Systematic study of tissue section thickness for MALDI MS profiling and imaging.

Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) has become a powerful method for studying the spatial distribution of molecules. Preparation of tissue sections is a critical step for obtaining high-quality imaging data. The thickness of the slice of tissue affects the feature quality of MALDI MSI. However, few studies involved in-depth and systematic examination of slice thickness. Herein, we investigate the effect of tissue slice thickness on MALDI MSI detection. We found that the thicker the slice, the worse the results obtained by MALDI MS, which we attributed to the charging effect. The optimal slice thickness of brain tissue obtained in this work is 2-6 µm. Comparisons of the effects of slice thickness on atmospheric pressure and vacuum MALDI assays indicated that the ion signals and imaging quality of vacuum MALDI were more seriously affected by the thickness, with atmospheric pressure (AP) MALDI having a greater tolerance for slice thickness than vacuum MALDI. The MALDI MSI of peptides after enzymatic digestion of tissue sections of different thicknesses was also studied, revealing that the most suitable tissue thickness for enzyme digestion is about 10 µm. Finally, we optimized the slice thicknesses of six tissues in mice to provide a reference for MALDI MSI studies. It is worth mentioning that in our study the values of slice thickness range from the nanometer level (400 nm) at the minimum to 150 µm at the maximum, values which were unprecedented. Detailed in-depth and systematic studies of slice thickness will promote the development of sample preparation technology of AP and vacuum MALDI MSI, which will provide important references for the selection of tissue section thickness.

Fluorinated covalent organic frameworks as a stationary phase for separation of fluoroquinolones by capillary electrochromatography.

A fluorinated covalent organic framework composed of 2,3,5,6-tetrafluoroterephthaldehyde (TFA) and 1,3,5-tri(4-aminophenyl)benzene (TAPB) is proposed for electrochromatographic separation. TFA-TAPB is for the first time regarded as the stationary phase of capillary electrochromatography. The TFA-TAPB-coated capillary columns exhibited satisfactory separation (resolution values > 1.5) and good reproducibility towards fluoroquinolones. The intraday relative standard deviations (RSDs) of retention time and peak areas were 0.54-0.68% and 1.69-2.82%, respectively. The interday RSDs of retention time and peak areas were less than 1.79% and 2.30%, respectively. The column-to-column RSDs of retention time and peak areas were 0.22-0.73% and 0.74-1.86%, respectively. And, the inter-batch RSDs of retention time and peak areas were less than 0.39% and 1.67%, respectively. Moreover, the possible separation mechanism was discussed, and it was found that the π-π stacking effect, hydrophobic interaction, hydrogen bonding, and fluorous interactions were the main factors. Overall, these results demonstrated that TFA-TAPB has high prospect for CEC separation.

Molybdenum disulfide-graphene oxide composites as dispersive solid-phase extraction adsorbents for the enrichment of four paraben preservatives in cosmetics.

Molybdenum disulfide-graphene oxide composite (MoS2/GO) was synthesized and used as the adsorbent in dispersive solid-phase extraction. Four paraben preservatives, namely, methylparaben, ethylparaben, propylparaben, and butylparaben, were enriched with MoS2/GO and determined by ultra-high-performance liquid chromatography. Molybdenum disulfide was intercalated into graphene oxide layers to reduce self-aggregation by using the solvothermal method. The experimental results indicated that the as-prepared MoS2/GO composite exhibited great enrichment capability toward those four paraben preservatives, and the adsorption time was 10 min and the elution time was as short as 1 min. The mechanism of MoS2/GO composite and parabens is attributed to hydrogen bonding and electrostatic attraction. The relative standard deviation (RSD, n = 9) of this method was below 7.6%. Limits of detection and limits of quantification were in the range 0.4-2.3 ng/mL and 1.4-7.6 ng/mL, respectively. The recoveries obtained from the parabens of cosmetic sample were in the range 91.3-124% with RSDs below 10%. The developed method has great potential for the determination of emerging contaminants with low cost and high sensitivity.

Ambient temperature fabrication of a covalent organic framework from 1,3,5-triformylphloroglucinol and 1,4-phenylenediamine as a coating for use in open-tubular capillary electrochromatography of drugs and amino acids.

A covalent organic framework (COF) named TpPa-1 was designed and synthesized at ambient temperature by an ultrasound-assisted method from 1,3,5-triformylphloroglucinol (Tp) and 1,4-phenylenediamine (Pa-1). It was utilized as a stationary phase in open-tubular capillary electrochromatography (OT-CEC). The column was coated with TpPa-1 using a covalent bonding strategy. The coated capillary was characterized by morphology, crystallography, and mesoporous analysis to confirm the successful fabrication. The OT-CEC method was utilized for the analysis of tetracyclines, sulfonamides, cephalosporins and amino acids with high-resolution (Rs > 1.81) and good precision (RSD < 4.9%). It takes about 12 h from COF preparation to OT-CEC separation. Graphical abstract A covalent organic framework (COF) named TpPa-1 was synthesized at ambient temperature by an ultrasound-assisted method from 1,3,5-triformylphloroglucinol (Tp) and 1,4-phenylenediamine (Pa-1). COF-TpPa-1 modified capillary column was utilized for the analysis of tetracyclines, sulfonamides, cephalosporins and amino acids with high-resolution and good precision.

Open-tubular capillary electrochromatographic determination of ten sulfonamides in tap water and milk by a metal-organic framework-coated capillary column.

In this study, a metal-organic framework (MOF), [Mn(cam)(bpy)], was synthesized and characterized by thermogravimetric analysis, scanning electron microscopy, and Fourier transform infrared spectrometry. An open-tubular capillary column was fabricated from [Mn(cam)(bpy)] via the amide coupling method. Ten types of sulfonamides were separated through the fabricated capillary column, which showed a good limits of detection (<0.07 µg/mL) and linear ranges (1-100 or 5-100 µg/mL) with a high correlation coefficients (R2 > 0.9987). The intra-day, inter-day and column-to-column relative standard deviations (RSDs) in the migration times ranged from 0.44 to 4.87%, and the peak area RSDs ranged from 0.80 to 7.28%. The developed capillary electrochromatography method can be successfully utilized for the determination of sulfonamides in tap water and milk samples.

Recent advances in metal-organic frameworks and covalent organic frameworks for sample preparation and chromatographic analysis.

In the field of analytical chemistry, sample preparation and chromatographic separation are two core procedures. The means by which to improve the sensitivity, selectivity and detection limit of a method have become a topic of great interest. Recently, porous organic frameworks, such as metal-organic frameworks (MOFs) and covalent organic frameworks (COFs), have been widely used in this research area because of their special features, and different methods have been developed. This review summarizes the applications of MOFs and COFs in sample preparation and chromatographic stationary phases. The MOF- or COF-based solid-phase extraction (SPE), solid-phase microextraction (SPME), gas chromatography (GC), high-performance liquid chromatography (HPLC) and capillary electrochromatography (CEC) methods are described. The excellent properties of MOFs and COFs have resulted in intense interest in exploring their performance and mechanisms for sample preparation and chromatographic separation.

Covalent bonding of homochiral metal-organic framework in capillaries for stereoisomer separation by capillary electrochromatography.

In this work, a [Cu(mal)(bpy)]⋅H2O (mal, L-(-)-malic acid; bpy, 4,4'-bipyridyl) homochiral metal-organic frameworks (MOFs) was synthesized and used for modifying the inner walls of capillary columns by utilizing amido bonds to form covalent links between the MOFs particles and capillary inner wall. The synthesized [Cu(mal)(bpy)]⋅H2 O and MOFs-modified capillary column were characterized by X-ray diffraction, thermogravimetric analysis, particle size distribution analysis, nitrogen absorption characterization, FTIR spectroscopy, SEM, and energy-dispersive X-ray spectroscopy (EDX). The MOFs-modified capillary column was used for the stereoisomer separation of some drugs. The LODs and LOQs of six analytes were 0.1 and 0.25 µg/mL, respectively. The linear range was 0.25-250 µg/mL for ephedrine, 0.25-250 µg/mL for pseudoephedrine, 0.25-180 µg/mL for D-penicillamine, 0.25-120 µg/mL for L-penicillamine, 0.25-180 µg/mL for D-phenylalanine, and 0.25-160 µg/mL for L-phenylalanine, all with R(2) > 0.999. Finally, the MOFs-modified capillary column was applied for the analysis of active ingredients in a real sample of the traditional Chinese medicine ephedra.

Cytokinins can act as suppressors of nitric oxide in Arabidopsis.

Maintaining nitric oxide (NO) homeostasis is essential for normal plant physiological processes. However, very little is known about the mechanisms of NO modulation in plants. Here, we report a unique mechanism for the catabolism of NO based on the reaction with the plant hormone cytokinin. We screened for NO-insensitive mutants in Arabidopsis and isolated two allelic lines, cnu1-1 and 1-2 (continuous NO-unstressed 1), that were identified as the previously reported altered meristem program 1 (amp1) and as having elevated levels of cytokinins. A double mutant of cnu1-2 and nitric oxide overexpression 1 (nox1) reduced the severity of the phenotypes ascribed to excess NO levels as did treating the nox1 line with trans-zeatin, the predominant form of cytokinin in Arabidopsis. We further showed that peroxinitrite, an active NO derivative, can react with zeatin in vitro, which together with the results in vivo suggests that cytokinins suppress the action of NO most likely through direct interaction between them, leading to the reduction of endogenous NO levels. These results provide insights into NO signaling and regulation of its bioactivity in plants.

Determination of dopamine, epinephrine, and norepinephrine by open-tubular capillary electrochromatography using graphene oxide molecularly imprinted polymers as the stationary phase.

A novel capillary electrochromatography method was developed for the determination of dopamine (DA), epinephrine (EP), and norepinephrine (NE) by using a graphene oxide (GO) molecularly imprinted polymers (MIPs) coated capillary. In this article, GO was introduced as supporting matrix to synthesize MIPs in the presence of DA as template molecule. Then GO MIPs were used as the stationary phase in electrochromatography for the determination of DA, EP, and NE. The separation of these three analytes was achieved under the optimal conditions with a satisfactory correlation coefficients (R(2) ) > 0.9957 in the range of 5.0-200.0 µg/mL for EP and NE, and 20.0-200.0 µg/mL for DA, respectively. The RSDs for the determination of three analytes were <6.19%, and the detection limits were 1.25 µg/mL for EP and NE, and 10.0 µg/mL for DA, respectively. Finally, this method was used for the determination of DA, EP, and NE in human serum and DA hydrochloride injection.

Molybdenum disulfide nanosheet induced reactive oxygen species for high-efficiency luminol chemiluminescence.

BACKGROUND: Luminol chemiluminescence (CL) sensing system remains an excellent candidate for application in bioanalysis due to its good water solubility. However, traditional luminol CL system usually requires the addition of oxidizing agents and sensitizers to obtain high efficiency for the improvement of detection sensitivity. Although numerous studies on the nanomaterial-enhanced luminol CL systems have been carried out, there is still great potential to develop inexpensive, readily available and easily handled catalysts to construct simple and effective CL platform for biomolecular sensing. RESULTS: Few-layered MoS2 nanosheets (NS) prepared by sonication-assisted exfoliation of commercially available bulk MoS2 were found to significantly enhance the CL of luminol‒dissolved oxygen in the absence of additional oxidants. The mechanism study demonstrated that exfoliated MoS2 NS could catalyze the decomposition of dissolved oxygen by virtue of its exposed active sites on the surface, generating increased reactive oxygen intermediates, which then oxidize luminol and produce intense CL emission. The proposed high-efficiency luminol CL system was then employed for the extremely sensitive identification of dopamine based on the quenching of CL by dopamine. The limit of detection (LOD) for dopamine can be as low as 2.07 nM. Besides, it also works well in the actual urine sample with good recoveries (99.6-100.6 %), confirming the practicability of the method. SIGNIFICANCE: As a new type of CL catalyst, MoS2 NS developed in this work are easy to obtain, simple to prepare and can be produced in large quantities, which lays a foundation for extending applicability of MoS2 NS in the CL field, and provides a new idea for developing simple and highly sensitive CL sensing system.

A ratiometric fluorescent dark box and smartphone integrated portable sensing platform based on hydrogen bonding induction for on-site determination of enrofloxacin.

Enrofloxacin (ENR) residues in animal-derived food and water threaten human health. Simple, low-cost and on-site detection methods are urgently needed. Blue emitting carbon quantum dots (CQDs) and orange rhodamine B (RhB) were used as recognition and reference signals, respectively, to construct a ratiometric fluorescence sensor. After the addition of ENR, the color of the sensor changed from orange to blue because hydrogen bonding induced a considerable increase in CQDs fluorescence. Based on this mechanism, a simple and low cost on-site portable sensing platform was constructed, which integrated a stable UV light strip and a smartphone with voice-controlled phototaking function and an RGB app. The t-test results of spiked ENR recoveries for diluted milk, honey and drinking water revealed no significant differences between the ratiometric fluorescent sensor and portable sensing platform. Thus, this portable sensing platform provides a novel strategy for on-site quantification of quinolone antibiotics in foodstuffs and environmental water.

Graphene oxide coated capillary for chiral separation by CE.

This article describes a new application of graphene oxide (GO) in CE based on the coating of fused silica capillary for chiral separation. The coated capillary was characterized by SEM, energy dispersive X-ray spectroscopy, and Raman spectra. The results indicated that the capillary was coated with GO. Chiral separations were carried out in the GO-coated capillary for the ephedrine-pseudoephedrine (E-PE) isomers and ß-methylphenethylamine (ß-Me-PEA) isomers at the optimal buffer conditions without any chiral selector by CE. The precision and reproducibility of GO-coated capillary were investigated, and the RSDs of migration time (n = 6) for the E-PE isomers were 1.35-1.41%, and 0.97-3.50% for ß-Me-PEA isomers, respectively. The LOD for E-PE isomers and ß-Me-PEA isomers was 3 µM and 18 µM, respectively.

Simultaneous determination of atropine, scopolamine, and anisodamine in Flos daturae by capillary electrophoresis using a capillary coated by graphene oxide.

A novel CE method was developed for the separation and determination of three main tropane alkaloids in Flos daturae with a capillary coated by graphene oxide (GO). The GO-coated capillary was characterized by SEM, energy dispersive X-ray spectroscopy, and Raman spectroscopy, and the results indicated that the inner surface of the capillary was partially coated by GO. A phosphate solution (40 mM, pH7.0) containing 20% v/v methanol and 30% v/v acetonitrile was used as the running buffer for the analysis of the atropine, scopolamine, and anisodamine. The linear ranges of atropine, scopolamine, and anisodamine was 0.5-200 µg/mL with satisfactory correlation coefficients (R(2)) > 0.9987, and this novel method provided an efficient separation for three tropane alkaloids as well as a good reproducibility and stability. Finally, the method was successfully applied for the determination of these three tropane alkaloids in plant extracts.

Recent Advances in Nanomaterial-Based Chemiluminescence Probes for Biosensing and Imaging of Reactive Oxygen Species.

Reactive oxygen species (ROS) play important roles in organisms and are closely related to various physiological and pathological processes. Due to the short lifetime and easy transformation of ROS, the determination of ROS content in biosystem has always been a challenging task. Chemiluminescence (CL) analysis has been widely used in the detection of ROS due to its advantages of high sensitivity, good selectivity and no background signal, among which nanomaterial-related CL probes are rapidly developing. In this review, the roles of nanomaterials in CL systems are summarized, mainly including their roles as catalysts, emitters, and carriers. The nanomaterial-based CL probes for biosensing and bioimaging of ROS developed in the past five years are reviewed. We expect that this review will provide guidance for the design and development of nanomaterial-based CL probes and facilitate the wider application of CL analysis in ROS sensing and imaging in biological systems.

Three-dimensional fluorinated covalent organic frameworks coated capillary for the separation of fluoroquinolones by capillary electrochromatography.

In this work, a three-dimensional fluorinated covalent organic frameworks (3D FCOFs) JUC-515 was synthesized from tetra(4-aminophenyl)methane (TAM) and 2,3,5,6-tetrafluoroterephthalol (TFA) by an ionic liquid method. JUC-515 was introduced into the capillary column and bonded to the inner wall of the capillary column by chemical bonding. Through a variety of characterization results, JUC-515 was successfully synthesized and introduced into the capillary column. The effects of buffer solution concentration, organic additive content and pH of the buffer solution on the separation of fluoroquinolones (FQs) were investigated in detail. The JUC-515-coated capillary column showed good resolution (>1.5) and reproducibility. The relative standard deviations (RSDs) of the retention time for intraday, interday, column-to-column and interbatch precision were less than 0.88%, 2.45%, 2.74% and 3.32%, respectively. The RSDs of the peak area for intraday, interday, column-to-column and interbatch precision were less than 3.79%, 4.31%, 3.33% and 5.62%, respectively. The JUC-515-coated capillary column could be used no less than 150 times. The results showed that the JUC-515-coated capillary column had good separation performance. In addition, by separating fluorinated ß-phenylalanine analogs, ß-phenylalanine and trifluoromethyl ß-phenylalanine analogs, the separation mechanism based on fluorine interactions was discussed. In conclusion, JUC-515 had good potential as a stationary phase for capillary electrochromatography.

Label-free microRNA detection using a locked-to-unlocked transforming system assembled by microfluidics.

MicroRNA (miRNA) is a potential biomarker for the early screening and diagnosis of cancers and is widely present in human blood, urine and saliva. Here, we report a microfluidics-assembled tool for miRNA detection based on the regulation of DNA locked and unlocked states and explore its application in complex samples. Microfluidic techniques are used to continuously assemble the locked-to-unlocked transforming system using a rapid one-step method. It only takes 2 min to produce enough locked-to-unlocked systems for a miRNA detection experiment. DNA molecules with a recognition sequence and a G-rich reporter sequence (G4m) are locked by attaching both ends to the surface of magnetic beads (MBs) in microchannels. The presence of the target miRNA can initiate the specific cleavage of one end of G4m by duplex-specific nuclease, resulting in the transition of G4m from a locked state to an unlocked state. This transition enables G4m to freely fold into a G-quadruplex, which can participate in the catalysis of ABTS oxidation and result in a turquoise color. During the whole process, the target miRNA remains intact and continuously initiate specific cleavage, facilitating signal amplification. Magnetic separation steps are employed to assist in miRNA enrichment and interference reduction. As a proof of concept, we quantified miRNA-21 using the locked-to-unlocked system. The assay allows specific detection of miRNA-21 in the range of 3.2-570 pM with a detection limit of 2.01 pM (S/N = 3). Furthermore, the locked-to-unlocked system is used to analyze miRNA-spiked urine, saliva and serum samples and shows robust performance in different matrices.

Adsorption behavior and mechanism of sulfonamides on controllably synthesized covalent organic frameworks.

In this work, four kinds of covalent organic framework (COF) materials (TpPa-1, TpBD, TpDT, and TFBBD) with different pore sizes or functional groups were synthesized by an ultrasonic method for the adsorption of five sulfonamides. Optimization experiments regarding the adsorption time, vortex speed, and pH were carried out to improve adsorption efficiency. In addition, kinetic and thermodynamic experiments were conducted to explore the adsorption mechanism of the sulfonamides on the different COFs. The adsorption processes of the five sulfonamides on the four COFs fit the pseudo-second-order kinetic model and Langmuir adsorption isotherm model. Additionally, pore filling, hydrogen bond interactions, and electrostatic attraction were found to be the main adsorption mechanisms.

Capillary coated with three-dimensional covalent organic frameworks for separation of fluoroquinolones by open-tubular capillary electrochromatography.

The Schiff-base reaction of 1,3,5-triformylphloroglucinol (Tp) and tetra(4-aminophenyl)methane (TAM) was performed for the synthesis of a three-dimensional covalent organic framework named 3D TpTAM, which was obtained by an ultrasound-assisted method for the first time. The morphology and structure of the synthesized TpTAM were characterized through various methods. Then, TpTAM-coated capillary columns were subsequently prepared by a covalent bonding method within a short time and applied for the separation of fluoroquinolones by capillary electrochromatography (CEC) with good resolution and reproducibility. The intraday relative standard deviations (RSDs) of the retention time and peak areas were 0.88%-0.95% and 2.27%-3.81%, respectively. The interday RSDs of retention time and peak areas were 0.71%-0.89% and 0.88%-3.60%, respectively. The column-to-column RSDs of retention time and peak areas were less than 1.90% and 13.56%, respectively. The interbatch RSDs of retention time and peak areas were less than 3.48% and 3.89%, respectively. The TpTAM-coated capillary columns could be used for no less than 100 runs with no observable changes in the separation efficiency. The separation mechanism was also studied, which indicated that π-π stacking effects, hydrophobic interactions and hydrogen bonding were the main factors. The results revealed that 3D TpTAM should have superior potential as the stationary phase in CEC for chromatographic separation.

A graphene oxide-molybdenum disulfide composite used as stationary phase for determination of sulfonamides in open-tubular capillary electrochromatography.

A graphene oxide-molybdenum disulfide (GO-MoS2) composite was synthesized and utilized as the highly efficient stationary phase of open-tubular capillary electrochromatography (OT-CEC). The characterization results indicated that GO-MoS2 composite was successfully synthesized. The GO-MoS2-coated capillary column was prepared by covalent immobilization method for the determination of seven sulfonamides. The baseline separation of seven sulfonamides was achieved by GO-MoS2-coated capillary column. The linear range was 0.05-100 µg/mL for sulfisomidine, sulfathiazole, sulfamerazine, phthalylsulfathiazole and sulfacetamide, 0.1-100 µg/mL for sulfamonomethoxine and sulfachloropyridazine with a satisfactory correlation coefficients (R2) > 0.9994. This developed OT-CEC method was successfully applied to determinate of seven sulfonamides in environmental water and milk samples with good recoveries of 85.77% - 109.10% and 80.03% - 109.97%, respectively. These results indicated that GO-MoS2-coated capillary column possessed good stability and repeatability.

(1R,3S)-Methyl 3-[(S)-2-(hydroxy-diphenyl-meth-yl)pyrrolidin-1-ylmeth-yl]-2,2-dimethyl-cyclo-propane-carboxyl-ate.

The asymmetric unit of the title compound, C(25)H(31)NO(3), prepared from (-)-1R-cis-caronaldehyde, contains three independent mol-ecules with similar conformations. The hydr-oxy groups are involved in intra-molecular O-H⋯N hydrogen bonds. The crystal packing exhibits weak inter-molecular O-H⋯O and C-H⋯O hydrogen bonds.