Enzymatic analysis by means of the air-gap electrode determination of urea in blood

The newly developed air-gap electrode has been used for enzymatic assay of urea in serum and whole blood; analyses can be done accurately, reliably, simply and quickly. The determination is highly selective because the electrode senses only the ammonium ion, which is selectively released from urea by urease in a preliminary rapid incubation step. The reproducibility of the determination (standard deviation less than 2.4%) is sufficient for clinical purposes; the linear range of the method is 10^-2–lO^-4M urea. Since the electrode actually never touches the sample solution, the problems caused by the presence of proteins, blood cells etc. do not arise.     

Voltammetric-enzymatic determination of ethanol in whole blood by flow injection analysis

Summary Two automatic methods are proposed for the voltammetric determination of ethanol in whole blood. They are based on the use of the enzymatic systems alcohol dehydrogenase/ nicotinamide adenine dinucleotide (method A) and alcohol dehydrogenase/nicotinamide adenine dinucleotide/ diaphorase/2,6-dichlorophenolindophenol (method B). The determination ranges are 2.5–30.0 g/ml and 0.5–30.0 g/ml with sampling rates of 60 and 30 samples/h, respectively. The coefficients of variation are about ±4%.

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Voltammetrisch-enzymatische Bestimmung von Ethanol in Vollblut durch Fließinjektionsanalyse

     

Direct determination of fluoride in aluminium reduction materials by using an ion-selective electrode

Macro amounts of fluoride in aluminium reduction materials are successfully determined with a fluoride electrode. Except for anhydrous aluminium fluoride, which requires fusion with sodium hydroxide, samples are dissolved in aqueous media. Cryolite and sodium fluorosilicatc are dissolved in boiling sodium hydroxide solution. Other materials containing fluoride, such as fluorspar and the reduction cell bath and pot-lining, require dissolution in a hydrochloric acid solution of aluminium chloride. Potential interference from large amounts of aluminium (and calcium, if present) is eliminated and pH control attained by using ammoniacal sulphosalicylate (and EDTA). The procedures are reasonably rapid. Relative errors of less than 2% and a relative standard deviation of 1% are achieved.     

Rapid and selective determination of ammonium by fluorimetric flow injection analysis

Selective and sensitive procedures for the determination of ammonium in river water and diluted urine were developed by using flow injection analysis equipment. The methods are based on the derivatization of ammonia with o-phthaldehyde (OPA) and thioglycolate under alkaline conditions. The formed isoindole derivative is detected fluorimetrically at an excitation wavelength of 415 nm and an emission wavelength of 485 nm. The derivatization only takes 15 to 20 s at room temperature to achieve the maximum sensitivity. The optimized OPA reagent shows a surprisingly high selectivity for ammonium in the presence of many primary amines. With respect to the analysis of turbid and fluorescent sample solutions the selectivity can be improved by separating the ammonia through a microporous membrane from the OPA reagent. Without this separation step ammonia can be detected in the range between 0.05 and 100 μM with excellent linearity. After the insertion of an optimized membrane separation cell ammonia can be determined in the linear range between 0.2 μM and 20 mM.     

Potentiometric and conductometric determination of ammonium by gas-diffusion flow injection analysis

Summary Considerable enhancement of selectivity in the potentiometric and conductometric determination of ammonium is provided by gas-diffusion separation in flow injection analysis. Ammonium and potassium selective liquid membrane electrodes can be used for determinations in the concentration range 10^–7–10^–2 mol/l with high precision and fast sample throughput. No interferences are encountered in the presence of ionic species and molecules that likely adsorb when the sensors are in direct contact with the sample. The selectivity over volatile amines is enhanced due to kinetic discrimination. Conductometric detection is shown to be as sensitive as the potentiometric detection. A major advantage, however, is the linear rather than logarithmic relationship between concentration and conductivity.     

Rapid and selective determination of ammonium by fluorimetric flow injection analysis

Selective and sensitive procedures for the determination of ammonium in river water and diluted urine were developed by using flow injection analysis equipment. The methods are based on the derivatization of ammonia with o-phthaldehyde (OPA) and thioglycolate under alkaline conditions. The formed isoindole derivative is detected fluorimetrically at an excitation wavelength of 415 nm and an emission wavelength of 485 nm. The derivatization only takes 15 to 20 s at room temperature to achieve the maximum sensitivity. The optimized OPA reagent shows a surprisingly high selectivity for ammonium in the presence of many primary amines. With respect to the analysis of turbid and fluorescent sample solutions the selectivity can be improved by separating the ammonia through a microporous membrane from the OPA reagent. Without this separation step ammonia can be detected in the range between 0.05 and 100 microM with excellent linearity. After the insertion of an optimized membrane separation cell ammonia can be determined in the linear range between 0.2 microM and 20 mM.     

Determination of creatinine in undiluted blood serum enzymatic flow injection analysis with optosensing

A specific enzymatic assay for creatinine in undiluted serum samples is described, exploiting the generation of ammonia from creatinine by immobilized creatinine iminohydrolase. The ammonia produced is separated from the sample matrix by gas diffusion into an acceptor stream containing a pH-sensitive indicator. The creatinine content is quantified by monitoring the resulting colour change of the indicator by means of reflectance measurement via optical fibers, the hydrophobic gas-permeable membrane serving as a diffuse reflector. Two approaches are used to overcome the interference caused by endogenous ammonia. The first is based on enzymatic abatement of endogenous ammonia by immobilized glutamate dehydrogenase. In the second, preferable, approach, endogenous ammonia, itself a parameter of clinical interest, is measured separately prior to the enzymatic degradation by creatinine iminohydrolase. Each assay requires only 30 μl of sample solution, and the sampling frequency is 60 h^?1. The relative standard deviation is approximately 3%.     

New flow injection potentiometric graphite coated ion-selective electrode for the determination of VO ions

A new coated ion-selective electrode for the determination of trace vanadyl ions (VO²⁺) by flow injection potentiometry (FIP) with a home-made flow cell has been developed. The PVC-based membrane was coated on a graphite electrode with an effective area of 4.90 mm². The optimum membrane contains 5 wt.% 1,8-diaminonaphtalene as ionophore, 35 wt.% plasticizer 2-nitrophenyl octyl ether, 55 wt.% PVC and 5 wt.% additive potassium tetrakis (p-chlorophenyl) borate. The electrode in flow injection potentiometry resulted in well defined peaks for vanadyl ions with a very high sampling rate (180 injections/h). Linear calibration was obtained from 1.14×10⁻⁷ to 1.14×10⁻¹ M vanadyl ions, with a slope of 28.3±0.3 mV per decade change in vanadyl concentration, and very low detection limit of 1.14×10⁻⁷ M and the electrode can be used for at least 1 months without any considerable change in potential response. Selectivity coefficients for several ions were obtained by the matched potential method with respect VO²⁺ ions. The flow cell is simple to construct and free from memory effect problems over long periods of use. The sensor was used for the recovery of trace VO²⁺ ions from tap water and the determination of VO²⁺ in synthetic sample.     

The fluoride ion-selective electrode in flow injection analysis : Part 3. Applications

Flow-injection potentiometry with a combination fluoride-selective electrode is used to determine fluoride in tap water, beverages and urine. Excellent sensitivity (down to 1 μg l^?1) and long-term stability are obtained, with a sample throughput of 30–40 h^?1, based on triplicate injections at 120 h^?1. The commonly used buffer TISAB-III is unsuitable for the analysis of undiluted tea and urine samples. The application of a modified citrate-containing TISAB overcomes interferences caused by high natural ionic strength and avoids complexation of fluoride. Recoveries after spiking tap water, tea and urine with fluoride concentration ranging from 0.01 to 1 mg l^?1 are in the range 91–106%. The equipment used provides a flexible system allowing fast changes between different buffers and carrier streams depending on the samples presented.     

Fluoride ion-selective electrode in flow injection analysis : Part 2. Interference Studies

The influence of the sample composition on the response characteristics of the fluoride ion-selective electrode in flow injection analysis is described. Sample parameters such as ionic strength, viscosity and pH affect the response time of the electrode and cause transient signals when limiting values are exceeded. The respective limiting values depend on the total ionic-strength adjustment buffer (TISAB) used and these interferences can be minimized by proper choice of the TISAB. The complex formation of fluoride by several elements in the presence of TISAB containing CDTA is discussed. Aluminium and magnesium were found to interfere when present at levels above 1 and 100 mg l^?1, respectively. The signal decrease in the presence of iron, calcium and silicon can be attributed to ionic strength effects rather than complexation. Provided that the ionic strength is taken into account and corrected for, no influence occurs even in the presence of 0.5, 2 and 5% of iron, calcium and silicon, respectively.     

Flow injection determination of bromide ion in a developer using bromide ion-selective electrode detector

A potentiometric flow injection determination method for bromide ion in a developer was proposed, by utilizing a flow-through type bromide ion-selective electrode detector. The sensing membrane of the electrode was Ag(2)S-AgBr membrane. The response of the electrode detector as a peak-shape signal was obtained for injected bromide ion in a developer. A linear relationship was found to exist between peak height and the concentration of the bromide ion in a developer in a concentration range from 1.0x10(-3) to 1.0x10(-2) mol l(-1). The relative standard deviation for 10 injections of a 6x10(-3) mol l(-1) bromide ion in a developer was 1.3% and the sampling rate was ca 17-20 samples h(-1). The present method was free from the interference of an organic reducing reagent, an organic substance in a developer sample solution for the determination of bromide ion in a developer.     

Enzymatic substrate determination by flow injection analysis in stopped flow modus using a single 5-port-valve for sample and reagent injection

Summary The determination of ethanol with alcohol dehydrogenase is described as an example of enzymatic determination with the flow injection analysis system (FIAS). Both, sample and reagent, are successively injected into the carrier stream by using only one valve. Compared with other techniques, the principle described is more economical with regard to reagent consumption and analysis time. Basic experiments about this kind of reagent addition (dispersion, reproducibility, possibility for gradient dilution) were made by simulation with dye solution. The determination of ethanol is carried out using the stopped flow technique. The peristaltic pumps are stopped when the reaction zone is located in the flow cell, and the change of absorbance with time is monitored. Thus background signals and other matrix influences can be minimized. The method is tested under real conditions for the determination of alcohol in several beverages.     

The fluoride ion-selective electrode in flow injection analysis: Part 1. General Methodology

A simple flow-injection system for determination of traces of fluoride by means of the fluoride-selective electrode is presented. A comparison of several flow-cell arrangements confirmed the advantages of a well-jet design. Systematic investigations of the parameters affecting response times (i.e., polishing procedure, flow rate, carrier composition) established the optimal experimental conditions for measurements down to 1 μg l^?1 fluoride. Calibration plots in the lower μg l^?1 range were neither Nernstein nor linear, but good precision (0.5–5%) was obtained even when the potential differences for concentration steps of one decade were as small as 3 mV.     

Potentiometric flow injection system for determination of reductants using a polymeric membrane permanganate ion-selective electrode based on current-controlled reagent delivery

A polymeric membrane permanganate-selective electrode has been developed as a current-controlled reagent release system for potentiometric detection of reductants in flow injection analysis. By applying an external current, diffusion of permanganate ions across the polymeric membrane can be controlled precisely. The permanganate ions released at the sample-membrane interface from the inner filling solution of the electrode are consumed by reaction with a reductant in the sample solution thus changing the measured membrane potential, by which the reductant can be sensed potentiometrically. Ascorbate, dopamine and norepinephrine have been employed as the model reductants. Under the optimized conditions, the potential peak heights are proportional to the reductant concentrations in the ranges of 1.0×10(-5) to 2.5×10(-7)M for ascorbate, of 1.0×10(-5) to 5.0×10(-7)M for dopamine, and of 1.0×10(-5) to 5.0×10(-7)M for norepinephrine, respectively with the corresponding detection limits of 7.8×10(-8), 1.0×10(-7) and 1.0×10(-7)M. The proposed system has been successfully applied to the determination of reductants in pharmaceutical preparations and vegetables, and the results agree well with those of iodimetric analysis.     

Flow injection analysis of chloride in tap and sewage water using ion-selective electrode detection

Summary The application of a chloride-selective electrode in flow injection analysis of chloride in tap and sewage water samples is described. With an extremely simple experimental set-up, from 60 to 120 samples per hour can be analysed. The standard deviation lies at 5%, when the chloride content is above 100 ppm, and 10%, if 10 ppm are present. Using a AgCl single crystal ion-selective electrode and high carrier solution flow rates, interfering anions like iodide, bromide, etc. are kinetically discriminated against and interfere only at orders of magnitude less than usual.

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Fließinjektionsanalyse von Chlorid in Leitungs- und Abwasser mit Hilfe der Detektion durch eine ionen-selektive Elektrode

Zusammenfassung Es wird der Einsatz einer ionenselektiven Chloridelektrode in Verbindung mit der Fließinjektionsanalyse von Chlorid in Leitungs- und Abwasserproben beschrieben. Mittels einer extrem einfachen Anordnung lassen sich so zwischen 60 und 120 Proben stündlich analysieren. Die Standardabweichung beträgt bei Chloridgehalten >100 ppm ca. 5% und bei Gehalten um 10 ppm ca. 10%. Wenn eine AgCl-Einkristallmembranelektrode in Verbindung mit einer hohen Flußrate benutzt wird, stören die üblichen Störionen, wie Iodid, Bromid usw., um Größenordnungen weniger als üblich.

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Dedicated to Prof. Dr. H. Monien on the occasion of his 60th birthday

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1983 on leave from Analytical Chemistry Department, Higher Chemical Technological Institute, 8, Kliment-Ochridsky-Str., Sofia 1156, Bulgaria     

Flow injection determination of ethanol in whole blood using immobilized enzymes

Enzymatic methods for the determination of ethanol in whole blood are proposed. They use different types of detection and flow injection analysis (FIA) modes: fluorometric detection (use of normal FIA and stopped-flow/FIA); amperometric detection by monitoring of NADH (use of normal amperometric and pulse mode) and with the aid of a coupled enzymatic reaction (2,6-dichlorophenolindophenol/diaphorase). Determination ranges between 0.1 and 30.0 μg/ml are obtained (which in all cases comprise the legal range of ethanol in blood), with good precision and sampling frequency. The sensitivity of the methods can be manipulated by changing the injected sample volume or the pH.     

Flow injection determination of iodide ion in a photographic developing solution using iodide ion-selective electrode detector

A potentiometric flow injection determination method for iodide ion in a photographic developing solution was proposed by utilizing a flow-through type iodide ion-selective electrode detector. The sensing membrane of the electrode was Ag₂S–AgI membrane. The response of the electrode detector as a peak-shape signal was obtained for injected iodide ion in a photographic developing solution. A linear relationship in the subnernstian zone was found to exist between peak height and the concentration of the iodide ion in a photographic developing solution in a concentration range from 0 to 6.0×10⁻⁵ mol l⁻¹. The relative standard deviation for ten injections of 2×10⁻⁵ mol l⁻¹ iodide ion in a photographic developing solution was 0.96% and the sampling rate was approximately 12–13 samples h⁻¹. The iodide ion could be determined under coexisting of an organic reducing reagent and inorganic electrolytes of high concentration in a photographic developing solution sample solution by the present method.