP in sunflower seed oil, cured meat, and grilled fish samples, achieving recoveries from 79.3 to 107%, and relative standard deviations below 10%. This molecularly imprinted fluorescence assay improves the selectivity of BaP in complex mixtures and could be extended to other analytes.

Molecular Mechanisms of Tebuconazole Affecting the Social Behavior and Reproduction of Zebrafish.

The aim of this study was to explore the underlying mechanism of adverse effects caused by tebuconazole (TEB) on the reproduction of aquatic organisms In the present study, in order to explore the effects of TEB on reproduction, four-month-old zebrafish were exposed to TEB (0, DMSO, 0.4 mg/L, 0.8 mg/L, and 1.6 mg/L) for 21 days. After exposure, the accumulations of TEB in gonads were observed and the cumulative egg production was evidently decreased. The decline of fertilization rate in F1 embryos was also observed. Then the changes in sperm motility and histomorphology of gonads were discovered, evaluating that TEB had adverse effects on gonadal development. Additionally, we also found the alternations of social behavior, 17ß-estradiol (E2) level, and testosterone (T) level. Furthermore, the expression levels of genes involved in the hypothalamic-pituitary-gonadal (HPG) axis and social behavior were remarkably altered. Taken together, it could be concluded that TEB affected the egg production and fertilization rate by interfering with gonadal development, sex hormone secretion, and social behavior, which were eventually attributed to the disruption of the expressions of genes associated with the HPG axis and social behavior. This study provides a new perspective to understanding the mechanism of TEB-induced reproductive toxicity.

Titanium dioxide nanoparticles decreases bioconcentration of azoxystrobin in zebrafish larvae leading to the alleviation of cardiotoxicity.

Interactions between titanium dioxide nanoparticles (n-TiO2) and pollutants in the aquatic environment may alter the bioavailability of pollutants, and thus altering their toxicity and fate. In order to investigate the bioconcentration of azoxystrobin (AZ) and its mechanism of cardiotoxicity in the presence of n-TiO2, the experiment was divided into control, n-TiO2 (100 µg/L), AZ (40, 200 and 1000 µg/L) and AZ (40, 200, 1000 µg/L) + n-TiO2 groups, and the zebrafish embryos were exposed to the exposure solution until 72 h post-fertilization. Results suggested the presence of n-TiO2 notably reduced the accumulation of AZ in larvae compared with exposure to AZ alone, thereby significantly decreasing AZ-induced cardiotoxicity, including heart rate changes, pericardium edema, venous thrombosis, increased sinus venosus and bulbus arteriosus distance and changes in cardiac-related gene expression. Further studies showed that AZ + n-TiO2 together restrained total-ATPase and Ca2+-ATPase activities, while the activity of Na+K+-ATPase increased at first and then decreased. Furthermore, there were significant changes in the expressions of oxidative phosphorylation and calcium channel-related genes, suggesting mitochondrial dysfunction may be the potential mechanism of cardiotoxicity induced by AZ and n-TiO2. This study supplies a new perspective for the joint action of AZ and environmental coexisting pollutants and provides a basis for ecological risk management of pesticides.

Microbial Degradation of Zearalenone by a Novel Microbial Consortium, NZDC-6, and Its Application on Contaminated Corncob by Semisolid Fermentation.

A novel microbial consortium (NZDC-6) was screened and characterized to detoxify the estrogenic mycotoxin zearalenone (ZEA), which commonly contaminates maize and is a major threat to food and health security. We found NZDC-6 to be thermophilic and highly effective, with a 90.3% ZEA degradation ratio at an optimum temperature of 60 °C. NZDC-6 was also effective at degrading the more estrogenic ZEA cognates, α-zearalenol (α-ZAL) and ß-zearalenol (ß-ZAL), with >90% degradation ratios. To evaluate a practical application, ZEA-contaminated corncobs were treated with NZDC-6 via semisolid fermentation. Measurements of physicochemical parameters and 16S microbial diversity and redundancy analysis (RDA) indicated that ZEA removal was most efficient at a low corncob solid content (< 5%), as a high solid content overwhelmed the microbial metabolic load, leading to increased dissolved oxygen and lowered pH. Our results demonstrate that the control of environmental variables is crucial for effective ZEA microbial removal in practical applications.

Simultaneous degradation of aflatoxin B1 and zearalenone by a microbial consortium.

Mycotoxins are toxic secondary metabolites mainly produced by filamentous fungal species that commonly contaminate staple foods and feeds. They cause significant economic losses and greatly harm food security. Simultaneous contamination of multiple mycotoxins and the accompanying additive and synergistic effects may cause even more serious harm. To develop a microbial consortium with the ability to degrade multiple mycotoxins, a previously identified consortium with aflatoxin B1 (AFB1) degradation ability, TADC7, was domesticated by co-culturing with 500 µg l-1 AFB1 and 500 µg l-1 zearalenone (ZEA), yielding the derived microbial consortium TMDC. After 168 h of co-culture with TMDC, 2000 µg l-1AFB1 and 2000 µg l-1 ZEA were degraded by 98.9% and 88.5%, respectively. The proteins or enzymes in the TADC7 cell-free supernatant played a major role in mycotoxins degradation. The degradation ratios of 5000 µg l-1 AFB1 and 5000 µg l-1 ZEA by 48 h TMDC cell-free supernatant were 93.8% and 90.3% at 72 h, respectively. Based on 16S rRNA sequencing, Geobacillus and Tepidimicrobium might play important roles in mycotoxin degradation by TMDC, and the TMDC community composition was stable, irrespective of mycotoxin. This study established the biodegradation of different categories of mycotoxins, and will facilitate the practical application of microbial consortia in mycotoxin degradation.

Rapid and sensitive detection of the phenoxy acid herbicides in environmental water samples by magnetic solid-phase extraction combined with liquid chromatography-tandem mass spectrometry.

Phenoxy acid herbicides are widely used herbicides that play an important role in improving the yield and quality of crops. However, some research has shown that this kind of herbicide is poisonous to human and animals. In this study, a rapid and sensitive method was developed for the detection of seven phenoxy acid herbicides in water samples based on magnetic solid-phase extraction followed by liquid chromatography and tandem mass spectrometry. Magnetic amino-functionalized multiwalled carbon nanotubes were prepared by mixing bare magnetic Fe3 O4 nanoparticles with commercial amino-functionalized multiwalled carbon nanotubes in water. Then the amino-functionalized multiwalled carbon nanotubes were used to enrich phenoxy acid herbicides from water samples based on hydrophobic and ionic interactions. The effects of experimental variables on the extraction efficiency have been studied in detail. Under the optimized conditions, the method validation was performed. Good linearities for seven phenoxy acid herbicides were obtained with squared regression coefficients ranging from 0.9971 to 0.9989. The limits of detection ranged from 0.01 to 0.02 µg/L. The method recoveries of seven phenoxy acid herbicides spiked at three concentration levels in a blank sample were from 92.3 to 103.2%, with inter- and intraday relative standard deviations less than 12.6%.