Characteristics, influence factors, and health risk assessment of volatile organic compounds through one year of high-resolution measurement at a refinery.

From January 2020 to December 2020, high-resolution data of volatile organic compound (VOC) concentrations were monitored by online instruments at a petroleum refinery. The measurement results showed that the external contaminants, meteorological conditions and photochemical reactions had a great influence on the VOC data measured in the petroleum refineries. Some significant differences were observed in the emission composition of different refineries, while propene (34.2%), propane (10.2%), n-butane (5.6%), i-pentane (5.0%) were the dominant species emitted from the refinery in this study. The correlations between compounds with similar atmospheric lifetimes were strong (R2 > 0.9), which indicated that the diagnostic ratios of these compounds could be used as indicators to identify the refinery emission source. Chronic health effects of non-carcinogenic risk results showed that acrolein had the highest non-carcinogenic risk and other compound-specific health risks may be of less concern in the refining area. Halogenates and aromatics accounted for 97.4% of the total carcinogenic risk values, while 1,2-dibromoethane, chloromethane, benzene, trichloromethane, 1,2-dichloroethane contributed approximately 80% of the total carcinogenic risk assessment values. This research has recorded valuable data about the VOC emission characteristics from the perspective of the high-resolution monitoring of the petroleum refinery. The results of this work will provide a reference to accurately quantify and identify the emission of petroleum refineries and further throw some light on effective VOC abatement strategies.

Computer-aided profiling of a unique broad-specific antibody and its application to an ultrasensitive fluoroimmunoassay for five N-methyl carbamate pesticides.

Pollution of N-methyl carbamate (NMC) pesticides is threatening the non-target organisms' survival. Thus, broad-specific antibodies and class-selective immunoassays are demanding for multiple NMCs determination. In this study, we employed a molecular docking-based virtual screening strategy to fast profile antibody spectrum, based on a designed chemical pool containing 17 compounds. A monoclonal antibody (mAb)-6G against carbofuran was used as the objective. The recombinant full-length IgG was successfully expressed to validate the antibody sequences for homology modeling. After docking, we manually categorized the antibody-chemical binding strength into three groups. Non-competitive surface plasmon resonance (SPR) demonstrated the mAb-6G affinitive binding toward five NMCs (carbofuran, isoprocarb, propoxur, carbaryl and carbosulfan), which were classified into strong and moderate binding categories. Antibody binding properties were confirmed again by ic-ELISA and lateral flow immunochromatographic strip. Subsequently, an ultrasensitive indirect competitive fluoromicrosphere-based immunoassay (ic-FMIA) was established with the IC50 (half-maximal inhibitory concentration) values of 0.08-3.37 ng/mL. This portable assay presented a 30-230-fold improved sensitivity than traditional ic-ELISA and was applied in European surface water analysis. Overall, our work provides an efficient platform integrating in-silico and experimental methodologies to accelerate the characterization of hapten-specific antibody binding properties and the development of high-sensitive immunoassays for multi-pollutants monitoring.

Recent Trends in Nanomaterial-Based Biosensors for Point-of-Care Testing.

In recent years, nanomaterials of different shape, size, and composition have been prepared and characterized, such as gold and silver nanoparticles, quantum dots, mesoporous silica nanoparticles, carbon nanomaterials, and hybrid nanocomposites. Because of their unique physical and chemical properties, these nanomaterials are increasingly used in point-of-care testing (POCT) to improve analytical performance and simplify detection process. They are used either as carriers for immobilizing biorecognition elements, or as labels for signal generation, transduction and amplification. In this commentary, we highlight recent POCT technologies that employ nanotechnology for the analysis of disease biomarkers, including small-molecule metabolites, enzymes, proteins, nucleic acids, cancer cells, and pathogens. Recent advances in lateral flow tests, printable electrochemical biosensors, and microfluidics-based devices are summarized. Existing challenges and future directions are also discussed.

Characterization of Variable Region Genes and Discovery of Key Recognition Sites in the Complementarity Determining Regions of the Anti-Thiacloprid Monoclonal Antibody.

Sequence-defined recombinant antibodies (rAbs) have emerged as alternatives to hybridoma-secreted monoclonal antibodies (mAbs) for performing immunoassays. However, the polyploidy nature of hybridomas often leads to the coexistence of aberrant or non-specific functional variable region (VR) gene transcripts, which complicates the identification of correct VR sequences. Herein, we introduced the use of LC-MS/MS combined with next-generation sequencing to characterize VR sequences in an anti-thiacloprid mAb, which was produced by a hybridoma with genetic antibody diversity. The certainty of VR sequences was verified by the functional analysis based on the recombinant antibody (rAb) expressed by HEK293 mammalian cells. The performance of the rAb was similar to that of the parental mAb, with IC50 values of 0.73 and 0.46 µg/L as measured by ELISAs. Moreover, molecular docking analysis revealed that Ser52 (H-CDR2), Trp98, and Trp93 (L-CDR3) residues in the complementarity determining regions (CDRs) of the identified VR sequences predominantly contributed to thiacloprid-specific recognition through hydrogen bonds and the CH-π interaction. Through single-site-directed alanine mutagenesis, we found that Trp98 and Trp93 (L-CDR3) showed high affinity to thiacloprid, while Ser52 (H-CDR2) had an auxiliary effect on the specific binding. This study presents an efficient and reliable way to determine the key recognition sites of hapten-specific mAbs, facilitating the improvement of antibody properties.

The expression of soluble functional α7-nicotinic acetylcholine receptors in E. coli and its high-affinity binding to neonicotinoid pesticides.

Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels mediating fast cholinergic synaptic transmission in nervous system. In insects, nAChRs are the target sites for several naturally occurring and synthetic compounds, including the neonicotinoid insecticides. So far, one of the major strategies to explore the interaction of nAChR and ligands is based on the heterologous expression of nAChR, which is tough, and needs to be explored. In this study, we expressed and purified extracellular domain of rat a7 subunit (Rα7-ECD), the binding site of the ligands in E. coli and determined the interactions and kinetic constants of neonicotinoid insecticides with Rα7-ECD. The recombinant Rα7-ECD is water-soluble and appears to be correctly folded. The interactions of three neonicotinoid pesticides with Rα7-ECD were assessed by surface plasmon resonance (SPR) biosensor. The results revealed a fast association and fast disassociation binding mode of Rα7-ECD/pesticides complexes with the KD value of clothianidin (6.414E-9 M) > imidacloprid (9.030E-9 M) > acetamiprid (2.874E-6 M), respectively. This study demonstrated that the nAChR expressed from E. coli was functional, and SPR biosensor technology would be a good alternative for characterizing members of nAChR receptor family.

Up-Converting Nanoparticle-Based Immunochromatographic Strip for Multi-Residue Detection of Three Organophosphorus Pesticides in Food.

Organophosphorus (OP) pesticides are widely used to control pests because of their high activity. This study described a rapid and sensitive lateral flow immunochromatographic (LFIC) assay based on up-converting nanoparticles (UCNPs) for multi-residue detection of three OP pesticides. The developed assay integrated novel fluorescent material UCNPs labeled with a broad-specific monoclonal antibody. Based on the competitive platform by immobilized antigen in the test zone, the optimized UCNPs-LFIC assay enabled sensitive detection for parathion, parathion-methyl, and fenitrothion with IC50 of 3.44, 3.98, and 12.49 ng/mL (R 2 ≥ 0.9776) within 40 min. The detectable ability ranged from 0.98 to 250 ng/mL. There was no cross-reactivity with fenthion, phoxim, isocarbophos, chlorpyrifos, or triazophos, even at a high concentration of 500 ng/mL. Matrix interference from various agricultural products was also studied in food sample detection. In the spiked test, recoveries of the three OP pesticides ranged from 67 to 120% and relative standard deviations were below 19.54%. These results indicated that the proposed strip assay can be an alternative screening tool for rapid detection of the three OP pesticides in food samples.

An amino-modified metal-organic framework (type UiO-66-NH2) loaded with cadmium(II) and lead(II) ions for simultaneous electrochemical immunosensing of triazophos and thiacloprid.

A method is described for simultaneous voltammetric determination of the pesticides triazophos (TRS) and thiacloprid (THD). A glassy carbon electrode (GCE) was modified with a metal-organic framework (type UiO-66-NH2) which has a large specific surface (1018 m2·g-1) and contains large amounts of Cd(II) and Pb(II) ions, with adsorption capacities of 230 and 271 mg·g-1, respectively. The antigen-loaded particles were then bound to antibody, magnetically separated, and analyzed by square wave voltammetry to give signals for Cd(II) and Pb(II) at -0.82 and - 0.56 V (vs. Ag/AgCl) for TRS and THD, respectively. Under optimized conditions, the method has a wide linear range (0.2-750 ng·mL-1) and low detection limits (0.07 and 0.1 ng·mL-1 at a S/N of 3 for TRS and THD, respectively). It is perceived that this assay represents a useful tool for simultaneous determination of multiple pesticide residues. The method has a wide scope in that may be extended to monitoring of other small organic pollutants by changing the types of metal ions and by using other antibodies. Graphical abstract Schematic presentation of an amino-modified metal-organic framework (type UiO-66-NH2) loaded with Cd(II) and Pb(II) ions for simultaneous electrochemical immunosensing of triazophos (TRS) and thiacloprid (THD). It is based on the fabrication of antigen (Ab)-immobilized UiO-66-NH2-based signal tags (a), and of an antibody (Ab)-immobilized magnetic bead (MB-COOH)-based capture probes (b).

Binding properties of broad-specific monoclonal antibodies against three organophosphorus pesticides by a direct surface plasmon resonance immunosensor.

In this study, heterologous indirect competitive enzyme-linked immunosorbent assay (icELISA) was introduced into the screening of hybridomas for the development of broad-specific monoclonal antibodies (mAbs) against organophosphorus (OP) pesticides. After immunization, two formats of icELISA based on the homologous hapten antigen and four heterologous hapten antigens were conducted for hybridoma screening. Two mAbs 2G6 and 7B2 with good recognition toward three OP pesticides (parathion, methyl-parathion, and fenitrothion) were produced. Results of the icELISA showed that the two mAbs exhibited high sensitivity against three OP pesticides, with IC50 ranging from 2.93 to 19.71 ng mL-1. Moreover, a non-competitive surface plasmon resonance (SPR) immunosensor was used for characterizing the binding properties of the mAbs to OP pesticides. After kinetic analysis, equilibrium dissociation constant (KD) values of mAbs 2G6 and 7B2 were calculated as 1.45 × 10-9 M and 4.26 × 10-9 M for parathion, 6.75 × 10-9 M and 4.17 × 10-9 M for methyl-parathion, and 2.44 × 10-8 M and 1.19 × 10-8 M for fenitrothion, respectively. Whereas, both icELISA and SPR-based immunoassay indicated that the two mAbs could not recognize other five OP analogs. Since SPR-based immunoassay provides comprehensive information of two molecules directly interacting with each other, it is a valuable tool during the development and characterization of broad-specific mAbs. Graphical abstract ᅟ.

A highly selective electrochemical sensor based on molecularly imprinted polypyrrole-modified gold electrode for the determination of glyphosate in cucumber and tap water.

An electrochemical sensor based on molecularly imprinted polypyrrole (MIPPy) was developed for selective and sensitive detection of the herbicide glyphosate (Gly) in cucumber and tap water samples. The sensor was prepared via synthesis of molecularly imprinted polymers on a gold electrode in the presence of Gly as the template molecule and pyrrole as the functional monomer by cyclic voltammetry (CV). The sensor preparation conditions including the ratio of template to functional monomers, number of CV cycles in the electropolymerization process, the method of template removal, incubation time, and pH were optimized. Under the optimal experimental conditions, the DPV peak currents of hexacyanoferrate/hexacyanoferrite changed linearly with Gly concentration in the range from 5 to 800 ng mL-1, with a detection limit of 0.27 ng mL-1 (S/N = 3). The sensor was used to detect the concentration of Gly in cucumber and tap water samples, with recoveries ranging from 72.70 to 98.96%. The proposed sensor showed excellent selectivity, good stability and reversibility, and could detect the Gly in real samples rapidly and sensitively. Graphical abstract Schematic illustration of the experimental procedure to detect Gly using the MIPPy electrode.

Multi-residue detection of pesticides using a sensitive immunochip assay based on nanogold enhancement.

This paper describes the development of a new multiplex immunoassay for simultaneous detection of seven pesticides (triazophos, methyl-parathion, fenpropathrin, carbofuran, thiacloprid, chlorothalonil, and carbendazim). Sixteen pairs of pesticide antibodies and antigens were screened for reactivity and cross-reaction. A microarray chip consisting of seven antigens immobilized on a nitrocellulose membrane was then constructed. Nanogold was employed for labeling and signal amplification to obtain a sensitive colorimetric immunoassay. The direct and indirect detection formats were further compared using primary antibody-gold and secondary antibody-gold conjugates as tracers. An integrated 7-plex immunochip assay based on the indirect model was established and optimized. The detection limits for the pesticides were 0.02-6.45 ng mL(-1), which meets detection requirements for pesticide residues. Naked-eye assessment showed the visual detection limits of the assay ranged from 1 to 100 ng mL(-1). Spiked recovery results demonstrated that the immunochip assay had potential for multi-analysis of pesticide residues in vegetables and fruits. The proposed microarray methodology is a flexible and versatile tool, which can be applied to other competitive multiplex immunoassays for small molecular compounds.

Residual behavior and risk assessment of the mixed formulation of benzene kresoxim-methyl and fluazinam in cucumber field application.

Benzene kresoxim-methyl (BKM) is a new strobilurin fungicide mixed with fluazinam (Flu) into 40 % suspension concentrate (SC) formulation to improve fungicidal efficacy and to reduce the risk of resistance on cucumber. However, the fate of the fungicide residues in a cucumber plantation remains unclear. Thus, an efficient method of ultra-performance liquid chromatography combined with a modified quick, easy, cheap, effective, rugged, and safe sample preparation was developed to simultaneously determine the BKM and Flu residues in cucumber and soil samples to investigate their residual behavior and risk assessment in the cucumber plantation. This analytical method revealed that the detection limits of BKM and Flu were 1.64 × 10(-3) and 1.82 × 10(-3) mg L(-1), respectively, and their average recoveries in the cucumber and soil samples were 77.5-106.9 %. The respective half-lives of BKM and Flu were 2.2-3.4 and 1.0-2.5 days in cucumber; in soil, the half-lives of BKM and Flu were 2.6-5.0 and 2.4-5.3 days, respectively. Seven days after application, the terminal residues of BKM and Flu in cucumber were less than 0.02 mg kg(-1). The residual profiles of BKM and Flu suggested that these fungicides could rapidly degrade in cucumber plantations. Their hazard quotient values were all less than 1 on the preharvest intervals of 3, 5, and 7 days, indicating that the dietary risk of BKM and Flu 40 % SC with the recommended usage on cucumber is negligible.

Enzyme-linked immunosorbent assay for the determination of five organophosphorus pesticides in camellia oil.

A matrix solid-phase dispersion and direct competitive enzyme-linked immunosorbent assay (MSPD-ELISA) was developed for five organophosphorus pesticides (OPs) in camellia oil. Seven haptens with different substituents in the aromatic ring were used to prepare different competitors; the ELISA showed highest sensitivity and specificity to OPs when the competitor had moderate heterology to the immunizing hapten. Several assay conditions were optimized to increase the ELISA sensitivity. The optimized ELISA for five OPs had 50% inhibitory concentrations of 6.3 ng/ml (parathion), 18.9 ng/ml (methyl parathion), 120.7 ng/ml (fenitrothion), 110.4 ng/ml (fenthion), and 20.7 ng/ml (phoxim). The average recoveries of five OPs in camellia oil ranged from 75.7 to 105.3%, with the interassay coefficient of variations ranging from 6.0 to 13.4%. Compared with the results previously reported, the ELISA that was developed in the present study showed a much higher sensitivity. Additionally, MSPD was used in the sample preparation to minimize the matrix effect. Recoveries from the method developed here were in agreement with those obtained by gas chromatography, which indicated that the detection performance of the MSPD-ELISA could meet the regulatory requirements of different governments and international organizations.

Development of a Mab-based heterologous immunoassay for the broad-selective determination of organophosphorus pesticides.

A broad-selective monoclonal antibody (Mab) for organophosphorus (OP) pesticides was raised using heterologous indirect enzyme-linked immunosorbent assay (ELISA) to screen hybridomas. On the basis of this Mab, five coating antigens were used to develop homologous and heterologous indirect competitive ELISAs. With the most suitable competitor, a sensitive and broad-selective ELISA was developed. The IC(50) values were estimated to be 20.32 ng/mL for parathion, 21.44 ng/mL for methyl-parathion, 42.15 ng/mL for fenitrothion, and 58.85 ng/mL for isocarbophos. Spike recoveries were between 70.52 and 103.27% for the detection of single pesticide residues of the four OP pesticides in purple-clayed paddy soil. Moreover, the chosen ELISA was then applied to the detection of mixtures of parathion and methyl-parathion in soil samples. The average recovery and coefficient of variation were 80.91 and 4.82%, respectively. Results proved that this broad-selective ELISA would be useful for the multiresidue determination of OP pesticides.