A selective catalytic voltammetric determination of vitamin C in pharmaceutical preparations and complex matrices of fresh fruit juices

A simple, selective and precise voltammetric method for the determination of ascorbic acid in pharmaceutical preparations and fresh fruit juices-complex matrices containing various reducing compounds-is described. The method is based on the electrocatalytic oxidation of ascorbic acid in homogeneous solution using electrogenerated ferriciniumcarboxylic acid as mediator. The pH and mediator concentration affecting the performance of the electrocatalytic oxidation of the analyte were optimized. The method was applied to determine vitamin C in deeply coloured, viscous and turbid fruit juice samples with ascorbic acid contents ranging from 15-45 mg per 100 ml, without further dilution, concentration or other pre-treatment of the samples. The amount of mediator used varied depending on the ascorbic acid concentration in the samples. The method was also used for pharmaceutical analysis using a calibration graph. For fruit juice samples the standard addition technique was adopted to prevent the matrix affecting the accuracy of the determination. The relative standard deviation for the analysis of vitamin C in fruit juices ranged from 1.5-5%. The reliability of the method was established by parallel determination against the official methods.     

Measurement of ascorbic acid and dehydroascorbic acid in gastric juice by HPLC

New methods are presented for measuring total vitamin C and the ascorbic acid/dehydroascorbic acid ratio in gastric juice. Extracts are prepared from a gastric juice which are suitable for direct injection onto a Waters Nova-pak C18 Radial-pak cartridge for high performance liquid chromatography (HPLC) using ultraviolet absorbance at 270 nm for detection. Both enable removal of interfering mucus and mucopolysaccharide breakdown products in a novel way. The first uses mini-columns of Sephadex G-50, run in acidic conditions to remove large molecular weight material while maintaining the ascorbic acid/dehydroascorbic acid ratio as it was in the fresh sample. Addition of dithiothreitol converts the dehydroascorbic acid quantitatively to ascorbic acid, thus enabling measurement of both components. The second method converts all the dehydroascorbic acid to ascorbic acid at the outset. A perchloric acid extract is neutralized and passed through a Sep-Pak C18. A new internal standard, reductic acid, is introduced for ascorbic acid analysis which behaves identically on Sep-Pak C18. Samples are analysed by ion-pair chromatography using 0.02 M NH4H2PO4 buffer (pH 7.1): methanol (80:20 v/v) containing 0.62 g/L tetrapentylammonium bromide. The detection limit was 1 ng ascorbic acid, and chromatography was completed in 5 min. The values obtained by the two independent HPLC methods were in good agreement with each other and with those obtained by the 2,4-dinitrophenylhydrazine colorimetric method.     

Determination of glutathione, glutathione disulphide, ascorbic acid and dehydroascorbic acid in tissues by reversed-phase liquid chromatography with electrochemical detection

Summary High-performance liquid chromatography with electrochemical detection was applied to the estimation of glutathione, glutathione disulphide, ascorbic acid and dehydroascorbic acid in various tissues of man, animal, and plant. The simultaneous determination of glutathione and ascorbic acid in tissues was done by a coulometric method. Separation of glutathione and ascorbic acid and unequivocal substance identifications were performed on a 100×4.6 mm RP-18 Spheri 5 column. As mobile phase 0.015 mol/l o-phosphoric acid, pH 2.3 was used. Retention time of ascorbic acid was 5.0 min and of glutathione 10.0 min. Dehydroascorbic acid was determined after reduction to ascorbic acid with dithiothreitol. Glutathione disulphide was reduced at pH 7.5 by -nicotinamide-dinucleotide phosphate and glutathione reductase, EC 1.6.4.2., to regenerate glutathione. To exclude interfering substances, several other compounds present in tissues and foods were investigated. This coulometric method is highly sensitive, specific and simple. Very low concentrations of ascorbic acid, glutathione, dehydroascorbic acid, and glutathione disulphide (<500 pg/injection) could be analysed using this HPLC-ECD method.(on leave to Mexico)     

Routine analysis of ascorbic acid in citrus juice using capillary electrophoresis.

A procedure to monitor citrus juice samples was established to quantitate vitamin C by capillary electrophoresis using a previously developed method. Dilution and filtration were the only preparation requirements and separation was achieved with an uncoated capillary using a 35mM sodium borate buffer (pH 9.3) containing 5% (v/v) acetonitrile at 21 kV and 23 degrees C. Detection was performed by high speed scanning between 200 and 360 nm. From the multiwave length scan, the electropherogram at 270 nm was extracted and used to quantitate ascorbic acid. The ascorbic acid concentration was calculated with an internal standard method, with ferulic acid as internal standard. The level of ascorbic acid during analysis was stabilized with ethylenediaminetetraacetic acid and dithiothreitol was used to reduce dehydroascorbic acid to ascorbic acid to estimate the total vitamin C level. Results were similar to those obtained by liquid chromatography and the method is now used to determine routinely the level of ascorbic acid in citrus juices.     

Selective spectrophotometric determination of ascorbic acid in drugs and foods

A new analytical method was developed for the determination of ascorbic acid. The method is based on the reaction of ascorbic acid with 4-chloro-7-nitrobenzofurazane (NBD-Cl) in the presence of 0.2M sodium hydroxide, where a bluish green colour (lambda(max) 582 nm) is developed after dilution with 50% (v/v) aqueous acetone solution. Beer's law was obeyed in a concentration range of 5-20 microg ascorbic acid/ml with a good correlation coefficient (r = 0.9990). The method was found to be highly specific for the determination of ascorbic acid in the presence of dehydro-ascorbic acid, all other vitamins and minerals possibly present in multivitamin preparations, rutin, salicylamide, acetyl salicylic acid, paracetamol, caffeine, phenylephrine hydrochloride and dipyrone. Moreover, the proposed procedure was also successfully applied for the determination of ascorbic acid in some canned and fresh fruit juices, some vegetables and infant milk products without interference from coloured and other substances present in the plant extracts.     

Spectrophotometric determination of ascorbic acid in pharmaceutical preparations and fruit juices

Conditions were established for the determination of ascorbic acid using phsophovanadotungstic acid as reagent. The method was applied to the determination of ascorbic acid in pure form, pharmaceutical preparations and fruit juices. The method is sensitive (2-24 micrograms ml-1 of ascorbic acid) and rapid and tolerates the presence of common ingredients usually found in fruit juices. The results obtained with the proposed method showed good agreement with those given by the standard method.     

Switch-on fluorescence sensor for ascorbic acid detection based on MoS2 quantum dots-MnO2 nanosheets system and its application in fruit samples

In this work,molybdenum disulfide quantum dots(MoS_2 QDs) were firstly prepared by hydrothermal method using sodium molybdate and glutathione as precursors,and applied in ascorbic acid detection.When joining MnO_2 nanosheets into MoS_2 QDs solution,they produced an obvious fluorescence quenching,which should be due to inner filter effect(IFE).Meanwhile,the fluorescent probe was formed,Interestingly,we found that this quenching phenomenon disappeared with the addition of ascorbic acid,In other words,the fluorescence gradually restored.This recovery phenomenon is mainly due to the reduction effect of ascorbic acid for MnO_2 nanosheets.Under the optimum conditions,the limit of detection(LOD) of 39 nmol/L for ascorbic acid was achieved with a linear range of 0.33-5.00 μmol/L.The repeatability was better than 5.0% for ascorbic acid in both standard and fruit samples(n = 3).Moreover,the as-fabricated fluorescent sensing system was successfully employed to detect the ascorbic acid levels in hawthorn and jujube with satisfactory results.     

Kinetic spectrophotometric determination of ascorbic acid by reduction of toluidine blue

A simple kinetic method is described for the determination of ascorbic acid. The procedure is based on the reduction of toluidine blue with ascorbic acid. The rate of reaction is followed by measuring the decrease in absorbance of toluidine blue (lambda(max) = 600 nm) as a result of its decolorization upon reduction by ascorbic acid. Ascorbic acid in the range of 3-35 microg/ml was determined using slope and fixed time methods of analysis, while the variable time method allowed the determination of 5-50 microg/ml of ascorbic acid. The percent relative standard deviation of the method varied from 0.78 to 1.32% depending on the kinetic method used. The high sensitivity of the method also allows determination of low levels of ascorbic acid in some fruits and vegetables such as dew melon, water melon, parsley and coriander.     

Simultaneous dual wavelength spectrophotometric determination of citric and ascorbic acids using an artificial neural network

In this study, a very simple spectrophotometric method for the simultaneous determination of citric and ascorbic acid based on the reaction of these acids with a copper(II)-ammonia complex is presented. The Cu²⁺-NH₃ complex (with λ_max = 600 nm) was decomposed by citrate ion and formed a Cu²⁺-citrate complex (with λ_max = 740 nm). On the other hand, during the reaction of ascorbic acid with copper(II)-ammonia complex, ascorbic acid is oxidized and the copper(II)-ammonia complex is reduced to the copper(I)-ammonia complex and the absorbance decreases to 600 nm. Although there is a spectral overlap between the absorbance spectra of complexes Cu²⁺-NH₃ and Cu²⁺-citrate, they have been simultaneously determined using an artificial neural network (ANN). The absorbances at 600 and 740 nm were used as the input layer. The ANN architectures were different for citric and ascorbic acid. The output of the citric acid ANN architecture was used as an input node for the ascorbic acid ANN architecture. This modification improves the capability of the ascorbic acid ANN model for the prediction of ascorbic acid concentrations. The dynamic ranges for citric and ascorbic acid were 1.0–125.0 and 1.0–35.0 mM, respectively. Finally, the proposed method was successfully applied to the determination of citric and ascorbic acids in vitamin C tablets and some powdered drink mixes. The text was submitted by the authors in English.     

Determination of ascorbic acid in human plasma with a view to stability using HPLC with UV detection

A method for the measurement of ascorbic acid using HPLC with UV detection and investigation into the protein precipitation techniques with regard to stability and recovery are described. The effectiveness of various protein precipitants was tested. Stability of ascorbic acid samples for analysis was investigated over 10 h. Ascorbic acid samples extracted with metaphosphoric acid were stable on a cooled autosampler (4 degrees C) for at least 10 h (with a decline of 1.8% for ascorbic acid solution and 2.8% for plasma). Perchloric acid as protein precipitant for ascorbic acid was unsuitable (with a decline of 36.0% for ascorbic acid solution and 7.3% for plasma). Analytical performance of this method is satisfactory. The intra- and interassay coefficients of variation were 2.1% (n = 10) and 5.8% (n = 12), respectively. The calibration curve was linear with the tested range of 2.0-250.0 micromol/L. The recovery was 96.1% with CV = 4.8% (n = 6) and the LOD was 3 micromol/L. The preliminary reference ranges of ascorbic acid in a group of blood donors are 50.8 +/- 22.4 micromol/L. This assay is a highly sensitive and reproducible HPLC method for the determination of ascorbic acid in human plasma.     

聚乙烯二茂铁修饰电极测定抗坏血酸

化学修饰电极是近几年来活跃于电化学及电分析化学领域的一个新的研究课题,成功地应用于表面配位化学、光电转化、表面催化反应及痕量元素检测等方面。具有电活性的聚乙烯二茂铁修饰电极对抗坏血酸的氧化有催化作用。     

Flow injection amperometric detection of ascorbic acid using a Prussian Blue film-modified electrode

The PB film-modified electrode was used as an amperometric detector for flow injection analysis of ascorbic acid. The modified electrode detector showed good sensitivity, stability and reproducibility. The calibration curve for ascorbic acid was linear over the concentration range from 5.0x10(-6) to 1.0x10(-3) mol l(-1) with a slope of 19.9 mA mol(-1) per litre and a correlation coefficient of 0.999. The detection limit of this method was 2.49x10(-6) mol l(-1). The relative standard deviation of six replicate injections of 2.5x10(-4) mol l(-1) ascorbic acid was 2.5%. The results obtained for ascorbic acid determination in pharmaceutical products are in good agreement with those obtained by using the procedure involving the reaction between triiodide and ascorbic acid.     

Determination of ascorbic acid in pharmaceuticals and urine by reverse flow injection.

Two reverse flow injection (FI) methods, using spectrophotometric detection, are proposed for the determination of ascorbic acid. Both methods are based on its reaction with the ethylenediaminetetraacetic acid-CoIII complex in a medium of 5% diethylamine. In the first method, using the peak-height FI technique, ascorbic acid is determined over the range from 2 x 10(-4) to 5 x 10(-3) mol dm-3 and in the second, using the peak-width FI method, the working range is extended (2 x 10(-3)-5 x 10(-2) mol dm-3). Both FI methods were applied to the determination of ascorbic acid in pharmaceuticals while the peak-height FI technique was also used to determine ascorbic acid in urine.     

方波极谱法间接测定抗坏血酸

利用Ｃｕ２＋与抗坏血酸（Ｖｃ）的氧化还原反应,建立了一种间接测定Ｖｃ的方波极谱法。在ＮＨ３－ＮＨ４Ｃｌ缓冲溶液中,Ｃｕ２＋在约－０．２４ Ｖ（ｖｓ．ＳＣＥ）处出现一个可逆的 Ｃｕ２＋－Ｃｕ＋的还原峰,加入Ｖｃ可将Ｃｕ２＋还原,根据 Ｃｕ２＋还原峰峰高的降低量间接测定Ｖｃ,线性范围为２．０×１０－６～ ３．０×１０－４ｍｏｌ／Ｌ,该法还应用于实际样品Ｖｃ片剂的含量测定,ＲＳＤ为２．９％。     

Simultaneous determination of ascorbic acid and acetylsalicylic acid in pharmaceutical formulations

A direct, simple, and practical first-derivative spectrophotometric method is described for simultaneous determination of ascorbic acid and acetylsalicylic acid. The effects of the solvent, excipients, and spectral variables on the analytical signal were investigated. The drugs were determined simultaneously with a 0.01 M methanolic hydrochloric acid solution as the solvent, and the signals were evaluated directly by using the zero-crossing method at 245.0 and 256.0 nm for acetylsalicylic acid and ascorbic acid, respectively. The method allows the simultaneous determinations of acetylsalicylic acid and ascorbic acid in the ranges of 6.6 x 10(-6) to 1.5 x 10(-4)M and 3.4 x 10(-6) to 2.0 x 10(-4)M, respectively, with standard deviation of <2.0%. The proposed method was applied to determinations of these drugs in tablets.     

二芳基汞的合成

将氯化重氮苯的溶液加入氯化汞、维生素C、丙酮和少量氯化铜的混合物中,得到80％的氯化苯汞;如在反应混合物中补加维生素C、氯化铜和浓氨水,则得到二苯汞。用同样的方法制备了其他的二芳基汞,并讨论了反应可能的机理。     

Kinetic spectrofluorimetric determination of trace ascorbic acid based on its inhibition on the oxidation of pyronine Y by nitrite

A highly sensitive spectrofluorimetric method is proposed for the determination of trace amount of ascorbic acid using a new indication. The method is based on the inhibition of ascorbic acid on the oxidation of pyronine Y (PRY) by nitrite. The detection limit for ascorbic acid is 0.012 microg ml(-1), the linear range of the determination is 0.02-0.36 microg ml(-1). Analytical parameters, such as reagent concentration, pH, reaction temperature and time, were optimized. The relative standard deviations of eleven replication determinations of 0.12 and 0.24 microg ml(-1) ascorbic acid were 1.4 and 0.72%, respectively. This method has been used to determine ascorbic acid in pharmaceuticals, vegetables, fruits and soft drink with satisfactory results.     

New tetrazolium method for phosphatase assay using ascorbic acid 2-phosphate as a substrate

A new method to assay alkaline and acid phosphatases assay using ascorbic acid 2-phosphate (AsA-P) and ditetrazolium salt nitroblue tetrazolium chloride (NBT) was developed. AsA-P is hydrolyzed in the presence of phosphatase to yield ascorbic acid. In turn, the ascorbic acid reduces NBT directly or indirectly, opening the tetrazole ring to produce an insoluble formazan as a colored precipitate. The proposed method for alkaline phosphatase was compared with a conventional method in which 5-bromo-4-chloro-3-indolyl phosphate (BCIP) is used in combination with NBT in the dot blots of a dilution series of β-lactoglobulin. AsA-P reduced NBT more effectively than BCIP in the presence of alkaline phosphatase. AsA-P could be also used as the chromogenic substrate for an acid phosphatase assay in the presence of phenazinium methylsulfate and NBT.     

Determination of ascorbic acid in elemental diet by high-performance liquid chromatography with electrochemical detection.

Determination of ascorbic acid in a multi-component elemental diet was performed by high-performance liquid chromatography with electrochemical detection. This method is suitable for the routine determination of ascorbic acid in elemental diet because it is simple, rapid, sensitive, highly selective and reproducible. The calibration graph of ascorbic acid was linear in the range 0-1.0 micrograms. The recovery of ascorbic acid was over 95% by the standard addition method. There was good agreement between the concentrations of ascorbic acid stated and found.     

Genetic‐Algorithm‐Based Wavelength Selection in Multicomponent Spectrophotometric Determination by PLS: Application on Ascorbic Acid and Uric Acid Mixture

Genetic algorithm (GA) is a suitable method for selecting wavelengths for partial least squares (PLS) calibration of mixtures with almost identical spectra without loss of prediction capacity using the spectrophotometric method. In this study, the concentration model is based on absorption spectra in the range of 200‐320 nm for 25 different mixtures of ascorbic acid (AA) and uric acid (UA). The calibration curve was linear over the concentration range of 1‐15 and 2‐16 μg mL^?1 for ascorbic acid and uric acid, respectively. The root mean square deviation (RMSD) for ascorbic acid and uric acid with GA and without GA were 0.3071 and 0.3006, 0.3971 and 0.7063, respectively. The proposed method was successfully applied to the simultaneous determination of both analytes in human serum and urine samples.