Electrochemiluminescence System Research Articles

Synthesis and ECL performance of highly efficient bimetallic ruthenium tris-bipyridyl complexes

In order to find the ideal carbon chain linkage number n for achieving the highest ECL in bimetallic ruthenium tris-bipyridyl complexes, a series of novel complexes [(bpy)(2)Ru(bpy')(CH(2))(n)(bpy')Ru(bpy)(2)](4+) (, where bpy is 2,2'-bipyridyl, n = 10, 12, 14) for a coreactant electrochemiluminescence (ECL) system have been synthesized. Their ECL properties at a Au electrode have been studied in 0.1 M phosphate buffer by using tripropylamine (TPrA), 2-(dibutylamino)ethanol (DBAE) and melamine as the coreactant, to compare with that of the previously reported bimetallic ruthenium analogous complex [(bpy)(2)Ru(bpy')(CH(2))(8)(bpy')Ru(bpy)(2)](4+). The results demonstrate that the ECL intensity depends largely on the length of the saturated carbon chain linkage number n. The highest ECL is reached when n = 10, suggesting that a synergistic effect on ECL enhancement co-exists between the two intramolecular linked ruthenium activating centers. Density functional theory (DFT) calculation demonstrated that the optimized bond distances between Ru and N(bpy') are the longest both in the ground and the excited triplet states in the case of n = 10, while those for Ru and N(bpy) are the shortest in the excited triplet states. All these factors may be responsible for the above mentioned results. This study provided a methodology to further improve and tune ECL efficiency by using bimetallic ruthenium complexes linked by a flexible saturated carbon chain.

A novel method to determine ciprofloxacin by enhanced electrochemiluminescence of Tb(bpy)23+–K2S2O8 system in aqueous solution

A simple and rapid electrochemiluminescence (ECL) method is developed by using Tb(bpy)23+–K2S2O8 system for the determination of trace amounts of ciprofloxacin (CPFX). Conditions for ECL emission were optimized. Under optimized condition, the linear range of ECL intensity versus concentrations of CPFX is 6 × 10−8 to 1.0 × 10−6 mol dm−3 with a relative standard deviation (R.S.D.) of 3.8% (n = 11) and the detection limit is 1.4 × 10−9 mol dm−3 for CPFX. The ECL emission spectra (the peaks are at 490, 545, 585 and 620 nm) match the characteristic emission spectrum of Tb3+, indicating that the emission is from the excited state of Tb3+. The enhanced mechanism of Tb(bpy)23+–K2S2O8 ECL system was proposed. This method was successfully applied to determine CPFX in tablets.

Self-quenching in the electrochemiluminescence of tris(2,2′-bipyridyl) ruthenium(ii) using metabolites of catecholamines as co-reactants

Electrochemiluminescence (ECL) of Ru(bpy)(3)(2+) using metabolites of catecholamines: homovanillic acid (HVA) and vanillylmandelic acid (VMA) as co-reactants were investigated in aqueous solution for the first time. When HVA and VMA were co-existent in the buffer solution containing Ru(bpy)(3)(2+), ECL peaks were observed at a potential corresponding to the oxidation of Ru(bpy)(3)(2+), and the ECL intensity was increased noticeably when the concentrations of HVA and VMA were at lower levels. The linear calibration range was from 8.0 × 10(-5) to 1.0 × 10(-9) M for HVA and VMA. The detection limit (S/N = 3) of HVA and VMA was 4.0 × 10(-10) M. The formation of the excited state Ru(bpy)(3)(2+*) was confirmed to result from the reaction between Ru(bpy)(3)(3+) and the intermediates of HVA or VMA radicals. Moreover, it was found that the ECL intensity was quenched significantly when the concentrations of HVA and VMA were relatively higher. The mechanism of self-quenching processes involved in the Ru(bpy)(3)(2+)-HVA and -VMA ECL systems are proposed in this study.

Development of electrochemiluminescent inhibition method for determination of gentian violet in aquatic water

Gentian violet (GV) was found to quench the electrochemiluminescence (ECL) of the tris(2,2′-bipyridyl)ruthenium(II)/tris-n-propylamine (Ru(bpy)32+-TPA) system at a glass carbon electrode (GCE). Based on the ECL signal changes, a simple and ultrasensitive detection method for GV in aquatic water was established. Under the optimized conditions, the quenched ECL intensity versus the logarithm of the concentration of GV was linear over a concentration range from 1.0×10−10 to 5.0×10−7molL−1, and the limit of detection (LOD) was found to be 4.5×10−12molL−1 (S/N=3). The results obtained by the ECL system were better than other reported methods in literatures in terms of sensitivity or linear response range. The method was successfully applied to determine GV in aquatic water, and the relative standard deviations (RSDs) were found less than 6.3%, and the recoveries were obtained from 98.7 to 111.0%. Moreover, a possible mechanism of the quenching effect was primarily discussed based on UV–visible absorption spectra, cyclic voltammograms and IECL–E curves.

Three-dimensional paper-based electrochemiluminescence immunodevice for multiplexed measurement of biomarkers and point-of-care testing

In this work, electrochemiluminescence (ECL) immunoassay was introduced into the recently proposed microfluidic paper-based analytical device (μPADs) based on directly screen-printed electrodes on paper for the very first time. The screen-printed paper-electrodes will be more important for further development of this paper-based ECL device in simple, low-cost and disposable application than commercialized ones. To further perform high-performance, high-throughput, simple and inexpensive ECL immunoassay on μPAD for point-of-care testing, a wax-patterned three-dimensional (3D) paper-based ECL device was demonstrated for the very first time. In this 3D paper-based ECL device, eight carbon working electrodes including their conductive pads were screen-printed on a piece of square paper and shared the same Ag/AgCl reference and carbon counter electrodes on another piece of square paper after stacking. Using typical tris-(bipyridine)-ruthenium (Ⅱ) - tri-n-propylamine ECL system, the application test of this 3D paper-based ECL device was performed through the diagnosis of four tumor markers in real clinical serum samples. With the aid of a facile device-holder and a section-switch assembled on the analyzer, eight working electrodes were sequentially placed into the circuit to trigger the ECL reaction in the sweeping range from 0.5 to 1.1 V at room temperature. In addition, this 3D paper-based ECL device can be easily integrated and combined with the recently emerging paper electronics to further develop simple, sensitive, low-cost, disposable and portable μPAD for point-of-care testing, public health and environmental monitoring in remote regions, developing or developed countries.

The levels of bone turnover markers in Chinese postmenopausal women

The aim of this study was to evaluate serum N-aminoterminal propeptide of type I collagen (P1NP), C-terminal telopeptide of type I collagen (β-CTX), and vitamin D status in healthy Chinese postmenopausal women. The study was also designed to investigate their possible relationships with osteoporosis phenotypes. A community-based population of 1,724 postmenopausal women in Beijing was randomly selected. Serum bone turnover markers and 25-hydroxyvitamin D [25(OH)D] were tested by an automated Roche electrochemiluminescence system. Dual-energy x-ray absorptiometry was used to measure bone mineral density (BMD). The mean (SD) values of serum β-CTX and P1NP were 0.439 (0.210) and 56.7 (27.9) ng/mL, respectively. The 25(OH)D level of postmenopausal women in Beijing was remarkably low (13.2 ± 5.4 ng/mL). Serum β-CTX and P1NP levels were negatively correlated with BMDs of lumbar spine, femoral neck, and total hip (all P < 0.01). The cubic regression model better fitted the relationships of BMD and bone turnover markers. Serum β-CTX levels were significantly higher in women with sustained osteoporotic fracture or vertebral fracture (P = 0.006 and 0.012, respectively). No association between P1NP and fracture or vertebral fracture was detected. The same situation applied to 25(OH)D. 25(OH)D was negatively correlated with β-CTX and P1NP (r = -0.073 and -0.088, P = 0.002 and <0.001, respectively). Serum β-CTX and P1NP levels were negatively correlated with BMD. β-CTX was significantly higher in postmenopausal women with sustained fracture or vertebral fracture. Vitamin D deficiency was highly prevalent in postmenopausal women in Beijing.

Electrochemiluminescence Behavior of Ru(bpy)32+/Carbofuran System on an Electrically Heated Microelectrode Chip

AbstractAn electrically heated microelectrode chip (HMEC) was designed and the Ru(bpy)32+/carbofuran electrochemiluminescence (ECL) systems were applied to characterize the performance of the HMEC. The ECL intensities increase at elevated electrode temperature, and the detection limit at 60°C (electrode surface temperature) was about 10 times lower than that at 30°C. The results indicate that new heated electrode can be handled easily and can be mass produced, the difference between the electrodes was little. The stability of the HMEC was good since the electrode surface can hardly be destroyed during detection and storage.

High Electron Transfer Efficiency of Titania Dioxide Nanotube for Low Potential Electrochemiluminescent Biosensing

AbstractStrong electrochemiluminescence (ECL) of titania dioxide nanotubes at physiological condition was observed, with relatively low cathodic potential due to its large surface area and high electron transfer efficiency. The coreactant of the ECL emission was proved to be H2O2, produced from the reduction of dissolved oxygen, leading to a mild ECL system for biosensing. Based on “coreactant inhibition” mechanism, the ECL quenchers could be measured by inhibiting the transformation of O2 to H2O2. Using hemoglobin as a model, the quenched ECL emission followed SternVolmer equation in a wide linear range. Thus a novel methodology for detection of “coreactant inhibition”‐related quencher could be developed with acceptable sensitivity.

Analytical applications of the electrochemiluminescence of tris(2,2′-bipyridyl)ruthenium(II) coupled to capillary/microchip electrophoresis: A review

A comprehensive review on the development of analytical methods, by coupling electrochemiluminescence (ECL) detection with capillary electrophoresis (CE) and microchip electrophoresis (ME), is presented. After the description of the basic mechanism of ECL, the addition mode of luminescence reagent in CE–ECL system has been discussed. The analytical applications of the CE–ECL technique in terms of different analytes are also given. Due to the importance of ME as a separation method for the present and future, the ME detection methods based on ECL are considered in a relatively detailed way. Finally, possible trends for CE/ME-ECL in the near future are discussed.

A highly sensitive method for detection of protein based on inhibition of Ru(bpy) 32+/TPrA electrochemiluminescent system

The detection of proteins is very important in the study of biochemistry procedure, and electrochemiluminescence method is a very practical tool for the study of protein folding, structure and quantification. In this work, bovine serum albumin (BSA) and casein were found to be able to significantly quench the electrochemiluminescence (ECL) of Ru(bpy) 3 2+/TPrA system, based on which a highly sensitive approach for the detection of protein was proposed. Under the optimized conditions, the logarithmic plot of the inhibited ECL versus the concentration of BSA and casein were linear over the ranges of 1–30 μg/L and 0.1–1.6 μg/L, respectively. The corresponding limit of detection (LOD) was 0.45 μg/L for BSA and 0.026 μg/L for casein (S/N = 3). UV, ECL and fluorescence methods were used to investigate the mechanism of the inhibited ECL of Ru(bpy) 3 2+/TPrA/BSA system. A mechanism based on the formation of protein-Ru(bpy) 3 2+ super molecule was proposed, which would prevent Ru(bpy) 3 2+ from reaching the working electrode surface so that induced ECL quenching.

A luminol-based micro-flow-injection electrochemiluminescent system to determine reactive oxygen species

A flow injection analysis (FIA) system with electrochemiluminescent (ECL) detection has been established. Based on a specially designed flow-through ECL cell with a very simple structure, the system possesses rapid response and high sensitivity. With luminol as the ECL reagent, the response of hydrogen peroxide (H2O2) was investigated on the developed FIA-ECL system. After optimizing the experimental conditions, such as the electric parameters, the buffer condition and the flow rate, it was demonstrated that the developed FIA-ECL system works well for quantified assays. Compared with reported works, the present results indicate that the developed FIA-ECL system has the lowest limit of detection (S/N=3) of 3.0×10−9mol/L for H2O2, which is equal to the level of chemiluminescence (CL). The developed system was successfully used to monitor the yield of reactive oxygen species (ROSs) in water vapour during the work of an ultrasonic humidifier with H2O2 as index. And the amount of ROSs in some other real samples, including tap water, drinking water and river water was detected with recoveries from 92.0% to 106%.

Efficient electrochemiluminescent cyclometalated iridium(III) complexes: Synthesis, photophysical and electrochemiluminescent properties

A series of new cyclometalated iridium(III) complexes for electrochemiluminescence (ECL) system were synthesized and fully characterized. Using tri-n-propylamine (TPA) as an oxidative–reductive co-reactant, their ECL properties were studied in acetonitrile (CH 3CN) and mixed CH 3CN/H 2O (50:50, v/v) solutions, respectively. Meanwhile, the influencing factors of ECL efficiencies, including working electrode, pH, and surfactant were investigated. A remarkable ECL enhancement (up to about 13.5 times), in comparison with the commonly used Ru(bpy) 3 2+ (2,2′-bipyridyl) ruthenium(II), is observed from Ir(FPP) 2(acac) (where FPP is 2-(4-fluorophenyl)-4-phenylpyridine, acac = acetylacetone) at Pt disk electrode. At the same time, an increase in ECL efficiency is also observed in surfactant media. This study provided a new method for further improving and tuning the ECL efficiency by designing new iridium complexes with the appropriate cyclometalated or ancillary ligands.

Development of an ultrasensitive electrochemiluminescence inhibition method for the determination of tetracyclines

An electrochemiluminescence (ECL) inhibition method is developed for quantitative determination of four tetracyclines (TCs) in honey samples, including tetracycline (TC), oxytetracycline (OTC), chlortetracycline (CTC) and doxycycline (DC). It was found that the four TCs strongly inhibited the ECL signal of the Ru(bpy) 3 2+/DBAE system. Based on the ECL signal changes, a simple and ultrasensitive detection method for TCs was thus established. The optimum experimental conditions including the scan mode and scan rate of the applied potential, the type of the buffer solution and its pH, and the concentration of Ru(bpy) 3 2+ and DBAE for the ECL inhibition method, were investigated in detail. Under the optimized conditions, the quenched ECL intensity versus the logarithm of the concentration of TCs is in good linear relationship over a concentration range from 4.0 × 10 −11 to 4.0 × 10 −9 g mL −1. The detection limits were found to be 2.0 × 10 −12 g mL −1. The results obtained by the proposed ECL system, in terms of sensitivity, were much better than those of previously reported methods. In addition, the method was applied successfully to determine the total residuals of the four TCs in honey samples. The relative standard deviations were found in a range of 4.9–14.3%, and the recoveries were obtained from 87.5% to 115.0%. A possible mechanism for the quenching effects of Ru(bpy) 3 2+/DBAE system was also proposed.

Electrochemiluminescence analysis of folate receptors on cell membrane with on-chip bipolar electrode

In this paper we report a transparent bipolar electrode based microfluidic chip-electrochemiluminescence (ECL) system for sensitive detection of folate receptors (FR) on cell membranes. This integrated system consists of a poly(dimethylsiloxane) (PDMS) layer containing a microchannel and a glass bottom sheet with indium tin oxide (ITO) strips as bipolar detectors. The ITO strips are fabricated using a PDMS micromold with carbon ink as a protective layer in place of traditional photoresist. The configuration of the bipolar electrode has great influence on the ECL intensity of Ru(bpy)(3)(2+)/tripropylamine(TPA) system. Further studies show that folic acid (FA) can strongly inhibit the ECL of the Ru(bpy)(3)(2+)/TPA system. Based on specific recognition between FA and FR on cell membrane, this microfluidic chip-ECL system is successfully applied for detecting the level of FR on human cervical tumor (HL-60) cells and MEF cells. It is found that the ECL intensity increases with the number of HL-60 cells in the range of 21 to 3.28 × 10(4) cells/mL. The average level of FR on HL-60 cells is calculated to be 8.05 ± 0.75 × 10(-18) mol/cell. While for MEF cells, it shows a much slower ECL increment than HL-60 cells due to the much lower FR level on MEF cells (5.30 ± 0.61 × 10(-19) mol/cell). Moreover, exocytosis of FA after FR mediated endocytosis was observed according to the change of the ECL signal with the incubation time of HL-60 cells in the FA- Ru(bpy)(3)(2+)/TPA system.

Potential controlling highly-efficient catalysis of wheat-like silver particles for electrochemiluminescence immunosensor labeled by nano-Pt@Ru and multi-sites biotin/streptavidin affinity

The potential controlling silver catalysis for Ru(bpy)(3)(2+) electrochemiluminescence (ECL) signal at a special potential -0.4∼1.25 V was newly developed as the new ECL signal amplification strategy for ultrasensitive protein detection. Firstly, the wheat-like deposited silver (DpAg) particles were modified on the bare glass carbon electrode (GCE) surface by cyclic voltammetry deposition to capture the primary antibodies and then bind the antigen analytes. Secondly, as a sandwich immunoreaction format, the secondary antibodies conjugated with the Ru(bpy)(3)(2+)-doped Pt (Pt@Ru) nanoparticles by the multi-sites biotin/streptavidin (SA) affinity can be captured onto the electrode surface to generate ECL signal. In the proposed Ru(bpy)(3)(2+) ECL system without any co-reactant, the detected ECL signal was amplified due to following multiple amplification strategies: (1) the ECL catalysis for Ru(bpy)(3)(2+) was performed by electro-inducing the DpAg particles to generate Ag(+) ion and controlled by the special potential. The catalyzer Ag(+) was produced near the electrode surface and reproduced by cyclic potential scan, which improved the catalytic efficiency. (2) The amount of the ECL signal probes linked to secondary antibodies were amplified by the adsorption of Pt nanoparticles and the multiple sites bridge linkage of biotin/SA. These new multiple signal amplification strategies made the proposed ECL immunosensor achieve ultrasensitive detection for model protein human IgG with a detection limit down to 3 pg mL(-1), which can be further extended to the detection of disease biomarkers.

Enhanced electrogenerated chemiluminescence of [formula omitted]/tripropylamine system on indium tin oxide nanoparticle modified transparent electrode

Electrochemiluminescence (ECL) systems using Ru ( bpy ) 3 2 + (bpy = 2,2′-bipyridyl) and tripropylamine (TPA) on metal and carbon electrodes are widely used in immunoassay and DNA hybridization detections. The large overpotential of TPA oxidation on indium tin oxide (ITO) electrodes has limited their ECL application. In this work, significant enhancement of ECL signal from the Ru ( bpy ) 3 2 + /TPA system was achieved on ITO nanoparticle-modified electrode. Due to the electrocatalytic activity of ITO nanoparticle electrodes, TPA oxidation potential was found to shift negatively by 600 mV, leading to 24-fold increases in ECL peak intensity over the conventional ITO electrode, and 2-fold increases in the integrated signal over the conventional gold electrode. As a result, the detection limit for Ru ( bpy ) 3 2 + in solution was 20-fold lower than the value on the conventional ITO electrode. Because the transmittance of ITO at 610 nm (the wavelength of Ru ( bpy ) 3 2 + emission maximum) was reduced only slightly after nanoparticle modification, it may offer a better alternative than metal and carbon electrodes in ECL measurements.

Same-Day Detection ofEscherichia coliO157:H7 from Spinach by Using Electrochemiluminescent and Cytometric Bead Array Biosensors

Contamination of fresh produce with Escherichia coli O157:H7 and other pathogens commonly causes food-borne illness and disease outbreaks. Thus, screening for pathogens is warranted, but improved testing procedures are needed to allow reproducible same-day detection of low initial contamination levels on perishable foods, and methods for detecting numerous pathogens in a single test are desired. Experimental procedures were developed to enable rapid screening of spinach for E. coli O157:H7 by using multiplex-capable immunological assays that are analyzed using biosensors. Detection was achieved using an automated electrochemiluminescent (ECL) assay system and a fluorescence-based cytometric bead array. Using the ECL system, less than 0.1 CFU of E. coli O157:H7 per gram of spinach was detected after 5 h of enrichment, corresponding to 6.5 h of total assay time. Using the cytometric bead array, less than 0.1 CFU/g was detected after 7 h of enrichment, with a total time to detection of less than 10 h. These results illustrate that both biosensor assays are useful for rapid detection of E. coli O157:H7 on produce in time frames that are comparable to or better than those of other testing formats. Both methods may be useful for multiplexed pathogen detection in the food industry and other testing situations.

Formula omitted] electrochemiluminescence in the presence of formaldehyde or formic acid

Two new electrochemiluminescence (ECL) systems, the Ru (bpy) 3 2 + /formaldehyde system and the Ru (bpy) 3 2 + /formic acid system, have been reported. Intense ECL was observed from Ru (bpy) 3 2 + in the presence of formaldehyde or formic acid at glassy carbon electrodes, but not at platinum and gold electrodes. The ECL reactions may involve the oxidation of formaldehyde and formic acid by electrogenerated OH to form the strong reducing intermediates CHO and CO 2 − that react with Ru (bpy) 3 3 + to emit light. The ECL intensity increases linearly with the formaldehyde or formic acid concentration over the range of concentration of 3 μM–1 mM. The detection limit is 1 μM for formaldehyde and formic acid.

DNA sensor by using electrochemiluminescence of acridinium ester initiated by tripropylamine

It was found that tripropylamine (TPA) could be used as a coreactant to initiate the electrochemiluminescence (ECL) of acridinium NHS ester (AE NHS) labels attached to DNA. The radicals generated in the electro-oxidation process of TPA reacted with AE NHS to form the excited N-methylacridone, giving rise to light emission. The AE/TPA ECL system was for the first time used as the detection system for developing an ECL-based DNA sensor. In the protocol, streptavidin-modified gold nanoparticles were firstly immobilized onto a thiol-treated gold electrode. The streptavidin could specifically interact with the biontinylated capture DNA. Afterwards, the target DNA and the AE-labeled report DNA were conjugated onto the electrode step by step due to the hybridization reactions, and a sandwich-type sensor was fabricated. The ECL signals of the sensor were obtained under pulse potential condition in alkaline solution containing 50.0 mmol L(-1) TPA. Under optimized experimental conditions, the linear range of the DNA sensor for the determination of the target DNA was from 5.0 × 10(-15) to 5.0 × 10(-12) mol L(-1). The detection limit (S/N = 3) was 3.0 × 10(-15) mol L(-1). Moreover, the sensor could specifically recognize the target DNA against one base-pair mismatched sequences, two base-pair mismatched sequences, and the noncomplementary sequences. It is of great application potential in clinic analysis.

Rapid and highly sensitive detection of mercury ion (Hg 2+) by magnetic beads-based electrochemiluminescence assay

A novel and highly sensitive electrochemiluminescence (ECL) assay based on magnetic beads separation/collection process and thymine–Hg 2+–thymine (T–Hg 2+–T) coordination chemistry has been designed to detect Hg 2+ ions in aqueous solution. In this protocol, two amine-terminated complementary DNA probes with five thymine–thymine (T–T) mismatches were introduced. One was coupled with carboxyl-modified magnetic beads and the other was labeled with tris (2, 2-bipyridine) ruthenium (II) (TBR). The couple of DNA probes, in the presence of Hg 2+, can form double-stranded structure via the Hg 2+-mediated coordination of T–Hg 2+–T base pair. Therefore, they can be collected on the surface of electrode using a magnetic field. On the electrode, TBR labels and tripropylamine (TPA) could react to emit photons, which were detected by a custom-built ECL system. The detection limit for Hg 2+ in this assay is 5 nM, which is below the upper limit of Hg 2+ for drinkable water mandated by United States Environmental Protection Agency (EPA), 10 nM. To the best of our knowledge, this is the first example of ECL assay applying to detect Hg 2+ with highly sensitivity and selectivity.