Utilization of Pyronine B as Voltammetric Probe for the Determination of DNA

Abstract

The electrochemical behaviors of the interaction of pyronine B (PB) with DNA were investigated on the mercury drop working electrode. In pH 2.0 Britton‐Robinson (B‐R) buffer solution, PB can be easily reduced on the mercury electrode and had a well‐defined voltammetric reductive wave at ?0.86 V (vs. saturated calomelelectrode, SDE). On the addition of DNA into the PB solution, the reductive peak current of PB decreased with the positive movement of the peak potential and without the appearance of new peaks. The result showed that a new supramolecular complex was formed via intercalation of PB with DNA, which can't be reduced on the Hg electrode. The conditions of interaction and the electrochemical detection were carefully investigated. Under the optimal conditions the decrease of peak current was proportional to the concentration of DNA in the range of 1.0～30.0 mg/L with the linear regression equation as ΔIp″(nA)=51.84C (mg/L)–94.97 (n=13, γ=0.993) and the detection limit was 0.90 mg/L. The interaction mechanism was discussed with the aggregation of DNA‐PB supramolecular complex and the stoichiometry of the supramolecular complex was calculated with the binding number as 3 and the binding constant as 1.61×10¹⁵.     

Voltammetric and spectroscopic studies on the interaction of anti-cancer herbal drug rutin with an anti-tuberculosis agent rifampicin

In this paper, the interaction of rutin (Rt) with rifampicin (RIF) has been investigated using voltammetric and spectroscopic techniques. With the addition of RIF into the Rt solution, both the reduction and oxidation currents of rutin decrease with few changes in the peak potentials and no new peaks appeared. The binding of Rt with RIF forms a kind of supramolecular complex Rt-RIF, which is electrochemically non-active. The experimental data showed that the formation of supramolecular complex is due to the electrostatic attraction of Rt with RIF. The binding reaction resulted in the decrease of the concentration of free Rt in the solution, and the decrease of the cathodic peak current of Rt. The stoichiometry of the Rt-RIF biocomplex was determinated to be 1: 1 by means of voltammetric data. The effect of pH on the binding reaction has been also studied. Based on the peak current values of Rt in the presence and absence of RIF, the binding constants of Rt-RIF at pHs 4, 7 and 9 were calculated as 0.72 × 10⁴ M⁻¹, 1.28 × 10⁴ M⁻¹ and 0.19 × 10⁴ M⁻¹, respectively. The experimental results indicate that there is a strength interaction between Rt and RIF in neutral pH. This binding reaction was supported by UV-Vis. and IR spectroscopy techniques.     

A New Electrochemical Method for the Determination of Proteins with Cupferron‐Cadmium(II) Complex

A new electrochemical method was proposed for the determination of trace amounts of proteins based on the cupferron (Cup) and cadmium(II) complex [Cup‐Cd(II)] as the voltammetric probe. In the selected pH 6.5 Britton–Robinson (B–R) buffer solution, Cup can interact with Cd(II) to form a stable complex of [Cup‐Cd(II)], which had a sensitive linear sweep voltammetric reductive peak at ?0.654 V (vs. SCE). The addition of human serum albumin (HSA) into [Cup‐Cd(II)] complex solution could greatly decrease the reductive peak current without the change of the reductive peak potential, which indicated that HSA could interact with [Cup‐Cd(II)] complex to form a supramolecular biocomplex. The interaction mechanism was discussed and the decrease of reductive peak current was proportional to the concentration of HSA, which could be further used for the proteins determination. The optimal conditions of the binding reaction and the electrochemical detection were carefully investigated. Under the optimal conditions a new quantitative determination method for different kinds of proteins such as HSA, bovine serum albumin (BSA) and bovine hemoglobin (BHb) etc. was developed. The proposed method was simple, practical and relatively free from the interferences of coexisting substances, and it was further applied to the samples determination with satisfactory results. The binding constant (β_s) and the binding number (m) of HSA with [Cup‐Cd(II)] complex were calculated by the voltammetric data with the results as β_s=1.12×10⁶ and m=1.     

Docosyltrimethylammonium chloride modified glassy carbon electrode for simultaneous determination of dopamine and ascorbic acid

A glassy carbon electrode (GCE) modified with docosyltrimethylammonium chloride (DCTMACl) is used for simultaneous determination of dopamine (DA) and ascorbic acid (AA) using differential pulse voltammetry (DPV) technique in 0.10 mol·L^?1 phosphate buffer solution of pH 5.0. The cationic surfactant DCTMACl modified film has a positive charge. DA exists as the positively charged species, whereas AA is the negatively charged one in the solution. Thus, at DCTMACl film-modified GCE, the oxidation peak potential of AA shifts toward less negative potential and the peak current of AA increases a little, while the oxidation peak potential of DA shifts toward more positive potential and peak current decreases greatly in comparison with that on bare electrode. The two anodic peaks are separated around 200 mV. Under optimal conditions, the catalytic peak currents obtained from DPV increase linearly with concentrations of DA and AA in the ranges of 1.0?×?10^?5 to 1.0?×?10^?3?mol·L^?1. This electrode has good reproducibility, high stability in its voltammetric response, and low detection limit (micromolar) for both AA and DA. The modified electrode has been applied to the determination of DA and AA in injection.     

Voltammetric Studies on Chromotrope 2B‐Protein Interaction and Its Analytical Application

In this paper the interaction of chromotrope 2B (Ch2B) with proteins was studied by the electrochemical method. Ch2B is an azo dye and shows irreversible electrochemical responses on the mercury electrode in a pH 3.0 Britton‐Robinson (B‐R) buffer solution. After the addition of human serum albumin (HSA) into the Ch2B solution, an interaction took place, and a supramolecular complex was formed in the mixed solution. The electrochemical parameters of the Ch2B‐HSA interaction system were calculated and compared. The results showed that in the absence and presence of HSA in Ch2B solution, the electrochemical parameters such as the formal potential E⁰, the electrode reaction standard rate constant k_s, etc. showed no significant changes, which indicated that an electro‐inactive supramolecular biocomplex was formed. The free concentration of Ch2B in reaction solution was decreased, and this resulted in the decrease of the peak current. The binding constant and the binding ratio were calculated as 7.85 × 10⁹ and 1:2, respectively, and the interaction mechanism was discussed. Based on the decrease of the peak current, this new electrochemical method was proposed for the determination of HSA in the concentration range of 2.0?25.0 mg/L with the linear regression equation as ΔIp′ (nA) = 50.56C (mg/L) — 6.72 (γ = 0.995). This method was further used to determine other different kinds of proteins, such as bovine serum albumin (BSA), oval albumin, etc‥ The new method was successfully applied to detect the content of albumin in healthy human serum samples with the results in good agreement with the traditional Coomassie Brilliant Blue G‐250 spectrophotometric method.     

Microdetermination of double-stranded DNA by linear sweep voltammetry with phenosafranine.

A novel voltammetric method for the determination of microamounts of fish sperm double-stranded (ds) DNA based on its interaction with phenosafranine (PSF) is proposed in this paper. In a pH 3.5 Britton-Robinson (B-R) buffer solution, PSF had a well-defined second-order derivative linear-sweep voltammetric reductive peak at -0.32 V (vs. SCE) on a mercury electrode. After the addition of dsDNA into the PSF solution, the reductive peak current decreased significantly without a shift of the peak potential, and no new peak appeared. The experiment results showed that a new supramolecular complex was formed after the interaction of dsDNA with PSF, which resulted in a decrease of the diffusion coefficient, and then a decrease of the reductive peak current. The interaction conditions and the electrochemical detection conditions were carefully investigated. Under the optimal conditions, the decrease of the peak current was proportional to the dsDNA concentration in the range 1.0 - 40.0 microg/mL with the linear regression equation DeltaI(p)'(nA) = 32.59C(microg/mL) - 4.03 (n = 13, gamma = 0.998) and a detection limit of 0.25 microg/mL (3 sigma). The interaction mechanism was considered based on the aggregation of the dsDNA-PSF supramolecular complex; the stoichiometry of this supramolecular complex was calculated based on voltammetric data with a binding number of 3 and a binding constant of 2.76 x 10(12). This method was successfully applied to the determination of synthetic samples and the polymerase chain reaction (PCR) product of the nopaline synthase gene (NOS) DNA from genetically modified organisms (GMOs) with satisfactory results.     

Electrochemical studies of the interaction of tetraphenylporphyrin tetrasulfonate (TPPS) with albumin

An electrochemical investigation of the interaction of TPPS with BSA on a Hg electrode is reported for the first time. The addition of BSA to TPPS solution results in a decrease of both the reduction and the oxidation current with no change of the peak potentials. In presence of BSA, no new peaks appear, and the standard rate constant k _s is not significantly changed. The reaction of TPPS with BSA yields a kind of supramolecular complex TPPS-BSA, which is electrochemically non-active. The equilibrium constant for the complex has been calculated. The decrease of the peak current can be used to determine BSA concentrations. Received: 2 August 1998 / Revised: 21 September 1998 / Accepted: 24 September 1998     

Electrochemical studies of the interaction of tetraphenylporphyrin tetrasulfonate (TPPS) with albumin

An electrochemical investigation of the interaction of TPPS with BSA on a Hg electrode is reported for the first time. The addition of BSA to TPPS solution results in a decrease of both the reduction and the oxidation current with no change of the peak potentials. In presence of BSA, no new peaks appear, and the standard rate constant k _s is not significantly changed. The reaction of TPPS with BSA yields a kind of supramolecular complex TPPS-BSA, which is electrochemically non-active. The equilibrium constant for the complex has been calculated. The decrease of the peak current can be used to determine BSA concentrations. Received: 2 August 1998 / Revised: 21 September 1998 / Accepted: 24 September 1998     

Application of arsenazo III for the polarographic detection of proteins

In this paper, a diazo dye of arsenazo III (AAIII) was selected as a new electrochemical probe for the determination of proteins. In Britton-Robinson (B-R) buffer solution of pH 2.4, AAIII had a sensitive second order derivative linear sweep voltammetric reductive peak at ?0.39 V (vs. SCE). After the addition of human serum albumin (HSA) into AAIII solution, an interaction was taken place in the mixed solution and a biosupramolecular complex was formed, which resulted in the decreased reductive peak currents of AAIII. Based on the observed decrease in peak current, a sensitive electrochemical method was proposed for the determination of different proteins such as HSA, bovine serum albumin (BSA) and bovine hemoglobin (BHb). The optimal conditions for the interaction and the interfering effects of coexisting substances on the detection were investigated. The proposed method was successfully applied to the determination of HSA in synthetic samples with the recoveries in the range of 99.13–100.50%. The stoichiometry of HSA-AAIII biocomplex was calculated by voltammetric data with a binding number of 2 and a binding constant of 7.53 × 10⁹.     

Electrochemical studies of the interaction of tetraphenylporphyrin tetrasulfonate (TPPS) with an antibody

A voltammetric study of the interaction of tetraphenylporphyrin tetrasulfonate (TPPS) with its polyclonal antibody on Hg electrode in Britton–Robinson buffer solution is described. The addition of antibody to TPPS solution can result in a decrease of both the reduction and the oxidation current with no change of the peak potential. In presence of antibody, no new peaks appear. The standard rate constant k_s and the transfer coefficient α are not significantly changed. The reaction of TPPS with antibody yields kind of supramolecular complexes, which are non-electrochemically active. The binding rate as well as the equilibrium constant for the complex has been calculated.     

A simple linear sweep voltammetric method for the determination of double-stranded DNA with malachite green

In pH 3.5 Britton—Robinson buffer solution double-stranded (ds) DNA can react with malachite green (MG) to form an interaction complex, which resulted in the decrease of the electrochemical response of MG, MG had a well-defined second-order derivative linear sweep voltammetric peak at −0.73 V (vs. SCE). After the addition of dsDNA into MG solution, the reductive peak current decreased with the positive shift of peak potential, which was the typical characteristic of intercalation. Based on the interaction, an indirect electrochemical determination method for dsDNA was established. The optimum conditions for the reaction were investigated and there were little or no interferences from the commonly coexisting substances. The decrease of peak current was linear with the concentration of dsDNA over the range of 0.8–12.0 μg cm⁻³ with the linear regression equation as ΔI _p″/nA = 91.70 C/(μg cm⁻³) + 74.55 (n = 10, γ = 0.990). The detection limit was calculated as 0.46 μg cm⁻³ (3σ). The method had high sensitivity and was further applied to the dsDNA synthetic samples with satisfactory result. The interaction mechanism was discussed with the intercalation of DNA-MG to form a supramolecular complex and the stoichiometry of the supramolecular complex was calculated by electrochemical method with the binding number 3 and the binding constant 2.35 × 10¹⁵ (mol dm⁻³)⁻³.     

Electrochemical sensor for heparin based on a poly(thionine) modified glassy carbon electrode

A poly(thionine) thin film modified electrode was successfully assembled on the surface of the glassy carbon electrode by means of electrochemical polymerization, which was carried out with cyclic voltammetric sweeping in the potential range 0 to +1.4 V (vs. Ag/AgCl) in perchloric acid solution containing 0.1 mmol L^?1 thionine. The film modified electrode exhibited a couple of well-defined redox peaks, and the redox peaks decreased correspondingly without a shift of the peak potential after the addition of heparin. The conditions of the binding reaction and the electrochemical detection were optimized. Under the optimum conditions the decrease of the peak current was proportional to the concentration of heparin in the range 4.0 to 22.0 μg mL^?1 and the detection limit was 0.28 μg mL^?1. The relative standard deviation (RSD) for five parallel determinations of 10.0 μg mL^?1 heparin was 0.93%. The effects of potentially interfering species were investigated and the method was successfully applied to the determination of heparin in a pharmaceutical formulation.     

Voltammetric Determination of Dopamine in Human Serum and Urine at a Glassy Carbon Electrode Modified by Cysteic Acid Based on Electrochemical Oxidation of L ‐cysteine

Abstract

The voltammetric behavior of dopamine was studied at a glassy carbon electrode modified by cysteic acid, based on electrochemical oxidation of L ‐cysteine. The modified electrode showed strong electrocatalytic activity towards dopamine and good selectivity. In a phosphate buffer solution (pH 7.4), the anodic peak current obtain from the differential pulse voltammetry of dopamine was linearly dependent on its concentration in the range of 5×10^?9 to 4.0×10^?6mol · L^?1, with a detection limit of 2×10^?9mol · L^?1. The low‐cost modified electrode had been applied to the determination of dopamine in human serum and urine samples with satisfactory results.     

Electrochemical monitoring of the interaction of cisplatin with cystine and ascorbic acid

The interaction of cisplatin with cystine (RSSR) and ascorbic acid (AA) has been investigated in aqueous solution at physiological pH, using drug concentration of 0.835×10⁻⁶ M to 46.76×10⁻⁶ M with RSSR concentration of 6.5×10⁻⁵ M by voltammetric techniques. Voltammetric results show that at low drug concentration (<0.835×10⁻⁶ M), the interaction between cisplatin and RSSR occurs slightly, while at higher drug concentrations (≥0.835×10⁻⁶ M), cisplatin binds to RSSR and gives an unseparated-reduction peak at −0.610 V. Based on the voltammetric behaviour of RSSR, it has been suggested that binding of cisplatin to RSSR occurs mainly at the disulphide bond. It has been shown that the interaction of cisplatin with RSSR in the presence of AA is very rapid with regards to the interaction without AA, and a new peak, probably corresponding to the interaction of cisplatin with RSSR and AA is appeared at −0.970 V. The fractional coefficient for the binding of cisplatin to RSSR with or without AA was calculated to be 0.031 and 0.055, respectively.     

Preparation of Polypyrrole/Nuclear Fast Red Films on Gold Electrode and Its Application on the Electrocatalytic Determination of Methyl‐dopa and Ascorbic Acid

A novel voltammetric method using the Ppyox/NFR/Au (poly pyrrole – nuclear fast red – gold) modified electrode was developed for simultaneous measurement of various combinations of ascorbic acid (AA) and methyldopa (MDA). Polypyrrole film was prepared by incorporation of nuclear fast red (NFR) as doping anion, during the electropolymerization of pyrrole onto a gold (Au) electrode in aqueous solution using cyclic voltammetric (CV) method, and then it was overoxidized at constant potential. Differential pulse voltammetry was utilized for the measurement of both analytes using modified electrode. Well‐separated voltammetric peaks were observed for ascorbic acid (AA) and methyldopa at the Ppyox/NFR/Au modified electrodes with peak separation of 0.210 V. It has been found that under optimum condition (pH 3.0), the oxidation of AA and MDA at the surface of the electrode occurs at a potential about 260 and 50 mV less positive than unmodified Au electrode respectively. The current catalytic oxidation peaks showed a linear dependent on the concentration of AA and MDA in the range of 9.0×10^?6 to 1.0×10^?3 and 1.0×10^?7 to 2.0×10^?5 mol L^?1 respectively. The detection limit of 5.8×10^?6 and 5.0×10^?8 mol L^?1 (S/N=3) were obtained for AA and MDA respectively. The modified electrode was used for determination of AA and MDA in some real samples such as human serum and tablet.     

A Linear Sweep Voltammetric Determination of DNA with Methyl Violet

A new quantitative method for the determination of DNA in aqueous solution based on the interaction of methyl violet (MV) with fsDNA by linear sweep voltammetric technique is proposed in this paper. The interaction of MV with fsDNA caused the decrease of the reductive peak current of MV at ?0.650 V (vs. SCE) on a mercury electrode in pH 1.5 Britton‐Robinson (B‐R) buffer solution. Factors including the acidity of the buffer, the reaction time, the optimal concentration of MV, the reaction temperature, the influences of the foreign substances, etc. were all carefully investigated. Under the optimal conditions, the decrease of the reductive peak current of MV was proportional to the concentration of fsDNA in the linear range of 0.90～20.0 mg/L and the detection limit for fsDNA was 0.25 mg/L with the RSD of 4.00%. Synthetic samples were determined with satisfactorily results. The stoichiometry of MV with DNA was calculated by voltammetric data and the results showed that the binding ratio of DNA:MV was 1:2 with the binding constant β as 7.36 × 10⁸.     

Single-Sweep Voltammetric Determination of Tamoxifen at Carbon Paste Electrode

Abstract

A single-sweep voltammetric method was proposed for the determination of tamoxifen. The proposed method took advantage of both the accumulation of carbon paste electrode toward tamoxifen and the rapidity of single-sweep voltammetry. In HAc-NaAc (pH 4.1) buffer/methanol (85:15 v/v) mixed solution, an irreversible oxidation peak of tamoxifen was observed at 1.1 V (versus SCE). The second-order derivative peak current of tamoxifen and its concentration plots were rectilinear over the range of 7.0 × 10^?10 ～ 3.0 × 10^?8 mol · l^?1 with a detection limit of 1.0 × 10^?10 mol · l^?1 without any preconcentration. The proposed method was evaluated by analyzing tamoxifen citrate tablets, which was characterized by rapidity and higher sensitivity.     

Determination of Tyrosine in the Presence of Sodium Dodecyl Sulfate Using a Gold Nanoparticle Modified Carbon Paste Electrode

A carbon paste electrode modified with gold nanoparticles (AuMCPE) was used as a highly sensitive sensor for determination of Tyrosine (Tyr), in the presence of an anionic surfactant, sodium dodecyl sulfate (SDS), in aqueous solution. The measurements were carried out by using of differential pulse voltammetry (DPV), cyclic voltammetry (CV), amd chronocoulometry and chronoamperometry methods. The prepared electrode shows voltammetric responses with high sensitivity and selectivity for Tyr in the presence of SDS. The relationship between the oxidation peak current of Tyr and its concentration was obtained linearly and it was 1.0 × 10^?7 to 1.0 × 10^?5 M with a detection limit of 5.5 × 10^?8 M in the absence of SDS. On the other hand the oxidation peak current of Tyr increased significantly at AuMCPE in the presence of SDS and its detection limit was reduced to 2.7 × 10^?9 M. The proposed voltammetric approach was also applied to the determination of Tyr concentration in human serum.     

Simultaneous Voltammetric Detection of Salsolinol and Uric Acid in the Presence of High Concentration of Ascorbic Acid with Gold Nanoparticles/Functionalized Multiwalled Carbon Nanotubes Composite Film Modified Electrode

This work demonstrates gold nanoparticles (AuNPs)/functionalized multiwalled carbon nanotubes (f‐MWCNT) composite film modified gold electrode via covalent‐bonding interaction self‐assembly technique for simultaneous determination of salsolinol (Sal) and uric Acid (UA) in the presence of high concentration of ascorbic acid (AA). In pH 7.0 PBS, the composite film modified electrode exhibits excellent voltammetric response for Sal and UA, while AA shows no voltammetric response. The oxidation peak current is linearly increased with concentrations of Sal from 0.24–11.76 μmol L^?1 and of UA from 3.36–96.36 μmol L^?1, respectively. The detection limits of Sal and UA is 3.2×10^?8 mol L^?1 and 1.7×10^?7 mol L^?1 , respectively.     

Determination of Ultra Trace Amounts of Uranium (VI) by Adsorptive Stripping Voltammetry Using L-3-(3, 4-dihydroxy phenyl) Alanine as a Selective Complexing Agent

Abstract

The spectrophotometric behavior of uranium (VI) with L-3-(3, 4-dihydroxy phenyl) alanine (LDOPA) reagent revealed that the uranium can form a ML₂ complex with LDOPA in solution. Thus a highly sensitive adsorptive stripping voltammetric protocol for measuring of trace uranium, in which the preconcentration was achieved by adsorption of the uranium-LDOPA complex at hanging mercury drop electrode (HMDE), is described. Optimal conditions were found to be a 0.02 M ammonium buffer (pH 9.5) containing 2.0 × 10^?5 M (LDOPA), an accumulation potential of ? 0.1 V (versus Ag/AgCl) and an accumulation time of 120 sec.

The peak current and concentration of uranium accorded with linear relationship in the range of 0.5–300 ng ml^?1. The relative standard deviation (at 10 ng ml^?1) is 3.6% and the detection limit is 0.27 ng ml^?1. The interference of some common ions was studied. Applicability to different real samples is illustrated. The attractive behavior of this reagent holds great promise for routine environmental and industrial monitoring of uranium.