Development of a sensitive,competitive, indirect ELISA for the detection of fumonisin B1 in corn originating from Anhui province,China

Fumonisin B₁ (FB₁) is a secondary metabolite produced by Fusarium verticillioides or Fusarium proliferatum, which present in food and feed. It causes hazardous effects on human and animal health. A monoclonal antibody (mAb) against FB₁ was produced and a simple, reliable and sensitive, competitive, indirect enzyme-linked immunosorbent assay (ci-ELISA) for detection of FB₁ was developed and the experiment conditions were optimized. The coating concentration of FB₁-ovalbumin (FB₁-OVA) was 500 ng mL^?1, the action concentrations of anti-FB₁ mAb and goat anti-mouse IgG were 1.28 × 10⁴ and 1:5000, respectively. The 50% inhibitory concentration (IC₅₀) was 11 ng mL^?1, with a detectable range of 1.25–250 ng mL^?1, and a limit of determination (LOD) of 1.15 ng mL^?1. The cross-reactivity (CR) of the antibody against fumonisin B₂ (FB₂) was 60.4, and <1% against deoxynivalenol (DON), aflatoxin B₁ (AFB₁), ochratoxin A (OTA) or zearalenone (ZEN). In spiked samples (250 ng g^?1, 500 ng g^?1, 1000 ng g^?1), the mean recoveries ranged from 86.7 ± 5% to 102 ± 4%, and the coefficient of variation (CV) ranged from 3% to 10%. A survey of 96 corn samples from Bozhou, Fuyang, Bengbu, and Hefei, in Anhui province, China, was performed. Frequencies of FB₁ contamination were 83.3%, 95.8%, 20.8% and 91.7%, and the mean concentrations of positive samples were 0.702 μg kg^?1, 0.883 μg kg^?1, 0.074 μg kg^?1, and 0.276 μg kg^?1, respectively. The results of this study suggest that the ci-ELISA developed in this study can be used to identify FB₁ in corn, furthermore, further study is needed to investigate FB₁ contamination in food and feed to prevent its harmful health effects.     

Comparison of in-house-developed ELISA with HPLC techniques for the analysis of atrazine residues

In-house developed ELISA was standardized to monitor atrazine residues in different environmental samples. The standard curve was linear, indicating an increase in log concentration with decrease in absorbance (%B/B₀ = 1.075–0.042 Log C; r = ?0.966). The middle of the test was at 75 ng/L and the lowest detection limit at 4 ng/L. ELISA significantly correlated with the high performance liquid chromatography (HPLC) (r = 0.990). Internal validation showed good accuracy and precision. Maximum atrazine residues were present in Jehlum River water/sediments and maize/sugarcane plant roots. Most of the food samples were found to be contaminated. ELISA required less clean-up steps than HPLC, but showed matrix effect in soil/colored extracts.     

Application of an enzyme-linked immunosorbent assay for the detection of clenbuterol residues in swine urine and feeds

The present study outlines applications of an enzyme-linked immunosorbent assay (ELISA) for the analysis of clenbuterol residues. Antisera were raised from rabbits immunized with diazotized clenbuterol-bovine serum albumin (BSA) conjugate. The assay was specific to clenbuterol with a half-maximum inhibition concentration (IC₅₀) of 1.8 ng/mL and 2.5 ng/mL in blank swine urine and phosphate buffer solution, respectively. The assay had high cross-reactivity (86%) with mabuterol, but low with other adrenergic agonists and antagonists. The average recovery of clenbuterol, as measured with the ELISA, ranged from 90% to 112% in swine urine samples and from 86% to 95% in feeds, respectively. This new assay was compared with commercial ELISA test kits. An excellent correlation (r ² = 0.98) between the two methods and satisfactory recoveries suggest that the new assay can be suitable for the determination of clenbuterol residues in real samples. The assay was used to analyze clenbuterol residues in 103 swine urine samples and 68 feed samples collected from northern China. Approximately 50% of the urine samples and 25% of the feed samples analyzed were found positive (concentration of clenbuterol ≥ 1 ppb). The results indicate that clenbuterol was misused in some of the areas surveyed.     

Development of IC-ELISA for detection of organophosphorus pesticides in water

Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC₅₀ of 0.428 μ g/mL with a detection limit of 0.0125 μ g/mL to parathion. The assay also indicated that the IC₅₀ values of pirimiphos and dichlorvos were 0.331 μ g/mL and 1.25 μ g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 μ g/mL and 0.048 μ g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples.     

Dissipation of pendimethalin in the soil of field pea (Pisum sativum L.) and detection of terminal residues in plants

Dissipation of pendimethalin in the soil of field peas (Pisum sativum L.) at 0 to 110 days, and terminal residues in green and mature pea were studied under field conditions. Pendimethalin was applied as pre-emergence herbicide at 750, to 185 g a.i. ha^?1 in winter, in field peas. Dissipation of pendimethalin in the soil at 0 to 110 days followed first-order kinetics showing a half-life of 19.83 days averaged over all doses. Low pendimethalin residues were found in mature pea grain (0.004, 0.003, <0.001 μg g^?1), and straw (0.007, 0.002, <0.001 μg g^?1) at 750, 350 and 185 g a.i. ha^?1 treatments, respectively. The study indicated that residues of pendimethalin in green and mature pea were within the prescribed MRL limits.     

Application of nanobalance technique and principal component analysis for detection of the soil fumigant Telone residues in the air

Quartz crystal nanobalance (QCN) technique is considered as a powerful mass sensitive sensor for monitoring of materials in the sub-nanogram level. In the current study, a method based on QCN technique developed to determine Telone in air. Various coating materials including methyl phenyl silicon, 75% phenyl (OV25) and molecularly imprinted polymer (MIP) were employed. The frequency shift of OV25-modified quartz crystal was found to be linear against organohalogen compounds [Telone (soil fumigant), Koril (Herbicide), Endosulfan (organochlorine insecticide) and Chloroform (solvent)] concentrations in the range of 2.4 to 48 mg L^?1 for Telone vapor and 4.8–24 mg L^?1 for three other vapors. The correlation coefficients for Telone, Koril, Endosulfan and Chloroform were 0.992, 0.996, 0.989 and 0.991, respectively. The principal component analysis was also utilized to process the frequency response data of the organic vapors. Using principal component analysis, it was found that more than 93.85% of the data variance could still be explained by use of two principal components (PC1 and PC2). Subsequently, the successful discrimination of Telone and other compounds was quite possible through the principal component analysis of the transient responses of the OV25-modified electrode. In the second method, a molecularly imprinted polymer-coated sensor for Telone was developed. Molecularly imprinted polymer coated quartz crystal (MIP-QCN) showed a selective response to Telone and gave a linear relationship between frequency shift and amount of Telone from 1 to 48 mg L^?1. In this investigation, the proficiency of MIP-QCN and OV25-modified QCN sensors were compared.     

Development of enzyme-linked immunosorbent assay (ELISA) for glutathione S-transferase (GST-S) protein in the intertidal copepod Tigriopus japonicus and its application for environmental monitoring

To utilize the GST-S protein as a useful biomarker for environmental contamination, we developed a polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) in the intertidal copepod Tigriopus japonicus. Two polyclonal antibodies, TJ-GST-S1 and TJ-GST-S2, were raised against two TJ-GST-S synthetic peptides. Also a recombinant TJ-GST-S protein was purified as a standard for ELISA development. Each polyclonal antibody was tested by Western blot analysis and indirect ELISA. Of two polyclonal antibodies, TJ-GST-S2 ELISA was further employed due to its wide range of detection and the limit of specificity compared to those of TJ-GST-S1 ELISA system. After exposure to 4 metals (Ag, As, Cd, and Cu) to T. japonicus, the amount of TJ-GST-S protein was significantly elevated in a concentration-dependent manner. Also, TJ-GST-S protein was upregulated at relative high concentrations of B[α]P, PCB, and TBT. In this paper, we suggest that T. japonicas ELISA for TJ-GST-S2 is useful as a potential indicator system for marine contaminants.     

响应面法优化化学沉淀螯合生物絮凝处理含镍废水

采用化学沉淀螯合自制生物絮凝的处理工艺处理含镍的模拟废水.首先,考察了不同的提取方法对脱水污泥提取的生物絮凝剂的絮凝效果的影响,进而确定生物絮凝剂的提取方法.然后,考察了不同的化学沉淀剂与生物絮凝剂组合处理含镍废水的能力,并采用Plackett-Burman设计法结合响应曲面法筛选优化了化学沉淀螯合生物絮凝处理含镍废水...     

Evidence for the Stepwise Behavioral Response Model (SBRM): the effects of Carbamate Pesticides on medaka (Oryzias latipes) in an online monitoring system

The Stepwise Behavioral Response Model (SBRM), which is a conceptual model, postulated that an organism displays a time-dependent sequence of compensatory Stepwise Behavioral Response (SBR) during exposure to pollutants above their respective thresholds of resistance. In order to prove the model, in this study, the behavioral responses (BRs) of medaka (Oryzias latipes) in the exposure of Arprocarb (A), Carbofuran (C) and Methomyl (M) were analyzed in an online monitoring system (OMS). The Self-Organizing Map (SOM) was utilized for patterning the obtained behavioral data in 0.1 TU (Toxic Unit), 1 TU, 2 TU, 5 TU, 10 TU and 20 TU treatments with control. Some differences among different Carbamate Pesticides (CPs) were observed in different concentrations and the profiles of behavior strength (BS) on SOM were variable depending upon levels of concentration. The time of the first significant decrease of BS (SD-BS) was in inverse ratio to the CP concentrations. Movement behavior showed by medaka mainly included No effect, Stimulation, Acclimation, Adjustment (Readjustment) and Toxic effect, which proved SBRM as a time-dependence model based on the time series BS data. Meanwhile, it was found that SBRM showed evident stress-dependence. Therefore, it was concluded that medaka SBR was both stress-dependent and time-dependent, which supported and developed SBRM, and data mining by SOM could be efficiently used to illustrate the behavioral processes and to monitor toxic chemicals in the environment.     

Analysis of pesticide residues using the Quick Easy Cheap Effective Rugged and Safe (QuEChERS) pesticide multiresidue method in traditional Chinese medicine by gas chromatography with electron capture detection

The Quick Easy Cheap Effective Rugged and Safe multiresidue method (QuEChERS) has been validated for the extraction of 15 organochlorine pesticides (OCPs) in completely different matrices of traditional Chinese medicine (TCM). The method employed a rapid, simple and cost-effective procedure. The spiking levels for the recovery experiments were 0.1, 0.5 and 2.0 mg kg⁻¹. Mean recoveries mostly ranged between 76.0% and 114.0% (96.0% on average), and relative standard deviations (RSD) were generally below 10% (4.72% on average). Based on these results, the methodology has been proven to be highly efficient and robust and thus suitable for monitoring the MRL compliance of a wide range of commodity. The contamination status of 15 OCPs on 80 different TCMs marketed in China, and a total of 400 samples were also performed using a previously validated method. 15 OCPs were benzene hexachloride (BHC, including α-BHC, β-BHC, γ-BHC, δ-BHC), hexachlorobenzene, heptachlor, heptachlor epoxide, aldrin, p, p′-DDE, p, p′-DDD, o, p′-DDT, p, p′-DDT, mirex, endrin, dieldrin. This study indicates that the proposed method is useful for analyzing OCPs in TCM.     

铅蓄电池落实生产者责任延伸制度成效与展望

铅蓄电池生产者责任延伸制度被写入新修订的《中华人民共和国固体废物污染环境防治法》,并确定了生产者的回收义务,弥补了相关政策实施的上位法空白。以2016年以来我国废铅蓄电池规范收集处理量为依据,梳理分析了我国推进落实铅蓄电池生产者责任延伸制度、建立规范的废铅蓄电池收集处理体系方面所取得的成效。同时,针对目前面临的一些实际问题探讨了相应的解决方向,以期对《中华人民共和国固体废物污染环境防治法》修订实施后,完善相关管理政策制度提供借鉴参考。     

A multi-residue method for the analysis of organophosphorus residues in cooked and polished rice using accelerated solvent extraction and dispersive-solid phase extraction (D-SPE) technique and uncertainty measurement

Quick, simple and efficient multi-residue analytical methods were developed and validated for the determination of organophosphorous insecticides from polished and cooked rice. Polished rice was extracted using a simple, automated technique namely accelerated solvent extraction (ASE) using dichloromethane as the extraction solvent. Cooked rice was extracted with acetone and cleaned up using dispersive-solid phase extraction (D-SPE) technique. The single step extraction method adopted for polished rice using accelerated solvent extractor provided satisfactory recovery for eight organophosphorus pesticides (OPPs) which ranged from 85.5–116.7%; 90.0–110.3% and 93.5–118.8% at 1, 5 and 10 limit of quantification (LOQ) levels, respectively. The recovery of cooked rice was in the range of 74–124%; from 75–100% and from 73–87% for 1, 5 and 10 level of fortification, respectively. The total uncertainty was evaluated, taking four main independent sources viz., weighing, purity of the standard, GC calibration curve and repeatability under consideration. The expanded uncertainty was found to be in the range of 5–20%.     

Effects of organic ligands on fractionation of rare earth elements (REEs) in hydroponic plants: an application to the determination of binding capacities by humic acid for modeling

Previous studies have revealed the fractionation processes of rare earth elements (REEs) in hydroponic plants, with a heavy REE (HREE, the elements from Gd to Lu) enrichment in leaves. In this study, effects on the HREE enrichment in soybean leaves with additions of carboxylic acids (acetate, malate, citrate, NTA, EDTA and DTPA) and two soil humic acids (HAs) were investigated. REE speciation in carboxylic acid and HA solutions was simulated using Visual MINTEQ and Model V, respectively. The results showed that the effects caused by carboxylic acids were strongly dependent on the differences between their binding strengths for light REEs (LREEs, the elements from La to Eu) and those for HREEs. A good correlation existed between these effects and the changes of free REE ions in solutions. This relationship was also observed for the HA treatments, provided that the intrinsic equilibrium constants of REEs for cation–proton exchange with HA (i.e., pK_MHA) in Model V were estimated using a free-energy relationship with the stability constants for REE complexation with lactic acid. It is suggested that this set of pK_MHA values is more suitable for use in Model V for the simulation of REE complexation with HA.