Development of an on-chip sample injection system with a 6-port valve incorporated in a microchip

Micro-flow-injection analysis (μFIA) is amenable to high-throughput systems with lower consumption of sample and reagent volumes. On-chip sample injection methods are important to prevent reduced analytical performance associated with dead volumes and diffusion of sample solutions. In this study, we have developed an on-chip sample injection system with a small-sized 6-port valve incorporated on a microchip. The valve is made with a 3D printer and is a simple structure that can be easily operated manually. A sample solution in a loading channel can be injected by switching the valve from the load to injection position. Sample injection tests using resorufin solutions revealed that samples can be injected below 100 μL min⁻¹, and the performance of the sample injection system is comparable to that of a commercially available injector. In addition, the sample injection system was successfully applied to a flow-based assay for hydrogen peroxide. The detection limit (3σ) of hydrogen peroxide was estimated to be 0.5 μM, and the assay time after sample injection was approximately 100 s. The developed sample injection system will be useful for various microfluidic-based analyses including μFIA.

We demonstrate on-chip sample injection using a 6-port valve incorporated in a microchip.     

A selective hybrid fluorescent sensor for fructose detection based on a phenylboronic acid and BODIPY-based hydrophobicity probe

Fructose is widely used in the food industry. However, it may be involved in diseases by generating harmful advanced glycation end-products. We have designed and synthesized a novel fluorescent probe for fructose detection by combining a phenylboronic acid group with a BODIPY-based hydrophobicity probe. This probe showed a linear fluorescence response to d-fructose concentration in the range of 100–1000 μM, with a detection limit of 32 μM, which is advantageous for the simple and sensitive determination of fructose.

We have designed and synthesized a novel fluorescent probe for fructose detection through hydrophobic interactions by combining a phenylboronic acid group and a BODIPY-based hydrophobicity probe with a detection limit of 32 μM.     

The colorimetric and microfluidic paper-based detection of cysteine and homocysteine using 1,5-diphenylcarbazide-capped silver nanoparticles

We have prepared a microfluidic paper-based analytical device (μPAD) for the determination of cysteine and homocysteine based on 1,5-diphenylcarbazide-capped silver nanoparticles. The μPAD was developed to identify and quantify the levels of cysteine and homocysteine. The proposed μPAD enabled the detection of cysteine and homocysteine using a colorimetric reaction based on modified silver nanoparticles. The color of the modified AgNPs in the test zone immediately changed after the addition of cysteine and homocysteine. Based on this change, the quantification of these two amino acids was achieved using an RGB color model and ImageJ software. Under optimized conditions, the proposed device enabled the determination of cysteine in the 0.20–20.0 μM concentration range with a limit of detection (LOD) of 0.16 μM. In addition, the LOD of homocysteine was calculated to be 0.25 μM with a linear range of 0.50–20.0 μM. In this work, we focused on the use of the μPAD for the analysis of a series of human urine samples.

A simple and novel portable method for the quantitative measurement of cysteine and homocysteine in human urine samples is presented.     

A sandwich-type bacteriophage-based amperometric biosensor for the detection of Shiga toxin-producing Escherichia coli serogroups in complex matrices

Immuno-based biosensors are a popular tool designed for pathogen screening and detection. The current antibody-based biosensors employ direct, indirect, or sandwich detection approaches; however, instability, cross-reactivity, and high-cost render them unreliable and impractical. To circumvent these drawbacks, here we report a portable sandwich-type bacteriophage-based amperometric biosensor, which is highly-specific to various Shiga toxin-producing Escherichia coli (STEC) serogroups. Environmentally isolated and biotinylated bacteriophages were directly immobilized onto a streptavidin-coated screen-printed carbon electrode (SPCE), which recognized and captured viable target cells. Samples (50 μL) were transferred to these bacteriophage-functionalized SPCEs (12 min, room temp) before sequentially adding a bacteriophage–gold nanoparticle solution (20 μL), H₂O₂ (40 mM), and 1,1′-ferrocenedicarboxylic acid for amperometric tests (100 mV s⁻¹) and analysis (ANOVA and LSD, P < 0.05). The optimum biotin concentration (10 mM) retained 94.47% bacteriophage viability. Non-target bacteria (Listeria monocytogenes and Salmonella Typhimurium) had delta currents below the threshold of a positive detection. With less than 1 h turn-around time, the amperometric biosensor had a detection limit of 10–10² CFU mL⁻¹ for STEC O157, O26, and O179 strains and R² values of 0.97, 0.99, and 0.87, respectively, and a similar detection limit was observed in complex matrices, 10–10² CFU g⁻¹ or mL⁻¹ with R² values of 0.98, 0.95, and 0.76, respectively. The newly developed portable amperometric biosensor was able to rapidly detect viable target cells at low inoculum levels, thus providing an inexpensive and improved alternative to the current immuno- and laboratory-based STEC screening methods.

Sandwich-type bacteriophage-based amperometric biosensor for the detection of Shiga toxin-producing Escherichia coli serogroups in complex matrices.     

Preparation of polymethacrylate monolith modified with cysteine for the determination of Cr(iii) ions

In this study, a simple and rapid polymer monolith microextraction procedure was developed for the determination of Cr(iii) ions by inductively coupled plasma-atomic emission spectrometry. A monolithic column modified with cysteine was synthesized and characterized by scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, thermal gravimetric analysis, specific surface area analysis and pore size distribution analysis. The influences of analytical parameters such as sample pH, adsorption time, eluent type, and coexisting ions were examined. The limit of detection (LOD) and limit of quantification (LOQ) for Cr(iii) ions were 0.005 μg mL⁻¹ and 0.017 μg mL⁻¹, and the relative standard deviation (RSD) was 7.4% (n = 5). The prepared cysteine functionalized monolithic column displayed good enrichment capacity and was successfully applied to the determination of Cr(iii) ions in real samples.

In this study, a simple and rapid polymer monolith microextraction procedure was developed for the determination of Cr(iii) ions by inductively coupled plasma-atomic emission spectrometry.     

Highly selective and sensitive detection of amaranth by using carbon dots-based nanosensor

In this work, a novel fluorescence nanosensor for selective and sensitive determination of amaranth was constructed using carbon dots (C-dots). Water soluble C-dots with strong fluorescence were obtained by a simple microwave-assisted method using urea and glycine as raw materials. It was found that amaranth can efficiently and sensitively quench the C-dots fluorescence by the inner filter effect (IFE) and non-radiative energy transfer (NRET) mechanisms. The fluorescence quenching efficiency (F₀/F) was strongly correlated with the concentration of amaranth in the 0.2–30 μM range. The detection limit (LOD) is 0.021 μM. There was no significant change in the fluorescence intensity of C-dots when other potentially interfering substances were present in the system. Our C-dots-based nanosensor was successfully utilized for the analysis of amaranth in drinks and showed rapid, sensitive and accurate responses. It indicates that the novel C-dots-based nanosensor has great potential in amaranth detection for real-life applications.

Illustration of the synthesis of C-dots and the determination of amaranth based on the fluorescence quenching of C-dots.     

Determination of berberine hydrochloride using a fluorimetric method with silica nanoparticles as a probe

The interaction of silica nanoparticles (SiO₂NPs) with berberine hydrochloride (BRH) was studied in aqueous solution at pH 9.0 and room temperature by using fluorophotometry. Based on a significant enhancement of the fluorescence intensity of the SiO₂NPs–BRH aggregates, a spectrofluorimetric method which was simple, sensitive and green was developed for the determination of BRH in aqueous solution. The linear range of the method was from 2.0–50.0 μg L⁻¹ with a detection limit of 0.73 μg L⁻¹. There was no interference from the compounds normally used to formulate pharmaceutical tablets. The proposed method was applied to the determination of BRH in tablets with satisfactory results and good consistency with the results obtained by standard methods.

A simple, green and sensitive spectrofluorimetric method was proposed for the determination of berberine hydrochloride (BRH) in aqueous solution.     

A simple preparation method of carbon dots by weak power bathroom lamp irradiation and their application for nimesulide detection and bioimaging

In this work, a novel strategy for synthesizing carbon dots (CDs) with a quantum yield of approximately 15.36% has been established by employing a bathroom lamp as a light source. Compared with other current protocols, the method described here displayed various advantages such as environmentally friendly manipulations and low power and cost. Subsequently, we applied the CDs as a fluorescence probe for the detection of nimesulide (Nim) firstly under the optimal conditions. A linear relationship between ln(F₀/F) and the concentration of Nim was obtained in the range from 0.5 μM to 75 μM with a detection limit of 100 nM. In addition, the as-prepared CDs showed excellent biocompatibility and were applied for cell imaging, which presented great potential applications in cell imaging.

This work reported the simple preparation method of carbon dots using weak power bathroom lamp irradiation, and explored their potential application in cell imaging and as a fluorescent sensor for the determination of nimesulide.     

Simultaneous detection of acetaminophen and 4-aminophenol with an electrochemical sensor based on silver–palladium bimetal nanoparticles and reduced graphene oxide

In this paper, Ag–Pd bimetallic nanoparticles uniformly distributed on reduced graphene oxide (rGO) were synthesized by redox reaction between Pd²⁺, Ag⁺and GO, and were characterized by X-ray diffractometry, field emission scanning electron microscopy, electrochemical impedance spectroscopy and thermal gravimetric analyses. A novel electrochemical sensor was constructed based on these nanocomposites using glassy carbon as a substrate. Under optimal conditions, the linear ranges were 0.50–300.00 μM for PA and 1.00–300.00 μM for 4-AP, with the detection limits of 0.23 μM for PA and 0.013 μM for 4-AP, respectively. This sensor was successfully applied to the determination of PA in pharmaceutical formulations and gave satisfactory results with a lower detection limit, wider linear range and good reproducibility.

Simultaneous detection of acetaminophen and 4-aminophenol with a highly sensitive electrochemical sensor based on silver–palladium bimetal nanoparticles and reduced graphene oxide.     

A new strategy for the sensitive electrochemical determination of nitrophenol isomers using β-cyclodextrin derivative-functionalized silicon carbide

In the present study, thiol β-cyclodextrin (SH-CD) and ethylenediamine β-cyclodextrin (NH₂-β-CD) were simultaneously grafted on the same interface of an Au NP deposited carboxyl SiC (Au@CSiC) nanocomposite. An electrochemical sensor for the simultaneous determination of nitrophenol isomers (o-nitrophenol, o-NP; p-nitrophenol, p-NP) using SH-CD and NH₂-β-CD functionalized Au@SiC (Au@CSiC-SH/NH₂-CD) nanocomposite was successfully constructed. Differential pulse voltammetry was used to quantify o-NP and p-NP within the concentration range of 0.01–150 μM under the optimal conditions. The detection limit (S/N = 3) of the sensor was 0.019 and 0.023 μM for o-NP and p-NP, respectively, indicating a low detection limit. Interference study results demonstrated that the sensor was not affected in the presence of similar aromatic compounds during the determination of NP isomers, showing high selectivity. The proposed electrochemical sensing platform was successfully used to determine NP isomers in tap water. The low detection limit and high selectivity of the proposed electrochemical sensor were caused by the high surface area, the excellent conductivity, and the more recognized (enriched) NP isomer molecules by SH-β-CD and NH₂-β-CD of the Au@CSiC-SH/NH₂-CD nanocomposite.

An illustration of simultaneous electrochemical determination of nitrophenol isomers using β-cyclodextrin derivative-functionalized silicon carbide.     

Novel single excitation dual-emission carbon dots for colorimetric and ratiometric fluorescent dual mode detection of Cu2+ and Al3+ ions

In this study, dual-emission carbon dots (D-CDs) are synthesized via a simple one-step solvothermal treatment of red tea. The obtained D-CDs are characterized by XPS, IR, TEM, XRD, fluorescence and UV-vis spectroscopy techniques. It is found that D-CDs present a strong red fluorescence emission peak at 671 nm and weak blue fluorescence emission peak at 478 nm under the excitation wavelength of 410 nm. The unique dual-emission properties of D-CDs provide great opportunities in ratiometric fluorescence sensing applications. The results show that Cu²⁺ ions can quench the fluorescence of the red emission band of D-CDs effectively, resulting in the disappearance of red fluorescence ultimately. Upon the addition of Al³⁺ ions, the fluorescence of blue emission band at 478 nm grows apparently, and the fluorescence color transforms gradually from red to orange, then to yellow-green. Based on these findings, a novel ratiometric fluorescence and colorimetric dual mode nanosensor is developed for simultaneous detection of Cu²⁺ and Al³⁺ ions. Regarding Cu²⁺ ions, the fluorescent detection linear range is 0.1–50 μM with detection limit of 0.1 μM, and the colorimetric detection limit is estimated as 25 μM. With regard to Al³⁺ ions, the fluorescent detection linear range is 0–20 μM and 25–100 μM with detection limit of 0.5 μM, and the colorimetric detection limit is 20 μM. Furthermore, the fluorescence response mechanisms of Cu²⁺ and Al³⁺ ions were discussed detailed. To the best of our current knowledge, this will be the first research work on the simultaneous determination of Cu²⁺ and Al³⁺ using D-CDs as fluorescent probes.

D-CDs with strong red emission and weak blue emission as an effective colorimetric and ratiometric fluorescence sensing probe are employed to realize the simultaneous detection of Cu²⁺ and Al³⁺ ions without any interference effect.     

Cobalt metal–organic framework-based ZIF-67 for the trace determination of herbicide molinate by ion mobility spectrometry: investigation of different morphologies

Co-MOF-based zeolitic imidazolate frameworks (ZIF-67) with various morphologies were prepared via an innovative way under distinct reaction conditions. By changing the reaction conditions, including the cobalt source, solvent, time, temperature, and linking agent to the cobalt ions, the morphological evolution of Co-MOF-based ZIF-67 was investigated. The Co-MOF-based ZIF-67 was applied as an adsorbent fiber in the solid-phase microextraction (SPME) technique for extracting a herbicide, namely molinate (as a test compound), in aqueous samples. For recognizing the molinate molecules, drift tube ion mobility spectrometry (IMS) was employed as a sensitive, rapid, and simple detection technique. Two essential parameters, namely extraction temperature and extraction time, influenced the extraction efficiency, and these parameters were also analyzed and optimized. The linear dynamic range (LDR) and the determination coefficient were found to be 0.5–20.0 μg L⁻¹ and 0.9990, respectively. In this regard, the limit of quantification (LOQ) and the detection limit (LOD) were calculated and found to be 0.5 μg L⁻¹ and 0.15 μg L⁻¹, respectively. Finally, the effect of the adsorbent with different morphologies on the extraction efficiency was compared.

Co-MOF-based zeolitic imidazolate frameworks (ZIF-67) with various morphologies were prepared via an innovative way under distinct reaction conditions.     

Simultaneous voltammetric detection of dopamine,ascorbic acid and uric acid using a poly(2-(N-morpholine)ethane sulfonic acid)/RGO modified electrode

A poly(2-(N-morpholine) ethane sulfonic acid)/reduced graphene oxide (RGO) modified glassy carbon electrode (GCE) was prepared using an electropolymerization method, and was characterized by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). The electrochemical behaviors and simultaneous detection of ascorbic acid (AA), dopamine (DA) and uric acid (UA) at this electrode were studied by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Tests showed that this electrode exhibited excellent electrocatalytic activity towards the oxidation of AA, DA and UA. The oxidation peak currents of AA, DA and UA were proportional with their concentrations in the ranges 1.0 μM–30 μM (30 μM–100 μM), 0.05 μM–100 μM and 0.1 μM–100 μM, with detection limits of 0.43 μM, 0.0062 μM and 0.056 μM, respectively. In addition, this electrode exhibited an excellent selectivity, reproducibility and stability, and has been successfully used to determine real samples with satisfactory results.

A sensitive electrochemical sensor for simultaneously detecting dopamine, ascorbic acid and uric acid.     

Co2+ detection,cell imaging,and temperature sensing based on excitation-independent green-fluorescent N-doped carbon dots

Green-fluorescent N-doped carbon dots (N-CDs) have been successfully fabricated using hydrothermal treatment of tyrosine and urea. The N-CDs obtained showed excitation-independent emission, superior stability and strong photoluminescence with a quantum yield of ca. 9.8%. Based on these striking behaviors, the as-prepared N-CDs have been utilized in Co²⁺ detection and temperature sensing. Due to an inner filter effect, the N-CDs obtained were dramatically quenched by Co²⁺ with linear ranges of 0.1 μM–10 μM, 25 μM–275 μM and 300 μM–400 μM, and they had a detection limit of 0.15 μM. The use of the as-prepared N-CDs has been extended to visualize Co²⁺ fluctuations in living cells. Additionally, the N-CDs obtained have also been applied for use as a temperature sensor with a linear range of 25–80 °C.

Green-fluorescent N-doped carbon dots (N-CDs) have been successfully fabricated using hydrothermal treatment of tyrosine and urea.     

Nanozymes with reductase-like activities: antioxidant properties and electrochemical behavior

Nanozymes (NZs) as stable cost-effective mimics of natural enzymes may be promising catalysts in food and environmental biotechnology, biosensors, alternative energy and medicine. The majority of known NZs are mimetics of oxidoreductases, although there are only limited data regarding mimetics of reductases. In the present research, a number of metal-based NZs were synthesized via chemical methods and screened for their antioxidant ability in solution. The most effective reductase-like Zn/Cd/Cu NZ was characterized in detail. Its antioxidant properties in comparison with several food products and Trolox, as well as substrate specificity, size and composition were studied. Zn/Cd/Cu NZ was shown to mimic preferentially selenite reductase. The amperometric sensor was constructed possessing a high sensitivity (1700 A M⁻¹ m⁻²) and a broad linear range (16–1000 μM) for selenite ions. The possibility to apply the fabricated sensor for selenite determination in commercial mineral water has been demonstrated.

Novel Zn/Cd/Cu^bd nanozymes possesses the ability to mimic coenzyme-dependent selenite reductase. A new amperometric biosensor for determination of selenite was constructed.     

Au–Ag core–shell composite nanoparticles as a selective and sensitive plasmonic chemical probe for l-cysteine detection in Lens culinaris (lentils)

The present work reported is a simple and selective method for the colorimetrical detection of l-cysteine in Lens culinaris (or lentils) using Au–Ag core–shell (Au core Ag shell) composite nanoparticles as a chemical probe. The phenomenon is based on the color change of composite nanoparticles from yellowish brown to light blue, followed by a shift of the localized surface plasmon resonance (LSPR) absorption band in the UV-visible region (i.e., 200–800 nm) with the addition of l-cysteine into the solution of bimetallic nanoparticles. The mechanism for the detection of l-cysteine is based on the electrostatic interaction of the metal ion with the thiol group of the amino acid, which causes the red shift of the LSPR band at 685 nm. The size distribution, morphology, composition and optical properties of the Au–Ag core–shell composite nanoparticles were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), energy dispersive X-ray diffraction (EDX), UV-visible spectrophotometer and Fourier transform infrared spectroscopy (FTIR) techniques. An excellent linearity range for the present method was observed in the range of 20–140 μg mL⁻¹ with a limit of detection at 1.95 μg mL⁻¹ and correlation coefficient (R²) of 0.986. A good% recovery of 4.0% showed the selectivity of the method for l-cysteine determination from sample matrices. The advantageous features of the present method are being simple, rapid, low cost and selectivity towards the determination of l-cysteine in lentils.

The present work reported is a simple and selective method for the colorimetrical detection of l-cysteine in Lens culinaris (or lentils) using Au–Ag core–shell (Au core Ag shell) composite nanoparticles as a chemical probe.     

Highly selective colorimetric determination of catechol based on the aggregation-induced oxidase–mimic activity decrease of δ-MnO2

Multiple enzyme-like activities of manganese oxides (MnO₂) have been reported and applied in catalysis, biosensors, and cancer therapy. Here, we report that catechol can be determined colorimetrically based on the 3,3′,5,5′-tetramethylbenzidine (TMB) oxidase-like activity of δ-MnO₂. The detection was based on pre-incubation of catechol containing water samples with δ-MnO₂, and then the residual TMB oxidase-like activity of reacted δ-MnO₂ was linearly dependent on the catechol concentration in the range of 0.5 to 10 μM. This determination method was stable at pH 3.73–6.00 and was not affected by ion strength up to 200 μM. Common co-solutes in water bodies (50 μM) have negligible effects and excellent selectivity of catechol among various phenolic compounds (15 μM) was facilitated. Both reduction and aggregation of δ-MnO₂ were observed during the incubation process with catechol, and aggregation-induced TMB oxidase–mimic activity decrease was the main factor for this colorimetric determination.

A new determination mechanism for catechol: aggregation-induced oxidase-mimic activity decrease of δ-MnO₂.     

A fungus-derived biomass porous carbon–MnO2 nanocomposite-modified electrode for the voltammetric determination of rutin

In this study, we designed a simple procedure for the synthesis of fungus-derived biomass porous carbon (FBPC), which was further used to prepare a MnO₂@FBPC composite by a hydrothermal method. The MnO₂@FBPC nanocomposite showed a porous structure, large specific surface area, and high conductivity, and was modified on the carbon ionic liquid electrode (CILE) to obtain a working electrode for the sensitive voltammetric determination of rutin. The electrochemical response of rutin was studied via cyclic voltammetry with electrochemical parameters calculated. Under the optimal conditions, the linear range for the rutin analysis was obtained by the differential pulse voltammetry from 0.008 to 700.0 μmol L⁻¹ with the detection limit of 2.67 nmol L⁻¹ (3σ). This MnO₂@FBPC/CILE was applied to directly detect the rutin concentration in drug and human urine samples with satisfactory results.

Fabrication process of MnO₂@FBPC composites and the electrochemical detection of rutin.     

Simultaneous determination of hydroquinone and catechol by a reduced graphene oxide–polydopamine–carboxylated multi-walled carbon nanotube nanocomposite

A reduced graphene oxide–polydopamine–carboxylated multi-walled carbon nanotube (RGO–PDA–cMWCNT) nanocomposite was fabricated via a facile, one-pot procedure and was characterized by a variety of techniques. A novel electrochemical sensor based on RGO–PDA–cMWCNT was constructed to determine hydroquinone (HQ) and catechol (CT) simultaneously. This newly prepared nanocomposite shows excellent electrocatalytic efficacy in the electrode reaction of the two isomers. Specifically, the peak-to-peak potential difference between the two dihydroxybenzenes is 115 mV for oxidation, which is obviously larger than similar electrochemical sensors. The established method displays a wide linear range from 0.5 to 5000 μM with a detection limit (S/N = 3) of 0.066 μM for HQ and 0.073 μM for CT. In addition, this electrochemical approach has been tested to measure the two dihydroxybenzenes in real samples and satisfactory results were recorded.

A novel reduced graphene oxide–polydopamine–carboxylated multi-walled carbon nanotube nanocomposite (RGO–PDA–cMWCNT) was fabricated for the sensitive and simultaneous determination of hydroquinone (HQ) and catechol (CT).     

Ultrasensitive detection of uric acid in serum of patients with gout by a new assay based on Pt@Ag nanoflowers

A ultrasensitive assay for the determination of uric acid (UA) based on Pt@Ag nanoflowers (Pt@Ag NFs) was constructed. H₂O₂ was formed by the reaction of uricase and UA and produced the hydroxyl radical (˙OH). The system was catalyzed by Pt@Ag NFs to change the color of 3,3′,5,5′-tetramethylbenzidine (TMB) from colorless to blue, and the morphology and chemical properties of Pt@Ag NFs were characterized by transmission electron microscopy and X-ray photoelectron spectroscopy. Under the optimized conditions, a linear relationship between the absorbance and UA concentration was in the range of 0.5–150 μM (R² = 0.995) with a limit of detection of 0.3 μM (S/N = 3). The method can be applied to detection of UA in actual samples with satisfactory results. The proposed assay was successfully applied to the detection of UA in human serum with recoveries over 96.8%. Thus, these results imply that the UA assay provides an effective tool in fast clinical analysis of gout.

A ultrasensitive assay for the determination of uric acid (UA) based on Pt@Ag nanoflowers (Pt@Ag NFs) was constructed.