A simple mesoporous silica Nanoparticle-based aptamers SERS sensor for the detection of acetamiprid.

In this study, we developed a novel, rapid, simple, and sensitive nano sensor based on the controlled release of 4-Aminothiophenol (4-ATP) signal molecules from aptamers (Apts) modified aminated mesoporous silica nanoparticles (MSNs-NH2) for the quantitative detection of acetamiprid (ACE). Firstly, we synthesized the positively charged MSNs-NH2 by one-pot method, then loaded 4-ATP signal molecules into the pore, and finally electrostatically adsorbed the Apts onto the MSNs-NH2, which acts as a gate to control the release of signal molecules. When ACE is added to the system, ACE preferentially and specifically binds to Apts, so the gate opens and 4-ATP signal molecules are released from the pore. Meanwhile, the silver-loaded mesoporous silica nanoparticles (Ag@SiO2) were prepared by one-pot method as surface-enhanced Raman spectroscopy (SERS) substrate to amplify the signal. The intensity of 4-ATP signal molecules at 1433 cm-1 position was observed to has a linear relationship with the concentration of ACE by SERS detection. Under the optimized detection conditions, a linear correlation was observed in the range of 5-60 ng/mL (R2 = 0.99749), and the limit of detection (LOD) was 2.66 ng/mL. The method has high sensitivity, good selectivity and reproducibility, and can be used for actual sample analysis with the recovery rate of 96.24-103.6 %. This study provides a reference for the rapid and convenient detection of ACE in agricultural products.

Fluorescent aptamer-modified mesoporous silica nanoparticles for quantitative acetamiprid detection.

Acetamiprid (ACE) is widely used to control aphids, brown planthoppers, and other pests in agricultural production. However, ACE is difficult to degrade in the environment, resulting in excessive residue, which causes acute and chronic toxicity to human beings and non-target organisms. Therefore, the development of a rapid, convenient, and highly sensitive method to quantify ACE is essential. In this study, aminated mesoporous silica nanoparticles (MSNs-NH2) were synthesized by one-pot method, and 6-carboxyl fluorescein modified aptamers (FAM-Apt) of ACE were adsorbed on the surface of MSNs-NH2 by electrostatic interaction. Finally, a simple and sensitive fluorescence analysis method for the rapid detection of ACE was established. In the absence of ACE, the negatively charged FAM-Apt was electrostatically bound to the positively charged MSNs-NH2, followed by centrifugation to precipitate MSNs-NH2@FAM-Apt, and no fluorescent signal was detected in the supernatant. In the presence of ACE, the specific combination of FAM-Apt with ACE was greater than its electrostatic interaction with MSNs-NH2, so that FAM-Apt was separated from MSNs-NH2, and the supernatant had strong fluorescence signal after centrifugation. For ACE detection, the linear concentration range was 50-1100 ng/mL, and the detection limit (LOD) was 30.26 ng/mL. The method exhibited high sensitivity, selectivity and reproducibility, which is suitable for practical sample analysis and provides guidance for rapid detection of pesticide residues.

Screening of anti-idiotypic domain antibody from phage library for development of Bt Cry1A simulants.

Anti-idiotypic antibody technique is a new approach for the rapid development of insecticidal protein. In this study, anti-Cry1A polyclonal antibodies were used as antigen to screen the anti-idiotypic antibody that can simulate Cry1A toxins from a phage display human domain antibody (DAB) library. After four rounds of panning, five positive clones that have binding activities with anti-Cry1A polyclonal antibodies were obtained. Indirect competitive ELISA (IC-ELISA) results showed that the positive clone D6 showed significant inhibition for the binding of Cry1A toxins with anti-Cry1A polyclonal antibodies, and the inhibition ratio increased with the increase of D6 content. While, B3, F4, G5, C7 and the controls showed no obvious inhibition to Cry1A toxins. The results suggest that D6 is the "ß" subtype anti-idiotypic antibody, which can simulate Cry1A toxins and competitive binding with anti-Cry1A polyclonal antibodies. Meanwhile, D6 had certain binding activity with the brush border membrane vesicles (BBMV) of p. xylostella, which was the receptor of Cry1A toxins. The results of bioassay showed that D6 had certain insecticidal activity, and the lethal concentration of 50% (LC50) was 976 ng/cm2. This study provides basic materials and experience for the development of Cry toxin simulants.

Production and characterization of a single-chain variable fragment antibody from a site-saturation mutagenesis library derived from the anti-Cry1A monoclonal antibody.

There are plenty of applications of Cry1A toxins (Cry1Aa, Cry1Ab, Cry1Ac) in genetically modified crops, and it is necessary to establish corresponding detection methods. In this study, a single-chain variable fragment (scFv) with high affinities to Cry1A toxins was produced. First, the variable regions of heavy (VH) and light chain (VL) were amplified from hybridoma cell 5B5 which secrete anti-Cry1A monoclonal antibody (mAb) and then spliced into scFv-5B5 by overlap extension polymerase chain reaction (SOE-PCR). Subsequently, site-saturation mutagenesis was performed after homology modeling and molecular docking, which showed that asparagine35, phenylalanine36, isoleucine104, tyrosine105, and serine196, respectively, located in VH complementarity-determining region (CDR1 and CDR3) and VL framework region (FR3) were key amino acid sites. Then, the mutagenesis scFv library (1.35 × 105 CFU/mL) was constructed and a mutant scFv-2G12 with equilibrium dissociation constant (KD) value of 9.819 × 10-9 M against Cry1Ab toxin, which was lower than scFv-5B5 (2.025 × 10-8 M) was obtained by biopanning. Then, enzyme-linked immunosorbent assay (ELISA) was established with limit of detection (LOD) and limit of quantitation (LOQ) of 4.6-9.2 and 11.1-17.1 ng mL-1 respectively for scFv-2G12, which were lower than scFv-5B5 (12.4-22.0 and 23.6-39.7 ng mL-1). Results indicated the promising prospect of scFv-2G12 used for the detection of Cry1A toxins.

Adsorption and Desorption Behaviors of Spirotetramat in Various Soils and Its Interaction Mechanism.

Spirotetramat is a pesticide with bidirectional systemicity and can effectively control pests by inhibiting the biosynthesis of fatty acids. In this study, adsorption and desorption behaviors of spirotetramat in six soils and its interaction mechanism were studied using the batch equilibrium method and infrared radiation. The results showed that the adsorption and desorption behaviors of spirotetramat conformed to the Freundlich isotherm model. The values of adsorption capacities KF-ads ranged from 2.11 to 12.40, and the values of desorption capacities KF-des varied from 2.97 to 32.90. From the hysteresis coefficient, spirotetramat was easily desorbed from the test soils. The adsorption capacity of the soil to spirotetramat enhanced with an increasing temperature. Moreover, the changes in pH values and exogenous addition of humic acid and surfactant could also affect soil adsorption capacity, but for desorption, there was no correlation.

Dissipation kinetics and risk assessments of tricyclazole during Oryza sativa L. growing, processing and storage.

Because of the increase of people's attention to food safety, monitoring the residue of pesticide in rice is becoming more and more important. Commercial and home processing techniques have been used to transform paddy rice into rice products for human or animal consumption, which may reduce the pesticide content in rice. The degradation of tricyclazole during different stages of commercial and home processing and storage was assessed in this paper. Many researches studying the occurrence and distribution of pesticide residues during rice cropping and processing have been reported. Rice samples were extracted with acetonitrile, the extracts were enriched, and then residues were analyzed by liquid chromatography/tandem mass spectrometry method. The dissipation dynamics of tricyclazole in rice plant, soil, and paddy water fitted the first-order kinetic equations. The dissipation half-lives of tricyclazole in the rice plant, water, and soil at dosage of 300~450 g a.i. hm -2 were 4.84~5.16, 4.64~4.85, and 3.57~3.82 days, respectively. The residue levels of tricyclazole gradually reduced with different processing procedures. What is more, decladding process could effectively remove the residues of tricyclazole in raw rice, and washing process could further remove the residues of tricyclazole in polished rice. Degradation dynamic equations of tricyclazole in the raw rice and polished rice were based on the first-order reaction dynamic equations, and the half-lives of the degradation of tricyclazole was 43.32~58.24 days and 46.83~56.35 days in raw rice and polished rice. These results provide information regarding the fate of tricyclazole in the rice food chain, while it provides a theoretical basis for systematic evaluation of the potential residual risk of tricyclazole.

Environmental behaviors of spirotetramat in water.

Spirotetramat is a pesticide with bidirectional systemicity in both xylem and phloem. Currently, researches show that spirotetramat has definite toxicity to aquatic organism. This paper aims to study the environmental behaviors of spirotetramat in water, in the hope of providing guidance for security evaluation of spirotetramat. The researches in this paper showed that under lighting condition, the half-life period of spirotetramat in water was 13.59 days. In water, spirotetramat could be degraded into B-enol and B-keto. As seen from the residual concentrations of two products, B-enol was the dominant degradation product. Under different temperatures, the hydrolysis products of spirotetramat remain B-enol and B-keto. The temperature has little effect on the residual concentration of spirotetramat in water. The residual concentration of B-enol in water gradually increased with the extension of time but B-keto had no significant change. In the buffer solution of different pH values, the degradation rate of spirotetramat was significantly enhanced with the increase of solution pH value. The hydrolysis products of spirotetramat in buffer solution of different pH values were still B-enol and B-keto, and pH exerted certain influence on the residual concentration of B-enol in water. The hydrolysis conversion of spirotetramat has theoretical and practical significance for the safe and reasonable usage of it, as well as for the further evaluation of spirotetramat's ecological risk in water.

Determination and Safety Assessment of Residual Spirotetramat and Its Metabolites in Amaranth (Amaranthus tricolor) and Soil by Liquid Chromatography Triple-Quadrupole Tandem Mass Spectrometry.

With the purpose of guaranteeing the safe use of spirotetramat and preventing its potential health threats to consumers, a QuEChERS extraction method coupled with LC triple-quadrupole tandem MS was applied in this study to determine residual spirotetramat metabolites in different tissues of amaranth (Amaranthus tricolor) and in soil. The results indicate that the spirotetramat degraded into different types of metabolites that were located in different tissues of amaranth and in soil. B-keto, B-glu, and B-enol were the three most representative degradation products in the leaf of amaranth, and B-glu and B-enol were the two major degradation products found in the stem of amaranth; however, only B-enol was detected in the root of amaranth. B-keto and B-mono were the two products detected in the soil in which the amaranth grew. The cytotoxicity results demonstrate that spirotetramat and its metabolite B-enol inhibited cellular growth, and the toxicity of spirotetramat and its metabolite B-enol exceeded than that of the metabolites B-keto, B-mono, and B-glu. This investigation is of great significance to the safe use of spirotetramat in agriculture.