DNA biosensor based on the electrochemiluminescence of Ru(bpy)3(2+) with DNA-binding intercalators

This paper reports a novel detection method for DNA hybridization based on the electrochemiluminescence (ECL) of Ru(bpy)(3)(2+) with a DNA-binding intercalator as a reductant of Ru(bpy)(3)(3+). Some ECL-inducible intercalators have been screened in this study using electrochemical methods combined with a chemiluminescent technique. The double-stranded DNA intercalated by doxorubicin, daunorubicin, or 4',6-diamidino-2-phenylindole (DAPI) shows a good ECL with Ru(bpy)(3)(2+) at +1.19 V (versus Ag/AgCl), while the non-intercalated single-stranded DNA does not. In order to stabilize the self-assembled DNA molecules during ECL reaction, we constructed the ECL DNA biosensor separating the ECL working electrode with an immobilized DNA probe. A gold electrode array on a plastic plate was assembled with a thru-hole array where oligonucleotide probes were immobilized in the side wall of thru-hole array. The fabricated ECL DNA biosensor was used to detect several pathogens using ECL technique. A good specificity of single point mutations for hepatitis disease was obtained by using the DAPI-intercalated Ru(bpy)(3)(2+) ECL.     

Glucose sensing by electrogenerated chemiluminescence of glucose-dehydrogenase produced NADH on electrodeposited redox hydrogel

In this work, we report a new sensing approach based on electrogenerated chemiluminescence (ECL) in an electrodeposited redox hydrogel using glucose dehydrogenase as a model system. The ECL-hydrogel films were electrodeposited by potential cycling of a PBS solution containing [poly(4-vinylpyridine)Ru(2,2'-bipyridine)(2)Cl(-)](+/2+). The film was easily prepared in a rapid, reproducible and well-controlled one-step procedure. The deposited hydrogel film is permeable to water-soluble chemicals and biochemicals, like enzyme substrates and coenzymes. Electrochemistry and ECL of NADH were studied at the level of the hydrogel film. Results indicate that ECL emission occurs at a relatively low anodic potential compared to the classical Ru(bipy)(3)(2+) complex. This is an important advantage since the measurements performed with the ECL hydrogel are thus less sensitive to interfering species. An ECL oxidative-reductive mechanism is presented for the ECL-hydrogel. Then we showed that the intensity of the ECL of NADH produced by the enzymatic activity varies with the enzyme substrate concentration. Such sensing approach combines enzymatic selectivity with the ECL advantages at low oxidation potential.     

Ligand and surfactant effects on a novel electrochemiluminescent reaction involving cadmium

Electrochemiluminescence (ECL) is the generation of light during a chemical reaction with at least one of the reagents being generated in situ at an electrode. Recently, a host of methods have been developed employing ECL reactions as an analytical technique where one of the ECL reagents is the analyte. Electrochemiluminescent reactions involving aqueous metal ions can offer an alternative method for quantitation of dissolved metals. Previous work developed an ECL system using 1,10-phenanthroline as an ECL reagent in conjunction with Cd²⁺ ions providing sufficient emission for reliable cadmium detection. This paper explores the effects of ligand modification, choice of surfactant, and the interaction between the surfactant and the co-reactant tripropylamine (TPA). The effectiveness of the reagents tested can be ranked as phenanthroline, bathophenanthroline salt, terpyridine, dimethylphenanthroline, and bipyridine in order of decreasing Cd²⁺ specific ECL emissions. The non-ionic surfactants Triton X-100, Thesit, and Nonidet P40 were surveyed.     

Carbon optically transparent electrodes for electrogenerated chemiluminescence

This study investigates pyrolyzed photoresist film (PPF)-based carbon optically transparent electrodes (C-OTEs) for use in electrogenerated chemiluminescence (ECL) studies. Oxidative-reductive ECL is obtained with a well-characterized ECL system, C8S3 J-aggregates with 2-(dibutylamino)ethanol (DBAE) as coreactant. Simultaneous cyclic voltammograms (CVs) and ECL transients are obtained for three thicknesses of PPFs and compared to nontransparent glassy carbon (GC) and the conventional transparent electrode indium tin oxide (ITO) in both front face and transmission electrode cell geometries. Despite positive potential shifts in oxidation and ECL peaks, attributed to the internal resistance of the PPFs that result from their nanoscale thickness, the PPFs display similar ECL activity to GC, including the low oxidation potential (LOP) observed for amine coreactants on hydrophobic electrodes. Reductive-oxidative ECL was obtained using the well-studied ECL luminophore Ru(bpy)(3)(2+), where the C-OTEs outperformed ITO because of electrochemical instability of ITO at very negative potentials. The C-OTEs are promising electrodes for ECL applications because they yield higher ECL than ITO in both oxidative-reductive and reductive-oxidative ECL modes, are more stable in alkaline solutions, display a wide potential window of stability, and have tunable transparency for more efficient detection of ECL.     

Quenching Behavior of the Electrochemiluminescence of Ru(bpy)32+/TPrA System by Phenols on a Smartphone-Based Sensor

Phenolic compounds such as vanillic and p-coumaric acids are pollutants of major concern in the agro-industrial processing, thereby their effective detection in the industrial environment is essential to reduce exposure. Herein, we present the quenching effect of these compounds on the electrochemiluminescence (ECL) of the Ru(bpy)₃²⁺/TPrA (TPrA=tri-n-propylamine) system at a disposable screen-printed carbon electrode. Transient ECL profiles are obtained from multiple video frames following 1.2 V application by a smartphone-based ECL sensor. A wide range of detection was achieved using the sensor with limit of detection of 0.26 μM and 0.68 μM for vanillic and p-coumaric acids, respectively. The estimated quenching constants determined that the quenching efficiency of vanillic acid is at least two-fold that of p-coumaric acid under the current detection conditions. The present ECL quenching approach provided an effective method to detect phenolic compounds using a low-cost, portable smartphone-based ECL sensor.     

Electrochemiluminescence bioassays based on carbon nitride nanomaterials and 2D transition metal carbides

Electrochemiluminescence (ECL) is a powerful technique for bioassays. To meet the growing demand for bioassays, it is necessary to develop new ECL emitters and co-reaction acceleration strategies to improve detection sensitivity and expand the application scope. Carbon nitride nanomaterials and 2D transition metal carbides, as newly emerging carbon-based nanomaterials, have been increasingly used for ECL bioassays due to their attractive optical and electrochemical properties as well as diversity. In this minireview, we summarized the latest advances in ECL bioassays using carbon nitride nanomaterials and 2D transition metal carbides in the past two years. Finally, we briefly discuss the future trends and challenges of carbon-based nanomaterials for ECL bioanalysis.     

Homogenous electrogenerated chemiluminescence immunoassay for human immunoglobulin G using N-(aminobutyl)-N-ethylisoluminol as luminescence label at gold nanoparticles modified paraffin-impregnated graphite electrode

A homogeneous electrogenerated chemiluminescence (ECL) immunoassay for human immunoglobulin G (hIgG) has been developed using a N-(aminobutyl)-N-ethylisoluminol (ABEI) as luminescence label at gold nanoparticles modified paraffin-impregnated graphite electrode (PIGE). ECL emission was electrochemically generated from the ABEI-labeled anti-hIgG antibody and markedly increased in the presence of hIgG antigen due to forming a more rigid structure of the ABEI moiety. The concentration of hIgG antigen was determined by the increase of ECL intensity at a gold nanoparticles modified PIGE. It was found that the ECL intensity of ABEI in presence of hydrogen peroxide was dramatically enhanced at gold nanoparticles modified PIGE in neutral aqueous solution and the detection limit of ABEI was 2 x 10(-14)mol/L (S/N=3). The integral ECL intensity was linearly related to the concentration of hIgG antigen from 3.0 x 10(-11) to 1.0 x 10(-9)g/mL with a detection limit of 1 x 10(-11)g/mL (S/N=3). The relative standard deviation was 3.1% at 1.0 x 10(-10)g/mL (n=11). This work demonstrates that the enhancement of the sensitivity of ECL and ECL immunoassay at a nanoparticles modified electrode is a promising strategy.     

Gold nanoparticle-modified ITO electrode for electrogenerated chemiluminescence: well-preserved transparency and highly enhanced activity

Although it is desirable to use transparent indium tin oxide (ITO)-coated glass substrates as working electrodes for electrogenerated chemiluminescence (ECL), their applications in ECL studies of the Ru(bpy)32+ (bpy, 2,2'-bipyridine)/tri-n-propylamine (TPrA) system have been limited because of the large overpotential of TPrA oxidation and the instability of the ITO surface at high anodic potentials. Here, we describe a simple method to achieve high ECL activity using ITO electrodes modified with gold nanoparticles (GNPs). The GNPs have been capped with fluorosurfactant ligands (i.e., Zonyl FSO). Much more facile TPrA oxidation was achieved by using the modified electrode, and an intense low-oxidation-potential (LOP) ECL signal was observed at approximately 0.88 V versus SCE. The electrode transmittance drop upon modification was generally less than 5% over the visible spectrum when small-sized GNPs (approximately 4 nm) were employed. The well-preserved transparency and highly enhanced activity make the modified electrode promising for ECL studies.     

Determination of nitrite based on its quenching effect on anodic electrochemiluminescence of CdSe quantum dots

A novel method for electrochemiluminescent (ECL) detection of nitrite was proposed based on its quenching effect on anodic ECL emission of CdSe quantum dots (QDs). The ECL emission could be greatly enhanced by sulfite and dissolved oxygen in a neutral system and occurred at a relatively low potential in comparison with traditional anodic ECL emitter, leading to high sensitivity and good selectivity. The quenching mechanism followed an “electrochemical oxidation inhibition” process, which was completely different from those of some analytes on the ECL emission of QDs. The coincidence of photoluminescence and ECL spectra of the QDs indicated that the ECL emission resulted from the redox process of QDs core and the sulfite acted as a coreactant. The nitrite quenched ECL emission could be analyzed according to the treatment of Stern-Volmer equation with a linear range from 1 μM to 0.5 mM for detection of nitrite. This work presented a new efficient ECL methodology for quencher-related detection.     

Ultrasensitive Imaging of Cells and Sub-Cellular Entities by Electrochemiluminescence

Here we report on a label-free electrochemiluminescence (ECL) microscopy using exceptionally low concentrations of the [Ru(bpy)₃]²⁺ luminophore. This work addresses the central point of the minimal concentration of the ECL luminophore required to image single entities. We demonstrate the possibility to record ECL images of cells and mitochondria at concentrations down to nM and pM. This is 7 orders of magnitude lower than classically-used concentrations and corresponds to a few hundreds of luminophores diffusing around the biological entities. Yet, it produces remarkably sharp negative optical contrast ECL images, as demonstrated by structural similarity index metric analyses and supported by predictions of the ECL image covering time. Finally, we show that the reported approach is a simple, fast, and highly sensitive method, which opens new avenues for ultrasensitive ECL imaging and ECL reactivity at the single molecule level.     

Ultrasensitive gaseous NH3 sensor based on ionic liquid-mediated signal-on electrochemiluminescence

This work reports that ammonia (NH(3)) can be used as an efficient co-reactant for tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)(3)(2+)) electrochemiluminescence (ECL) in ionic liquids (ILs), on the basis of which a signal-on ECL sensor for directly detecting gaseous NH(3) has been developed. The NH(3) ECL sensor has a very high sensitivity, with a detection limit of 10 ppt NH(3) (at signal-to-noise ratio of 3) without any preconcentration. The high sensitivity is mainly due to the zero ECL background of Ru(bpy)(3)(2+) in the ILs, strong co-reactant ECL activity of NH(3), and high solubility of NH(3) in imidazolium-based ILs. Additionally, the ECL sensor shows an excellent selectivity against common interfering gases and a wide linear response range from 10 ppt to 10 ppm.     

Electrochemiluminescence immunosensor for the determination of ag alpha fetoprotein based on energy scavenging of quantum dots

An electrochemiluminescence (ECL) immunosensor for ultrasensitive detection of alpha fetoprotein (AFP) was fabricated using graphene-CdS quantum dots-alginate (G-CdS QDs-AL) as the immobilizing support and CdSe/ZnS QDs as the label. CdSe/ZnS QDs could effectively scavenge the ECL of G-CdS QDs-AL composite, and the quenched ECL intensity depended linearly on the logarithm for AFP concentration in the range from 0.05 to 500 fg/mL. The detection limit was 20 ag/mL. The proposed ECL immunoassay protocol for AFP detection is stable, specific, highly sensitive and promising for clinical application.     

Electrogenerated chemiluminescence of luminol on a gold-nanorod-modified gold electrode

Electrogenerated chemiluminescence (ECL) of luminol on a gold-nanorod-modified gold electrode was studied, and five ECL peaks were obtained under conventional cyclic voltammetry in both neutral and alkaline solutions. Among them, four ECL peaks (ECL-1-4) were also observed on a gold-nanosphere-modified gold electrode, but the intensities of these ECL peaks were enhanced about 2-10-fold on a gold-nanorod-modified gold electrode in neutral solution. One new strong ECL peak (ECL-5) was obtained at -0.28 V (vs SCE) on a gold-nanorod-modified gold electrode in both neutral and alkaline solutions and enhanced with an increase in pH. In strong alkaline solutions, ECL-1 and ECL-2 on a gold-nanosphere-modified electrode were much stronger than those on a gold-nanorod-modified gold electrode, while ECL-3-5 appeared to only happen on a gold-nanorod-modified gold electrode. The emitter of all the ECL peaks was identified as 3-aminophthalate. The ECL peaks were found to depend on the scan direction, the electrolytes, the pH, and the presence of O(2) and N(2). The reaction pathways for ECL-4 have been further elucidated, and the mechanism of the new ECL peak (ECL-5) has been proposed. The results indicate that a gold-nanorod-modified gold electrode has a catalytic effect on luminol ECL different from that of a gold-nanosphere-modified gold electrode, revealing that the shape of the metal nanoparticles has an important effect on the luminol ECL behavior. The strong ECL of luminol in neutral solution obtained on a gold-nanorod-modified electrode may be used for the sensitive detection of biologically important compounds in physiological conditions.     

Enhanced electrochemiluminescent of lucigenin at an electrically heated cylindrical microelectrode

In this paper, an ECL detection system equipped with an electrically heating controlled cylindrical microelectrode (HME) was used to study the ECL behavior of lucigenin. The ECL intensity of lucigenin would be increased at elevated electrode temperature but the noise had not been increased. It was found that ECL intensity at higher temperature of electrode surface (80 °C) was more than two magnitudes stronger than that at the room temperature (22 °C). The detection limit for ECL of lucigenin on a HME is much lower than that on an electrode without heating, based on which, it is possible to establish a more sensitive method for measurement of ECL by using a HME. The heating of electrode has been used to renew the electrode, which avoid the tedious work for refreshing the electrode surface. The reproducibility of lucigenin ECL system at HME is satisfactory.     

A solid-state electrochemiluminescence sensing platform for detection of adenosine based on ferrocene-labeled structure-switching signaling aptamer

A solid-state electrochemiluminescence sensing platform based on ferrocene-labeled structure-switching signaling aptamer (Fc-aptamer) for highly sensitive detection of small molecules is developed successfully using adenosine as a model analyte. Such special sensing platform included two main parts, an electrochemiluminescence (ECL) substrate and an ECL intensity switch. The ECL substrate was made by modifying the complex of Au nanoparticle and Ruthenium (II) tris-(bipyridine) (Ru(bpy)₃²⁺-AuNPs) onto Au electrode. An anti-adenosine aptamer labeled by ferrocene acted as the ECL intensity switch. A short complementary ssDNA for the aptamer was applied to hybridizing with the aptamer, yielding a double-stranded complex of the aptamer and the ssDNA on the electrode surface. The introduction of adenosine triggered structure switching of the aptamer. As a result, the ssDNA was forced to dissociate from the sensing platform. Such structural change of the aptamer resulted in an obvious ECL intensity decrease due to the increased quenching effect of Fc to the ECL substrate. The analytic results were sensitive and specific.     

中性微乳液介质增敏鲁米诺电化学发光研究

磷酸盐缓冲的中性溴化十六烷基三甲基铵-正戊醇-正庚烷微乳液介质体系对鲁米诺的电化学发光有显著的增敏作用,本文对此增敏作用进行了研究,并在此介质体系中采用电化学发光淬灭法测定了生物活性分子褪黑素,此法可用于测定脑白金胶囊中的褪黑素。     

Electrochemiluminescence of metal-organic complex nanowires based on graphene-Nafion modified electrode for biosensing application

In this work, we chose tris(2,2′-bipyridyl)ruthenium(II)hexafluorophosphate(Ru(bpy)3(PF6)2), a metal-organic complex material,to prepare nanowires, which were subsequently applied for the construction of electrochemiluminescence(ECL) biosensor by immobilizing them onto a glassy carbon electrode(GCE) with graphene-Nafion composite films. The graphene therein, being a two-dimensional carbon nanomaterial with outstanding electronic properties, can obviously improve the conductivity of the Nafion film, as well as enhance the electrochemical signal and ECL intensity of the Ru(bpy)3(PF6)2 nanowires(RuNWs) at low graphene concentration. The developed biosensor exhibited excellent ECL stability with tripropylamine(TPrA) as co-reactant. The ECL biosensor exhibited high sensitive ECL response in a wide linear range and low detection limit for the detection of proline. It is considered that the oxidation products of proline would be responsible for the ECL enhancement. The large electro-active area of the nanowires and the enhancement effect of the graphene played critical roles in the high detection performance of the ECL biosensor. The results demonstrated herein may provide a useful enlightenment for the design of more sensitive ECL biosensors.     

[Ru(bpy)2(L-Trp)]ClO4-KCl水溶液体系的电致化学发光特性研究

合成了[Ru(bpy)2(L-Trp)]ClO4, 并对其电致化学发光特性进行研究。发现在KCl溶液中, 在三角波脉冲电压作用下, 该配合物在铂电极上有电致化学发光活性。其线性动力学响应范围为5.9×10^-^9～1.2×10^-^7mol . L^-^1。当信噪比为3时, 可检测浓度为5.9×10^-^9mol . L^-^1。浓度为5.9×10^-^8mol . L^-^1时, 10次测试的相对标准偏差为5.3%。根据电化学研究的结果, 提出了该配合物的电致化学发光反应机理。     

Surface-Bonding-Enhanced Self-Co-Reactant Electrogenerated Chemiluminescence for Sensitive and Selective Detection of Thioglycolic Acid in Cosmetics

Thioglycolic acid (TGA) is an organic compound widely used in cosmetics that cause a variety of health problems when overexposed to it. So far many attempts have been made to develop methods for TGA detection, but most of them need sophisticated instrumentations and are a little bit complicated. Therefore, a simple, cheap and sensitive detection method of TGA is highly desired. Herein, we demonstrated for the first time an Au−S bonding amplified, highly sensitive electrochemiluminescence (ECL) sensing method for TGA detection using tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)₃²⁺) as a luminophore and TGA as a self-co-reactant, via an anodic reaction at the Au electrode surface. Due to different molecular coordination environments of the TGA at the electrode surface, the ECL signal intensity of the developed ECL system gives much higher ECL signal in borate buffer than phosphate buffer of the same pH. Under the optimized experimental conditions, the ECL intensity has a direct relationship with the concentration of TGA in the range of 0.03 μM to 300 μM and a limit of detection of 0.013 μM (3σ/m). The reported ECL system has further been applied for the detection of TGA in cosmetics with acceptable recoveries.     

磺酸基卟啉的电化学发光研究及应用

以四(4-磺酸基苯基)卟啉（TSPP）为发光体,过硫酸钾（K₂S₂O₈）为共反应剂,构建了一个新的电化学发光（ECL）体系. 在扫描范围为0~-1.5V时,该体系出现两个阴极ECL峰,分别为TSPP的还原峰（-0.8V）和K₂S₂O₈ 的还原峰 (-1.2V). 亚甲基蓝能有效猝灭四(4-磺酸基苯基)卟啉的电化学发光,根据猝灭效率与亚甲基蓝浓度的线性关系,建立了一种测定亚甲基蓝含量的新方法.