Application of N-PLS calibration to the simultaneous determination of Cu(2+), Cd(2+) and Pb(2+) using peptide modified electrochemical sensors

The simultaneous determination of Cu(2+), Cd(2+) and Pb(2+) is demonstrated at four modified gold electrodes using N-PLS calibration. Three of the electrodes were modified with the peptides Gly-Gly-His, gamma-Glu-Cys Gly and human angiotensin I which were covalently attached to thioctic acid self-assembled monolayers and the fourth electrode was modified with thioctic acid only. Voltammetry at the modified electrodes in the presence of the three metal ions revealed one peak due to the reduction of copper and another due to the overlapping peaks of cadmium and lead which made quantification using conventional methods difficult. N-PLS was used to calibrate and predict trace concentrations (100 nM to 10 microM) of mixtures of Cu(2+), Cd(2+) and Pb(2+).     

Modified microelectrodes and multivariate calibration for flow injection amperometric simultaneous determination of ascorbic acid, dopamine, epinephrine and dipyrone

Flow injection amperometric quantification of ascorbic acid (AA), dopamine (DA), epinephrine (EP) and dipyrone (DI) in mixtures (in the microgram g-1 range) was successfully performed by using an array of microelectrodes with units modified by the electrodeposition of different noble metals, together with multivariate calibration analysis. The four groups of microelectrodes utilized included a pure gold electrode and electrodes modified by electrodeposition of platinum, palladium or a mixture of platinum + palladium. The array of microelectrodes was inserted in a flow cell and the amperometric data acquisition was performed with a four-channel potentiostat. The analysis of the resulting signals was carried out by a multivariate calibration method, using a group of 16 standard mixtures selected by a two-level factorial design. The analysis of synthetic samples and pharmaceutical compounds containing AA and DI led to very similar values to those obtained by the classical iodimetric analysis. The average absolute errors (in microgram g-1) calculated for each analyte were 0.3, 0.2, 0.4 and 0.4 for AA, DA, EP and DI, respectively.     

Sub part-per-million mass accuracy by using stepwise-external calibration in fourier transform ion cyclotron resonance mass spectrometry

A new external calibration procedure for FT-ICR mass spectrometry is presented, stepwise-external calibration. This method is demonstrated for MALDI analysis of peptide mixtures, but is applicable to any ionization method. For this procedure, the masses of analyte peaks are first accurately measured at a low trapping potential (0.63 V) using external calibration. These accurately determined (< 1 ppm accuracy) analyte peaks are used as internal calibrant points for a second mass spectrum that is acquired for the same sample at a higher trapping potential (1.0 V). The second mass spectrum has a approximately 10-fold improvement in detection dynamic range compared with the first spectrum acquired at a low trapping potential. A calibration equation that accounts for local and global space charge is shown to provide mass accuracy with external calibration that is nearly identical to that of internal calibration, without the drawbacks of experimental complexity or reduction of abundance dynamic range. For the 609 mass peaks measured using stepwise-external calibration method, the root-mean-square error is 0.9 ppm. The errors appear to have a Gaussian distribution; 99.3% of the mass errors are shown to lie within three times the sample standard deviation (2.6 ppm) of their true value.     

酒石酸修饰电极及其铜的溶出伏安法研究

本文研究了酒石酸修饰电极的制作,讨论了铜在酒石酸修饰电极上的反应机理,并确定了痕量铜的阳极溶出伏安法测定条件。     

Theoretical investigation of the peroxidase-oxidase chemical oscillator for quantitative enzyme analysis

The theoretical basis for quantitative enzyme determinations by using the features of chemical oscillations is developed. An existing model of the peroxidase-oxidase chemical oscillator, consisting of the enzyme horseradish peroxidase, oxygen and reduced nicotinamide adenine dinucleotide (NADH), is modified to include a competing (analyte) reaction. The competitive effect between the analyte and the peroxidase on the observed periodic and chaotic oscillations forms the basis of the modified model. Corresponding differential equations are numerically integrated to produce plots of dissolved oxygen concentration vs. time. The calculated oscillatory oxygen transient shows a sensitive dependence on the analyte concentration. Utilizing the property of period doubling, a theoretical calibration graph can be generated for the determination of an analyte enzyme concentration. Special properties of the technique offer a potential combination of wide dynamic range and selectable precision. This demonstrates that the oscillator should prove experimentally useful for quantitative analysis.     

Modified solid electrodes for stripping voltammetric determination of tin

The paper describes the determination of tin by ASV using modified thick film electrodes. Three different types of electrodes were developed: One modified with a mixture of Nafion and mercury(II)acetate, one modified with diethyldithiocarbamate (DDC) or pyrrolidinedithiocarbamate (PDC) and mercury(II)acetate, and one modified with calomel. The analyte was accumulated on the electrode surface after special electrochemical pretreatment of the modified electrode. After recording the voltammogram the electrodes were electrochemically regenerated. By virtue of their lifetime and their measurement reproducibility, we preferred the DDC and PDC modified electrodes. They can be used for months without changing their chemical characteristics. The linear range for tin determination with these electrodes is between 1 and 100 μg/L; the detection limit was calculated as 0.9 μg/L. The electrodes were applied to the direct determination of tin in different canned fruit juices without special sample pretreatment. Received: 20 December 1996 / Revised: 11 March 1997 / Accepted: 13 March 1997     

Modified solid electrodes for stripping voltammetric determination of tin

The paper describes the determination of tin by ASV using modified thick film electrodes. Three different types of electrodes were developed: One modified with a mixture of Nafion and mercury(II)acetate, one modified with diethyldithiocarbamate (DDC) or pyrrolidinedithiocarbamate (PDC) and mercury(II)acetate, and one modified with calomel. The analyte was accumulated on the electrode surface after special electrochemical pretreatment of the modified electrode. After recording the voltammogram the electrodes were electrochemically regenerated. By virtue of their lifetime and their measurement reproducibility, we preferred the DDC and PDC modified electrodes. They can be used for months without changing their chemical characteristics. The linear range for tin determination with these electrodes is between 1 and 100 μg/L; the detection limit was calculated as 0.9 μg/L. The electrodes were applied to the direct determination of tin in different canned fruit juices without special sample pretreatment. Received: 20 December 1996 / Revised: 11 March 1997 / Accepted: 13 March 1997     

Development of screen-printed carbon electrodes,chemically modified with cobalt phthalocyanine,for electrochemical sensor applications

A method is described for the production of screen-printed graphite electrodes and also for similar electrodes chemically modified with the electrocatalyst cobalt phthalocyanine. Using cyclic voltammetry, the electrochemical behaviour of these electrodes towards ascorbic acid, reduced glutatione and coenzyme A (CoA-SH) was investigated. The modified electrodes were found to give significant decreases in the over-potential required for the oxidation of these species at carbon electrodes. The useful electrochemical window for the unmodified carbon film electrodes was ?1.08 V to +0.85 V vs. SCE, using 1 μA background current cut-off points.Amperometry in stirred solutions was used to investigate the hydrodynamic behaviour of the electrodes and their calibration performance. The limits of detection for ascorbic acid and reduced glutathione at the modified films were 5 × 10^?8 and 1 × 10^?7 M, respectively. The calibration graphs were also linear up to 2 mM concentrations of both analytes. Using differential-pulse voltammetry, linear calibration graphs were obtained for both species up to 2.5 mM. This technique was also used to assess the reproducibility of the electrode manufacture; the coefficient of variation was 2.8% for 1.49 mM ascorbic acid and 6.9% for 0.92 mM reduced glutathione.     

High-pressure flow-injection assembly. Indirect determination of glycine by atomic absorption spectrometry

A procedure for the determination of glycine is described. The method is based on the reaction of the analyte with finely powdered, solid copper(II) carbonate in a continuous-flow assembly. The optimum experimental conditions of pH, temperature, sample volume, flow-rate, column length and internal diameter, and the linear range of calibration, were studied. Interference from foreign substances that accompany this amino acid in pharmaceutical formulations was studied, and the method was applied to the determination of glycine.     

Copper Oxide – Graphite Composite Electrodes: Application to Nitrite Sensing

A simple method for the modification of carbon powder with copper oxides is presented. Carbon powder is impregnated with copper(II) nitrate by stirring carbon powder in copper(II) nitrate solution for 1 hour and subsequently thermally treated at 823 K. The modified carbon powder was characterized using electrochemical and spectroscopic techniques. The existence of both copper(I) and copper(II) oxides have been established. The copper oxide modified carbon powder was used for preparation of composite electrodes, and the electrochemical and electrocatalytic behavior of the resulting composite electrodes was studied. The copper oxide modified carbon powder – epoxy composite electrodes showed a high electrocatalytic activity for the nitrite detection in aqueous media, with the detection limit comparable or lower than detection limits obtained with other electrochemical sensors.     

Chloroquine polymeric membrane electrodes: Development and applications

Three main types of PVC solvent polymeric membrane ion-selective electrodes for chloroquine are described. They are based on three ion-pairing agents namely dipicrylamine (DPA), tetraphenylborate (TPB) or tetrakis(4-chlorophenyl)borate (TCPB) with either dioctylphenyl phosphonate (DOPP) or trioctyl phosphate (TOP) solvent mediator. All electrodes exhibit Nernstian responses, fast dynamic response times and a wide useful pH range. The best all-round electrode is based on TPB and TOP plasticizing solvent mediators with a limit of detection of 7.1 × 10⁻⁶M and was utilized for the assay of chloroquine in tablets. Direct potentiometric determinations with either the analyte addition method or the normal calibration method gave results comparable to the official method.     

Effects of competitive binding on the amperometric determination of copper with electrodes modified with chromotrope 2B

Electrodes modified with Chromotrope 2B incorporated by ion-exchange into a polycationic film of electropolymerized [Ru(v-bpy)(3)](2+) (v-bpy = 4-vinyl-4'-methyl-2,2'-bipyridyl) have been employed in the amperometric determination of copper in solution and exhibit very high sensitivity as well as linear calibration curves in the concentration range 7 x 10(-8)-1 x 10(-4)M. The effects of competing ligands, including chloride, bromide, oxalate, ammonia, acetate, citrate, borate, humic and fulvic acids, or the presence of competing metal ions such as cobalt or nickel on the uptake of copper by the modified electrodes have also been studied. The presence of competing ligands or metal ions decreases the analytical signal due to copper incorporation. The magnitude of this effect is dependent on the relative strength of coordination of the competing ligands for copper ions or of Chromotrope 2B for the competing metals, and also on the concentration of the interferents. The relevance of this work to speciation studies is discussed.     

Exploring the use of the tripeptide Gly-Gly-his as a selective recognition element for the fabrication of electrochemical copper sensors

The modification of electrodes with the tripeptide Gly-Gly-His for the detection of copper in water samples is described in detail. The tripeptide modified electrode was prepared by first self-assembling 3-mercaptopropionic acid (MPA) onto the gold electrode followed by covalent attachment of the tripeptide to the self-assembled monolayer using carbodiimide coupling. The electrodes were characterized using electrochemistry, a newly developed mass-spectrometry method and quantum mechanical calculations. The mass spectrometry confirmed the modification to proceed as expected with peptide bonds formed between the carboxylic acids of the MPA and the terminal amine of the peptide. Electrochemical measurements indicated that approximately half the MPA molecules in a SAM are modified with the peptide. The peptide modified electrodes exhibited high sensitivity to copper which is attributed to the stable 4N coordinate complex the peptide formed around the metal ion to give copper the preferred tetragonal coordination. The formation of a 4 coordinate complex was predicted using quantum mechanical calculation and confirmed using mass spectrometry. The adsorption of the copper to the peptide modified electrode was consistent with a Langmuir isotherm with a binding constant of (8.1 +/- 0.4) 10(10) M(-1) at 25 degrees C.     

Studies on differential sensing of dopamine at the surface of chemically sensitized ormosil-modified electrodes

We report herein the preparation of few chemically sensitized organically modified sol–gel glass (ormosil) films and sensing of dopamine at the surface of the modified electrodes derived from these films. The chemical sensitization in ormosil-modified electrodes is introduced by incorporating: (a) potassium ferricyanide and (b) either Nafion, or dibenzo-18-crown-6 or in situ generated Prussian blue from potassium ferricyanide. Electrochemical sensing of dopamine on the surfaces of these modified electrodes have been investigated and found that: (i) the presence of dibenzo-18-crown-6 facilitate the magnitude of dopamine sensing, (ii) conversion of potassium ferricyanide into Prussian blue also enhances the magnitude of dopamine sensing as compared to that of control and Nafion sensitized modified electrodes, (iii) both dibenzo-18-crown-6 and Nafion sensitized ormosil-modified electrodes are found selective to dopamine in the presence of ascorbic acid present under physiological concentration range. These finding again directed our attention to investigate the sensing of dopamine: (a) on dibenzo-18-crown-6 incorporated within Prussian blue sensitized modified electrode and (b) in the presence of varying concentrations of dibenzo-18-crown-6 in the Prussian blue modified electrodes. The investigations made on these lines again suggested the following: (1) increase in dibenzo-18-crown-6 concentrations in the modified electrode increases the magnitude of dopamine sensing upto an optimum concentration of macrocycle; (2) the detection limit of dopamine sensing goes down to 30 nM as compared to that of dibenzo-18-crown-6 incorporated with potassium ferricyanide which was found to the order of 100 nM. Investigations of the interference of ascorbic acid revealed that the presence of dibenzo-18-crown-6 introduces selectivity in dopamine sensing in the presence such common interfering analyte like ascorbic acid.     

Spectrophotometric simultaneous determination of cobalt, copper and nickel using nitroso-R-salt in alloys by partial least squares

Partial least squares modeling as a powerful multivariate statistical tool applied to spectrophotometric simultaneous determination of cobalt, copper, and nickel in aqueous solutions. The concentration range for cobalt, copper and nickel were 0.4-2.6, 0.6-3.4, 0.5-5.5 ppm, respectively. The experimental calibration set was composed with 36 sample solutions using a mixture design for three component mixtures. The absorption spectra were recorded from 470 to 600 nm. The effect of pH on the sensitivity and selectivity was studied according to net analyte signal (NAS). The values of root mean square difference (RMSD) for cobalt, copper and nickel using partial least squares (PLS) were 0.0192, 0.0263 and 0.0446 ppm, respectively. The effects of various cations and anions were investigated. The method was used to determination of cobalt, copper and nickel in two sample alloys based on copper, nickel and cobalt (known as cunico) and based on cobalt, nickel and iron (known as conife).     

Electrochemically Generated Recognition Sites in Self‐doped Polyaniline Modified Electrodes for Voltammetric and Potentiometric Determination of Copper(II) Ion

Chemical recognition elements for copper(II) ion have been generated in electrodes modified with poly(aniline‐co‐metanilic acid), P(An‐co‐MA), membrane and the resulting electrodes were used as selective sensors for voltammetric and potentiometric determination of this ion in an extended pH range. The P(An‐co‐MA) membrane was electrodeposited from aqueous mixed monomer solutions of An and MA, without the presence of a supporting electrolyte. For generating the recognition elements, P(An‐co‐MA) modified electrodes were subjected to several consecutive reduction/oxidation potential steps in copper(II) ion solution. It seems that during these potential steps, the receptor sites of the membrane are adjusted to the size, complexing property and hard/soft nature of copper(II) ion. This electrochemically mediated templating process, provided a selective sensor for determination of copper(II) ion. The results of preconcentration/differential pulse anodic stripping voltammetry, indicated analytical relation between the peak current and concentration of copper(II) from 1.0×10⁻⁹ to 1.0×10⁻⁴ M. The interference effect of various metal ions was explored and it was found that only mercury and silver ions show a considerable interference. The sensor exhibited selective potentiometric response for copper(II) over a wide concentration range (1.0×10⁻⁸ to 1.0×10⁻³ M) with a Nernstian slope of 27.9±0.3 mV per decade of copper(II) ion activity.     

Liquid chromatography/tandem mass spectrometry methods for quantitation of mevalonic acid in human plasma and urine: method validation,demonstration of using a surrogate analyte,and demonstration of unacceptable matrix effect in spite of use of a stable isotope analog internal standard

Selective, accurate, and reproducible liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods were developed and validated for the determination of mevalonic acid, an intermediate in the biosynthesis of cholesterol and therefore a useful biomarker in the development of cholesterol lowering drugs, in human plasma and urine. A hepta-deuterated analog of mevalonic acid was used as the internal standard. For both methods, calibration standards were prepared in water, instead of human plasma and urine, due to unacceptably high levels of endogenous mevalonic acid. The lower quality control (QC) samples were prepared in water while the higher QC samples were prepared in the biological matrices. For the isolation/purification of mevalonic acid from the plasma and urine matrices, the samples were first acidified to convert the acid analyte into its lactone form. For the plasma samples, the lactone analyte was retained on and then eluted off a polymeric solid-phase extraction (SPE) sorbent. For the urine method, the sample containing the lactone analyte was passed through a C-18 SPE column, which did not retain the analyte, with the subsequent analyte retention on and then elution off a polymeric SPE sorbent. Chromatographic separation was achieved isocratically on a polar-endcapped C-18 analytical column with a water/methanol mobile phase containing 0.5 mM formic acid. Detection was by negative-ion electrospray tandem mass spectrometry. The standard curve range was 0.500-20.0 ng/mL for the plasma method and 25.0-1,000 ng/mL for the urine method. Excellent accuracy and precision were obtained for both methods at all concentration levels tested. It was interesting to note that for certain batches of urine, when a larger sample volume was used for analysis, a high degree of matrix effect was observed which resulted not only in the attenuation of the absolute response, but also in a change of analyte/internal standard response ratio. This demonstrated that, under certain conditions, the use of a stable isotope analog internal standard does not, contrary to conventional thinking, guarantee the constancy of the analyte/internal response ratio, which is a prerequisite for a rugged bioanalytical method. On the other hand, under conditions where the sample matrix does not have such a deleterious effect, we have found that a stable isotope analog could serve as a surrogate (substitute) analyte. Thus, we have shown that using calibration standards prepared by spiking plasma with tri-deuterated or tetra-deuterated mevalonic acid, instead of mevalonic acid itself (the analyte), plasma QC samples that contain mevalonic acid can be successfully analyzed for the accurate and precise quantitation of mevalonic acid. The use of a surrogate analyte provides the opportunity to gauge the daily performance of the method for the low concentration levels prepared in the biological matrix, which otherwise is not achievable because of the endogenous concentrations of the analyte in the biological matrices.     

Nanostructured metal particlemodified electrodes for electrocatalytic and sensor applications

Nanotechnology has become one of the most exciting frontier fields in analytical chemistry. The huge interest in nanomaterials, for example in chemical sensors and catalysis, is driven by their many desirable properties. Although metal is a poor catalyst in bulk form, nanometre-sized particles can exhibit excellent catalytic activity due to their relative high surface area-to-volume ratio and their interface-dominated properties, which significantly differ from those of the bulk material. The integration of metal nanoparticles into thin film of permselective membrane is particularly important for various applications, for example in biological sensing and in electrocatalysis. We have already established different techniques to design permselective membrane-coated chemically modified electrodes with incorporated redox molecules for electrocatalytic, electrochromic and sensor applications. Recently, we have prepared nanostructured platinum and copper (represented M_nano, M = Pt and Cu) modified GC/Nafion electrodes (GC/Nf/M_nano) and characterized by using AFM, XPS, XRD and electrochemical techniques. The nanostructured M_nano modified electrodes were utilized for efficient electrocatalytic selective oxidation of neurotransmitter molecules in the presence of interfering species such as ascorbic acid (AA) and uric acid (UA). It has been also shown that the modified electrodes could be used as sensors for the detection of submicromolar concentrations of biomolecules with practical applications to real samples such as blood plasma and dopamine hydrochloride injection solution. The GC/Cu_nano electrode has been used for catalytic reduction of oxygen.     

Parameters important in fabricating enzyme electrodes using self-assembled monolayers of alkanethiols.

The fabrication of enzyme electrodes using self-assembled monolayers (SAMs) has attracted considerable interest because of the spatial control over the enzyme immobilization. A model system of glucose oxidase covalently bound to a gold electrode modified with a SAM of 3-mercaptopropionic acid was investigated with regard to the effect of fabrication variables such as the surface topography of the underlying gold electrode, the conditions during covalent attachment of the enzyme and the buffer used. The resultant monolayer enzyme electrodes have excellent sensitivity and dynamic range which can easily be adjusted by controlling the amount of enzyme immobilized. The major drawback of such electrodes is the response which is limited by the kinetics of the enzyme rather than mass transport of substrates. Approaches to bringing such enzyme electrodes into the mass transport limiting regime by exploiting direct electron transfer between the enzyme and the electrode are outlined.     

Automated analysis of fluvoxamine in rat plasma using a column-switching system and ion-pair high-performance liquid chromatography

We have established a robust, fully automated analytical method for the analysis of fluvoxamine in rat plasma using a column-switching ion-pair high-performance chromatography system. The plasma sample was injected onto a precolumn packed with Shim-pack MAYI-ODS (50 microm), where the drug was automatically purified and enriched by on-line solid-phase extraction. After elution of the plasma proteins, the analyte was back-flushed from the precolumn and then separated isocratically on a reversed-phase C18 column (L-column ODS) with a mobile phase (acetonitrile-0.1% phosphoric acid, 36:64, v/v) containing 2 mM sodium 1-octanesulfonate. The analyte was monitored by a UV detector at a wavelength of 254 nm. The calibration line for fluvoxamine showed good linearity in the range of 5-5000 ng/mL (r > 0.999) with the limit of quantification of 5 ng/mL (RSD = 6.51%). Accuracy ranged from -2.94 to 4.82%, and the within- and between-day precision of the assay was better than 8% across the calibration range. The analytical sensitivity and accuracy of this assay is suitable for characterization of the pharmacokinetics of orally-administered fluvoxamine in rats.