Gold nanoparticle-based near-infrared fluorescent detection of biological thiols in human plasma

In this work, a new fluorescent method for sensitive detection of biological thiols in human plasma was developed using a near-infrared (NIR) fluorescent dye, FR 730. The sensing approach was based on the strong affinity of thiols to gold and highly efficient fluorescent quenching ability of gold nanoparticles (Au NPs). In the presence of thiols, the NIR fluorescence would enhance dramatically due to desorption of FR 730 from the surfaces of Au NPs, which allowed the analysis of thiol-containing amino acids in a very simple approach. The size of Au NPs was found to affect the fluorescent assay and the best response for cysteine detection was achieved when using Au NPs with the diameter of 24 nm, where a linear range of 2.5 x 10(-8) M to 4.0 x 10(-6) M and a detection limit of as low as 10 nM was obtained. This method also demonstrated a high selectivity to thiol-containing amino acids due to the strong affinity of thiols to gold.

Solid-state electrochemiluminescence sensor based on the Nafion/poly(sodium 4-styrene sulfonate) composite film

An effective electrochemiluminescence (ECL) sensor based on Nafion/poly(sodium 4-styrene sulfonate) (PSS) composite film-modified ITO electrode was developed. The Nafion/PSS/Ru composite film was characterized by atomic force microscopy, UV-vis absorbance spectroscopy and electrochemical experiments. The Nafion/PSS composite film could effectively immobilize tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)(3)(2+)) via ion-exchange and electrostatic interaction. The ECL behavior of Ru(bpy)(3)(2+) immobilized in Nafion/PSS composite film was investigated using tripropylamine (TPA) as an analyte. The detection limit (S/N = 3) for TPA at the Nafion/PSS/Ru composite-modified electrode was estimated to be 3.0 nM, which is 3 orders of magnitude lower than that obtained at the Nafion/Ru modified electrode. The Nafion/PSS/Ru composite film-modified indium tin oxide (ITO) electrode also exhibited good ECL stability. In addition, this kind of immobilization approach was simple, effective, and timesaving.

Electrochemiluminescence sensor based on partial sulfonation of polystyrene with carbon nanotubes

Herein, homogenously partial sulfonation of polystyrene (PSP) was performed. An effective electrochemiluminescence (ECL) sensor based on PSP with carbon nanotube (CNTs) composite film was developed. Cyclic voltammetry and electrochemical impendence spectroscopy were applied to characterize this composite film. The PSP was used as an immobilization matrix to entrap the ECL reagent Ru(bpy)(3)(2+) due to the electrostatic interactions between sulfonic acid groups and Ru(bpy)(3)(2+) cations. The introduction of CNTs into PSP acted not only as a conducting pathway to accelerate the electron transfer but also as a proper matrix to immobilize Ru(bpy)(3)(2+) on the electrode by hydrophobic interaction. Furthermore, the results indicated the ECL intensity produced at this composite film was over 3-fold compared with that of the pure PSP film due to the electrocatalytic activity of the CNTs. Such a sensor was verified by the sensitive determinations of 2-(dibutylamino)ethanol and tripropylamine.

Ionic liquids as selectors for the enhanced detection of proteins

Herein, we report an approach for protein detection enhanced by ionic liquid (IL) selectors in capillary electrophoresis (CE), with avidin as a model protein. Hydrophilic ILs were added into the running buffer of CE and acted as selectors for sample injection, enriching the positive target and excluding the negative from the capillary. When using 3% (v/v) IL selector, the detection sensitivity of avidin was improved by over one order of magnitude, while the interference from protein adsorption was effectively avoided, even in an uncoated capillary. The electrochemiluminescence method was initially used for IL-based CE with low noise that was independent of the IL concentration, making ILs almost transparent as additives in the electrophoresis buffer.

Adaptive recognition of small molecules by nucleic acid aptamers through a label-free approach

We report a novel label-free method for the investigation of the adaptive recognition of small molecules by nucleic acid aptamers using capillary electrophoresis analysis. Cocaine and argininamide were chosen as model molecules, and the two corresponding DNA aptamers were used. These single-strand DNAs folded into their specific secondary structures, which were mainly responsible for the binding of the target molecules with high affinity and specificity. For molecular recognition, the nucleic acid structures then underwent additional conformational changes, while keeping the target molecules stabilized by intermolecular hydrogen bonds. The intrinsic chemical and physical properties of the target molecules enabled them to act as indicators for adaptive binding. Thus any labeling or modification of the aptamers or target molecules were made obsolete. This label-free method for aptamer-based molecular recognition was also successfully applied to biological fluids and therefore indicates that this approach is a promising tool for bioanalysis.

A novel alcohol dehydrogenase biosensor based on solid-state electrogenerated chemiluminescence by assembling dehydrogenase to Ru(bpy)(3)(2+)-Au nanoparticles aggregates

Based on electrogenerated chemiluminescence (ECL), a novel method for fabrication of alcohol dehydrogenase (ADH) biosensor by self-assembling ADH to Ru(bpy)(3)(2+) -AuNPs aggregates (Ru-AuNPs) on indium tin oxide (ITO) electrode surface has been developed. Positively charged Ru(bpy)(3)(2+) could be immobilized stably on the electrode surface with negatively charged AuNPs in the form of aggregate via electrostatic interaction. On the other hand, AuNPs are favourable candidates for the immobilization of enzymes because amine groups and cysteine residues in the enzymes are known to bind strongly with AuNPs. Moreover, AuNPs can act as tiny conduction centers to facilitate the transfer of electrons. Such biosensor combined enzymatic selectivity with the sensitivity of ECL detection for quantification of enzyme substrate, and it displayed wide linear range, high sensitivity and good stability.

Tris(2,2 '-bipyridyl)ruthenium(II) electrochemiluminescent detection of coreactants containing aromatic diol group by the interaction between diol and borate anion

Previous studies show that aromatic diols inhibited Ru(bpy)(3)(2+) electrochemiluminescence (ECL), and all reported Ru(bpy)(3)(2+) ECL methods for the determination of aromatic diols-containing coreactants are based on inhibition of Ru(bpy)(3)(2+)/tripropylamine ECL. In this study, the interaction between diol and borate anion was exploited for Ru(bpy)(3)(2+) ECL detection of coreactants containing aromatic diol group using epinephrine as a model analyte. The interaction prevented from the inhibition of Ru(bpy)(3)(2+) ECL by aromatic diol group of epinephrine. As a result, epinephrine was successfully detected in the absence of tripropylamine simply by using borate buffer solution as the supporting electrolyte. Under the optimum conditions, the log of the ECL intensity increases linearly with the log of epinephrine concentrations over the concentration range of 1.0x10(-9)-1.0x10(-4) M. The detection limit is 5.0x10(-10) M at a signal-to-noise ratio of three. The proposed method exhibit wider dynamic range and better detection limit than that by inhibited Ru(bpy)(3)(2+) ECL method. The relative standard deviation for 14 consecutive determinations of 5 mu M epinephrine was 3.5%. The strategy by interaction with borate anion or boronate derivatives is promising for the determination of coreactants containing aromatic diol group or aromatic hydroxyl acid group. Such interaction can also be used to avoid interference from aromatic diols or aromatic hydroxyl acids.

Colloidal photonic crystals with a graded lattice-constant distribution

In this work, a gradient polystyrene colloidal photonic crystal was fabricated by annealing in a graded temperature field. The lattice constant of the gradient crystal gradually varied along the temperature-gradient direction. The positional bandgap wavelength as well as the attenuation of the bandgap wavelength could be tuned dependent on the position of the gradient colloidal crystal along the gradient direction because of the lattice-constant variation.

Analytical applications of the electrochemiluminescence of tris (2.2 '-bipyridyl) ruthenium and its derivatives

This article presents the state of the art of analytical applications of the electrochemiluminescence (ECL) of tris (2,2'-bipyridyl) ruthenium (Ru(bpy)(3)(2+)) and its derivatives. in the last seven years, Ru(bpy)(3)(2+) ECL has attracted much interest from analysts and been successfully exploited as a detector of flow injection analysis (FIA), high-performance liquid chromatography (HPLC), capillary electrophoresis (CE), and micro total analysis systems (TAS). Immobilization of Ru(bPY)(3)(2+) on a solid surface provides several advantages over the solution-phase ECL procedure, such as the simplicity of experimental design and cost-effectiveness. After a brief discussion of the mechanism of Ru(bpy)(3)(2+) ECL, we discuss its applications in FIA, HPLC, CE and TAS and give special attention to the design of Ru(bpy)(3)(2+) ECL cells and some immobilization techniques of Ru(bpy)(3)(2+); we focus on papers published after 1997.

Electrochemiluminescence detection with integrated indium tin oxide electrode on electrophoretic microchip for direct bioanalysis of lincomycin in the urine

In this article, an antibiotic, lincomycin was determined in the urine sample by microchip capillary electrophoresis (CE) with integrated indium tin oxide (ITO) working electrode based on electrochemiluminescence (ECL) detection. This microchip CE-ECL system can be used for the rapid analysis of lincomycin within 40 s. Under the optimized conditions, the linear range was obtained from 5 to 100 muM with correlation coefficient of 0.998. The limit of detection (LOD) of 3.1 muM was obtained for lincomycin in the standard solution. We also applied this method to analyzing lincomycin in the urine matrix. The limit of detection of 9.0 muM was obtained. This method can determine lincomycin in the urine sample without pretreatment, which demonstrated that it is a promising method of detection of lincomycin in clinical and pharmaceutical area.

Structural elucidation and identification of alkaloids in Rhizoma Coptidis by electrospray ionization tandem mass spectrometry

Simple, convenient, sensitive and accurate analytical methods are needed for the structural characterization and identification of alkaloid components in Rhizoma Coptidis in traditional Chinese herbal medicine, which has important bioactivity. In this work, the identification of alkaloid compounds in Rhizoma Coptidis was investigated by obtaining molecular mass information using electrospray ionization mass spectrometry (ESI-MS). Multi-stage tandem mass spectrometric (ESI-MSn) data for the alkaloid compounds were used for detailed structural characterization, then structure information was obtained by comparison of the fragmentation mechanisms of both alkaloids in Rhizoma Coptidis and standard samples of berberine, palmatine, coptisine and jatrorrhizine by MS. Based on the results obtained, the structure of a novel compound was elucidated. The results of the experiments demonstrate that ESI-MSn is a sensitive, selective and effective tool for the rapid determination of alkaloids in Rhizoma Coptidis.

Electrochemiluminescent detection based on solid-phase extraction at tris(2,2 '-bipyridyl)ruthenium(II)-modified ceramic carbon electrode

A sensitive electrochemiluminescent detection scheme by solid-phase extraction at Ru(bpy)(3)(2+)-modified ceramic carbon electrodes (CCEs) was developed. The as-prepared Ru(bpy)(3)(2+)-modified CCEs show much better long-term stability than other Nafion-based Ru(bpy)(3)(2+)-modified electrodes and enjoy the inherent advantages of CCEs. The log-log calibration plot for dioxopromethazine is linear from 1.0 x 10(-9) to 1.0 x 10(-4) mol L-1 using the new detection scheme. The detection limit is 6.6 x 10(-10) mol L-1 at a signal-to-noise ratio of 3. The new scheme improves the sensitivity by similar to 3 orders of magnitude, which is the most sensitive Ru(bpy)(3)(2+) ECL method. The scheme allows the detection of dioxopromethazine in a urine sample within 3 min. Since Ru(bpy)(3)(2+) ECL is a powerful technique for determination of numerous amine-containing substances, the new detection scheme holds great promise in measurement of free concentrations, investigation of protein-drug interactions and DNA-drug interactions, pharmaceutical analysis, and so on.

Synthesis of PtNPs/AQ/Ru(bpy)(3)(2+) colloid and its application as a sensitive solid-state electrochemiluminescence sensor material

The facile synthesis of the novel platinum nanoparticles/Eastman AQ55D/ruthenium(II) tris( bipyridine) (PtNPs/ AQ/Ru(bpy)(3)(2+)) colloidal material for ultrasensitive ECL solid-state sensors was reported for the first time. The cation ion-exchanger AQ was used not only to immobilize ECL active species Ru(bpy)(3)(2+) but also as the dispersant of PtNPs. Colloidal characterization was accomplished by transmission electron microscopy (TEM), X-ray photoelectron spectrum (XPS), and UV-vis spectroscopy. Directly coating the as-prepared colloid on the surface of a glassy carbon electrode produces an electrochemiluminescence (ECL) sensor. The electronic conductivity and electroactivity of PtNPs in composite film made the sensor exhibit faster electron transfer, higher ECL intensity of Ru(bpy)(3)(2+), and a shorter equilibration time than Ru(bpy)(3)(2+) immobilized in pure AQ film. Furthermore, it was demonstrated that the combination of PtNPs and permselective cation exchanger made the sensor exhibite excellent ECL behavior and stability and a very low limit of detection (1 x 10(-15) M) of tripropylamine with application prospects in bioanalysis. This method was very simple, effective, and low cost.

Facile separation and determination of Aconitine alkaloids in traditional Chinese medicines by CE with tris(2,2 '-bipyridyl) ruthenium(II)-based electrochemi-luminescence detection

A facile CE method coupled with tris(2,2'-bipyridyl) ruthenium(ll)-based electrochem iluminescence [Ru(bpy)(3)(2+)] detection was developed for simultaneous determination of Aconitum alkaloids, i.e., hypaconitine (HA), aconitine (AC), and mesaconitine (MA) in baseline separation. The optimal separation of these Aconitum alkaloids was achieved in a fused-silica capillary column (50 cm x 25 mu m id) with 30 mM phosphate solution (pH 8.40) as running buffer at 12 kV applied voltage. The three alkaloids can be determined within 10 min by a single run. The calibration curves showed a linear range from 2.0 x 10(-7) to 2.0 x 10(-5) M for HA, 3.4 x 10(-7) to 1.7 x 10(-5) M for AC, and 3.8 x 10(-7) to 1.9 x 10(-5) M for MA. The RSDs; for all analytes were below 3.01%. Good linear relationships were found with correlation coefficients for all analytes exceeding 0.993. The detection limits were 2.0 x 10(-8) M for HA, 1.7 x 10(-7) M for AC, and 1.9 x 10(-7) M for MA under optimal conditions. This method was successfully applied to determine the three alkaloids in Aconitum plants.