Improved matrix purification using a graphene oxide-coated melamine sponge for UPLC-MS/MS-based determination of 37 veterinary drugs in milks.

A rapid and highly sensitive method was established for the analysis of 37 veterinary drug residues in milk using a modified QuEChERS method based on a reduced graphene oxide-coated melamine sponge (rGO@MeS) coupled with UPLC-MS/MS. Under optimal chromatographic and mass spectrometric conditions, the effects of different dehydrated salts (MgSO4 and Na2SO4) and metal chelating agents (Na2EDTA) on extraction efficiency were first investigated. Next, the influence of a dynamic and static purification mode was evaluated in terms of drug recoveries. Calibration curves of 37 veterinary drugs were constructed in the range 0.6-500 µg kg-1, and good linearities were obtained with all determination coefficients (R2) ≥0.992. The limits of detection (LODs) and quantitation (LOQs) were in the range 0.3-1.1 µg kg-1 and 0.6-3.5 µg kg-1, respectively. The recoveries of all compounds were in the range 61.3-118.2% at three spiked levels (20, 100, and 200 µg kg-1) with RSDs ≤15.4% for both intra- and inter-day precisions. Compared to pristine melamine sponges and commercial adsorbents (C18, PSA, and GCB), rGO@MeS demonstrated an equal or even better purification performance in terms of recoveries, matrix effects, and matrix removal efficiency. This method is rapid, simple, efficient, and appropriate for the qualitative and quantitative analyses of 37 veterinary drug residues in milk, providing a new detection strategy and technical support for the routine analysis of animal-derived food.

The crucial role of Y109 and R162 as catalytic residues of nanoKAZ: insights from molecular docking, molecular dynamics simulation, and quantum chemical investigations.

CONTEXT: nanoKAZ is a compact luciferase that exhibits intense blue light emission when it catalyzes the substrate Furimazine (FMZ) as a luciferin, making it an excellent candidate as a reporter protein. However, the specific catalytic residues and mechanism of nanoKAZ have not been revealed. Recently, the structure of nanoKAZ was determined, and it was observed that the luminescent properties changed when FMZ analogs with naphthalene replacing benzene were used. It is speculated that the substituted naphthalene may influence the interaction between the catalytic residues and luciferins, thereby affecting the energy of the emitted light signal. METHOD: Therefore, the primary objective of this study is to analyze and compare the molecular recognition between nanoKAZ and FMZ along with its four activity-altered naphthalene analogs, with aiming to identify the catalytic residues. Molecular docking was employed to construct all nanoKAZ-luciferin models, followed by a 500 ns molecular dynamics simulation. The simulation trajectory was subjected to MM/PBSA analysis to identify crucial residues that contribute significantly to luciferin binding. In the result, two polar residues Y109, and R162 were identified as active residues as their notable contributions to the binding energy. Subsequently, an oxygen molecule was introduced into the local region of the nanoKAZ-FMZ complex and followed with quantum chemical calculations (semiempirical and DFT methods were used) to investigate the catalysis details. The results illustrated the involvement of Y109 and R162 in the oxygenation of FMZ, leading to the formation of dioxetanone, which has been suggested as an important intermediate in the oxidation process among various luciferins sharing the same functional group as FMZ.

Hydrophobic microporous extraction on polyurethane sponges for convenient and sensitive analysis of organic molecules in water.

RATIONALE: Various solvent supports have been developed to overcome solvent instability during liquid-phase microextraction. The hydrophobic polyurethane sponge (PS) possesses numerous cross-linked internal microchannels and terminal micropores that can facilitate steady solvent storage capacity, high extraction efficiency, extractant loading, and recycling convenience. METHODS: In this study, an easy, convenient, and efficient PS-supported liquid-phase microextraction (PS-LPME) coupled with gas chromatography-mass spectrometry (GC-MS) method was developed for the trace analysis of different organic compounds in aqueous solutions. Different extraction solvents, PS dosages, stirring speeds, and extraction times were first investigated by extracting eight polycyclic aromatic hydrocarbons (PAHs: naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, and pyrene), and then applied for the analysis of triazines, amides, chloroacetamides, and organophosphorus compounds. RESULTS: High enrichment factors (approximately 208-439) were observed for the monitored PAHs. Good linearities, with determination coefficients (r2 ) greater than 0.9992, were achieved in the concentration range of 0.01-50 µg L-1 . Low limits of detection and quantification were found in the ranges of 0.3-3 ng L-1 and 1-10 ng L-1 , respectively. At three spiked concentrations (0.1, 1, and 10 µg L-1 ), good recoveries were obtained in the range of 91.6-118.5% with intra- and inter-day relative standard deviations of less than 6.4% and 11.7%, respectively. CONCLUSIONS: The developed PS-LPME method coupled with GC-MS was successfully applied in the analysis of different organic compounds in aqueous solutions and has shown great convenience and satisfactory enrichment performance in microextraction analysis.

Mortality burden due to long-term exposure to ambient PM2.5 above the new WHO air quality guideline based on 296 cities in China.

OBJECTIVE: Quantifying the spatial and socioeconomic variation of mortality burden attributable to particulate matters with aerodynamic diameter ≤ 2.5 µm (PM2.5) has important implications for pollution control policy. This study aims to examine the regional and socioeconomic disparities in the mortality burden attributable to long-term exposure to ambient PM2.5 in China. METHODS: Using data of 296 cities across China from 2015 to 2019, we estimated all-cause mortality (people aged ≥ 16 years) attributable to the long-term exposure to ambient PM2.5 above the new WHO air quality guideline (5 µg/m3). Attributed fraction (AF), attributed deaths (AD), attributed mortality rate (AMR) and total value of statistical life lost (VSL) by regional and socioeconomic levels were reported. RESULTS: Over the period of 2015-2019, 17.0% [95% confidence interval (CI): 7.4-25.2] of all-cause mortality were attributable to long-term exposure to ambient PM2.5, corresponding to 1,425.2 thousand deaths (95% CI: 622.4-2,099.6), 103.5/105 (95% CI: 44.9-153.3) AMR, and 1006.9 billion USD (95% CI: 439.8-1483.4) total VSL per year. The AMR decreased from 120.5/105 (95% CI: 52.9-176.6) to 92.7/105 (95% CI:39.9-138.5) from 2015 to 2019. The highest mortality burden was observed in the north region (annual average AF = 24.2%, 95% CI: 10.8-35.1; annual average AMR = 137.0/105, 95% CI: 60.9-198.5). The highest AD and economic loss were observed in the east region (annual average AD = 390.0 thousand persons, 95% CI: 170.3-574.6; annual total VSL = 275.6 billion USD, 95% CI: 120.3-406.0). Highest AMR was in the cities with middle level of GDP per capita (PGDP)/urbanization. The majority of the top ten cities of AF, AMR and VSL were in high and middle PGDP/urbanization regions. CONCLUSION: There were significant regional and socioeconomic disparities in PM2.5 attributed mortality burden among Chinese cities, suggesting differential mitigation policies are required for different regions in China.

A facile fluorescent sensor based on nitrogen-doped carbon dots derived from Listeria monocytogenes for highly selective and visual detection of iodide and pH.

Sensitive and visual analysis of iodide (I-) and pH is significant in environmental and food applications. Herein, we present a facile fluorescent sensor for highly selective and visual detection of I- and pH based on nitrogen-doped carbon dots derived from Listeria monocytogenes (NCDs-LM). The NCDs-LM-based fluorescent sensor showed a good linear relationship to I- concentrations, and the detection limit was calculated as 20 nmol L-1. The developed sensor was successfully applied to the detection of I- in drinking water and milk samples. Meanwhile, the as-synthesized NCDs-LM sensor can be used to detect pH, achieving a wide linear pH range. Furthermore, fluorescent test papers based on NCDs-LM were designed for semi-quantitative detection of I- and pH via the naked-eye colorimetric assay. The present work indicates that the NCDs-LM-based fluorescent sensor has high potential for use in environmental monitoring and food analysis.

Application of silanized melamine sponges in matrix purification for rapid multi-residue analysis of veterinary drugs in eggs by UPLC-MS/MS.

Fast and convenient matrix purification is an important prerequisite for high-throughput analysis of drug multiresidues in food. In this study, a silanized melamine sponge was prepared and first applied in the rapid determination of multiclass veterinary drugs in eggs by ultrahigh-performance liquid chromatography-tandem mass spectrometry. Within five seconds, fast, convenient and efficient matrix separation could be achieved through simple soaking and squeezing. Compared to other matrix adsorbents, the developed material demonstrated equivalent or better purification performance. Good validation results were obtained in terms of drug recoveries (61.5%~97.0%, relative standard deviation (RSD) ≤ 10.8%), and linearities (R2 ≥ 0.999), as well as low limits of quantitation (0.3 ~ 10.9 µg·kg-1) and detection (0.1 ~ 3.8 µg·kg-1). By analyzing 52 egg samples, high concentrations of ofloxacin, trimethoprim, metronidazole, and dimetridazole were found at 542.9, 121.2, 66.1 and 58.0 µg·kg-1, respectively. The silanized melamine sponge has shown its great potential for rapid analysis of multiclass residues in food safety.

Development of a modified quick, easy, cheap, effective, rugged, and safe method based on melamine sponge for multi-residue analysis of veterinary drugs in milks by ultra-performance liquid chromatography tandem mass spectrometry.

The purpose of this study was to develop a modified QuEChERS method based on melamine sponge for rapid determination of multi-class veterinary drugs in milks by UPLC-MS/MS. Through simple infiltration and extrusion, fast and convenient matrix purification could be achieved within several seconds, and there was no need of extra phase separation operations. Good linearity with correlation coefficient (R2) ≥0.999 was obtained for all drugs in the range of 2~500 µg·kg-1. The obtained matrix effects were within ±20% for all monitored drugs. The recoveries of all monitored drugs ranged from 60.7% to 116.0% at three spiked levels (50, 100, and 200 µg·kg-1), with relative standard deviations less than 7.4%. Comparatively low LODs and LOQs were obtained in the ranges of 0.1~3.8 µg·kg-1 and 0.2~6.3 µg·kg-1, respectively. Compared with conventional purification adsorbents, melamine sponge yielded an equal or higher purification performance with matrix removal rate as high as 52.5% and acceptable recoveries in range of 60%-120% for all monitored drugs. The satisfactory results have demonstrated the good potential of melamine sponge in matrix purification for rapid determination of multiclass residues in food safety.

Flow controllable three-dimensional paper-based microfluidic analytical devices fabricated by 3D printing technology.

In most cases, three-dimensional paper-based microfluidic analytical devices (3D-µPADs) were fabricated manually by stacking or folding methods. For the first time, digital light processing stereolithography (DLP-SLA) 3D printing technology was adopted to automatically make 3D-µPADs. In the fabrication process, a printing pause was set between two layers to allow paper to be placed in the resin tank. The resin on the fresh paper spontaneously bonded to the former cured paper layer during curing, thus realizing the automatic bonding and alignment between different layers of paper and avoiding the human participation and errors as in stacking and folding methods. There was a gap between two vertical aligned flow paths, therefore the liquid did not flow spontaneously from the upper layer to the lower layer. Most of the fluid flow in 3D-µPADs was spontaneous or manually activated, which was not conducive to complex assays that require different regents to be delivered sequentially. Herein, we used an electric field or airflow to trigger the fluid flow and demonstrated the flow controllability by a proof-of-concept colorimetric assay. The limits of detection of glucose and albumin were 0.8 mM and 3.5 µM, respectively, which were sufficient for clinical requirements. Given the characteristic of flow controllability, we believe that the proposed 3D-µPADs have great potential to make paper-based complex assays automated and programmable.

Low-cost and convenient ballpoint tip-protected liquid-phase microextraction for sensitive analysis of organic molecules in water samples.

Simplification and miniaturization in analytical procedures are highly preferred by analysts and other researchers. In this study, a low-cost, convenient and efficient liquid-phase microextraction method, termed ballpoint tip-protected liquid-phase microextraction (BT-LPME), was established. The bullet-shaped BT possessed a hollow cavity of several microliters for solvent storage and an opening tail for solute extraction. Magnetic field-induced BT spinning significantly accelerated the extraction process. By virtue of the adhesion between a stainless steel sheath and an organic solvent, along with cave protection, finely stable storage of extractant was achieved even under high spinning speeds (>1000 rpm). The BT-LPME performance was evaluated by extracting five polycyclic aromatic hydrocarbons (PAHs) from aqueous solution, followed by gas chromatography-mass spectrometry (GCMS) analysis. Compared to static single drop microextraction (SDME), the BT-LPME method provided higher enrichments (128-173-fold) for the five PAHs. Good linearities (from 0.01∼0.05 µg L-1 to 50 µg L-1) with a regression coefficient (r2) ≥0.9993, as well as low limits of detection (LODs, 0.002-0.011 µg L-1) and limits of quantification (LOQs, 0.007-0.023 µg L-1), were obtained. Relative recoveries varied from 92.3% to 103.4% at three spiked levels of 0.1, 1 and 10 µg L-1. The BT-LPME technique was also successfully applied to the enrichment of other organic compounds, such as organophosphorus compounds, organochlorines and triazines.

Analysis of Compounds Dissolved in Nonpolar Solvents by Electrospray Ionization on Conductive Nanomaterials.

Electrospray ionization mass spectrometry (ESI-MS) technique has limitations in analysis of compounds that are dissolved in nonpolar solvents. In this study, ambient ionization of compounds in solvents that are not "friendly" to electrospray ionization, such as n-hexane, is achieved by conductive nanomaterials spray ionization (CNMSI) on nanomaterial emitters, including carbon nanotubes paper and mesodendritic silver covered metal, which applies high voltages to emitters made of these materials without the assistance of polar solvents. Although the time intensity curves (TIC) commonly vary from 4.5% to 23.7% over analyses, protonated molecular ions were found to be the most abundant species, demonstrating good reproducibility of the technique in terms of ionized species. Higher mass spectrometric responses are observed in analyzing nonpolar systems than polar systems. 2-Methoxyacetophenone, 4-methylacetophenone, benzothiazole, quinolone, and cycloheptanone as low as 2 pg in n-hexane can be directly detected using the developed method. The developed technique expands the analysis capability of ESI-MS for direct, online analysis of nonpolar systems, such as low polarity extracts, normal phase liquid chromatography eluates, and synthetic mixtures. Graphical Abstract.

Determination of Sulfonamides in Chicken Muscle by Pulsed Direct Current Electrospray Ionization Tandem Mass Spectrometry.

A simple and rapid approach for the simultaneous detection of trace amounts of six sulfonamides in chicken muscle was developed using pulsed direct current electrospray ionization tandem mass spectrometry (pulsed-dc ESI-MS/MS). The pretreatment of chicken muscle samples consisted of two steps: acetonitrile extraction and n-hexane delipidation. Sulfonamides do not need to be derivatized or chromatographed prior to pulsed-dc ESI-MS/MS. The factors affecting the performance of pulsed-dc ESI-MS/MS were studied. Under optimum conditions, the quantitative performance of pulsed-dc ESI-MS/MS was validated according to European Union Decision 2002/657/EC, and the sensitivity of pulsed-dc ESI-MS/MS was 3 times higher than that of ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The limits of detection obtained by pulsed-dc ESI-MS/MS were in the range of 0.07-0.11 µg/kg. The proposed method was simple, rapid, and sensitive, and was successfully used for quantitation and rapid screening of sulfonamides in real chicken muscle samples.

Application of fractionized sampling and stacking for construction of an interface for online heart-cutting two-dimensional liquid chromatography.

In this study, an efficient interface, based on a fractionized sampling and stacking (FSS) strategy, was developed for online heart-cutting two-dimensional liquid chromatography (2D LC). This interface consisted of a two-position 4-port valve, a two-position 6-port valve and a two-position 10-port valve equipped with two 450-µL stainless steel loops. Hydrophilic interaction chromatography (HILIC) and reversed phase liquid chromatography (RP LC) were used in the first and second dimensions, respectively. The peak compression efficiency of this interface was investigated by analysis of ten standards. Good peak shapes of the ten standards were observed when the dilution ratio was over five and the co-eluate plug volume was less than 10µL. The 2D LC system was further applied to analysis of a crude extract of Panax ginseng leaves. Seventeen major constituents in the extract were monitored, which could not be well separated by one-dimensional (1D) HILIC or RP LC method in a long separation gradient. The FSS interface successfully achieved the efficient combination of HILIC and RP LC, and the 17 constituents in ginseng extract got well separated under the optimized conditions. The FSS interface has shown great potential for 2D LC analysis of complex natural product samples.

Generating Electrospray Ionization on Ballpoint Tips.

In this study, we report a simple and economical ballpoint electrospray ionization mass spectrometry (BP-ESI-MS) technique. This combines a small ballpoint tip with a syringe pump for the direct loading and ionization of various samples in different phases (including solution, semisolid, and solid) and allows for additional applications in surface analysis. The tiny metal ball on the ballpoint tip exhibits a larger surface for ionization than that of a conventional sharp tip end, resulting in higher ionization efficiency and less sample consumption. The adamant properties of the ballpoint tip allow sampling by simply penetrating or scraping various surfaces, such as a fruit peel, paper, or fabric. Complex samples, such as fine herbal powders and small solid samples, could be stored in the hollow space in the ballpoint socket and subsequently extracted online, which greatly facilitated MS analysis with little to no sample preparation. Positive ion mode was attempted, and various compounds, including amino acids, carbohydrates, flavonoids, and alkaloids, were detected from different types of samples. The results demonstrated that the special and excellent physical characteristics of ballpoint tips allowed for fast and convenient sampling and ionization for mass spectrometry analysis by the BP-ESI-MS method.

Development and characterization of a nanodendritic silver-based solid-phase extraction sorbent for selective enrichment of endocrine-disrupting chemicals in water and milk samples.

In this study, 4-[4-phenylazo-phenoxy] butyl-1-thiol (AzSH) functionalized nanodendritic silver (AzS@AgNDs) materials were prepared as a solid-phase extraction (SPE) sorbent for the selective extraction of estrogens. AzS@AgNDs possess an extremely large surface-to-volume ratio and a small average particle size. The performance of the material was evaluated by selective enrichment of hexestrol, diethylstilbestrol, dienestrol and bisphenol A in water and milk samples followed by rapid ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) analyses. The results exhibited that AzS@AgNDs had excellent adsorption capability for the targeted estrogens. The limits of detection of the four estrogens ranged from 0.1 to 5.0 pg/mL. The recoveries of the estrogens spiked into tap water were over the range of 83.6-105.3% with relative standard deviations of 2.8-6.0%. The results indicated the capability of this method for the rapid determination of estrogens in milk and other environmental water samples. In addition, this method would be useful for the determination of human exposure and health risk assessments trace level of endocrine-disrupting compounds (EDCs) in drinking water.

Strategy to rapidly discriminate trace isomeric lignan compounds from Gymnotheca chinensis by probe electrospray ionization tandem mass spectrometry.

Probe electrospray ionization (PESI) is a recently developed ionization technique based on electrospray ionization (ESI) that generates electrospray from the tip of a solid needle. High tolerance to salts, requirements of a trace amount of sample and direct ambient sampling- are major advantages of PESI compared with conventional ESI. In this report, three pairs of isomeric lignans bearing tetra-hydrofuran with variable conformations from Gymnotheca chinensis were investigated by probe electrospray tandem mass spectrometry (PESI-MS/MS) in the positive ion mode. The diagnostic characteristics of these compounds were obtained and the isomers could be successfully distinguished by comparison with their breakdown curves, even though the isomers differed only in the conformation of some groups of the isomer pairs. This report provides a rapid and reliable method for the identification of trace amounts of isomeric lignans by PESI-MS/MS. Furthermore, application of PESI and breakdown curves should have value in mass spectrometry studies of isomeric natural products compounds.