The present study demonstrates cation-driven optical properties of artificial luciferases (ALucs) from copepod luciferases, as an optical readout for bioanalysis. An assignment of the supersecondary structure code (SSC) of ALucs revealed that ALucs carry a helix-loop-helix structure, which appears at the same sites of the EF-hands of typical Ca2+-binding proteins. A mutagenesis study shows that the EF-hand-like structure is a pivotal site for enzymatic activity. The effects of 20 kinds of mono- and multivalent cations on ALuc activities were estimated with column-purified ALuc16. High pH values boost the ALuc activities with both the native coelenterazine and an analog called 6-pi-OH-CTZ. Multivalent cations, Ca(II), Mg(II), and Cr(VI), elevate and prolong the ALuc activities, whereas Co(II), Cu(II) and Pb(II) greatly hamper the ALuc activities. Ca(II) greatly prolongs the optical intensities, suggesting a contribution to the structural robustness of ALucs. The inhibitory effect of multivalent cations on the ALuc activities was utilized for creating dose-response curves. The intrinsic cation-driven selectivity and optical intensity of ALucs enable researchers to constitute de novo biosensors for multivalent cations.
Novel, sensitive and rapid electrochemical methods for the determination of phenothiazine and azaphenothiazine derivatives were developed. A boron-doped diamond (BDD) electrode was used for electrochemical oxidation of levomepromazine, promazine and prothipendyl. The electrooxidation of these substances demonstrated reversible peaks of oxidation at the potential range 0.55 – 0.75 V vs. SCE. Examining the influence of scan rate allowed is to demonstrate that the currents registered typical of the diffusion-controlled process. Determinations of the studied analytes were carried out by means of a square wave voltammetry (SWV) method and a differential pulse voltammetry (DPV) method. Linear ranges of determination with the use of the BDD electrode and the SWV method were obtained in the ranges: from 4 × 10−7 to 1.38 × 10−4 mol L−1 for levomepromazine, from 4 × 10−7 to 1.17 × 10−5 mol L−1 for promazine and from 4.95 × 10−7 to 4.54 × 10−5 mol L−1 for prothipendyl. The influence of interferences on the voltammetric signal of the studied analytes was also checked. The proposed procedures were used for quantitative determination of the studied compounds in pharmaceutical preparations. The measurements showed high accuracy. The recovery values obtained ranged from 98.52 to 99.57%. The developed procedures were compared with pharmacopoeial reference methods.
In this paper, nitrogen-doped carbon dots (N-CDs) with high quantum yield (QY) of 40.5% were prepared through a facile and straightforward hydrothermal route. The as-prepared N-CDs exhibited excellent photoluminescence properties, good water-solublity and photostability, negligible cytotoxicity and favourable biocompatibility. Such N-CDs were found to serve as an effective fluorescent sensor for selective and sensitive detection of Hg2+ in a wide linear response concentration range of 0 – 8 μM with a limit of detection (LOD) of 0.087 μM and could be applied to the determination of Hg2+ in environmental water samples. The corresponding mechanisms were discussed in detail. Moreover, another attractive finding was that the N-CDs showed satisfactory performance in bioimaging before and after the addition of Hg2+ in human lung cancer PC14 cells. With excellent sensitivity, selectivity and biocompatibility, such cheap carbonmaterials are potentially suitable for monitoring of Hg2+ in environmental applications and promising for biological applications.
Changes in the interfacial tension of a lipid monolayer membrane formed at the water/chloroform interface upon DNA addition were measured using the quasi-elastic laser scattering (QELS) method. A cationic lipid, N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP), as well as zwitterionic lipids, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), were used to form lipid monolayer membranes at different calcium ion concentrations. A rapid decrease of the interfacial tension resulting from electrostatic interactions between DOTAP and DNA was observed within 10 s. However, such rapid decreases were not observed for DOPE or DOPC. A decrease in the interfacial tension was exhibited by DOPE after 1000 s from the addition of DNA, which may be due to an overall structural change in the DOPE membrane. A DOTAP/DOPE complex system showed behaviors attributable to both DOTAP and DOPE, whereas the behavior of the DOTAP/DOPC system resembled that of DOPC alone. The current results provide a model for the so-called lipoplex carriers used in gene therapy.
A new sensitive method for total mercury determination in reference materials using a 5-phase digitally controlled rotating field plasma source (RFP) for optical emission spectrometry (OES) was developed. A novel synergic effect of ultrasonic nebulization (USN) and ultraviolet-visible light (UV-Vis) irradiation when used in combination was exploited for efficient Hg vapor generation. UV- and Vis-based irradiation systems were studied. It was found that the most advantageous design was an ultrasonic nebulizer fitted with a 6 W mercury lamp supplying a microliter sample to a quartz oscillator, converting liquid into aerosol at the entrance of the UV spray chamber. Optimal conditions involved using a 20% v/v solution of acetic acid as the generation medium. The mercury cold vapor, favorably generated from Hg2+ solutions by UV irradiation, was rapidly transported into a plasma source with rotating field generated within the five electrodes and detected by digitally controlled rotating field plasma optical emission spectrometry (RFP-OES). Under optimal conditions, the experimental concentration detection limit for the determination, calculated as the concentration giving a signal equal to three times the standard deviation of the blank (LOD, 3σblank criterion, peak height), was 4.1 ng mL−1. The relative standard deviation for samples was equal to or better than 5% for liquid analysis and microsampling capability. The methodology was validated through determination of mercury in three certified reference materials (corresponding to biological and environmental samples) (NRCC DOLT-2, NRCC PACS-1, NIST 2710) using the external aqueous standard calibration techniques in acetic acid media, with satisfactory recoveries. Mercury serves as an example element to validate the capability of this approach. This is a simple, reagent-saving, cost-effective and green analytical method for mercury determination.
A new approach for the development of a highly sensitive aluminium(III) ion sensor via the preconcentration of aluminium(III) ion with a self-assembled monolayer on a gold nanoparticles modified screen-printed carbon electrode and current mediation by potassium ferricyanide redox behavior during aluminium(III) ion binding has been attempted. A monolayer of mercaptosuccinic acid served as an effective complexation ligand for the preconcentration of trace aluminium; this led to an enhancement of aluminium(III) ion capture and thus improved the sensitivity of the sensor with a detection limit of down to the ppb level. Under the optimum experimental conditions, the sensor exhibited a wide linear dynamic range from 0.041 to 12.4 μM. The lower detection limit of the developed sensor was 0.037 μM (8.90 ppb) using a 10 min preconcentration time. The sensor showed excellent selectivity towards aluminium(III) ion over other interference ions.
One way to determine the pH at the air/water interface with a confocal fluorescence microscope has been proposed. The relation between the pH at the air/water interface and that in a bulk solution has been formulated in connection with the adsorption equilibrium and the dissociation equilibrium of the dye adsorbed. Rhodamine B (RhB) is used as a surface-active fluorescent pH probe. The corrected fluorescence spectrum of RhB molecules at the air/water interface with the surface density of 1.0 nmol m−2 level shows pH-dependent shifts representing an acid-base equilibrium. Two ways to determine the unknown acid-base equilibrium constant of RhB molecules at the air/water interface have been discussed. With surface-tension measurements, the adsorption properties, maximum surface density, and adsorption equilibrium constants were estimated for both cationic and zwitterionic forms of RhB molecules at the air/water interface.
This paper describes a convenient and rapid fluorescence sensor for determination of paraquat (PA) based on glutathione-capped CdS quantum dots (QDs). The methodology enabled the use of a simple synthesis procedure for water solubilization of CdS QDs via a fast route using glutathione as a capping agent within 15 min. The resulting water-soluble QDs exhibit a strong fluorescence emission at 536 nm with high and reproducible photostability. PA is an important class of electron acceptors for QDs. Thus, the fluorescence intensity of the glutathione-capped CdS QDs probe could be dramatically quenched by PA due to the electron transfer mechanism. The fluorescence intensity of the CdS QDs system was proportional to PA concentration in the range of 0.025 to 1.5 μg mL−1, with a detection limit of 0.01 μg mL−1. The time of analysis sample, including preparation of QDs and fluorescent measurement for PA, was only 20 min. Most of the potentially coexisting substances did not interfere with the PA-induced quenching effect except diquat. Furthermore, the analytical applicability of the proposed method was demonstrated by analyzing PA in water, rice and cabbage samples, and the recoveries were between 86 and 105% which satisfied the requirement of detection for PA. These results showed that the proposed method was simple in design and fast in operation, and could be used as a sensitive tool for detecting PA in environmental and agricultural samples.
We developed moment analysis of affinity kinetics by chromatographic capillary electrophoresis (MKCCE) method for the kinetic study of intermolecular interactions. Association and dissociation rate constants of the interaction in a micellar electrokinetic chromatography (MEKC) system between thymol and sodium dodecylsulfate micelle were determined by the MKCCE method. It is a method based on the moment theory for the kinetic study of intermolecular interactions under the conditions that neither immobilization nor chemical modification of molecules is required. In CCE mode, experimental conditions are controlled so that the migration of solute-micelle complex is stopped and only solute molecules migrate in a capillary. Mass transfer behavior of solute molecules in the CCE system is analogous to that in a chromatographic system. However, because it was difficult in practice to really perform CE experiments under the CCE conditions, CE data were measured with changing experimental conditions, i.e., applied pressure, under the conditions that the migration velocity of solute-micelle complex was around zero. The rate constants could be analytically determined from the CE data. In the MKCCE method, it is not necessary to fit elution curves numerically calculated to those experimentally measured for the determination of the rate constants. Regarding the interaction between thymol and SDS micelles, association equilibrium constant and association and dissociation rate constants were determined as 6.35 × 103 dm3 mol−1, 5.6 × 104 dm3 mol−1 s−1, and 8.7 s−1, respectively. It was demonstrated that the MKCCE method was effective for the kinetic study of intermolecular interactions.
A novel biosensor was developed by immobilizing hemoglobin (Hb) on a glassy carbon electrode (GCE) modified with a composite of ZnO nano-rods and carbon nanofiber (CNF), a strong reducer, hydrazine, was firstly used to evaluate the electrochemical behavior of Hb on Hb/ZnO/CNF/GCE. UV-vis and circular dichroism (CD) spectra indicated the conformational structure of Hb interaction with ZnO/CNF was predominantly an α-helical structure. The modified electrodes were characterized by scanning electron microscopy (SEM), electron impedance spectroscopy (EIS), and cyclic voltammetry. Electrocatalytic mechanism of Hb to oxidation reaction of hydrazine was suggested. The bioelectrocatalytic activity, kinetic parameters of Michaelis–Menten constant (Km), stability and reproducibility were also investigated. A linear dependence of peak currents to the concentrations of hydrazine was observed in the range from 1.98 × 10−5 to 1.71 × 10−3 mol L−1 with a correlation coefficient of 0.998, and a detection limit (S/N = 3) of 6.60 μmol L−1 was estimated.
Metal-organic framework Fe-MIL-88NH2 as a dual colorimetric and fluorometric sensor has been designed for a wide range of dopamine quantitative detection. It is easy to implement the assay for visual detection of dopamine based on restraining the color change of the 3,3′,5,5′-tetramethylbenzidine-H2O2 system that is catalyzed by Fe-MIL-88NH2 with intrinsic peroxidase-like catalytic activity. The linear range is from 50 nM to 30 μM. In addition, the Fe-MIL-88NH2 can exhibit a dramatic decrease of its fluorescent intensity when exposed to dopamine, which may be attributed to the electron transfer from the Fe-MIL-88NH2 to the oxidized dopamine-quinone. The linear response range is from 30 μM to 4 mM. Meanwhile, both colorimetric and fluorometric methods exhibit higher selectivity for DA over a number of possible interfering substances. Furthermore, the proposed method has been successfully applied to detect DA in human serum samples, which suggests its great potential for analytical applications.
Herein, we developed a sensing strategy for the label-free detection of Zn2+ based on G-quadruplex. In the absence of Zn2+, there was a fluorescence enhancement of thioflavin T by interaction with human telomere sequence. On the addition of Zn2+, Zn2+ induced a more compact G-quadruplex structure to release thioflavin T, resulting in a fluorescence decrease. This simple “mix-then-detect” method gave the detection limit of 0.91 μM with linear dynamic ranges from 0 to 10 μM. Because it does not require the use of expensive and unstable DNAzyme systems, or need synthesis and modification of nanomaterials, this label-free biosensor is simple, fast, cost-effective and applicable for real samples taken from lake water.
The use of a microwave assisted solvent-free technique for silica coating of iron magnetic nanoparticles (Fe3O4-MNPs) and their functionalization with three aliphatic diamines: 1,2-ethylenediamine (1,2EDA), 1,5-pentanediamine (1,5PDA) and 1.8-octanediamine (1,8-ODA), were successfully achieved in a very short time. Only 60 min were needed for the nano-adsorbent modification as compared with more than 1000 min using conventional methods under reflux conditions. Their surface characteristics (observed by TEM, XRD and FT-IR), in addition to Cu(II) adsorption capacities (1.805, 1.928 and 2.116 mmol g−1) and time of equilibration (5 s) were almost the same. Thus, the time required to accomplish the solid phase extraction process is greatly reduced. On the other hand, the phenomenon of the fast equilibration kinetics was successfully extended on using the functionalized aliphatic diamines magnetic nano-adsorbents as precursors for further microwave treatment. Three selective magnetic nano-adsorbents (Fe3O4-MNPs-SiO2-1,2EDA-3FSA, Fe3O4-MNPs-SiO2-1,5PDA-3FSA and Fe3O4-MNPs-SiO2-1,8ODA-3FSA) were obtained via the reaction with 3-formayl salicylic acid (3FSA) as a selective reagent for Fe(III). At 5 s contact time, they exhibited maximum Fe(III) uptake equal to 4.512, 4.987 and 5.367 mmol g−1, respectively. Furthermore, modeling of values of metal uptake capacity obtained at different shaking time intervals supports pseudo-second order kinetics.
A novel chemosensor L based on coumarin Schiff-base was synthesized and investigated. Sensor L showed remarkable selectivity for Al3+ in Tris–HCl aqueous buffer solution (pH 7.2), and the selectivity was not affected by the presence of a large excess of other competitive ions. The sensor responded rapidly to Al3+ in aqueous solutions with a 2:1 stoichiometry. Meanwhile, it indicated significant improvement of quantum efficiency and ideal fluorescent lifetime.
A simple and reliable enzymatic system for organophosporus pesticide detection was successfully developed, by exploiting the synergy between the magnetic beads collection capacity and the outstanding electrochemistry property of boron-doped diamond electrodes. The determination of an organophosphate pesticide, chlorpyrifos (CPF), was performed based on the inhibition system of the enzyme acetylcholinesterase bonded to magnetic beads through a biotin-streptavidin complex system. A better sensitivity was found for a system with magnetic beads in the concentration range of 10−9 to 10−5 M. The estimated limits of detection based on IC10 (10% acetylcholinesterase (AChE) inhibition) have been detected and optimized to be 5.7 × 10−10 M CPF. Spiked samples of water of Yokohama (Japan) have been measured to validate the efficiency of the enzymatic system. The results suggested that the use of magnetic beads to immobilize biomolecules or biosensing agents is suitable to maintain the superiority of BDD electrodes.
In the present study, we examined the reversible thermal deformation of the membrane protein light-harvesting complex LH2 adsorbed on mesoporous silica (MPS) supports. The LH2 complex from Thermochromatium tepidum cells was conjugated to MPS supports with a series of pore diameter (2.4 to 10.6 nm), and absorption spectra of the resulting LH2/MPS conjugates were observed over a temperature range of 273 – 313 K in order to examine the structure of the LH2 adsorbed on the MPS support. The experimental results confirmed that a slight ellipsoidal deformation of LH2 was induced by adsorption on the MPS supports. On the other hand, the structural stability of LH2 was not perturbed by the adsorption. Since the pore diameter of MPS support did not influence the structural stability of LH2, it could be considered that the spatial confinement of LH2 in size-matches pore did not improve the structural stability of LH2.
Graphite material is abundantly available from recyclable sources. It possesses a good electrical conductance property, which makes it an attractive material as a working electrode. However, due to a high activation overpotential it has limited applications as compared to other solid metal electrodes. In this present work, we obtained a graphite rod from a used battery, and carried out electrochemical improvements by electro-deposition with gold nanoparticles (AuNPs). The heterogeneous electron transfer rate and electron transfer resistance of the fabricated electrode were improved. The electrode overpotential has shown improvement by 50 mV, and the effective surface area has increased by 2 fold. To determine the practicability of the AuNPs/graphite electrode, we used the electrode in the analysis of myricetin. A square-wave voltammetry was used in the analysis, and the detection response increased by 2.5 fold, which suggested an improvement in the electrode sensitivity.
The aim of this study was to develop and validate a simple liquid-chromatography method, with good accuracy, reproducibility and sensitivity, for the quantification of norfloxacin in β-cyclodextrin inclusion complexes. In the method validation, the parameters evaluated were linearity, limits of detection and quantification, specificity, accuracy, precision and robustness. The stability-indication property of the method was evaluated through studies on the degradation under stress conditions. A method employing a simple mobile phase consisting of phosphate buffer (pH 3.0) and acetonitrile (86:14 v/v) was developed. Fluorescence detection was employed to minimize the influence of degradation products, due to its high sensitivity, selectivity and specificity. The method was specific, linear in the concentration range of 1 – 30 μg/mL, robust, precise and accurate. The proposed method was successfully applied in the determination of norfloxacin in inclusion complexes, thus aiding quality-control analysis in the future development of drug delivery systems.
Amylose derivatives chiral stationary phases (CSPs), some of which are commercially available, are well known for their powerful enantioseparation performance. However, due to the dissolution or swelling properties of amylose derivatives, this type of CSPs prepared by coating method exhibit poor solvent tolerance and stability. In order to overcome the defect as well as maintain the chiral recognition capability of amylose tris(3,5-dimethylphenylcarbamate), chitin bis(3-chloro-4-methylphenylcarbamate) with good stability and good chiral recognition capability was blended with the amylose derivative at different ratios, and the resulting blends were further coated onto 3-aminopropyl silica gel to obtain three biselector CSPs. Meanwhile, the corresponding individual selector CSPs were also prepared, respectively, for the sake of comparison with biselector CSPs. The chiral recognition capacity and solvent tolerance of five CSPs were systematically investigated. In addition, the influence of composition and interaction of amylose tris(3,5-dimethylphenylcarbamate) and chitin bis(3-chloro-4-methylphenylcarbamate) in the biselector CSPs on the chiral recognition and the elute orders of enantiomers was also discussed in detail.
An advanced stopped-in-loop flow analysis (SILFA) is proposed for the catalytic determinations of vanadium and iron. The chemistry relies on a vanadium- or iron-catalyzed oxidative reaction of p-anisidine by bromate or hydrogen peroxide in the presence of an activator (Tiron or 1,10-phenanthlorine) to form a red dye (510 nm). Reagents for the vanadium- or iron-catalyzed reaction are well mixed by a reagents-merging zones technique. A sample solution is loaded together with well-mixed reagents into a loop in the SILFA configuration, followed by spectrophotometric detection. The advanced SILFA system provides a selective method for the trace determination of vanadium and iron.
Edited and published by : The Japan Society for Analytical Chemistry Produced and listed by : Sobun Print Co., Ltd. (Vol.25 No.6-) Chuo Printing Co., Ltd. (Vol.14-Vol.18, Vol.20-Vol.25 No. 5) The Japan Society for Analytical Chemistry (Vol.19)