HRMS analysis of pesticides in vegetables from Shanghai and risk assessment.

A rapid analytical method for the simultaneous determination of 550 pesticide residues in vegetable samples was developed based on ultra-high performance liquid chromatography-tandem Q/Orbitrap high-resolution mass spectrometry (UPLC-Q/Orbitrap-HRMS). To investigate the risk of exposure to pesticide residues through vegetable consumption, 704 leafy vegetable samples from Shanghai were analysed for multiple residues using this method. A total of 54 pesticide residues were identified in these vegetable samples and 302 samples contained one or more pesticide residue. The levels of the detected pesticides did not pose a health risk in the long term and were acceptable according to the results of the chronic dietary risk assessment. Risk rankings displayed that most of the pesticides were low to medium risk. The findings of this study provide a reference for future pesticide monitoring programmes.

Integrative transcriptome and metabolome analysis reveals the mechanisms of light-induced pigmentation in purple waxy maize.

Introduction: Waxy maize, mainly consumed at the immature stage, is a staple and vegetable food in Asia. The pigmentation in the kernel of purple waxy maize enhances its nutritional and market values. Light, a critical environmental factor, affects anthocyanin biosynthesis and results in pigmentation in different parts of plants, including in the kernel. SWL502 is a light-sensitive waxy maize inbred line with purple kernel color, but the regulatory mechanism of pigmentation in the kernel resulting in purple color is still unknown. Methods: In this study, cyanidin, peonidin, and pelargonidin were identified as the main anthocyanin components in SWL502, evaluated by the ultra-performance liquid chromatography (UPLC) method. Investigation of pigment accumulation in the kernel of SWL502 was performed at 12, 17, and 22 days after pollination (DAP) under both dark and light treatment conditions via transcriptome and metabolome analyses. Results: Dark treatment affected genes and metabolites associated with metabolic pathways of amino acid, carbohydrate, lipid, and galactose, biosynthesis of phenylpropanoid and terpenoid backbone, and ABC transporters. The expression of anthocyanin biosynthesis genes, such as 4CL2, CHS, F3H, and UGT, was reduced under dark treatment. Dynamic changes were identified in genes and metabolites by time-series analysis. The genes and metabolites involved in photosynthesis and purine metabolism were altered in light treatment, and the expression of genes and metabolites associated with carotenoid biosynthesis, sphingolipid metabolism, MAPK signaling pathway, and plant hormone signal transduction pathway were induced by dark treatment. Light treatment increased the expression level of major transcription factors such as LRL1, myc7, bHLH125, PIF1, BH093, PIL5, MYBS1, and BH074 in purple waxy maize kernels, while dark treatment greatly promoted the expression level of transcription factors RVE6, MYB4, MY1R1, and MYB145. Discussion: This study is the first report to investigate the effects of light on waxy maize kernel pigmentation and the underlying mechanism at both transcriptome and metabolome levels, and the results from this study are valuable for future research to better understand the effects of light on the regulation of plant growth.

Multi-Residue Screening of Pesticides in Aquatic Products Using High-Performance Liquid Chromatography-Tandem High-Resolution Mass Spectrometry.

Pesticide residues in aquatic products are of great concern due to the risk of environmental transmission and their extensive use in aquaculture. In our work, a quick screening approach was developed for the qualitative and semi-quantitative screening of 87 pesticide residues in aquatic products. The sample preparation was investigated, including extract solvent, extract methods, buffer salts, lipid removal, cleanup materials and filter membranes for aquatic products. Samples were extracted using a modified QuEChERS procedure, and two clean-up procedures were developed for UHPLC-Q/Orbitrap MS analysis based on the fat content of the aquatic products. The screening detection limits for all studied pesticides were distributed between 1 and 500 µg/kg in the three representative matrices. Seventy-one pesticides could be analyzed with a screening limit between 1 and 25 µg/kg in grass carp and crayfish, sixty-one pesticides could be screened for limits between 1 and 50 µg/kg in crab. The accuracy results showed that recoveries ranged from 50 to 120% for 60, 56 and 52 pesticides at medium-level for grass carp, crayfish and crab, respectively. At high spiking levels, 74, 65 and 59 pesticides were recovered within the range of 50-120% for the three matrices, respectively. The relative standard deviations of most compounds in different matrices were less than 20%. With this method, the local farmed aquatic products were tested for pesticide residues. In these samples, ethoxyquinoline, prometryn and phoxim were frequently detected. The majority of these confirmed compounds did not exceed 2.00 µg/kg. A grass carp with trichlorfon at 4.87 µg/kg and two carps with ethoxyquinoline at 200 µg/kg were detected, indicating the potential dietary risk.

Improved LC/MS/MS Quantification Using Dual Deuterated Isomers as the Surrogates: A Case Analysis of Enrofloxacin Residue in Aquatic Products.

Extensive and high residue variations in enrofloxacin (ENR) exist in different aquatic products. A novel quantitative method for measuring ENR using high-performance liquid chromatography-tandem mass spectrometry was developed employing enrofloxacin-d5 (ENR-d5) and enrofloxacin-d3 (ENR-d3) as isotope surrogates. This reduced the deviation of detected values, which results from the overpass of the linear range and/or the large difference in the residue between the isotope standard and ENR, from the actual content. Furthermore, high residue levels of ENR can be directly diluted and re-calibrated by the corresponding curve with the addition of high levels of another internal surrogate without repeated sample preparation, avoiding the overflow of the instrument response. The validation results demonstrated that the method can simultaneously determine ENR residues from MQL (2 µg/kg) to 5000 × MQL (method quantification limit) with recoveries between 97.1 and 106%, and intra-precision of no more than 2.14%. This method realized a wide linear calibration range with dual deuterated isomers, which has not been previously reported in the literature. The developed method was successfully applied to the analysis of ENR in different aquatic products, with ENR residue levels varying from 108 to 4340 µg/kg and an interval of precision in the range of 0.175~6.72%. These results demonstrate that batch samples with a high variation in ENR residues (over the linear range with a single isotope standard) can be detected by the dual isotope surrogates method in a single sample preparation process.

H-beta zeolite-based dispersive solid-phase strategy for the multi-residue determination of pesticides.

In this work, electrostatic interactions, hydrogen bonding and other interactions between a variety of pesticides and H-beta zeolite are discussed based on thermodynamics (Gibbs free energy, enthalpy, and entropy). Combined with the physical properties of the target compounds (octanol-water partition coefficient and molar volume), the characteristics of the physical adsorption on the synthetic zeolite were analyzed, the results of H-beta zeolite adsorption showed that the adsorption rate had a nonlinear relationship with the physical properties, and the adsorption reaction was dominated by the adsorption on the surface and in the pores of the H-beta zeolite. Based on the principle of zeolite adsorption, the parameters of absorbent amount, adsorption time, sample pH, ion strength, sample volume and desorption solvent were optimized, a vortex-assisted dispersive solid-phase extraction procedure coupled with liquid chromatography-tandem mass spectrometry was developed for the analysis of multiple pesticides in surface water. The sample was adjusted by phosphate buffer or hydrochloric acid to pH 4, then adsorbed by 50 mg of zeolite within 10 min, adding 2% (g/v) sodium chloride to the solution. The absorbent was eluted with a mixture of acetonitrile-methanol containing 0.5% formic acid after separation, then filtered prior to analysis. The optimized vortex-assisted dispersive solid-phase extraction method simultaneously achieved a low adsorbent dosage, a high enrichment factor of 25, rapid pre-concentration for 30 min, a low solvent usage of 2 mL and multi-target simultaneous analysis. Method performance in terms of accuracy, precision, limit of detection, and limit of quantitation was performed to validate the reliability of the procedure. The proposed protocol achieved acceptable accuracy (recoveries between 62% and 107%), and precision (relative standard deviation < 20%). The limit of detection of the method was below 0.1 µg/L, and the limit of quantitation was 0.08 µg/L or 0.2 µg/L. The results indicated that zeolite is a promising adsorbent with high adsorption capacity. It has the advantages of being a fast, simple, green and economical method for determining residual pesticides in water.

Pesticides and risk assessment in Shanghai fruit and raw eaten vegetables.

A database of 392 pesticides established by an Ultrahigh performance liquid chromatography-tandem Q/Orbitrap high-resolution mass spectrometer (UPLC-Q/Orbitrap-HRMS) was used to screen multiple residues of pesticides in fruit and raw eaten vegetables from planting farms in Shanghai. Risk assessment was conducted with the screened results of the determined pesticides as to evaluate food safety. In 95% of the samples, one or more pesticides had a content below the maximum residue limits (MRLs) as set in the national Chinese standard. The co-occurrence of multi-residues of pesticides was more severe in peach and muskmelon, when compared with other food. All hazard index values of different groups were in the range of 0.19% to 12.3%, demonstrating that chronic dietary risk of studied fruits and raw eaten vegetables is low and the studied food samples were safe for human consumption in terms of these detected pesticides.

Determination and dietary risk assessment of 284 pesticide residues in local fruit cultivars in Shanghai, China.

The presence of pesticide residues has become one of the main risk factors affecting the safety and quality of agro-food. In this study, a multi-residue method for the analysis of 284 pesticides in five local fruit cultivars in Shanghai was developed based on ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS). The limits of determination and the limits of quantitation of pesticides were 0.6-10 and 2-30 µg/kg, respectively. A total of 44, 10, 10, 18, and 7 pesticides were detected in strawberries, watermelons, melons, peaches, and grapes, respectively. The pesticide levels in 95.0% of the samples were below the maximum residual limits (MRLs) prescribed by China, and in 66.2% of the samples below the EU MRLs. The dietary risk assessment study showed big differences in the chronic and acute exposure risk values among different Chinese consumer groups. Through fruit consumption, children/females showed higher exposure risks than adults/males. But both the risk values were less than 100%, indicating that potential dietary risk induced by the pesticides was not significant for Chinese consumers. Nevertheless, certain measures are needed for both growers and the government in order to decrease the MRL-exceeding rate of pesticide residues and ensure the quality and safety of fruits for consumers.

Melatonin enhances the cadmium tolerance of mushrooms through antioxidant-related metabolites and enzymes.

Many wild mushroom species are known to immune heavy metals. However, the mechanism by which this occurs remains largely unsolved. Melatonin (MT) has been proven to play an important defensive role against various abiotic stresses in plants and animals. This study reports on the presence of MT in edible fungi and its role in the response to cadmium (Cd)-induced stress. We found that melatonin was widely distributed in all experimental species and could also relieve Cd-induced damage in the Volvariella volvacea. Comparative metabolic and proteome analyses reveal that tryptophan/proline/tyrosine metabolism, the citrate cycle, nitrogen and glutathione metabolism, and oxidation-reduction processes were enriched after Cd and/or MT addition, indicating an antioxidant mechanism was aroused. Finally, different MT and cadmium treatments were studied for their effects on the expression and activity of oxidation-related enzymes (superoxide dismutase, catalase, peroxidase, etc), which further verified the ameliorative influence of MT on Cd-induced oxidative stress.

Determination of type A trichothecenes in coix seed by magnetic solid-phase extraction based on magnetic multi-walled carbon nanotubes coupled with ultra-high performance liquid chromatography-tandem mass spectrometry.

Magnetic solid-phase extraction (m-SPE) is a promising sample preparation approach due to its convenience, speed, and simplicity. For the first time, a rapid and reliable m-SPE approach using magnetic multi-walled carbon nanotubes (m-MWCNTs) as the adsorbent was proposed for purification of type A trichothecenes including T-2 toxins (T2), HT-2 toxins (HT-2), diacetoxyscirpenol (DAS), and neosolaniol (NEO) in coix seed. The m-MWCNTs were synthesized by assembling the magnetic nanoparticles (Fe3O4) with MWCNTs by sonication through an aggregation wrap mechanism, and characterized by transmission electron microscope. Several key parameters affecting the performance of the procedure were extensively investigated including extraction solutions, desorption solvents, and m-MWCNT amounts. Under the optimal sample preparation conditions followed by analysis with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), high sensitivity (limit of quantification in the range of 0.3-1.5 µg kg(-1)), good linearity (R (2) > 0.99), satisfactory recovery (73.6-90.6 %), and acceptable precision (≤2.5 %) were obtained. The analytical performance of the developed method has also been successfully evaluated in real coix seed samples. Graphical Abstract Flow chart of determination of type A trichothecenes in coix seed by magnetic solid-phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry.

An efficient method for the synthesis of pyranoid glycals.

A simple and efficient procedure was designed for the preparation of pyranoid glycals. In a novel fashion, a series of protected glycopyranosyl bromides underwent reductive elimination in the presence of zinc dust and ammonium chloride in CH3CN at 30-60 °C. The corresponding glycals were obtained with excellent isolated yields (72-96%) in a short time (20-50 min). Furthermore, the transformation was compatible with different protection patterns and conveniently scalable (86% for 45 g acetobromoglucose) which made it very applicable in organic synthesis.

Ion-exchange solid-phase extraction combined with liquid chromatography-tandem mass spectrometry for the determination of veterinary drugs in organic fertilizers.

The analysis of veterinary drugs in organic fertilizers is crucial for an assessment of potential risks to soil microbial communities and human health. We develop a robust and sensitive method to quantitatively determine 19 veterinary drugs (amantadine, sulfonamides and fluoroquinolones) in organic fertilizers. The method involved a simple solid-liquid extraction step using the combination of acetonitrile and McIlvaine buffer as extraction solvent, followed by cleanup with a solid-phase extraction cartridge containing polymeric mixed-mode anion-exchange sorbents. Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to separate and detect target analytes. We particularly focused on the optimization of sample clean-up step: different diluents and dilution factors were tested. The developed method was validated in terms of linearity, recovery, precision, sensitivity and specificity. The recoveries of all the drugs ranged from 70.9% to 112.7% at three concentration levels, with the intra-day and inter-day relative standard deviation lower than 15.7%. The limits of quantification were between 1.0 and 10.0µg/kg for all the drugs. Matrix effect was minimized by matrix-matched calibration curves. The analytical method was successfully applied for the survey of veterinary drugs contamination in 20 compost samples. The results indicated that fluoroquinolones had higher incidence rate and mean concentration levels ranging from 31.9 to 308.7µg/kg compared with other drugs. We expect the method will provide the basis for risk assessment of veterinary drugs in organic fertilizers.

Multi-walled carbon nanotubes as solid-phase extraction sorbents for simultaneous determination of type A trichothecenes in maize, wheat and rice by ultra-high performance liquid chromatography-tandem mass spectrometry.

A solid-phase extraction (SPE) procedure using multi-walled carbon nanotubes (MWCNTs) as sorbents coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed for simultaneous determination of four type A trichothecenes in maize, wheat and rice for the first time. Several key parameters including the composition of sample loading solutions, washing and elution solvents were thoroughly investigated to achieve optimal SPE recoveries and efficiency. Performance of the MWCNTs materials was significantly affected by pH, and after optimization, n-hexane and 5% methanol aqueous solution as the washing solutions and methanol containing 1% formic acid as the elution solvent presented an excellent purification efficiency for the four targets in the different matrices. The method was validated by determining the linearity (R(2)≥0.992), recovery (73.4-113.7%), precision (1.2-17.1%) and sensitivity (limit of quantification in the range of 0.02-0.10µg/kg), and was further applied for simultaneous determination of type A trichothecenes in 30 samples. Although low contamination levels of type A trichothecenes in wheat, maize and rice were observed revealing mitigated risks to consumers in Shanghai, China, the developed method has proven to be a valuable tool for type A trichothecenes monitoring in complex crop matrices.

A highly efficient deprotection of the 2,2,2-trichloroethyl group at the anomeric oxygen of carbohydrates.

Commercially available zinc dust in the presence of ammonium chloride in acetonitrile at reflux removes the 2,2,2-trichloroethyl (TCE) group at anomeric centers with excellent yields (>95%) in short reaction times. This present method is easily implemented on substrates containing acyl and benzyl groups and large-scale reactions also proceed in high yield.