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Fatty acids (FAs) play a vital physiological role in lipid metabolism, which is reported as potential diagnostic biomarker for various diseases. Thus, it is urgent to develop a credible method that can profile FA metabolism with a holistic view. Here, a targeted strategy to screen FAs was developed by parallel labeling with d 0 /d 6 -dansylhydrazine (d 0 /d 6 -DnsHz) and using ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC–MS/MS) in data-dependent MS/MS (ddMS2) mode. The simple and mild derivatization procedure within 3 h allowed for a significant improvement in sensitivity. Additionally, the characteristic product ions introduced by the derivatization reagent assist to identify the unknown FA species. A quantitation method was established by multiple reaction monitoring (MRM) and the d 6 -DnsHz tagged standards for each analyte were used as internal standards to overcome the matrix effects. By applying the method to determine FA levels in plasma collected from the esophageal squamous cell carcinoma (ESCC) patients and healthy controls, 65 FA metabolites were characterized and six FAs were found to be altered by the invasion of tumors. The parallel derivatization strategy provides insights into the identification of unknown FAs and paves a new way for targeted metabolomics. Also, this novel method is a powerful tool for characterization and quantification of FAs in biological samples, which shows a great potential application in clinical diagnosis and investigation of disease mechanisms. [Display omitted] • A novel parallel derivatization strategy was developed to identify FAs in biosample. • Dansylhydrazine-based derivatization enhanced ionization efficiency of FAs. • The isotope-labeled standard provides one-to-one internal standard for FA analysis. • Six FAs were screened out as potential diagnostic biomarkers of ESCC. [ABSTRACT FROM AUTHOR]

We report here for the first time a self-powered biosensing platform based on graphene/graphdiyne/graphene (GDY-Gr) heterostructure substrate material for ultrasensitive hepatocarcinoma marker (microRNA-21) detection in both electrochemical and colorimetric test modes. The dual-mode signal intuitively displayed on a smartphone fundamentally improves the detection accuracy. In electrochemical mode, the calibration curve is established in the linear range of 0.1–10000 fM, and the detection limit is as low as 0.333 fM (S/N = 3). Simultaneously, colorimetric analysis of the miRNA-21 is realized by using ABTS as an indicator. The detection limit is confirmed as 32 fM (S/N = 3), and miRNA-21 of concentration from 0.1 pM to 1 nM exhibit a linear relationship with R2 = 0.9968. Overall, the combination of GDY-Gr and multiple signal amplification strategy significantly improved the sensitivity by 310 times compared with traditional enzymatic biofuel cells (EBFCs) based detection platform, showing broad application prospects for on-site analysis and future mobile medical services. [Display omitted] • Ultra-sensitive self-powered biosensor platform based on graphdiyne/graphene heterogeneous is designed for liver cancer markers detection. • Dual-mode signal intuitive displayed on a smartphone and fundamentally improve the detection accuracy. • Detection limit are as low as 0.33 fM and 32 fM (S/N = 3) in electrochemical and colorimetric analysis mode, respectively. [ABSTRACT FROM AUTHOR]

Abstract The pseudo-targeted metabolomics approach was developed recently which combined the advantages of untargeted and targeted analysis. However, the current pseudo-targeted analysis method has limitations due to the technical characteristics. In this study, a novel metabolic pathway-based pseudo-targeted approach was proposed for urine metabolomics analysis using an ultra-high-performance liquid chromatography (UPLC)-MS/MS system operated in the multiple reaction monitoring (MRM) mode. MRM ion pairs were acquired from urine samples through untargeted analysis using UPLC-HRMS, as well as by searching for metabolites in related pathways in relevant databases and from previous relevant research, including amino acids, fatty acids, nucleosides, carnitines, glycolysis metabolites, and steroids. This improved pseudo-targeted method exhibited good repeatability and precision, and no complicated peak alignment was required. As a proof of concept, the developed novel method was applied to the discovery of urine biomarkers for patients with esophageal squamous cell carcinoma (ESCC). The results showed that ESCC patients had altered acylcarnitines, amino acids, nucleosides, and steroid derivative levels et al. compared to those of healthy controls. The novelty of this study lies in the fact that it provides an approach for acquiring MRM ion pairs not only from untargeted MS analysis but also from targeted searching for metabolites in related metabolic pathways. By improving the detection limit of low-abundance metabolites, it enlarges the range for the discovery of potential biomarkers. Our work provides a foundation for achieving pseudo-targeted metabolomics analysis on the widely used LC-MS/MS MRM platform. Graphical abstract fx1 Highlights • Ion pairs were from untargeted analysis and targeted searching in metabolic pathways. • It improves the detection limit for low-abundance metabolites. • It enlarges the range for the discovery of potential biomarkers. • A low-abundance potential biomarkers for ESCC was newly discovered by this method. [ABSTRACT FROM AUTHOR]

A new selective probe with a quinoxalinone structure, QP-1 , has been developed for detection of Cys from biothiols. QP-1 features superb selectivity to Cys and a wide pH range. QP-1 has selectivity to Cys over Hcy, GSH, other amino acids and ions. HRMS spectra confirmed that the detection process was a conjugate addition-addition-elimination reaction. Moreover, QP-1 has been successfully applied in the imaging of Cys in living cells. Finally, QP-1 has been used to detect Cys in rat urine samples. [ABSTRACT FROM AUTHOR]

Asymptomatic infection of COVID-19 is a global threat for public health. Unfortunately, the study about metabolic dysregulation of asymptomatic infection is barely investigated. Here, we performed carboxylic submetabolome profiling of serum from 62 asymptomatic and 122 control individuals, by a highly sensitive chemical isotope labelling method. Twenty-one discriminative carboxylic features, including 12-hydroxyeicosatetraenoic acid, cholic acid, glycoursodeoxycholic acid and 15,16-dihydroxyoctadeca-9,12-dienoic acid were discovered to be dysregulated in asymptomatic patients. This panel containing 21 carboxylic features could accurately identify asymptomatic patients based on a random forest model, providing an accuracy of 85.7% with only 3.6% false positive rate and 7.1% false negative rate. The dysregulated metabolites found in asymptomatic patients covered several important pathways, such as arachidonic acid metabolism, synthesis of bile acid, β-oxidation of fatty acids, activation of macrophage and platelet aggregation. This work provided valuable knowledge about serum biomarkers and molecular clues associated with asymptomatic COVID-19 patients. [Display omitted] • Carboxylic submetabolome of COVID-19 asymptomatic patients were performed using chemical isotope labelling-LC-MS method. • A total of 21 potential carboxylic biomarkers of COVID-19 asymptomatic infection were identified. • The potential biomarkers cover pathways including arachidonic acid metabolism, bile acid synthesis, β-oxidation of fatty acids, activation of macrophage and platelet aggregation. • A machine learning method was developed for risk assessment of asymptomatic infection. [ABSTRACT FROM AUTHOR]

Abstract: A light-emitting diode-induced fluorescence detector (LED-FD) for capillary electrophoresis was constructed and evaluated. A lamp LED with an enhanced emission spectrum and a band pass filter was used as the excitation light source. Refractive index matching fluid (RIMF) was used in the detection cell to reduce scattering light and the noise level. The limit of detection (LOD) for fluorescein was 1.5nM (SNR=3). The system exhibited linear responses in the range of 1×10−8 to 5×10−6 M (R =0.999). Application of the lamp LED-FD for the analysis of FITC-labeled ephedra herb extract by capillary electrophoresis was demonstrated. [Copyright &y& Elsevier]

Abstract: A glycerol assisted light-emitting diode (LED)-induced fluorescence detector (IF) for capillary flow systems was constructed and evaluated. A blue LED was used as the excitation source, and optical fibers (OF) were used to transmit the excitation light and collect the fluorescence. A commercial available 5-port manifold was used as detection cell, where the capillary tube and the OF were fixed into the manifold. The precision of the holes on the manifold ensured a self-alignment of optical path. A refractive index matching fluid (RIMF)-glycerol was used to eliminate the interfaces between the OF and the LED, as well as between the fused silica capillary and the transmitting/collecting fiber. The enhancement of excitation light led to 2.8-folds improvement on the signal-to-noise ratio. The use of RIMF also eliminates focusing effect of the capillary wall and reduces both the excitation light directed to the detection cell and background signal, resulting in reduction in the fluorescence intensity and noise level. The intensity was reduced to 47–63% for laser and 60–77% for LED, respectively, for capillaries with i.d. from 50 to 250μm; while the noise level was reduced to 1/3 when RIMF was used for both laser and LED on the tested capillaries. About 5.6-fold enhancement in signal-to-noise ratio was obtained in total. The detection limit of the LED-IF for fluorescein isothiocyanate (FITC) was 4nM. Application of the LED-IF for the analysis of FITC-labeled amino acids by electrophoresis was demonstrated. [Copyright &y& Elsevier]

Abstract: The effect of axial temperature gradient (ATG) along a microcolumn on the separation performance at both isocratic and gradient elution mode was investigated. A thermostat system was designed to form an ATG along the packed column. Polycyclic aromatic hydrocarbons (PAHs) were separated on a 0.53mm ×150mm i.d. 5μm C18 microcolumn, with water and acetonitrile as mobile phase. The separation results obtained at mobile phase gradient (MPG) and ATG in microcolumn HPLC were compared with the results performed at ambient conditions. Extrapolated curves of peak width at half height versus lnk showed that is narrower at the same retention time when ATG was applied in addition to MPG. The column efficiency was enhanced 20–30% and the resolution was slightly reduced because of reduction of selectivity at elevated temperature at ATG condition. The RSD of retention time in ATG mode was less than 2.5%. [Copyright &y& Elsevier]

Abstract: Tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)3 2+)–Zirconia–Nafion composite modified glassy carbon disk electrode as a solid-state electrochemiluminescence (ECL) detector is successfully applied to an electrophoretic microchip system with a wall-jet configuration. Pharmaceuticals such as tramadol, lidocaine and ofloxacin were selected to characterize the performance of this microchip capillary electrophoresis (CE)–ECL detection system. Voltammetric and ECL behaviors of immobilized Ru(bpy)3 2+ were investigated in lidocaine system. Influences of the separation electric field to cyclic voltammograms (CVs) of the immobilized Ru(bpy)3 2+ were also investigated. Tramadol, lidocaine and ofloxacin can be baseline separated without any additives. The detection limits (S/N=3) were 2.5×10−5 molL−1 for tramadol, 5.0×10−6 molL−1 for lidocaine, 1.0×10−5 molL−1 for ofloxacin under the sample injection of picoliters, and the linear ranges were from 5.0×10−5 to 2.5×10−3 molL−1 for tramadol, 1.0×10−5 to 1.0×10−3 molL−1 for lidocaine, and 1.0×10−5 to 2.5×10−3 molL−1 for ofloxacin, respectively. [Copyright &y& Elsevier]

Abstract: An easy and universal wall-jet configuration for microchip CE-ECL detection system was constructed and investigated in this work. Two detection modes of pre-column and post-column were applied to the above system. TPA, tramadol and lidocaine were chosen as model analytes to estimate the system in both modes. The important operational parameters such as the concentration of luminescent reagent and the distance between the separation outlet and the working electrode were optimally obtained and compared for the first time. [Copyright &y& Elsevier]

Abstract: In this work, a new kinetic method was proposed for quantification phenoxyl radicals generated in enzyme reaction. Instead of direct detecting the spectral signals of phenoxyl radicals, a molecular probe, the reduced form of nicotinamide adenine dinucleotide (NADH), was employed to indicate the formation of phenoxyl free radicals. It was found that the reactions of NADH and phenoxyl radicals are very fast, but can be followed by using stopped-flow fast scanning spectrophotometric technique. The initial rate of accelerated-oxidation of NADH represents the reactivity of phenoxyl free radical, which is proportional in a certain range to the initial concentration of the parent chlorophenols of the radicals. With this method, the phenoxyl radicals generated in oxidation reaction of chlorophenols (2-CP; 4-CP; 2,4-DCP; 2,4,6-TCP and 2,3,4,6-Tetra-CP) with hydrogen peroxide, catalyzed by horseradish peroxidase, were investigated. The method is highly sensitive. Phenoxyl radicals generated from as low as 1×10−8 M 2,4-DCP, for example, can be readily detected with the proposed method. The results show that the reactivity of various phenoxyl radicals are in the following order: 2,4-DCP>4-CP>2-CP>2,4,6-TCP>2,3,4,6-Tetra-CP. A mechanism is proposed to explain the possible pathway of the probe reaction. The feasibility of this method was assessed by the determination of enzymatic generation of phenoxyl radicals in lake water samples. [Copyright &y& Elsevier]

Abstract: Poly(dimethylsiloxane) (PDMS) microfluidic channels modified by citrate-stabilized gold nanoparticles after coating a layer of linear polyethylenimine (LPEI) were successfully used to separate dopamine and epinephrine, which were difficult to be separated from baseline in native and hybrid PDMS microchannels. In-channel amperometric detection with a single carbon fibre cylindrical electrode was employed. Experimental parameters of separation and detection processes were optimized in detail. The analytes were well separated within 100s in a 3.7cm long separation channel at a separation voltage of +800V using a 30mM phosphate buffer solution (PBS, pH 7.0). Linear responses of them were obtained both from 25 to 600μM with detection limits of 2μM for dopamine and 5μM for epinephrine, respectively. The modified PDMS channels have a long-term stability and an excellent reproducibility within 2 weeks. [Copyright &y& Elsevier]

Abstract: A novel solid-contact potentiometric sensor for ascorbic acid based on cobalt phthalocyanine nanoparticles (NanoCoPc) as ionophore was fabricated without any need of auxiliary materials (such as membrane matrix, plasticizer, and other additives). The electrode was prepared by simple drop-coating NanoCoPc colloid on the surface of a glassy carbon electrode. A smooth, bright and blue thin film was strongly attached on the surface of the glassy carbon electrode. The electrode showed high selectivity for ascorbic acid, as compared with many common anions. The influences of the amount of NanoCoPc at the electrode surface and pH on the response characteristics of the electrode were investigated. To overcome the instability of the formal potential of the coated wire electrode, a novel electrochemical pretreatment method was proposed for the potentiometric sensor based on redox mechanism. This resulting sensor demonstrates potentiometric response over a wide linear range of ascorbic acid concentration (5.5×10−7 to 5.5×10−2 M) with a fast response (<15s), lower detection limit (ca. 1.0×10−7 M), and a long-term stability. Furthermore, microsensors based on different conductors (carbon fiber and Cu wire) were also successfully fabricated for the determination of practical samples. [Copyright &y& Elsevier]

Abstract: The interactions between proteins and Ponceau 4R (PR) in aqueous solution have been studied by the techniques of resonance light scattering (RLS) spectroscopy, the absorption spectroscopy, zeta potential assay and circular dichroism (CD) spectrum. The dry PR can assemble on the surface of protein via electrostatic and hydrophobic forces to produce an associated compound of protein-PR, this compound can enhance the RLS of protein. Based on this fact, a simple, rapid, and sensitive method has been developed for the determination of proteins at nanogram level by RLS technique with a common spectrofluorimeter. Under optimum conditions, the linear range is 0.10–39.2μgmL−1 for the determination of both bovine serum albumin (BSA) and human serum albumin (HSA). The detection limits (S/N=3) are 6.96ngmL−1 for BSA and 5.71ngmL−1 for HSA, respectively. There is almost no interference from amino acids, most of the metal ions, and other coexistent substances. The method has been satisfactorily applied to the direct determination of the total protein in human serum. [Copyright &y& Elsevier]

In this work, carbon nanosphere (CNS)-based fluorescence “turn off/on” aptasensor was developed for targeted detection of breast cancer cell MCF-7 by conjugation with FAM (a dye)-labeled mucin1 (MUC1) aptamer P0 (P0-FAM), which can recognize MUC1 protein overexpressed on the surface of MCF-7. Different from other carbon based fluorescence quenching materials, CNSs prepared by the carbonization of glucose not only have the high fluorescence quenching efficiency (98.8%), but also possess negligible cytotoxicity (in the concentration range of 0–1 mg/mL, which is 10 times higher than that of traditional carbon nanotubes or graphene oxide (0–100 µg/mL)). As for the detection of the mimic of the tumor antigen MUC1, the resulting fluorescence intensity increases nearly linearly in the range of 0–6 μM with the limit of detection (LOD) of 25 nM. [ABSTRACT FROM AUTHOR]

A simple yet powerful polymerase chain reaction (PCR)-free strategy for visualized assay of human telomerase activity was reported in this work. Gold nanoparticles (AuNPs) based colorimetric strategy was applied with well-designed enzyme-aided cyclic amplification. Briefly, the detection relies on the elongated primers of telomerase substrate (TS) induced by telomerase, which open the hairpin DNA and hybridize with linker DNA, the trigger of AuNPs aggregation. Nicking endonuclease was added in the sensing system, which cleaved linker DNA after hybridization and released complimentary strand for cyclic hybridization with linker DNA, resulted in high sensitivity for the detection of telomerase. Down to 25 HeLa cells with high expression of telomerase could be recognized. The proposed strategy provides a good platform for the determination of telomerase activity, differentiation of cancer cell lines from normal cell line and screening of telomerase-targeted anticancer drugs. [ABSTRACT FROM AUTHOR]

Some non-coding RNAs are abnormally expressed during the occurrence and development of diseases, so it is necessary to develop analytical methods that can specifically and sensitively detect them. In typical CRISPR/Cas12a system, a complete crRNA that can recognize single-stranded or double-stranded DNA is necessary to activate its trans -cleavage activity, which limits its direct application in RNA detection. Here, we prospectively find that slicing the facilitated crRNA in the typical CRISPR/Cas12a system at a fitted site did not affect its trans -cleavage activity, and a m ini c rRNA- m ediated CRISPR/Cas12a system (MCM-CRISPR/Cas12a) was proposed based on this. This system can detect non-coding RNA to pM-level (10 pM for miRNA-21). To expand the application of this system, we combined it with HCR and CHA to establish a detection platform for non-coding RNA. The results show that the proposed method can specifically detect RNA to fM-level (2.5 fM for miRNA-21, 8.98 fM for miR-128–3p, and 81.6 fM for lncRNA PACER). The spiked recovery rates of miRNA-21, miR-128–3p, and lncRNA PACER in normal human serum were in range from 104.7 to 109.4 %, indicating the proposed method owns good applicability. In general, this MCM-CRISPR/Cas12a system further breaks the limitations of the typical CRISPR/Cas12a system that cannot be directly used for non-coding RNA detection. Besides, its combination with HCR and CHA achieves highly sensitive detection of non-coding RNA. [Display omitted] • Finding slicing the facilitated crRNA suitably did not affect the trans -cleavage activity of CRISPR/Cas12a system. • Proposing MCM-CRISPR/Cas12a system to directly detect RNA. • Constructing RNA detection platform by combining MCM-CRISPR/Cas12a system, HCR, and CHA. [ABSTRACT FROM AUTHOR]

Abstract: In this work, a rapid, sensitive and low-cost colorimetric method for detection of quaternary ammonium surfactants using citrate-capped silver nanoparticles (Ag NPs) was developed. The quaternary ammonium surfactants induce the aggregation of Ag NPs through the hydrophobic effect, which is a novel aggregation mechanism of Ag NPs. The addition of cationic surfactant results in color change of Ag NPs solution from yellow to red and finally to colorless, which is due to the broadening of the surface plasmon band. The color change was monitored using a UV–vis spectrophotometer. The LOD of different cationic surfactants was in the range of 0.5–5µM. More importantly, this detection method was successfully utilized to the disinfectant residual sample. [Copyright &y& Elsevier]

Abstract: In this paper, gold nanoparticle–thionine–reduced graphene oxide (GNP–THi–GR) nanocomposites were prepared to design a label-free immunosensor for the sensitive detection of carcinoembryonic antigen (CEA). The nanocomposites with good biocompatibility, excellent redox electrochemical activity and large surface area were coated onto the glassy carbon electrode (GCE) surface and then CEA antibody (anti-CEA) was immobilized on the electrode to construct the immunosensor. The morphologies and electrochemistry of the formed nanocomposites were investigated by using scanning electron microscopy (SEM), ultraviolet–visible (UV–vis) spectrometry, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). CV and differential pulse voltammetry (DPV) studies demonstrated that the formation of antibody–antigen complexes decreased the peak current of THi in the GNP–THi–GR nanocomposites. The decreased currents were proportional to the CEA concentration in the range of 10–500pg/mL with a detection limit of 4pg/mL. The proposed method was simple, fast and inexpensive for the determination of CEA at very low levels. [Copyright &y& Elsevier]

Abstract: We report a simple and novel method of stirring-only-driven accumulation and electrochemical determination of arsenite (As(III)) with both of the oxidation and reduction peaks associated with As(0)/As(III) using a gold nanofilm electrode in neutral solution. Under stirring, a large amount of As(III) was deposited on the modified electrode and the electrochemical response was greatly amplified. The accumulated As(III) on the electrode showed well-defined redox couple in 0.1M blank phosphate buffer solution (pH 7.0), which could be used for the measurement of As(III). Under optimal conditions, As(III) could be detected in the range from 0.20 to 375ppb with a detection limit of 0.04ppb. In particular, with the use of the reduction peak of As(III) the modified electrode exhibits excellent performance for As(III) determination even in the presence of abundant Cu(II). The regeneration of the electrodes is facile with good reproducibility. The electrochemical system was applied to analyze As(III) in lake water, As(III) spiked tap water and drinking water. [Copyright &y& Elsevier]

Abstract: A simple, rapid, and sensitive identification method of fenamithion and acetamiprid is developed using supramolecular nano-sensitizers combining of CdTe quantum dots (QDs) and p-sulfonatocalix[4]arene as additive by fluorescent spectroscopic technique in water. Depending on p-sulfonatocalix[4]arene, the selectivity of CdTe QDs is tuned between fenamithion and acetamiprid. The luminescence of free CdTe QDs is quenched selectively to fenamithion. While in the presence of p-sulfonatocalix[4]arene, it shows that the fluorescence intensity of QDs is enhanced selectively to acetamiprid due to the cooperation of QDs and p-sulfonatocalix[4]arene. Based on the response characteristics of the QDs, a fluorescent method is performed for tuning selective determination of the pesticides. Under optimum conditions described, it is found that the pesticides effect on the luminescence of the CdTe QDs in concentrations dependence are described by a Stern–Volmer-type equation or a Langmuir binding isotherm equation in the range of 0–10−4 M (fenamithion) and 0–10−3 M (acetamiprid), with the corresponding detection limits (3σ) of 1.2×10−8 M (fenamithion) and 3.4×10−8 M (acetamiprid), respectively. The possible mechanism is discussed. [Copyright &y& Elsevier]

Abstract: In this work, chloride, chlorate and perchlorate are fast separated on PDMS microchip and detected via in-channel indirect amperometric detection mode. With PDMS/PDMS microchip treated by oxygen plasma, anions chloride (Cl−), chlorate (ClO3 −), and perchlorate (ClO4 −) are separated within 35s. Some parameters including buffer salt concentration, buffer pH, separation voltage and detection potential are investigated in detail. The separation conditions using 15mM (pH 6.12) of 2-(N-morpholino)ethanesulfonic acid (MES)+ l-histidine (l-His) as running buffer, −2000V as separation voltage and 0.7V as detection potential are optimized. Under this condition, the detection limits of Cl−, ClO3 −, and ClO4 − are 1.9, 3.6, and 2.8μM, respectively. [Copyright &y& Elsevier]

Abstract: It was found that gold nanoparticles with different sizes could enhance the chemiluminescence (CL) of the luminol–NaIO4 system in alkaline solution. The most intensive CL signals were obtained with gold nanoparticles in diameter of 4nm and the CL intensity increased linearly with the concentration of gold nanoparticles. The studies of UV–vis spectra, CL spectra, effects of concentrations of luminol and periodate solution were carried out to explore the CL enhancement mechanism. Catechol, hydroquinone and resorcinol were found to inhibit the CL signals of the luminol–NaIO4 reaction catalyzed by gold nanoparticles, which made it applicable for the determination of these polyphenols. Under the selected experimental conditions, the detection limits (3σ) were in the range of 2.1×10−9 to 1.0×10−10 gml−1, the relative standard deviation (R.S.D., n =11) were in the range of 1.7–2.9%. The method has been successfully applied to the determination of catechol in tap water and synthesized samples with satisfactory results. [Copyright &y& Elsevier]

Abstract: In-channel indirect amperometric detection mode for microchip capillary electrophoresis with positive separation electric field is successfully applied to some heavy metal ions. The influences of separation voltage, detection potential, the concentration and pH value of running buffer on the response of the detector have been investigated. An optimized condition of 1200V separation voltage, −0.1V detection potential, 20mM (pH 4.46) running buffer of 2-(N-morpholino)ethanesulfonic acid (MES)+ l-histidine (l-His) was selected. The results clearly showed that Pb2+, Cd2+, and Cu2+ were efficiently separated within 80s in a 3.7cm long native separation PDMS/PDMS channel and successfully detected at a single carbon fibre electrode. The theoretical plate numbers of Pb2+, Cd2+, and Cu2+ were 1.2×105, 2.5×105, and 1.9×105 m−1, respectively. The detection limits for Pb2+, Cd2+, and Cu2+ were 1.3, 3.3 and 7.4μM (S/N=3). [Copyright &y& Elsevier]

Abstract: An integrated light emitting diode (LED)-induced fluorescence detector was described and evaluated. The LED and its related components including lens and interference filter, the optical fiber used to collect fluorescence, and the capillary column are integrated into a substrate block, which eliminates the need of align procedure of the fiber and the capillary. Forty-fold enhancement of sensitivity was obtained compared with our previous work and the detection limit for fluorescein was 5nM. Application of the detector for the analysis of FITC-labeled Ephedrine extract was demonstrated. [Copyright &y& Elsevier]

The DNA nanomachines as excellent synthetic biological tools have been widely used for the sensitive detection of intracellular microRNA (miRNA) and DNAzyme-involved gene silencing. However, intelligent DNA nanomachines which have the ability to sense intracellular specific biomolecules and respond to external information in complex environments still remain challenging. Herein, we develop a miRNA-responsive DNAzyme cascaded catalytic (MDCC) nanomachine to perform multilayer cascade reactions, enabling the amplified intracellular miRNA imaging and miRNA-guided efficient gene silencing. The intelligent MDCC nanomachine is designed based on multiple DNAzyme subunit-encoded catalyzed hairpin assembly (CHA) reactants sustained by the pH-responsive Zeolitic imidazolate framework-8 (ZIF-8) nanoparticles. After cellular uptake, the MDCC nanomachine degrades in acidic endosome and releases three hairpin DNA reactants and Zn2+, and the latter can act as an effective cofactor for DNAzyme. In the presence of miRNA-21, a catalytic hairpin assembly (CHA) reaction is triggered, which produces a large number of Y-shaped fluorescent DNA constructs containing three DNAzyme modules for gene silencing. The construction of Y-shaped DNA modified with multisite fluorescence and the circular reaction realizes ultrasensitive miRNA-21 imaging of cancer cells. Moreover, miRNA-guided gene silencing inhibits the cancer cell proliferation through the DNAzyme-specific recognition and cleavage of target EGR-1 (Early Growth Response-1) mRNA, which is one key tumor-involved mRNA. The strategy may provide a promising platform for highly sensitive determination of biomolecules and accurate gene therapy of cancer cells. [Display omitted] • A miRNA-responsive intelligent DNA nanomachine is successfully constructed. • The nanomachine with well-designed control center can respond to external information in complex environment. • Amplified miRNA detection and precise gene therapy in cancer cells are achieved. [ABSTRACT FROM AUTHOR]

In this paper, a novel prototype liquid-air dual gradient chip is introduced, which has paved the way for effective synergic effect bio-evaluation of air pollutant. The chip is composed of an array of the agarose liquid-air interfaces, top air gradient layer and bottom liquid gradient layer. The novel agarose liquid-air interface allows for non-biased exposure of cells to all the substances in the air and diffusive interactions with the liquid phase; while the dual liquid-air gradient provides powerful screening abilities, which well reduced errors, saved time and cost from repeated experiment. Coupling the two functions, the chip subsequently facilitates synergic effect evaluation of both liquid and air factors on cells. Here cigarette smoke was taken as the model air pollutant, and its strong synergic effects with inflammatory level of A549 lung cancer cells on their fate were successfully quantified for the first time. These results well testified that the proposed dual-gradient chip is powerful and indispensable for bio-evaluation of air pollutant. [ABSTRACT FROM AUTHOR]

A paper-based SERS test strip combining strengths of paper chip and Raman active substrate was demonstrated to overcome challenges in spectroscopic sensing of complicated samples and realize quantitative detection of disease markers in whole blood. The precisely controlled Au NPs were not only capable of generating condensed hot spots on the fibers, but also enhanced the size exclusion effect of paper, resulting in the novel performance on both SERS detection and sample pretreatment. A biosensor for Mucin-1 is developed by equipping the Au NPs with aptamer. Combining all these merits, this small, cheap and portable test strip might find wide application in clinical diagnosis and health evaluation. [ABSTRACT FROM AUTHOR]

In this work, a miniaturized optical sensor was developed for fluoride determination in tea samples to evaluate their specific risks of fluorosis for public health based on evanescent-wave interaction. The sensor design was integrated on the optical fiber by utilizing the evanescent wave produced on the fiber surface to react with sensing reagents. According to the absorption change at 575 nm, fluoride could be determined by colorimetric method and evaluated by Beer's law. With improved performances of small detection volume (1.2 μL), fast analysis (0.41 min), wide linear range (0.01–1.4 mg L −1 ), low detection limit (3.5 μg L −1 , 3σ) and excellent repeatability (2.34%), the sensor has been applied to fluoride determination in six different tea samples. Conventional spectrophotometry and ion chromatography were employed to validate the sensor's accuracy and potential application. Furthermore, this sensor fabrication provided a miniaturized colorimetric detection platform for other hazardous species monitoring based on evanescent wave interaction. [ABSTRACT FROM AUTHOR]

The direct separation and analysis of chiral drugs in the complex matrix systems are meaningful and challenging. As the most common broad-spectrum antibiotic, levofloxacin has a strong antibacterial ability, but its enantiomer, dextrofloxacin can cause serious harm to human health. In this work, we reported a rapid on-line extraction/ionization device coupled with Electrospray Mass Spectrometry (ESI-MS) for chiral analysis of ofloxacin enantiomers in complex matrix of milk. Since ofloxacin is difficult to dissolve in water and most organic solvents, the procedure of separating ofloxacin in complex system is often complicated. Using the homemade apparatus, the sample pretreatment process was greatly simplified. Milk sample was directly injected and chiral ofloxacin in the sample was extracted at PTFE membrane for further ionization. It took less than 10 s to finish all the procedures including sampling, extraction, reagents mixing, ionization and mass analysis. Utilizing reaction thermodynamics method, trimeric cluster ion [Ni ΙΙ (ref) 2 Ofloxacin-H] + was formed and collisionally dissociated to get chiral resolution of levofloxacin and dextrofloxacin due to the different relative stabilities of the two diastereomeric clusters produced through the dissociation of Ni ΙΙ bound trimeric clusters. With the proposed method, qualitative and quantitative chiral analysis of ofloxacin in milk was successfully achieved in a simple and fast way. [ABSTRACT FROM AUTHOR]

A highly sensitive colorimetric/fluorescent dual-mode sensor based on hybridization chain reaction (HCR) combining multifunctional Au NPs is presented for the detection of Hg 2+ in aqueous solution. In Hg 2+ absent solution, the surface of Au NPs was covered by hairpin auxiliary DNAs and a single strand DNA (ssDNA), which prevented Au NPs from salt-induced aggregation. At the same time, the fluorescence intensity of the dye-labeled hairpin probes was significantly quenched by Au NPs. In the presence of Hg 2+ , T-Hg 2+ -T coordination chemistry between helper DNA and the hairpin probes was induced which triggered the formation of extended double-stranded DNA (dsDNA) polymers via HCR. The formed dsDNA polymers were stiffer which couldn’t attach to Au NPs, resulting in a red-to-blue color change along with salt-induced aggregation of Au NPs for colorimetric sensing. Meanwhile, the fluorescence of dye-labeled DNA turns on. Due to the HCR amplification effect, a highly sensitive detection of Hg 2+ was achieved with detection limit of 0.1 nM. Colorimetry is suitable for the analysis in salt solution with concentration lower than 100 mM. It serves as an intuitive method that Hg 2+ down to 1.0 nM could be identified by naked eyes. For high-concentration salt solutions such as industrial effluent, fluorescent sensing acts as a better choice. Both the color and fluorescence changes of the proposed sensor exhibited high selectivity against other metal ions. Lake water was collected and analyzed using the dual-mode sensor, the results confirmed the practicability of the proposed approach. [ABSTRACT FROM AUTHOR]

A highly sensitive electrogenerated chemiluminescence (ECL) biosensor was developed on the basis of a closed bipolar electrode (BPE) apparatus for the analysis of prostate specific antigen (PSA). Bipolar modifications bring up two different stages of enhancement on the same electrode. Anodic enhancement was conducted by modifying gold nanoparticles (Au NPs) to catalyze the anodic ECL reaction between luminol and hydrogen peroxide. Cathodic introduction of thionine tagged PSA antibody led to a further pertinently enhancement synchronized with the PSA amount variation, because the existence of thionine greatly increased the rate of electron gains on cathode, leading to the corresponding acceleration of anodic ECL reaction. The more thionine modified target molecules were introduced, the faster luminol was oxidized, the higher faraday current approached, and sensitive quantification was realized in correlation with the responsive ECL intensity differences. The quantification resulted in a good determination range between 0.1 pg/mL and 0.1 µg/mL. This strategy mainly took advantage of the special structure of closed BPE to realize a simultaneous amplification on both ends of BPE. Moreover, the platform had a potential of providing a multi-functional strategy for the realization of other bio-detections by simply substituting the PSA sandwich structure with other bio-structures. [ABSTRACT FROM AUTHOR]

In this work, we report a novel microfluidic gas collecting platform aiming at simultaneous sample extraction and multiplex mass spectrometry (MS) analysis. An alveolar-mimicking elastic polydimethylsiloxane (PDMS) structures was designed to move dynamically driven by external pressure. The movement was well tuned both by its amplitude and rhythm following the natural process of human respiration. By integrating the alveolar units into arrays and assembling them to gas channels, a cyclic contraction/expansion system for gas inhale and exhale was successfully constructed. Upon equipping this system with a droplet array on the alveolar array surface, we were able to get information of inhaled smoke in a new strategy. Here, with cigarette smoke as an example, analysis of accumulation for target molecules during passive smoking is taken. Relationships between the breathing times, distances away from smokers and inhaled content of nicotine are clarified. Further, by applying different types of extraction solvent droplets on different locations of the droplet array, simultaneous extraction of nicotine, formaldehyde and caproic acid in sidestream smoke (SS) are realized. Since the extract droplets are spatially separated, they can be directly analyzed by MS which is fast and can rid us of all complex sample separation and purification steps. Combining all these merits, this small, cheap and portable platform might find wide application in inhaled air pollutant analysis both in and outdoors. [ABSTRACT FROM AUTHOR]

In this work, poly-thymine stabilized copper nanoclusters have been used as a fluorescence probe for melamine sensing for the first time. Melamine can bind to thymine through hydrogen bond, which could dramatically enhance the fluorescence intensity of poly-thymine stabilized copper nanoclusters. The enhancement factors ( I − I 0 )/ I 0 increase linearly with the lgC melamine over the melamine concentration range of 0.1 µM to 6 µM. The detection limit of melamine is 95 nM, which is 200 times lower than the US Food and Drug Administration estimate melamine safety limit 20 µM. Melamine in milk was detected with good recovery, which suggested that this novel fluorescence probe has great potential in practical application. [ABSTRACT FROM AUTHOR]

Here we have developed a sensitive cancer cell amplified detection method which combined Au NPs enhanced electrochemiluminescence (ECL) of CdS nanocrystals (NCs) film, with isothermal circular amplification reaction of polymerase. In DNA circular amplification detection system, hairpin DNA beacon/Au NPs composite modified CdS NCs film was used as an ECL emitter. Messenger DNA is hybridized with the aptamer modified on magnetic beads (MBs) to form MB–Au bioconjugates. In the presence of HL-60 cell, the aptamer would conjugate with the glycoprotein at cell surface and messenger DNA sequence would be released. The released messenger DNA sequence was then introduced into the cycle amplification system to trigger circular polymerizations. This assay allows us to determine the released messenger DNA equivalent to 10 cells and exhibits a significant specificity for HL-60 cells. [ABSTRACT FROM AUTHOR]

Helicobacter pylori infections are threats to public health due to their high infection rate and drug resistance. Identification of single-nucleotide variants (SNVs) in H. pylori is crucial for both diagnosis and therapy. Yet the clinical testing of resistant H. pylori mutants is still facing some challenges, such as the selectivity is not good enough for SNVs in abundant wild-type DNA, the lack of clinical validation and the economical burden on patients. Herein, an X-shaped DNA probe with a toehold initiator was designed, which could specifically hybridize with certain genotype DNA due to the thermodynamically driven reaction. A competitive reaction was developed to amplify the thermodynamic difference between wild-type DNA and SNVs, diminishing the interference of wild-type DNA. By this means, multiple SNVs in H. pylori were successfully identified and two SNVs related to clarithromycin resistance are chosen as model targets. A paper strip was fabricated for visual, fast screening of SNVs. Furthermore, the approach was validated using clinical samples, and a point-of-care (POCT) testing diagnosis was executed on saliva samples, demonstrating its potential for the prevention and cure of H. pylori infections. An X-shaped DNA probe with a toehold initiator was designed to identify multiple single-nucleotide variants for clinical evaluation of Helicobacter pylori drug resistance. [Display omitted] • An X-shaped DNA probe with a toehold initiator was designed aiming at drug resistance induced by single-nucleotide variants. • The strategy was validated in clinical samples collected from patients. • A competitive reaction was developed to amplify the thermodynamic difference between wild-type DNA and SNVs. • A POCT diagnosis was executed on saliva samples, demonstrating its potential for the prevention and cure of H. pylori infections. [ABSTRACT FROM AUTHOR]

Abstract: Low cost disposable working electrodes are specifically desired for practical applications of electrochemical detection considering maturity of electrochemical stations and data collection protocols. In this paper double-sided conductive adhesive carbon tape with nanostructure was applied to fabricate disposable working electrodes. Being supported by indium tin oxide glass, the prepared carbon tape electrodes were coated with bismuth film for stripping analysis of heavy metal ions. By integrating the bismuth modified electrodes with paper-based analytical devices, we were able to differentiate Zn, Cd and Pb ions with the sample volume of around 15μL. After the optimization of parameters, including modification of bismuth film and the area of the electrodes, etc., Pb ions could be measured in the linear range from 10 to 500μg/L with the detection limit of 2μg/L. Our experimental results revealed that the disposable modified electrodes could be used to quantify migrated lead from toys with the results agreed well with that using atomic absorption spectrometry. Although bismuth modification and stripping analysis could be influenced by the low conductivity of the carbon tape, the low cost disposable carbon tape electrodes take the advantages of large-scaled produced double-sided carbon tape, including its reproducible nanostructure and scaled-up fabrication process. In addition, the preparation of disposable electrodes avoids time-consuming pretreatment and experienced operation. This study implied that the carbon tape might be an alternative candidate for practical applications of electrochemical detection. [Copyright &y& Elsevier]

Abstract: Aiming to find an alternative electrochemiluminescence (ECL) quenching route of high efficiency, biocatalytic precipitation (BCP) was firstly coupled with ECL for investigating its insulating effect on ECL. Experimental data revealed that an insulating layer could be formed via BCP onto the electrode surface, inhibiting the reaction between the coreactant and luminophore and thereby impairing the ECL emission dramatically. Since the extent of insulating effect associated strongly with analyte concentration, a new ECL biosensor was successfully realized. In a model horseradish peroxidase (HRP)-based system, the fabricated biosensor possessed high sensitivity and wide linear range from 1.0×10−10 M to 1.0×10−6 M for H2O2 determination with a detection limit of 4×10−11 M (S/N =3). This method has great potential in extending the application of ECL biosensor for various bioanalytes. [Copyright &y& Elsevier]

Abstract: This paper presents a simple and cost-effective UV-ablation technique for fabrication of size-tunable nanofluidics devices via photochemical decomposition reaction. UV-irradiation through a PET photomask results in continuous decomposition of poly(carbonate) (PC), forming nanochannel and carboxyl groups on the surface of the etched PC. This photochemical decomposition process occurs at molecular scale, therefore, the depth of nanochannels can be controlled at nanometer level. The etching rate is estimated to be ca. 0.015nms−1. To demonstrate the potential application of the present UV-ablation technique, a nanochannel was fabricated and integrated with microchannels to form a micro/nanofluidics chip for protein concentration. Using this device, about 103–105 fold protein concentration can be achieved within 10min. The present approach offers a simple and practical solution to fabricate nanofluidics devices at low-cost, and the resulting device could provide ideal platforms for μTAS towards various applications in biology and chemistry. [Copyright &y& Elsevier]

Abstract: An electrochemical approach for the sensitive detection of sequence-specific DNA has been developed. Horseradish peroxidase (HRP) assembled on the Fe3O4 nanoparticles (NPs) were utilized as signal amplification sources. High-content HRP was adsorbed on the Fe3O4 NPs via layer-by-layer (LbL) technique to prepare HRP-functionalized Fe3O4 NPs. Signal probe and diluting probe were then immobilized on the HRP-functionalized Fe3O4 NPs through the bridge of Au NPs. Thereafter, the resulting DNA–Au–HRP–Fe3O4 (DAHF) bioconjugates were successfully anchored to the gold nanofilm (GNF) modified electrode surface for the construction of sandwich-type electrochemical DNA biosensor. The electrochemical behaviors of the prepared biosensor had been investigated by the cyclic voltammetry (CV), chronoamperometry (i–t), and electrochemical impedance spectroscopy (EIS). Under optimal conditions, the proposed strategy could detect the target DNA down to the level of 0.7 fmol with a dynamic range spanning 4 orders of magnitude and exhibited excellent discrimination to two-base mismatched DNA and non-complementary DNA sequences. [Copyright &y& Elsevier]

Abstract: Trenbolone (TRE) is a steroid used by veterinarians on livestock to increase appetite and body weight. The use of TRE has been restricted because of its harmful side effect for consumers. To effectively control TRE residue in food and food product, a rapid and convenient immunoassay was developed by preparing an anti-TRE monoclonal antibody. The immunogen and coating antigen were prepared by coupling TRE hapten with carrier proteins via 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC) method. The optimized method gave an average IC50 value of 0.323ngmL−1 towards TRE and an average detection limit (LOD) of 0.06ngmL−1, which is much lower than the maximum residue levels (2.0ngg−1) accepted by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). The specificity of the antibody was evaluated by measuring cross-reactivity of six structurally related compounds, including 19-nortestosterone (9.7%), testosterone (0.13%), methyltestosterone (<0.01%), methandrostenolone (<0.01%), (+)-dehydroisoandrosterone (<0.001%) and β-estradiol (<0.001%). The recovery rates of the test in detection of TRE-fortified animal tissue, urine and animal feed samples were in the range of 81.3–89.4%, while the intra- and inter-assay coefficients of variation were less than 12.0%. [ABSTRACT FROM AUTHOR]

Abstract: Selective extraction of low molecular weight (LMW) proteins and peptides from complex biological samples plays an important role in the discovery of useful biomarkers and signaling molecules. Various methods, such as solid-phase extraction (SPE), ultrafiltration, and size-exclusion chromatography have been developed for such extraction purpose. In this study, we present, to our knowledge, the first demonstration of alkyl-diol@SiO2 mesoporous material MCM-41 (alkyl-diol group on the external surface of mesoporous material) for selective extraction of LMW proteins and peptides from complex biological samples. The adsorption kinetics of LMW proteins, the influence of pH on adsorption and the desorption recovery by different elution solvents were investigated by using standard proteins as model samples. It was demonstrated that the modification of alkyl-diol group on the external surface could efficiently decrease the adsorption of HMW protein and increase the desorption recovery of LMW protein. Furthermore, the mesoporous materials were applied to selectively extract LMW proteins and peptides (<10kDa) from crude human plasma. And the modified MCM-41 material had much better extraction selectivity and efficiency for LMW proteins and peptides than unmodified one. [Copyright &y& Elsevier]

Abstract: The separation of three kinds of aminophenol isomers were achieved within 1min in polyelectrolytes multilayers modified PDMS microchips by layer-by-layer assembly with electrochemical detection (EC). Two polyelectrolytes, poly(dially dimethyl ammonium chloride) (PDDA) and poly(sodium-4-styrene-sulfonate) (PSS) were used to form polyelectrolyte multilayers (PEMs). The surface characteristic of the modified microchip was studied by XPS. The electroosmotic flow (EOF) on PEMs modified PDMS microchips was more stable than that of the native PDMS microchips and the adsorption of samples was greatly reduced on PEMs modified PDMS microchips during the electrophoretic process. The column efficiencies on PEMs modified microchip were increased by 100 times and the signals enhanced by 2 times compared with those of native microchips. The separation conditions such as running buffer pH, running buffer concentration and separation voltage were also optimized. [Copyright &y& Elsevier]

Abstract: A mixture of five amino acids including arginine, histidine, phenylalanine, serine and glutamic acid was successfully separated in microchip capillary electrophoresis and detected with laser-induced fluorescence (LIF) detector. These amino acids were labeled with 5-(4, 6-dichloro-s-triazin-2-ylamino) fluorescein (DTAF). The analyses were performed on two kinds of modified poly(dimethylsiloxane) (PDMS) microchips. One kind of chip was simply treated with oxygen plasma (OP-chip), and the other was further modified by coating double layers of non-ionic polymer poly(vinyl alcohol) (PVA) after plasma oxidization (PVA-chip). The derivatization condition of amino acids by DTAF was optimized. The properties of the two modified PDMS microchips were studied and separation conditions, such as the buffer pH, buffer concentration and separation voltage, were also optimized. The column efficiencies of the two microchips were in the range of 193,000–1,370,000 plates/m. The DTAF-labeled amino acids were sufficiently separated within 50s and 90s in 2.5cm channels on OP-chip and PVA-chip, respectively. [Copyright &y& Elsevier]