A new colorimetric lactate biosensor based on CUPRAC reagent using binary enzyme (lactate-pyruvate oxidases)-immobilized silanized magnetite nanoparticles.

A novel optical lactate biosensor is presented that utilizes a colorimetric interaction between H2O2 liberated by a binary enzymatic reaction and bis(neocuproine)copper(II) complex ([Cu(Nc)2]2+) known as CUPRAC (cupric reducing antioxidant capacity) reagent. In the first step, lactate oxidase (LOx) and pyruvate oxidase (POx) were separately immobilized on silanized magnetite nanoparticles (SiO2@Fe3O4 NPs), and thus, 2 mol of H2O2 was released per 1 mol of the substrate due to a sequential enzymatic reaction of the mixture of LOx-SiO2@Fe3O4 and POx-SiO2@Fe3O4 NPs with lactate and pyruvate, respectively. In the second step, the absorbance at 450 nm of the yellow-orange [Cu(Nc)2]+ complex formed through the color reaction of enzymatically produced H2O2 with [Cu(Nc)2]2+ was recorded. The results indicate that the developed colorimetric binary enzymatic biosensor exhibits a broad linear range of response between 0.5 and 50.0 µM for lactate under optimal conditions with a detection limit of 0.17 µM. The fabricated biosensor did not respond to other saccharides, while the positive interferences of certain reducing compounds such as dopamine, ascorbic acid, and uric acid were minimized through their oxidative removal with a pre-oxidant (NaBiO3) before enzymatic and colorimetric reactions. The fabricated optical biosensor was applied to various samples such as artificial blood, artificial/real sweat, and cow milk. The high recovery values (close to 100%) achieved for lactate-spiked samples indicate an acceptable accuracy of this colorimetric biosensor in the determination of lactate in real samples. Due to the increase in H2O2 production with the bienzymatic lactate sensor, the proposed method displays double-fold sensitivity relative to monoenzymatic biosensors and involves a neat color reaction with cupric-neocuproine having a clear stoichiometry as opposed to the rather indefinite stoichiometry of analogous redox dye methods.

Voltammetric determination of Molnupiravir used in treatment of the COVID-19 at magnetite nanoparticle modified carbon paste electrode.

To reduce the progression of the viral process in patients infected with COVID-19, new treatments and drug active substances are needed. One of these drugs is Molnupiravir (MNP) which has a direct antiviral effect and has also proven to be highly effective in reducing the azopharyngeal SARS-CoV-2 infectious virus and viral RNA. Due to the importance and frequent use of this drug in the treatment of COVID-19, its accurate, quick, and cheap detection in pharmaceutical or biological samples is crucial. In this work, electrochemical behavior and sensitive voltammetric determination of MNP are described using a magnetite nanoparticle modified carbon paste electrode (Fe3O4@CPE) for the first time. Fe3O4 nanoparticles (NPs) were characterized by recording their transmission electron microscopy (TEM) images, energy dispersive X-ray (EDX), and X-ray diffraction (XRD) spectra. Cyclic voltammetric measurements showed that MNP was irreversibly oxidized at Fe3O4@CPE at 760 mV in pH 2.0 Britton Robinson buffer solution (BRBS). The peak current of MNP was increased approximately threefold at Fe3O4@CPE compared to bare CPE due to a good electrocatalytic efficiency of Fe3O4 NPs. According to differential pulse voltammetric studies, the fabricated electrode exhibited a linear range (LR) between 0.25 and 750 µM with sensitivity and limit of detection (LOD) of 4591.0 µA mM-1 cm-2 and 0.05 µM, respectively. On the other hand, although lower sensitivity (327.3 µA mM-1 cm-2) was obtained from CV compared to DPV, a wider linear calibration curve between 0.25 and 1500 µM was obtained in CV. Studies performed in tablet samples confirmed that the Fe3O4@CPE exhibits high applicability for selective and accurate voltammetric determination of MNP in real samples.

A novel flow injection amperometric method for sensitive determination of total antioxidant capacity at cupric-neocuproine complex modified MWCNT glassy carbon electrode.

A novel amperometric method is presented for the determination of total antioxidant capacity in flow injection analysis (FIA) system using copper(II)-neocuproine complex modified on Nafion-functionalized multi-walled carbon nanotube-glassy carbon electrode ([Cu(Ncp)22+]/Nf@f-MWCNT/GCE). Cyclic voltammetric studies showed that the modified electrode exhibits a very well-formed reversible redox couple for Cu(II)-/Cu(I)-complex. In addition, the [Cu(Ncp)22+]/[Cu(Ncp)2+] redox pair shows very good electrocatalytic activity towards the oxidation of polyphenolic compounds (PPhCs) such as trolox, catechin, and quercetin due to the enhancement of the anodic peak current of the redox couple in the presence of these analytes. This electrocatalytic oxidation current at the [Cu(Ncp)22+]/Nf@f-MWCNT/GCE was used for flow injection (FI) amperometric determination of PPhCs. FI amperometric-time curves recorded under optimized conditions (applied potential: + 0.6 V vs. Ag/AgCl/KCl(0.10 M), flow rate: 2 mL/min) showed that the proposed electrode had a wide linear range (LR) with a very low detection limit (LOD) for PPhCs. LR and LOD were 0.5-800 and 0.2 µM for trolox, respectively and 0.50-250 and 0.14 µM, respectively, for both quercetin and catechin. This sensitive method was successfully applied to the amperometric measurement of total antioxidant capacity (TAC) of some herbal teas, giving compatible results with the spectrophotometric CUPRAC method. The proposed method gave higher rank to fast-reacting antioxidants; it was equally precise but had a wider linear range and lower LOD than the spectrophotometric CUPRAC assay (e.g., LOD for ascorbic acid and gallic acid were 0.07 and 0.08 µM, respectively), and similar electroanalytical methods using the CUPRAC reagent.

Sensitive Voltammetric Determination of Natural Flavonoid Quercetin on a Disposable Graphite Lead.

In this paper, a pencil graphite electrode was pretreated using chronoamperometry technique in phosphate buffer solution (pH=7.0) for sensitive determination of quercetin. Oxidation of quercetin was investigated using pretreated pencil graphite electrode and anodic stripping differential pulse voltammetry. Under optimal conditions, the anodic current of quercetin exhibited linear response to its concentration in the range from 0.001 to 1.5 µmol/L with the limit of detection of 0.3·10-3 µmol/L. The proposed method was successfully applied for the determination of quercetin in cranberry and blackcurrant juices with recovery rate from 93.2 to 94.7%. Solid-phase extraction was found to be necessary prior to voltammetric determination of quercetin in fruit juice samples using pretreated pencil graphite electrode.

A Self-Powered Enzymatic Glucose Sensor Utilizing Bimetallic Nanoparticle Composites Modified Pencil Graphite Electrodes as Cathode.

Enzymatic biofuel cells (EBFC) are promising sources of green energy owing to the benefits of using renewable biofuels, eco-friendly biocatalysts, and moderate operating conditions. In this study, a simple and effective EBFC was presented using an enzymatic composite material-based anode and a nonenzymatic bimetallic nanoparticle-based cathode respectively. The anode was constructed from a glassy carbon electrode (GCE) modified with a multi-walled carbon nanotube (MWCNT) and ferrocene (Fc) as a conductive layer coupled with the enzyme glucose oxidase (GOx) as a sensitive detection layer for glucose. A chitosan layer was also applied to the electrode as a protective layer to complete the composite anode. Chronoamperometry (CA) results show that the MWCNT-Fc-GOx/GCE electrode has a linear relationship between current and glucose concentration, which varied from 1 to 10 mM. The LOD and LOQ were calculated for anode as 0.26 mM and 0.87 mM glucose, respectively. Also the sensitivity of the proposed sensor was calculated as 25.71 µ A/mM. Moreover, the studies of some potential interferants show that there is no significant interference for anode in the determination of glucose except ascorbic acid (AA), uric acid (UA), and dopamine (DA). On the other hand, the cathode consisted of a disposable pencil graphite electrode (PGE) modified with platinum-palladium bimetallic nanoparticles (Nps) which exhibit excellent conductivity and electron transfer rate for the oxygen reduction reaction (ORR). The constructed EBFC was optimized and characterized using various electroanalytical techniques. The EBFC consisting of MWCNT-Fc-GOx/GCE anode and Pt-PdNps/PGE cathode exhibits an open circuit potential of 285.0 mV and a maximum power density of 32.25 µW cm-2 under optimized conditions. The results show that the proposed EBFC consisting of an enzymatic composite-based anode and bimetallic nanozyme-based cathode is a unique design and a promising candidate for detecting glucose while harvesting power from glucose-containing natural or artificial fluids.

The application of multi-walled carbon nanotubes modified pencil graphite electrode for voltammetric determination of favipiravir used in COVID-19 treatment.

This study describes the first application of an improved procedure on a pencil graphite electrode decorated with functionalized multi-walled carbon nanotubes (f-MWCNTs/PGE) for the determination of the COVID-19 antiviral drug, favipiravir (FVP). The electrochemical behavior of FVP at f-MWCNTs/PGE was examined by cyclic voltammetry and differential pulse voltammetry (DPV) methods, and it was noted that the voltammetric response significantly increased with the modification of f -MWCNTs to the surface. The linear range and limit of detection from DPV studies were determined as 1-1500 µM and 0.27 µM, respectively. In addition, the selectivity of the method was tested toward potential interferences, which can be present in pharmaceutical and biological samples, and it was found that f-MWCNTs/PGE showed high selectivity for the determination of FVP in the presence of probable interferences. The results with high accuracies and precisions from the obtained feasibility studies also revealed that the designed procedure can be used for accurate and selective voltammetric determination of FVP in real samples.

Fabrication of a Novel Optical Glucose Biosensor Using Copper(II) Neocuproine as a Chromogenic Oxidant and Glucose Dehydrogenase-Immobilized Magnetite Nanoparticles.

This study describes a novel optical glucose biosensor based on a colorimetric reaction between reduced nicotinamide adenine dinucleotide (NADH) and a copper(II) neocuproine complex ([Cu(Nc)2]2+) as a chromogenic oxidant. An enzymatic reaction takes place between glucose and glucose dehydrogenase (GDH)-chitosan (CS) immobilized on silanized magnetite nanoparticles (CS@SiO2@Fe3O4) in the presence of coenzyme NAD+. The oxidation of glucose to gluconolactone via the immobilized enzyme is coupled with the reduction of NAD+ to NADH at the same time. After the separation of GDH-immobilized SiO2@Fe3O4 with a magnet, the enzymatically produced NADH chemically reduces the chromogenic oxidant cupric neocuproine to the cuprous chelate. Thus, the glucose biosensor is fabricated based on the measurement of the absorbance of the formed yellow-orange complex ([Cu(Nc)2]+) at 450 nm. The obtained results show that the colorimetric biosensor has a wide linear response range for glucose, between 1.0 and 150.0 µM under optimized conditions. The limit of detection and limit of quantification were found to be 0.31 and 1.02 µM, respectively. The selectivity properties of the fabricated biosensor were tested with various interfering species. This biosensor was applied to various samples, and the obtained results suggest that the fabricated optical biosensor can be successfully used for the selective and sensitive determination of glucose in real samples.

Voltammetric Determination of Favipiravir Used as an Antiviral Drug for the Treatment of Covid-19 at Pencil Graphite Electrode.

This work describes the sensitive voltammetric determination of favipiravir (FAV) based on its reduction for the first time with a low-cost and disposable pencil graphite electrode (PGE). In addition, the determination of FAV was also performed based on its oxidation. Differential pulse (DP) voltammograms recorded in 0.5 M H2SO4 for the reduction of FAV show that peak currents increase linearly in the range of 1.0 to 600.0 µM with a limit of detection of 0.35 µM. The acceptable recovery values (98.9-106.0 %) obtained from a pharmaceutical tablet, real human urine, and artificial blood serum samples spiked with FAV confirm the high accuracy of the proposed method.

Adsorption and deposition-assisted anodic stripping voltammetry for determination of antimony(III) in presence of hematoxylin on glassy carbon electrode.

A validated simple, reliable and sensitive adsorptive anodic stripping voltammetric procedure is suggested for the accurate determination of antimony(III) on a glassy carbon electrode (GCE) in the presence of hematoxylin (HT), used as a chemical receptor in antimony analysis for the first time. Under optimized conditions, the plot of stripping peak current versus Sb(III) concentration showed two dynamic linear ranges, 1.0×10-4 - 0.01µm (R2 =0.9986) and 0.01 - 1.0µm (R2 =0.9973) with a detection limit of 9.5×10-11molL-1 (11.57ngL-1). The RSD values of intra-day precision and inter-day precision (n =5) for 5.0×10-7molL-1 Sb(III) were calculated as 1.63% and 2.27%, respectively, indicating that the proposed method has a good precision. Recoveries for Sb(III) in water samples were found to be between 97% and 102% for the 5.0×10-7molL-1 Sb(III), proving the reliability and accuracy of the method proposed.

Photoamperometric flow injection analysis of glucose based on dehydrogenase modified quantum dots-carbon nanotube nanocomposite electrode.

In this work, a core-shell quantum dot (QD, ZnS-CdS) was electrodeposited onto multiwalled carbon nanotube modified glassy carbon electrode (ZnS-CdS/MWCNT/GCE) and following glucose dehydrogenase (GDH) was immobilized onto QD modified electrode. The proposed electrode (GDH/ZnS-CdS/MWCNT/GCE) was effectively used for the photoelectrochemical biosensing of glucose in flow injection analysis (FIA) system using a home-made flow cell. Results from cyclic voltammetric and FI amperometric measurements have revealed that GDH/ZnS-CdS/MWCNT/GCE is capable of signaling photoelectrocatalytic activity toward NADH when the surface of enzyme modified electrode was irradiated with a light source (250W Halogen lamp). Thus, photoelectrochemical biosensing of glucose was monitored by recording current-time curve of enzymatically produced NADH at optimized conditions. The biosensor response was found linear over the range 0.010-2.0mM glucose with detection limits of 6.0 and 4.0µM for amperometric and photoamperometric methods, respectively. The relative standard deviations (n=5) for 0.5mM glucose were 5.8% and 3.8% for photoamperometric and amperometric results, respectively. The photoelectrochemical biosensor was successfully applied to the real samples. The results with this biosensor showed good selectivity, repeatability and sensitivity for monitoring glucose in amperometric and photoamperometric FIA studies.

Biosensing of glucose in flow injection analysis system based on glucose oxidase-quantum dot modified pencil graphite electrode.

A novel amperometric glucose biosensor was proposed in flow injection analysis (FIA) system using glucose oxidase (GOD) and Quantum dot (ZnS-CdS) modified Pencil Graphite Electrode (PGE). After ZnS-CdS film was electrochemically deposited onto PGE surface, GOD was immobilized on the surface of ZnS-CdS/PGE through crosslinking with chitosan (CT). A pair of well-defined reversible redox peak of GOD was observed at GOD/CT/ZnS-CdS/PGE based on enzyme electrode by direct electron transfer between the protein and electrode. Further, obtained GOD/CT/ZnS-CdS/PGE offers a disposable, low cost, selective and sensitive electrochemical biosensing of glucose in FIA system based on the decrease of the electrocatalytic response of the reduced form of GOD to dissolved oxygen. Under optimum conditions (flow rate, 1.3mL min(-1); transmission tubing length, 10cm; injection volume, 100µL; and constant applied potential, -500mV vs. Ag/AgCl), the proposed method displayed a linear response to glucose in the range of 0.01-1.0mM with detection limit of 3.0µM. The results obtained from this study would provide the basis for further development of the biosensing using PGE based FIA systems.

Differential pulse voltammetric determination of eugenol at a pencil graphite electrode.

In this study, the electrochemical behavior of eugenol, a widely used herbal drug, was investigated at a pencil graphite electrode (PGE). A low-cost, disposable, sensitive and selective electrochemical sensor is proposed for the determination of eugenol by recording its differential pulse voltammograms in Britton-Robinson buffer solution containing 0.1 M KCl with pH of 2.0 at the PGE. The PGE displayed a very good electrochemical behavior with significant enhancement of the peak current compared to a glassy carbon electrode. Under experimental conditions, the PGE had a linear response range from 0.3 µM to 50.0 µM eugenol with a detection limit of 0.085 µM (based on 3S(b)). Relative standard deviations of 2.4 and 4.8% were obtained for five successive determinations of 30.0 and 5.0 µM eugenol, respectively, which indicate acceptable repeatability. This voltammetric method was successfully applied for the direct determination of eugenol in real samples. The effect of various interfering compounds on the eugenol peak current was also studied.

Photocurrent generation from thylakoid membranes on osmium-redox-polymer-modified electrodes.

Thylakoid membranes (TMs) are uniquely suited for photosynthesis owing to their distinctive structure and composition. Substantial efforts have been directed towards use of isolated photosynthetic reaction centers (PRCs) for solar energy harvesting, however, few studies investigate the communication between whole TMs and electrode surfaces, due to their complex structure. Here we report on a promising approach to generate photosynthesis-derived bioelectricity upon illumination of TMs wired with an osmium-redox-polymer modified graphite electrode, and generate a photocurrent density of 42.4 µA cm(-2).

Electrocatalytic oxidation of NADH using a pencil graphite electrode modified with quercetin.

In the present study, the electrocatalytic oxidation of reduced ß nicotinamide adenine dinucleotide (NADH) was investigated using a pencil graphite electrode modified with quercetin (PGE/QH(2)). The PGE/QH(2) was prepared through two steps: (i) the pre-treatment of PGE at 1.40 V vs. Ag|AgCl|KCl((sat.)) in pH 7.0 phosphate buffer containing 0.1 M KCl for 60s and (ii) adsorption of QH(2) on the PGE via immersion of PGE into a 1.0mM QH(2) solution (in ethanol) for 60s. Cyclic voltammetric studies show that the peak potential of NADH oxidation shifts from +500 mV at bare PGE to +300 mV at PGE/QH(2). The electrocatalytic currents obtained from amperometric measurements at +300 mV vs. Ag|AgCl|KCl((sat.)) and in phosphate buffer solution at pH 7.0 containing 0.1M KCl were linearly related to the concentration of NADH. Linear calibration plots are obtained in the concentration range from 0.5 µM to 100 µM. The limit of detection was found to be 0.15 µM.

Flow injection amperometric detection of sulfide using a prussian blue modified glassy carbon electrode.

This study investigates a new approach for the electrocatalytic determination of sulfide in a flow injection analysis (FIA) system using a Prussian blue modified glassy carbon electrode (PB/GCE). The results from experiments show that PB/GCE significantly enhances the electrocatalytic activity towards sulfide oxidation. A homemade flow electrochemical cell was used to perform the electrocatalytic determination of sulfide in the FIA system. The currents obtained from amperometric measurements in the FIA system at optimum conditions (carrier solution, pH 8.0; Britton-Robinson buffer solution containing 0.1 M KCl; flow rate, 1.4 mL/min; transmission tubing length, 10 cm; injection volume, 100 µL; constant applied potential, +150 mV vs. Ag/AgCl/KClsat) were linearly correlated with the sulfide concentration. A calibration curve was obtained for sulfide concentrations in the range of 0.5 - 100 µM. The detection limit was found to be 0.3 µM for the amperometric method. The proposed method was successfully applied to wastewater sample. Finally, results from sulfide measurements by PB/GCE were in good agreement with those obtained from the spectrophotometric method.

Amperometric determination of sulfide based on its electrocatalytic oxidation at a pencil graphite electrode modified with quercetin.

This study describes a new approach for the investigation of electrocatalytic oxidation of sulfide using a pencil graphite electrode modified with quercetin (PGE/QH(2)). Adsorption procedure was used for the preparation of the modified electrodes. It was observed that PGE/QH(2) showed a significant electrocatalytic activity toward sulfide oxidation. Cyclic voltammetric studies show that the peak potential of sulfide shifts from +450 mV at bare PGE to +280 mV at PGE/QH(2). The electrocatalytic currents obtained from amperometric measurements at +300 mV vs. Ag/AgCl/KCl(sat) and at pH 8.0 BR buffer solution containing 0.1M NH(4)Cl were linearly related to the concentration of sulfide. The calibration graph consisted of two linear segments of 1-20 µM and 20-800 µM with a detection limit of 0.3 µM (based on 3s(b)). The proposed method was successfully applied to the determination of sulfide in waste waters and was compared with the spectrophotometric method.

Photoelectrocatalytic oxidation of NADH in a flow injection analysis system using a poly-hematoxylin modified glassy carbon electrode.

A stable electroactive thin film of poly-hematoxylin (poly-HT) was successfully prepared on a glassy carbon electrode (GCE) surface by recording successive cyclic voltammograms of 0.3 mM HT, in a phosphate buffer solution (pH 7.0) containing 0.1 M NaNO3, in the potential range of -0.5 to +2.0 V vs. Ag/AgCl. The deposition of HT on GCE surface can be explained through the electropolymerization process. This poly-HT modified electrode exhibited a good electrocatalytic activity towards the NADH oxidation in a phosphate buffer solution (pH 7.0), and led to a significant decrease in the overpotential by more than 320 mV compared with the bare GCE. In order to perform the photoelectrocatalytic determination of NADH in a flow injection analysis (FIA) system, a home-made flow electrochemical cell with a suitable transparent window for irradiation of the electrode surface was constructed. Flow rate of carrier solution, transmission tubing length, injection volume and applied potential for the amperometric and photoamperometric FIA studies were optimized as 1.3 mL min(-1), 10 cm, 100 µL and +300 mV vs. Ag/AgCl, respectively. The currents obtained from amperometric and photoamperometric measurements in FIA system at optimum conditions were linearly dependent on the NADH concentration and linear calibration curves were obtained in the range of 1.0×10(-7)-1.5×10(-4) M and in the range of 1.0×10(-7)-2.5×10(-4) M NADH, respectively. The relative standard deviation (RSD) of six replicate injections of 6.0×10(-5) M NADH was calculated as 2.2% and 4.3% for the amperometric and the photoamperometric method, respectively. The limit of detection was found to be 3.0×10(-8) M for the photoamperometric determination of NADH.

Determination of heavy metal pollution with environmental physicochemical parameters in waste water of Kocabas Stream (Biga, Canakkale, Turkey) by ICP-AES.

Waste water pollution of industrial areas can answer for the serious consequences of one of the most important environmental threats to the future. In this study, inductively coupled plasma-atomic emission spectrometry method (ICP-AES) is proposed to determine heavy metals (Pb, Cu, Cd, Cr, Zn, Al, Fe, Ni, Co, Mn) and major elements (Ca, Mg) in waste water of Kocabas Stream. The concentration of metals in the waste water samples taken from 9 different stations (St.) in Biga-Kocabas Stream in November 2004 (autumn period) were determined after simple pretreatment of samples by the proposed ICP-AES method. An analysis of a given sample is completed in about 15 min for ICP-AES the method. The results of heavy metals concentrations in waste water were found between 0.00001-77.69610 mg l(-1) by the ICP-AES technique. The concentrations of Pb, Cd, Cu, Zn, Cr, Al, Fe, Mn, Ni, Co, Mg and Ca 0.00001 (St.3,6,7) - 0.0087 mg l(-1) (St.9), 0.00001 (St.4-7) - 0.0020 mg l(-1) (St.8), 0.00001 (St.1,3-7,9) - 0.0041 mg l(-1) (St.2), 0.0620 (St.2) - 0.2080 mg l(-1) (St.3), 0.0082 (St.6) - 0.2290 mg l(-1) (St.8), 0.3580 (St.2) - 1.7400 mg l(-1) (St.3), 0.2240 (St.1) - 0.6790 mg l(-1) (St.3), 0.0080 (St.1) - 1.5840 mg l(-1) (St.3), 0.0170 (St.3) - 0.0640 mg l(-1) (St.2), 0.0010 (St.1,4,5,8) - 0.0080 mg l(-1) (St.3), 5.0640 (St.9) - 5.2140 mg l(-1) (St.1) and 43.3600 (St.2) - 77.6961 mg l(-1) (St.9), respectively. Also we measured environmental physicochemical parameters such as temperature, salinity, specific conductivity, total dissolved solid (TDS), pH, oxidation and reduction potential (ORP), and dissolved oxygen (DO) in the waste water at sampling stations.

Fluorimetric determination of ascorbic acid in vitamin C tablets using methylene blue.

In this study, a simple and sensitive fluorimetric method was described for the determination of Ascorbic Acid (AA). The procedure is based on the reaction between AA and Methylene Blue (MB). The fluorescence intensity of MB was measured at excitation and emission of 664 and 682 nm, respectively. MB concentration was decreased as a function of decreasing fluorescence intensity due to forming colorless form of MB (Leuco-MB) in the reaction between AA and MB. A linear relationship was obtained between the decreasing fluorescence intensity and the concentration of AA in the range of 3.0 x 10(-7)-6.0 x 10(-6) mol.l(-1). The detection limit was 2.5 x 10(-7) mol.l(-1). The proposed method was applied successfully for the determination of AA in Vitamin C tablets.