Differential Pulse Voltammetry Signal Research Articles

Combination of electrochemistry with chemometrics to introduce an efficient analytical method for simultaneous quantification of five opium alkaloids in complex matrices

For the first time, an analytical methodology based on differential pulse voltammetry (DPV) at a glassy carbon electrode (GCE) and integration of three efficient strategies including variable selection based on ant colony optimization (ACO), mathematical pre-processing selection by genetic algorithm (GA), and sample selection (SS) through a distance-based procedure to improve partial least squares-1 (PLS-1, ACO-GA-SS-PLS-1) multivariate calibration (MVC) for the simultaneous determination of five opium alkaloids including morphine (MOP), noscapine (NOP), thebaine (TEB), codeine (COD), and papaverine (PAP) was used and validated. The baselines of the DPV signals were modeled as a smooth curve, using P-splines, a combination of B-splines and a discrete roughness penalty. After subtraction of the baseline we got a signal with a two-component probability density. One component was for the peaks and it was approximated by a uniform distribution on the potential axis. The other component was for the observed noise around the baseline. Some sources of bi-linearity deviation for electrochemical data were discussed and analyzed. The lack of bi-linearity was tackled by potential shift correction using correlation optimized warping (COW) algorithm. The MVC model was developed as a quinternary calibration model in a blank human serum sample (drug-free) provided by a healthy volunteer to regard the presence of a strong matrix effect which may be caused by the possible interferents present in the serum, and it was validated and tested with two independent sets of analytes mixtures in the blank and actual human serum samples, respectively. Fortunately, the proposed methodology was successful in simultaneous determination of MOP, NOP, TEB, COD, and PAP in both blank and actual human serum samples and its results were satisfactory comparable to those obtained by applying the reference method based on high performance liquid chromatography-ultraviolet detection (HPLC-UV).

Toward therapeutic effects evaluation of chronic myeloid leukemia drug: Electrochemical platform for caspase-3 activity sensing

In recent decades, advanced therapies and novel scientific drug evaluation systems for chronic myeloid leukemia (CML) treatment are very urgent due to its increasing morbidity. The combination of dasatinib with tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) was supposed to be effective for leukemia therapy. Taking full advantage of novel nano-biotechnology, we have developed a robust electrochemical cytosensing approach to profile the therapeutic effects of dasatinib and TRAIL by probing the activity of caspase-3 from apoptotic CML cells. The sensor was on a base of a glassy carbon electrode (GCE) modified with nano-materials composed of Au nanoparticles (AuNPs), poly(dimethyl diallyl ammonium chloride) (PDDA), and carbon nanotubes (CNTs). Then the platform immobilized the biotinylated DEVD-peptide (biotin-Gly-Asp-Gly-Asp-Glu-Val-Asp-Gly-Cys) via the strong bonding between AuNPs and the thiol group (Au–S bond). In particular, the sensor was then constructed with the environmentally friendly alkaline phosphatase (ALP) via the specific interaction between the biotin and streptavidin, and could retest detection indirectly for caspase-3 sensing by detecting the differential pulse voltammetry (DPV) signal of enzymatic catalysis product, ascorbic acid (AA). The results indicated that either dasatinib or TRAIL could successfully induce the apoptosis of CML cells, while the combination of dasatinib and TRAIL resulted in an improved therapeutic effect, suggesting a novel optimized strategy for CML therapy. This novel electrochemical sensing strategy exhibits attractive advantages of environmental benignity, simple performance, high stability, and may be readily expanded to evaluate other cancer therapeutic effects.

DNA-AuNPs based signal amplification for highly sensitive detection of DNA methylation, methyltransferase activity and inhibitor screening

A sensitive and selective electrochemical method was developed for the detection of DNA methylation, determination of DNA methyltransferase (MTase) activity and screening of MTase inhibitor. Methylene blue (MB) was employed as electrochemical indicator and DNA-modified gold nanoparticles (AuNPs) were used as signal amplification unit because the DNA strands in this composite have strong adsorption ability for MB. First, the thiolated single-stranded DNA S1 was self-assembled on gold electrode, hybridization between the lower portion of DNA S1 and its complementary DNA S2 formed an identical double-stranded tetranucleotide target sequence for both DNA adenine methylation (Dam) MTase and methylation-resistant endonuclease Mbo I, then the upper portion of DNA S1 was hybridized with its complementary DNA S3 modified on AuNPs to bring the DNA S3-AuNPs amplification units onto the electrode. The DNA S1/S2/S3-AuNPs bioconjugate has lots of DNA strands, and they can adsorb abundant MB. Mbo I endounuclease could not cleave the identical target sequence after it was methylated by Dam MTase. On the contrary, the sequence without methylation could be cleaved, which would decrease the amount of adsorbed MB. The presence of redox-active MB was detected electrochemically by differential pulse voltammetry (DPV). Thus, the activity of Dam MTase and methylation status were sensitively converted to the DNA S3-AuNPs amplified DPV signals. The DPV signal demonstrated a linear relationship with logarithm of Dam concentration ranging from 0.075 to 30U/mL, achieving a detection limit of 0.02U/mL (S/N=3). Also, screening of Dam MTase inhibitor 5-fluorouracil was successfully investigated using this fabricated sensor.

DNA Wrapped Metallic Single-walled Carbon Nanotube Sensor for Pb (II) Detection

Based on the interaction of Pb (II) ions with nucleic acid via phosphate groups and nucleic bases of the deoxyribonucleic acid structure, an electrochemical sensor with DNA wrapped metallic single-walled carbon nanotube (SWNT) has been fabricated for sensitive and selective detection of lead ion (Pb2+). The sensor shows a rapid differential pulse voltammetry signal response to the content of Pb2+, and a quantitative detection of Pb2+ has been established with an expanded linear responsive range from 1 × 10−9 to 1.0 × 10−8 mol L−1 (R = 0.9979) and a detection limit of 4.1 × 10−10 mol L−1 in buffer solution (pH 5.0). Furthermore, the sensor displayed a negligible differential pulse voltammetry signal response for other metal ions, indicating its high selectivity for Pb2+. The investigation of electrochemical impedance spectroscopy and cyclic voltammetry in buffer solution demonstrated that metallic SWNTs played an important role in enlarging current response and increasing sensitivity of the sensor.

A label-free electrochemical biosensor for trace uranium based on DNAzymes and gold nanoparticles

A novel and highly sensitive electrochemical DNAzymes biosensor was fabricated using Au nanoparticles (AuNPs) immobilized on the surface of Au electrode that had been previously modified with self-assembled monolayers of 1,6-hexanedithiol. Different modified electrodes were prepared and characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The AuNPs were found to have a large surface area to anchor a large number of negatively charged phosphate backbones of DNAzymes, which further absorbed the electroactive indicator of hexaammineruthenium(III) ([Ru(NH3)6]3+) to amplify the electrochemical signal. In the presence of target molecules, a large amount of DNA partly associated with [Ru(NH3)6]3+ were removed from the electrode surface, leading to a significant decrease in peak current. Differential pulse voltammetry signals of [Ru(NH3)6]3+ provided quantitative measures of the concentrations of uranyl ion (UO2 2+), with linear calibration ranging from 13 pM to 0.15 nM and a detection limit of 5 pM. The presence of other metal ions did not affect the detection of UO2 2+, which indicated the high specificity of UO2 2+. Therefore, a new electrochemical DNAzymes sensor was designed with specific DNAzymes and AuNPs as immobilization platform and signal amplifier.

Electrochemical biosensors for the assay of DNA damage initiated by ferric ions catalyzed oxidation of dopamine in room temperature ionic liquid

Oxidative DNA damage is one of the most significant and extensively studied mechanisms of neurodegenerative diseases such as Parkinson and Alzheimer. This study was devoted to proposing an electrochemical method for monitoring oxidative DNA damage by hydroxyl radical through dopamine reacted with the localization of ferric catalytic. A hydrophobic room temperature ionic liquid, 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]), was applied as nonaqueous solvent for the generation of free radicals. DNA damage could be detected by differential pulse voltammetry signals of the electroactive indicator [Co(bpy)3]3+ and electrochemical impedance spectroscopy signals of [Fe(CN)6]3−/4−. More dramatic damage degree of DNA film initiated by dopamine/Fe2(SO4)3 reaction in [bmim][PF6] compared with that in an aqueous solution indicated that ionic liquid was much more suitable for the investigation of radicals induced DNA damage. The influences of incubation time and the molar ratio of dopamine/Fe2(SO4)3 to current respond were also explored. The most likely mechanism of DNA damage here was that the hydroxyl radical generated from the ferric ions facilitated oxidation of dopamine disturbed DNA double helix structure or caused DNA strand break. Furthermore, it turned out that both ascorbic acid and rutin could protect DNA from oxidative damage efficiently under certain conditions.

Fabrication of β-cyclodextrin-coated poly (diallyldimethylammonium chloride)-functionalized graphene composite film modified glassy carbon-rotating disk electrode and its application for simultaneous electrochemical determination colorants of sunset yellow and tartrazine

We proposed a green and facile approach for the synthesis of β-cyclodextrin-coated poly(diallyldimethylammonium chloride)-functionalized graphene composite film (β-CD-PDDA-Gr) by using l-ascorbic acid (l-AA) as the reducing agent at room temperature. The β-CD-PDDA-Gr composite film modified glassy carbon-rotating disk electrode (GC-RDE) was then developed for the sensitive simultaneous determination of two synthetic food colorants: sunset yellow (SY) and tartrazine (TT). By cyclic voltammetry (CV), the peak currents of SY and TT increased obviously on the developed electrochemical sensor. The kinetic parameters, such as diffusion coefficient D and standard heterogeneous rate constant kb, were estimated by linear sweep voltammetry (LSV). Under the optimal conditions, the differential pulse voltammetry (DPV) signals of SY and TT on the β-CD-PDDA-Gr modified GC-RDE were significantly enhanced. The enhanced anodic peak currents represented the excellent analytical performance of simultaneous detection of SY and TT in the range of 5.0×10−8 to 2.0×10−5molL−1, with a low limit of detection (LOD) of 1.25×10−8molL−1 for SY and 1.43×10−8molL−1 for TT (SN−1=3). This proposed method displayed outstanding selectivity, good stability and acceptable repeatability and reproducibility, and also has been used to simultaneously determine SY and TT in some commercial soft drinks with satisfactory results. The obtained results were compared to HPLC of analysis for those two colorants and no significant differences were found. By the treatment of the experimental data, the electrochemical reaction mechanisms of SY and TT both involved a one-electron-one-proton-transfer process.

Simple and Sensitive Electrochemical DNA Detection of Primer Generation‐Rolling Circle Amplification

AbstractA new electrochemical sequence‐specific DNA detection platform based on primer generation‐rolling circle amplification (PG‐RCA), methylene blue (MB) redox indicator, and indium tin oxide (ITO) electrode is reported. In the presence of a specific target sequence, PG‐RCA, an isothermal DNA amplification technique, produced large amounts of amplicons in an exponential manner. In addition to the standard components, the reaction mixture contained MB, which bound with the PG‐RCA amplicons. End‐point electrochemical measurement by differential pulse voltammetry (DPV) was performed using ITO electrode. The amplicon‐bound MB resulted in a lower DPV signal than free MB due to a smaller diffusion coefficient as well as electrostatic repulsion between the negatively charged amplicon‐bound MB and ITO electrode. With simple assay design (recognition probe) and instrumentation (operating temperature at 37 °C and ITO electrode without the need for probe immobilization), this detection platform is well suited for point‐of‐care and on‐site testing. Real‐time measurement was also achieved by pretreating the ITO electrode with bovine serum albumin.

A novel electrochemical aptasensor for thrombin detection based on the hybridization chain reaction with hemin/G-quadruplex DNAzyme-signal amplification

In this work, a novel signal amplification electrochemical aptasensor for the sensitive and selective detection of thrombin was successfully fabricated. The amplification method was based on the hybridization chain reaction (HCR) and a pseudobienzyme electrocatalytic system. HCR-based double-stranded DNA (dsDNA) polymers not only constructed an effective carrier for anchoring larger amounts of electron mediator methylene blue (MB) into the DNA duplexes to produce a strong differential pulse voltammetry (DPV) signal, but also resulted in the formation of hemin/G-quadruplex DNAzymes nanowires by intercalating hemin into two induced single-stranded DNA (ssDNA). With the addition of NADH into the electrolytic cell, the hemin/G-quadruplex acting as an NADH oxidase and HRP-mimicking DNAzyme for the pseudobienzyme amplifying system could in situ biocatalyze the formation of H₂O₂ with local concentrations and low transfer loss resulting in dramatic signal enhancements. The binding event can be detected by a decrease in the integrated charge of MB which electrostatically absorbed onto dsDNA polymers. In the presence of thrombin, the dsDNA polymers associated with MB and hemin/G-quadruplex structures were removed from the electrode surface, leading to a significant decrease of redox current. DPV signals of MB provided quantitative measures of the concentrations of thrombin, with a linear calibration range of 0.01-50 nM and a detection limit of 2 pM. Moreover, the resulting aptasensor also exhibited good specificity, acceptable reproducibility and stability, indicating that the present strategy was promising for broad potential application in clinic assay and various protein analyses.

Voltammetric detection of uridin diphosphate glucuronosyl transferase 1A9 (UGT1A9) gene corresponding oligonucleotide covering promoter region from −268 to −280 including (A/T) polymorphism at position −275 and optimization of the detection factors

In this study, we developed a new peptide nucleic acid (PNA) biosensor for detection of a single nucleotide polymorphism (SNP) in the UGT1A9 gene promoter region via electrochemical assay. The sensor relies on the immobilization of a 13-mer single stranded PNA probe related to the UGT1A9 gene on the Au electrode (AuE). The hybridization between the probe and its complementary sequence (DcUG275) as the target was studied by differential pulse voltammetry (DPV) of methylene blue (MB) signal. In this approach the extent of hybridization is evaluated on the basis of the difference between DPV signals of MB accumulated on the probe-AuE and MB accumulated on the probe-target-AuE. Some experimental variables affecting the performance of the biosensor including oxygen interference during the assay, probe immobilization time, probe concentration and MB accumulation time were investigated. The PNA probe modified AuE in its optimum condition was shown to be an effective sensor for the detection of hybridization and point mutations. The obtained detection limit of the utilized biosensor has been calculated as 22 nm.

Multiwalled carbon nanotubes-ceramic electrode modified with substrate-selective imprinted polymer for ultra-trace detection of bovine serum albumin

This study describes the synthesis of a new class of substrate-selective molecularly imprinted polymer. This involved tetraethylene glycol 3-morpholin propionate acrylate (functional monomer) and bovine serum albumin (template) for polymerization in aqueous condition, using “surface grafting-from” approach directly on a vinyl exposed multiwalled carbon nanotubes-ceramic electrode. The analyte recapture at pH 6.8 in aqueous environment simultaneously involved hydrophobically driven hydrogen bonds and ionic interactions between negatively charged bovine serum albumin and positively charged imprinted nanofilm. The selectively encapsulated bovine serum albumin first gets reduced at −0.9V and then oxidized within the cavity, without getting stripped off, to respond a differential pulse voltammetry signal. The limit of detection [0.42ngmL−1 (3σ, RSD≤1.02%)] obtained was free from any cross-reactivity and matrix complications in aqueous, pharmaceutical, serum, and liquid milk samples. The proposed sensor can be used as a practical sensor for ultra-trace analysis of bovine serum albumin in clinical settings.

Fabrication of the Cu2O/polyvinyl pyrrolidone-graphene modified glassy carbon-rotating disk electrode and its application for sensitive detection of herbicide paraquat

A polyvinyl pyrrolidone (PVP) functionalized graphene nanosheets (GNs) and cuprous oxide (Cu2O) sub-microparticles (micro-Cu2O/PVP-GNs) modified glassy carbon-rotating disk electrode (GC-RDE) was successfully fabricated. The electrochemical sensor based on PVP-GNs and micro-Cu2O was then developed for the investigation of the electrochemical behavior and the sensitive detection of herbicide paraquat (PQ). By cyclic voltammetry (CV) and differential pulse voltammetry (DPV), it was found that the modified electrode provided an electron-transfer microenvironment to accelerate the electrochemical catalytic reaction of PQ. The DPV signals of PQ on the micro-Cu2O/PVP-GNs modified GC-RDE were significantly enhanced. The enhanced DPV signals represented the excellent analytical performance of PQ detection in the range of 1.0 × 10−6–2.0 × 10−4 mol L−1, with a low limit of detection (LOD) of 2.65 × 10−7 mol L−1 (S/N = 3), and the acceptable reproducibility (RSD = 2.02%, n = 5). The proposed method has been used to determine the residual amount of PQ in cabbage leaves samples. By the treatment of the experimental data, the electrochemical reaction mechanism of PQ involves a diffusion-controlled quasi-reversible process and a two-electron-transfer process.

Novel printed electrode immunosensors for Schistosoma japonicum.

To improve the sensitivity and the linear range of electrochemical immunosensor to detect Schistosoma japonicum (S. japonicum) antibody. Carbon inks and silver/silver chloride inks were printed on a polyethylene terephthalate (PET) board to make a two-electrode test strip, where carbon was the working electrode and S. japonicum soluble egg antigen (SEA) was fixed at one end of working electrode by different methods; silver/ silver chloride electrode was used as control. We tested the valency of the antibody by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in an electrochemistry workstation, and conducted comparison with the results of ELISA. Two new immunosensing electrodes have been developed, based on glutaraldehyde cross-linked (GA) or chitosan-glutaraldehyde cross-linked (Chit-GA) transducer fixing S. japonicum antigen. We tested the titer of the antibody by means of CV and DPV. Our experimental S. japonicum antigen (50 μg/L) is the optimal test concentration for the GA sensor, and 10 μg/L for Chit-GA sensors. The immune reaction time of both electrodes is all essentially complete in 1 minute. The linear range for S. japonicum antibody in human positive serum sample detection by the glutaraldehyde cross-linked immunosensor is 1:1000 to 1:400, and by the chitosan-glutaraldehyde cross-linked immunosensor is 1:1000 to 1:500. As the concentration of dilution ratio of S. japonicum antibody in human positive serum sample increased, the test value of DPV increased proportionally. GA sensor and Chit-GA cross-linked S. japonicum sensors have high sensitivity and broad linear range response, and both exhibited a good linear relationship between the DPV signal and the test antibody titer.

Highly sensitive electrochemical detection of potential cytotoxicity of CdSe/ZnS quantum dots using neural cell chip

Cell chip was recently developed as a simple and highly sensitive tool for the toxicity assessment of various kinds of chemicals or nano-materials. Here, we report newly discovered potential cytotoxic effects of CdSe/ZnS quantum dots (QDs) on intracellular redox environment of neural cancer cells at very low concentrations which can be only detected by cell chip technology. Green (2.1 nm in diameter) and red (6.3 nm in diameter) QDs capped with cysteamine (CA) or thioglycolic acid (TA) were found to be toxic at 100 μg/mL when assessed by trypan blue and differential pulse voltammetry (DPV). However, in case of concentration-dependent cytotoxicity, toxic effects of TA-capped QDs on human neural cells were only measured by DPV method when conventional MTT assay did not show toxicity of TA-capped QDs at low concentrations (1–10 μg/mL). Red-TA QDs and Green-TA QDs were found to decrease electrochemical signals from cells at 10 μg/mL and 5 μg/mL, respectively, while cell viability decreased at 100 μg/mL and 50 μg/mL when assessed by MTT assay, respectively. The relative decreases of cell viability determined by MTT assay were 15% and 11.9% when cells were treated with 5–50 μg/mL of Red-TA QDs and 5–30 μg/mL of Green-TA QDs, respectively. However, DPV signals decreased 37.5% and 39.2% at the same concentration range, respectively. This means that redox environment of cells is more sensitive than other components and can be easily affected by CdSe/ZnS QDs even at low concentrations. Thus, our proposed neural cell chip can be applied to detect potential cytotoxicity of various kinds of molecular imaging agents simply and accurately.

Electropolymerization of Thin Film Conducting Polymer and Its Application for Simultaneous Determination of Ascorbic Acid, Dopamine and Uric Acid

AbstractIn this paper electropolymerization of a thin film of para‐phenylenediamine (PPD) is studied at glassy carbon electrode (GCE) in sulfuric acid media by cyclic voltammetry. The results showed that this polymer was conducting and had a reproducible redox couple in the potential region from 0.0 to 0.4 V in phosphate buffer solution. This modified GCE (p‐PPD‐GCE) was applied for simultaneous determination of ascorbic acid (AA), dopamine (DA) and uric acid (UA) using differential pulse voltammetry (DPV). The p‐PPD‐GCE in 0.1 M phosphate buffer solution (pH 5.0) separated the DPV signals of AA, DA and UA with sufficient potential differences between AA–DA and DA–UA and also enhanced their oxidation peak currents. The oxidation currents were increased from 2.0 to 2000.0 µM for AA, 10.0 to 1250.0 µM for DA and 50.0 to 1600.0 µM for UA. The detection limits were evaluated as 0.4, 1.0 and 2.5 µM for AA, DA and UA, respectively (S/N=3).

An electrochemical aptasensor based on enzyme linked aptamer assay

An aptamer is an artificial functional oligonucleic acid, which can interact with its target molecule with high affinity and specificity. Enzyme linked aptamer assay (ELAA) is developed to detect cocaine using aptamer fragment/cocaine configuration based on the affinity interaction between aptamer fragments with cocaine. The aptasensor was constructed by cleaving anticocaine aptamer into two fragments: one was assembled on a gold electrode surface, while the other was modified with biotin at 3′-end, which could be further labelled with streptavidin-horseradish peroxidase (SA-HRP). Upon binding with cocaine, the HRP-labelled aptamer fragment/cocaine complex formed on the electrode would increase the reduction current of hydroquinone (HQ) in the presence of H2O2. The sensitivity and the specificity of the proposed electrochemical aptasensor were investigated by differential pulse voltammetry (DPV). The results indicated that the DPV signal change could be used to sensitively detect cocaine with the dynamic range from 0.1μM to 50μM and the detection limit down to 20nM (S/N=3). The proposed aptasensor has the advantages of high sensitivity and low background current. Furthermore, a new configuration for ELAA requiring only a single aptamer sequence is constructed, which can be generalized for detecting different kinds of targets by cleaving the aptamers into two suitable segments.

Simply amplified electrochemical aptasensor of Ochratoxin A based on exonuclease-catalyzed target recycling

A new “signal-on” aptasensor for ultrasensitive detection of Ochratoxin A (OTA) in wheat starch was developed based on exonuclease-catalyzed target recycling. To construct the aptasensor, a ferrocene (Fc) labeled probe DNA (S1) was immobilized on a gold electrode (GE) via Au–S bonding for the following hybridization with the complementary OTA aptamer, with the labeled Fc on S1 far from the GE surface. In the presence of analyte OTA, the formation of aptamer-OTA complex would result in not only the dissociation of aptamer from the double-strand DNA but also the transformation of the probe DNA into a hairpin structure. Subsequently, the OTA could be liberated from the aptamer-OTA complex for analyte recycling due to the employment of exonuclease, which is a single-stranded DNA specific exonuclease to selectively digest the appointed DNA (aptamer). Owing to the labeled Fc in close proximity to the electrode surface caused by the formation of the hairpin DNA and to the analyte recycling, differential pulse voltammetry (DPV) signal could be produced with enhanced signal amplification. Based on this strategy, an ultrasensitive aptasensor for the detection of OTA could be exhibited with a wide linear range of 0.005–10.0ngmL−1 with a low detection limit (LOD) of 1.0pgmL−1 OTA (at 3σ). The fabricated biosensor was then applied for the measurement of OTA in real wheat starch sample and validated by ELISA method.

Label-free electrochemical aptasensor for thrombin detection based on the nafion@graphene as platform

A sensitive label-free electrochemical aptasensor was successfully fabricated for thrombin detection with nafion@graphene as platform. With electrostatic interaction between nafion and methylene blue (MB), positive charged MB was successfully assembled on nafion@graphene modified electrode surface, which provided amounts of redox probes for electrochemical aptasensor. In the presence of thrombin, the thrombin aptamer (TBA) on the electrode surface would catch the target on the electrode interface, which made a barrier for electrons and inhibits the electro-transfer, resulting in the decreased differential pulse voltammetry signals of MB. As a result, the proposed approach showed a high sensitivity and a wider linearity to thrombin in the range 0.01-50 nM with a detection limit of 6pM.

Solid-State Probe Based Electrochemical Aptasensor for Cocaine: A Potentially Convenient, Sensitive, Repeatable, and Integrated Sensing Platform for Drugs

Aptamers, which are artificial oligonucleotides selected in vitro, have been employed to design novel biosensors (i.e., aptasensors). In this work, we first constructed a label-free electrochemical aptasensor introducing a probe immobilization technique by the use of a layer-by-layer (LBL) self-assembled multilayer with ferrocene-appended poly(ethyleneimine) (Fc-PEI) on an indium tin oxide (ITO) array electrode for detection of cocaine. The Fc-PEI and gold nanoparticles (AuNPs) were LBL assembled on the electrode surface via electrostatic interaction. Then, cocaine aptamer fragments, SH-C2, were covalently labeled onto the outermost AuNP layer. When the target cocaine and cocaine aptamer C1 were present simultaneously, the SH-C2 layer hybridized partly with C1 to bind the cocaine, which led to a decreased differential pulse voltammetry (DPV) signal of Fc-PEI. This DPV signal change could be used to sensitively detect cocaine with the lowest detectable concentration down to 0.1 microM and the detection range up to 38.8 microM, which falls in the the expected range for medical use of detecting drug abuse involving cocaine. Meanwhile, the sensor was specific to cocaine in complex biologic fluids such as human plasma, human saliva, etc. The sensing strategy had general applicability, and the detection of thrombin could also be realized, displayed a low detection limit, and exhibited worthiness to other analytes. The aptasensor based on the array electrode held promising potential for integration of the sensing ability in multianalysis for simultaneous detection.

Brilliant cresyl blue as electroactive indicator in electrochemical DNA oligonucleotide sensors

A new electrochemical DNA biosensor is presented based on carbon past electrode (CPE) for immobilization and detection of short DNA sequences with brilliant cresyl blue (BCB) as electroactive label. The interaction of BCB with DNA is electrochemically detected and BCB displays different signals in the interaction to ssDNA and dsDNA and variation in the BCB signal represents the extent of hybridization at the electrode surface. The effect of solution pH on electrochemical behavior of BCB was investigated. Additionally, the effect of solution pH on BCB accumulation on the CPE was studied. Furthermore, experiments showed that the solution pH could influence the differential pulse voltammetry (DPV) signal of BCB accumulated on the electrode and the highest BCB signal was obtained in pH 7.00. The effect of electrochemical pretreatment of CPE on the ability of electrode in probe adsorption, BCB accumulation and conditions of probe immobilization including potential and time was investigated and optimum conditions were suggested. The peak currents of BCB were linearly related to the concentration of the target oligonucleotide sequence in the range of 1.0 × 10 − 8 to 5.0 × 10 − 6 M. The detection limit of this approach was 9.00 nM. The selectivity of the biosensor was studied using noncomplementary oligonucleotide.