In situ growth of imine-based covalent organic framework as stationary phase for open-tubular capillary electrochromatographic separation.

Designing advanced stationary phases to improve separation efficiency is essential in capillary electrochromatography. Due to their outstanding performance, covalent organic frameworks have recently demonstrated considerable promise in the field of separation science. Herein, an open-tubular capillary electrochromatography method was reported using porous imine-based covalent organic framework with sufficiently available interaction sites as stationary phase. The imine-based covalent organic framework coated capillary was easily prepared via an in situ growth method at room temperature, and its separation performance was evaluated, indicating the high separation efficiency for three types of analytes, including herbicides, polybrominated dibenzofurans, and bisphenols. Moreover, the imine-based covalent organic framework coated capillary showed good reproducibility and stability, with intraday (n = 3), interday (n = 3), and column-to-column (n = 3) relative standard deviations of retention time and peak areas of less than 5%. The separation efficiency of the coated capillary remained unchanged even after 200 runs and the maximum theoretical plates reached up to 85 595 N/m for 4,4'-ethylidenebisphenol. It was predicted that the imine-based covalent organic framework stationary phase would be a strong contender for chromatographic separation with high efficiency.

BSA Capped gold Nanoclusters Modulated by Copper ion for Sensitive and Selective Detection of Histidine in Biological Fluid.

This research proposed a sample and environmentally sustainable technique for the synthesis of bovine serum albumin capped gold nanoclusters (BSA-AuNCs) with outstanding fluorescence. The synthesized BSA-AuNCs were investigated using various ways before being combined with Cu2+ to produce a fluorescent switch probe (BSA-AuNCs-Cu2+) for histidine determination. After adding Cu2+, the fluorescence of the BSA-AuNCs was quenched, the fluorescence intensity was enhanced after adding histidine due to good coordination between Cu2+ and histidine. The significant chelation of histidine with Cu2+ demonstrated the viability of developing a selective "switch on" probe for histidine detecting over other amino acids. Unlike existing fluorescent nanomaterial-based approaches for detecting histidine, this study promises good selectivity, high efficiency, and the avoiding of chemical solvents. The designed BSA-AuNCs-Cu2+ fluorescent probe demonstrated an acceptable linear detection range of 0 to 240 µM under optimum circumstances, with a detection limit of 0.9 µM. The BSA-AuNCs-Cu2+ system was investigated in rat serum and human urine, with recoveries ranging from 97.2 to 108.2%, demonstrating its potential applicability for histidine detection with favorable results.

Red Emission Carbon Nanoparticles Which Can Simultaneously Responding to Hypochlorite and pH.

Multi-targets detection has obtained much attention because this sensing mode can realize the detection of multi-targets simultaneously, which is helpful for biomedical analysis. Carbon nanoparticles have attracted extensive attention due to their superior optical and chemical properties, but there are few reports about red emission carbon nanoparticles for simultaneous detection of multi-targets. In this paper, a red emission fluorescent carbon nanoparticles were prepared by 1, 2, 4-triaminobenzene dihydrochloride at room temperature. The as-prepared red emission fluorescent carbon nanoparticles exhibited strong emission peak located at 635 nm with an absolute quantum yield up to 24%. They showed excellent solubility, high photostability and good biocompatibility. Furthermore, it could sensitively and selectively response to hypochlorite and pH, thus simultaneous detection of hypochlorite and pH was achieved by combining the red emission fluorescent carbon nanoparticles with computational chemistry. The formation mechanisms of red emission fluorescent carbon nanoparticles and their response to hypochlorite and pH were investigated, respectively.

Preparation of D-histidine modified zeolitic imidazolate framework-90 coated capillary column and its application in open-tube capillary electrochromatography enantioseparation.

Metal-organic framework materials are a class of novel crystalline porous materials with regular pore structures formed by covalent bonding between metal centers and organic functional groups. Metal-organic framework materials have attracted great interest in analytical chemistry due to their unique properties such as good stability and permanent porosity. In this work, D-histidine was used to carry out chiral modification of zeolitic imidazolate framework-90 under mild conditions, and the D-histidine modified zeolitic imidazolate framework-90 coated capillary column was prepared. This chiral capillary column was used to separate epinephrine, norepinephrine, terbutaline, and tryptophan enantiomers. Under optimum conditions, baseline separations were achieved. The intra-day, inter-day, and inter-column relative standard deviations (n = 3) of the four pairs of enantiomeric migration times were 0.15%-0.56%, 0.74%-2.40%, and 1.93%-3.18%, respectively. Moreover, the D-histidine modified zeolitic imidazolate framework-90 coated capillary could be reused for at least 150 runs without significant changes in the separation efficiency and migration time.

Development of a facile and sensitive method for detecting alkaline phosphatase activity in serum with fluorescent gold nanoclusters based on the inner filter effect.

In this work, a simple and sensitive method based on the inner filter effect (IFE) of p-nitrophenol (PNP) on the fluorescence of gold nanoclusters (AuNCs) has been developed for detecting alkaline phosphatase (ALP) activity. Bright orange fluorescent AuNCs were synthesized by one-pot synthesis and used directly as IFE fluorophores. p-Nitrophenyl phosphate (PNPP) is hydrolyzed by ALP to PNP, which quenches the fluorescence of AuNCs by the IFE. In the presence of ALP, PNPP was converted to PNP, and the absorption band shifted from 310 nm to 405 nm, which resulted in a certain degree of overlap between the absorption of PNP and the excitation of AuNCs. Due to the competitive absorption between AuNCs and PNP, the excitation of AuNCs was clearly diminished, leading to the quenching of the fluorescence of AuNCs. The IFE detection method exhibited a good linear relationship between 0.01 and 7.0 U L-1 (R2 = 0.9990) with the lowest detection limit of 0.003 U L-1 (the signal-to-noise ratio is 3). The proposed detection method was successfully applied for detecting ALP in serum samples and studying ALP inhibitors.

Characterization of the Ligand Exchange Reactions on CdSe/ZnS QDs by Capillary Electrophoresis.

The continuous development of semiconductor quantum dots (QDs) in biochemical research has attracted special attention, and surface functionalizing becomes more important to optimize their performance. Ligand exchange reactions are commonly used to modify the surface of QDs for their biomedical applications. However, the kinetics of ligand exchange for semiconductor QDs remain fully unexplored. Here, we describe a simple and rapid method to characterize the ligand exchange reactions on CdSe/ZnS QDs by capillary electrophoresis (CE). The results of ultraviolet-visible absorption spectra, fluorescence spectra, and Fourier transform infrared spectroscopy indicated the successful implementation of the ligand exchange process. The dynamics of ligand exchange of OA-coated CdSe/ZnS QDs with 4-mercaptobenzoic acid was monitored by CE, and the observed ligand exchange trends were fitted with logistic functions. When the ligand exchange reactions reached equilibrium, the ligand density of QDs can be quantified by CE. It is anticipated that CE will be a new powerful technique for quantitative analysis of the ligand exchange reactions on the surface of QDs.

Ratiometric Detection of Intracellular Lysine and pH with One-Pot Synthesized Dual Emissive Carbon Dots.

Recently, the development of new fluorescent probes for the ratiometric detection of target objects inside living cells has received great attention. Normally, the preparation, modification as well as conjugation procedures of these probes are complicated. On this basis, great efforts have been paid to establish convenient method for the preparation of dual emissive nanosensor. In this work, a functional dual emissive carbon dots (dCDs) was prepared by a one-pot hydrothermal carbonization method. The dCDs exhibits two distinctive fluorescence emission peaks at 440 and 624 nm with the excitation at 380 nm. Different from the commonly reported dCDs, this probe exhibited an interesting wavelength dependent dual responsive functionality toward lysine (440 nm) and pH (624 nm), enabling the ratiometric detection of these two targets. The quantitative analysis displayed that a linear range of 0.5-260 µM with a detection limit of 94 nM toward lysine and the differentiation of pH variation from 1.5 to 5.0 could be readily realized in a ratiometric strategy, which was not reported before with other carbon dots (CDs) as the probe. Furthermore, because of the low cytotoxicity, good optical and colloidal stability, and excellent wavelength dependent sensitivity and selectivity toward lysine and pH, this probe was successfully applied to monitor the dynamic variation of lysine and pH in cellular systems, demonstrating the promising applicability for biosensing in the future.

A label-free colorimetric biosensor for sensitive detection of vascular endothelial growth factor-165.

Sensitive detection of a low abundant protein is essential for biomedical research and clinical diagnostics. Herein, we develop a label-free colorimetric biosensor for the sensitive detection of recombinant human vascular endothelial growth factor-165 (VEGF165). This biosensor consists of an aptamer-based hairpin probe, an assistant DNA-trigger duplex and a linear template. In the presence of VEGF165, the specific binding of VEGF165 with the aptamer-based hairpin probe results in the opening of a hairpin probe and the opened hairpin probe subsequently hybridizes with the single-stranded region of the assistant DNA-trigger duplex to initiate the strand displacement amplification (SDA) to yield abundant triggers. The released triggers can further function as the primers to anneal with the hairpin probe and lead to the opening of the hairpin structure, which subsequently hybridizes with the assistant DNA-trigger duplex to initiate the next round of SDA reaction and generates more triggers. Large amounts of triggers could be generated by the synergistic operation of dual SDA reaction, and the obtained triggers can initiate a new round of SDA reaction to yield numerous G-quadruplex DNAzymes, which subsequently catalyze the conversion of ABTS2- to ABTS˙- by H2O2 to yield a color change with the assistance of a cofactor hemin. In contrast, in the absence of target VEGF165, the hairpin probe, the assistant DNA-trigger duplex and the linear template can stably coexist in solution, and thus no color change is observed because no trigger can initiate SDA to generate the G-quadruplex DNAzyme. This biosensor has a low detection limit of 1.70 pM and a dynamic range over 3 orders of magnitude from 24.00 pM to 11.25 nM. Moreover, the biosensor shows excellent specificity toward the target VEGF165 and the entire reaction can be carried out in an isothermal manner without the involvement of a high precision thermal cycler, making the current assay extremely cost effective.

Determination of enantiomers by FESI-sweeping with an acid-labile sweeper in nonaqueous capillary electrophoresis.

In this work, a facile and highly efficient on-line concentration strategy based on a coupling of field enhanced sample injection (FESI) and sweeping was developed for the determination of trace enantiomers (propranolol, PL) by nonaqueous capillary electrophoresis (NACE). In this FESI-sweeping method, the use of a sample of high acidity and low conductivity (pH* = 2.5, 4.0 µS cm(-1)) allowed for a large amount of analyte injection. Then, the concentration of the analytes was carried out by sweeping based on the interaction of an acid-labile anionic selector, di-n-butyl L-tartrate-boric acid complex acid, and cationic analytes. Simultaneously, the concentrated analytes were released and focused at the boundary of the acid sample solution and separation buffer due to the decomposition of the selector in the acid sample solution. Under the optimum conditions, a 21,000-fold sensitivity enhancement upon normal capillary zone electrophoresis (CZE) was achieved for PL enantiomers. The detection limits of R-propranolol and S-propranolol were 0.26 ng mL(-1) and 0.31 ng mL(-1), respectively. Eventually, the FESI-sweeping method was applied to detect PL enantiomers in plasma, saliva, and urine.

Solid-phase synthesis of highly fluorescent nitrogen-doped carbon dots for sensitive and selective probing ferric ions in living cells.

Carbon quantum dots (C-Dots) have drawn extensive attention in recent years due to their stable physicochemical and photochemical properties. However, the development of nitrogen-doped carbon quantum dots (N-doped C-Dots) is still on its early stage. In this paper, a facile and high-output solid-phase synthesis approach was proposed for the fabrication of N-doped, highly fluorescent carbon quantum dots. The obtained N-doped C-Dots exhibited a strong blue emission with an absolute quantum yield (QY) of up to 31%, owing to fluorescence enhancement effect of introduced N atoms into carbon dots. The strong coordination of oxygen-rich groups on N-doped C-Dots to Fe(3+) caused fluorescence quenching via nonradiative electron-transfer, leading to the quantitative detection of Fe(3+). The probe exhibited a wide linear response concentration range (0.01-500 µM) to Fe(3+) with a detection limit of 2.5 nM. Significantly, the N-doped C-Dots possess negligible cytotoxicity, excellent biocompatibility, and high photostability. All these features are favorable for label-free monitoring of Fe(3+) in complex biological samples. It was then successfully applied for the fluorescence imaging of intracellular Fe(3+). As an efficient chemosensor, the N-doped C-Dots hold great promise to broaden applications in biological systems.

A novel fluorescence sensor based on Al3+-mediated aggregation of gold nanoclusters for determination of citric acid in beverages.

This paper revealed a new strategy for citric acid (CA) detection using aggregation-induced emission (AIE)-based fluorescent gold nanoclusters (AuNCs). AuNCs was synthesized using glutathione (GSH) as the template and reducing agent and used as the fluorescent probe to detect CA under aluminum ion (Al3+) mediation. The fluorescence intensity of AuNCs increased about 4 times with the addition of Al3+, but the enhanced fluorescence was quenched after the addition of CA. Based on this fluorescence phenomenon, an "on-off" fluorescence strategy was designed for the sensitive determination of CA and a linear detection range for CA was achieved within 0-80.0 µM. In addition, the developed probe exhibited high selectivity and accuracy for determination of CA. The mechanism of fluorescence enhancement and quenching of AuNCs was explored in detail. The established probe was used successfully for CA detection in beverages. The spiked recoveries from 97.50% to 103.67% were gratifying, which indicated the probe had potential prospects for detecting CA in food.

In situ synthesis of MIL-100(Fe) in the capillary column for capillary electrochromatographic separation of small organic molecules.

Because of the unusual properties of the structure, the metal organic frameworks (MOFs) have received great interest in separation science. However, the most existing methods for the applications of MOFs in separation science require an off-line procedure to prepare the materials. Here, we report an in situ, layer-by-layer self-assembly approach to fabricate MIL-100(Fe) coated open tubular (OT) capillary columns for capillary electrochromatography. By a controllable manner, the OT capillary columns with a tailored MIL-100(Fe) coating have been successfully synthesized. The results of SEM, XRD, FT-IR, and ICP-AES indicated that MIL-100(Fe) was successfully grafted on the inner wall of the capillary. Some neutral, acidic and basic analytes were used to evaluate the performance of the MIL-100(Fe) coating OT capillary column. Because of the size selectivity of lattice aperture and hydrophobicity of the organic ligands, three types of analytes were well separated with this novel MIL-100(Fe) coating OT capillary column. For three consecutive runs, the intraday relative standard deviations (RSDs) of migration time and peak areas were 0.4-4.6% and 1.2-6.6%, respectively. The interday RSDs of migration time and peak areas were 0.6-8.0% and 2.2-9.5%, respectively. The column-to-column reproducibility of retention time was in range of 0.6-9.2%. Additionally, the 10 cycles OT capillary column (10-LC) could be used for more than 150 runs with no observable changes on the separation efficiency.

Highly sensitive detection of microRNA by chemiluminescence based on enzymatic polymerization.

We have developed a new methodology for miRNA assay using chemiluminescence imaging by poly(U) polymerase catalyzed miRNA polymerization. This method is very sensitive with a 50 fM limit of detection, which is comparable to or better than current assay methods. Multiplex detection for miRNA can be easily realized by introducing different capture probes onto the biosensor array, which will make it highly versatile for various research purposes.

Concise analysis of γ-hydroxybutyric acid in beverages and urine by capillary electrophoresis with capacitively coupled contactless conductivity detection using 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid as background electrolyte.

γ-Hydroxybutyric acid (GHB), a neurotransmitter or neuromodulator in the human central nervous system, is often abused in drug-facilitated sexual assaults due to its euphoric and sedative effects. While the analysis of GHB has received continuous attention, its inherent characteristics pose challenges. In the current study, capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C4D) was built, and Good's buffers were evaluated as the background electrolytes for CE separation and C4D detection. On this basis, a simple and efficient CE-C4D method was developed for GHB analysis. Through theoretical discussion and experimental optimization, the separation of GHB and related positional isomers α-hydroxybutyric acid (AHB) and ß-hydroxybutyric acid (BHB) was achieved within 4 min using 150 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) as the running buffer. Under the optimized condition, the relative standard deviations of migration time and peak area were less than 1.1% and 4.5%, indicating good precision. The C4D signal of GHB showed a good linear relationship with GHB concentration in the range of 3-300 µM with a determination coefficient of 0.9997, and the detection limit was calculated to be 0.37 µM based on the signal-to-noise ratio of three. Furthermore, liquid-liquid extraction (LLE) and solid-phase extraction (SPE) were comparatively studied for sample matrix purification. Combined with the optimized SPE procedure, the developed CE-C4D method has been successfully applied for the determination of exogenous GHB in spiked beverages and endogenous GHB in human urine.

Synthesis of a covalent organic framework with hydrazine linkages and its application in open-tubular capillary electrochromatography.

Recently, covalent organic frameworks (COFs), owning to their excellent and unique properties, are attracting the attention of numerous researchers in some areas, especially the domain of chromatographic separation. However, the application of hydrazine linkages COFs in open-tubular capillary electrochromatography (OT-CEC) lies in the early stage at present. Herein, a well-crystallized hydrazine-linked COF (Tf-DHzOH) was synthesized successfully from 2,5-dihydroxyterephtalohydrazide and 1,3,5-triformyl-benzene. Tf-DHzOH was firstly regarded as a stationary phase material to prepare the Tf-DHzOH coated capillaries with different coating thickness by covalent bonding in this work. The characterization results showed that Tf-DHzOH was successfully synthesized. The separation performance and stability of the Tf-DHzOH coated capillary were evaluated by considering amino acids, sulfonamides, tetracyclines and benzene compounds as analytes. The relative standard deviations (RSDs) of separation time in the intra-day (n = 9), inter-day (n = 6), column-to-column (n = 3) and batch-to-batch (n = 3) were 0.76-4.97%, 1.59-5.94%1.78-8.72% and 1.66%-8.23%, respectively, the RSDs of peak areas were 1.90-5.16%, 1.73-5.24%, 1.26-7.33% and 3.77%-11.24%, respectively. It was confirmed that there was no visible change of separation efficiency after the Tf-DHzOH-coated capillary was used more than 200 runs. The results make clear that 2D hydrazine-linked COF (Tf-DHzOH) has superior potential as the stationary phase in OT-CEC for chromatographic separation.

Combined effect of early life stress and acute stress on colonic sensory and motor responses through serotonin pathways: differences between proximal and distal colon in rats.

Clinically, adults who have experienced stresses in childhood present with episodes of serious symptoms of irritable bowel syndrome that are associated with acute stress, but the mechanism is not well understood. This study aimed to investigate the colonic sensory/motor responses to acute water avoidance stress (WAS) in male adult rats subjected to neonatal maternal separation (NMS), and the underlying mechanism of sensory/motor responses. Effects of the combined acute and early life stress on visceral sensation, colonic motility, and the tissue and luminal content of serotonin (5-hydroxytryptamine, 5-HT) in the proximal and distal colon were evaluated using the abdominal withdrawal reflex test, faecal pellet output measurement and capillary electrophoresis analysis, respectively. Results showed that WAS significantly increased not only visceral sensitivity but also colonic motility in NMS rats compared to the normal rats. These alterations were accompanied by significant increase in 5-HT content in the proximal but not the distal colonic tissues; these alterations were also associated with increased density of enterochromaffin (EC) cells in the proximal segment. In contrast, the faecal content of 5-HT increased similarly in both segments. Consecutive administration of parachlorophenylalanine to NMS rats was more potent at 500 mg kg⁻¹ day⁻¹ than at 150 mg kg⁻¹ day⁻¹ in suppressing colonic sensory/motor responses to WAS, corresponding to the greater reduction of the tissue and faecal content of 5-HT and of EC cell density in the colon. These data indicate that combined early life stress and acute stress effectively induce visceral hyperalgesia and motility disorder through 5-HT pathways in the colon of rats, and the proximal and distal colon have different responses towards the combined stressors.

Extremely large volume electrokinetic stacking of cationic molecules in MEKC by EOF modulation with strong acids in sample solutions.

In this report, a novel facile way of online preconcentration of trace levels of analytes in capillary electrophoresis is presented. The proposed strategy is based on the combination of strong acidic phosphate as sample buffers with borate separation buffer containing sodium dodecyl sulfate. When injection voltage is applied, the continuous introduction of low pH sample causes the apparent bulk flow inside the capillary gradually slows down. Finally at a certain point, it reaches the same magnitude as that of the oppositely migrating anionic micelles, thus the frontier of the micelle zone becomes stagnant. This steady state can be maintained for a very long time so that essentially extremely large volume of sample solutions can be injected into the capillary, and the cationic analytes may be efficiently stacked at the neutralized micelle zone. A theoretical model was proposed and preconcentration conditions of two model analytes, matrine and oxymatrine, were optimized with the aid of the model. Under optimized conditions, more than 1000-fold increase in sensitivity was obtained as compared with the normal hydrodynamic injection without sample stacking. The limits of detection for oxymatrine and matrine were 0.81 and 0.18 ng/mL, respectively, using photodiode array UV detection at wavelength 211 nm.

Quantification of luminally released serotonin in rat proximal colon by capillary electrophoresis with laser-induced fluorescence detection.

Serotonin (5-hydroxytryptamine, 5-HT) plays vital roles in regulating gastrointestinal functions. Thus, the detection of 5-HT in the gastrointestinal tract is of great importance for biomedical research, medical diagnosis, and pharmaceutical therapy. This paper presents a simple, sensitive, and fast method for the quantification of luminally released serotonin in the feces and tissues of the rat proximal colon by means of capillary electrophoresis with laser-induced fluorescence detection. 5-Carboxyfluorescein N-succinimidyl ester was used for precolumn derivatization of serotonin. The optimal separation and detection conditions were obtained with an electrophoretic buffer containing 60 mM borate (pH 8.90) and an air-cooled argon-ion laser (excitation at 488 nm, emission at 520 nm). The serotonin concentrations in the feces and tissues of proximal colons were analyzed with this method, and the average values of serotonin in the feces samples were 1.951 +/- 0.446 ng/mg (male) and 2.095 +/- 0.533 ng/mg (female) and 1.397 +/- 0.267 ng/mg in rat proximal colon tissues. The results demonstrate that this method can accurately determine luminally released 5-HT in rats.

Low-temperature bath/high-conductivity zone/stacking micellar electrokinetic chromatography for the analysis of phenolic acids in coffee drink.

Two stacking methods in micellar electrokinetic chromatography (MEKC) were investigated in this article in an attempt to increase the amount of sample injected, as well as to focus analytes onto a small zone. One employed a "high-conductivity zone", which was inserted between the sample zone and background solution to build an unequal conductivity gradient. The other employed a "low-temperature bath". A portion of the capillary was immersed in a low-temperature bath, which served as a "pseudo-low-conductivity zone". As a result, a large volume of sample injection can be achieved. Using three phenolic acids-chlorogenic acid (CGA), caffeic acid (CA) and ferulic acid (FA) in coffee as model compounds, the limit of detection (LoD) was determined to be 0.31microg/ml (S/N=3) for CGA by means of normal MEKC under suppressed electroosmotic flow (EOF). The LoD could be improved to 2.8 x10(-2), 5.3 x10(-3) and 6.0 x10(-3) microg/ml, respectively, when normal MEKC-stacking, high-conductivity zone MEKC-stacking and the low-temperature zone MEKC-stacking methods were applied. Furthermore, the high conductivity zone and the low-temperature bath were operated simultaneously on one capillary to investigate the synergism effect. The results showed that there did exist synergism effect for CGA and CA when the two were hyphenated. The stacking efficiency was higher than that of the single one used. However, there was not synergism effect for FA.