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Results for “Flow Injection Analysis”

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PubMedDate not supplied

Novel flow injection analysis systems for drug analysis.

Flow injection analysis (FIA) has become a versatile tool for rapid and automated analyses. As its capabilities have increased, so have the complexity and operation of the apparatus. We have been investigating ways to simplify both the apparatus and the application of injection techniques. A novel cam-driven syringe pump and the development of sequential injection analysis (SIA) are reviewed, and some applications presented. Flow injection coulometric titrations are presented as a means to further alleviate reagent stability and calibration problems. These systems have potential for automatically carrying out many drug assays.

Calibration↗

PubMed1992

Biosensors and flow injection analysis.

Combining flow injection analysis with a biosensor is a novel biosensing process which has allowed speedy and accurate analysis. Diagnostic analysis is the most important application for biosensing flow injection analysis, but other applications include bioprocess monitoring, analysis of food and agricultural products, as well as environmental analysis. In addition, the analysis of compounds, such as explosives and abused drugs, and monitoring of Salmonella, the microorganism that causes food poisoning, have been reported.

Animals↗

PubMedDate not supplied

The determination of the preservative, chlorocresol, in a pharmaceutical formulation by flow injection analysis.

A flow injection analysis (FIA) procedure is described for the determination of chlorocresol in a parenteral pharmaceutical formulation. The product is directly injected into a carrier stream of water and subsequently reacted with a reagent stream of nitrous acid. The resulting brown nitro-derivative is determined spectrophotometrically at 400 nm. The method has been validated and should be applicable to chlorocresol in other pharmaceutical products and to compounds containing a phenolic ring, assuming absence of matrix interference.

Chemistry, Pharmaceutical↗

PubMedDate not supplied

The determination of pentavalent antimony in sodium stibogluconate in a pharmaceutical formulation by flow injection analysis.

A flow injection analysis (FIA) procedure is described for the determination of pentavalent antimony (Sb5+) in the drug, sodium stibogluconate, in a parenteral pharmaceutical formulation. The sample solution is injected directly into a carrier stream of iodide ion which is then mixed with an acid stream in situ. Sb5+ is determined by the redox reaction with acidified iodide to liberate iodine, which is monitored spectrophotometrically at 350 nm. The closed conditions prevent interference from atmospheric oxygen and the rapid reaction time assists in minimizing interference from side reactions. The use of tartaric acid as a solvent for sample and standard solutions ensures obedience of Beer's law over the Sb5+ concentration range 0.01-0.2% (w/v). The method is specific for the higher oxidation state in an ionic mixture of Sb5+ and Sb3+, and has been fully validated for use in a pharmaceutical preparation. Assuming absence of matrix interference it is applicable to Sb5+ from other sources and should be applicable to other reducible ionic species.

Antimony↗

PubMed1990

Selective determination of histamine by flow injection analysis.

A flow injection analysis (FIA) method for the determination of histamine is described. Control of reaction timing allows exploitation of a transient, chemical-kinetic increase in selectivity that occurs when o-phthalaldehyde reacts with histamine. The molar fluorescence ratio (selectivity) of histamine/histidine reaches a maximum value of 800 in 32 s, precluding the need for separation of histamine from histidine, spermidine, and other potential interferences in biological samples. On-line dilution prevents matrix effects and affords a linear response up to approximately 4.45 mM histamine, or 500 mg of histamine free base/100 g. Under these conditions the detection limit (3 times peak-to-peak baseline noise) is 5.5 pg (corresponding to 0.60 mg of histamine free base/100 g of sample) and throughput is 60 injections per hour. The high sensitivity and high selectivity of the method allow the rapid determination of histamine in fish with minimal sample conditioning and will find application in the determination of endogenous histamine as well, such as in blood plasma and brain tissue.

Animals↗

PubMed1992

Determination of organophosphorous and carbamate insecticides by flow injection analysis.

A flow injection system, incorporating an acetylcholinesterase (AChE) single bead string reactor (SBSR), for the determination of some organophosphorous (azinphos-ethyl, azinphos-methyl, bromophos-methyl, dichlorovos, fenitrothion, malathion, paraoxon, parathion-ethyl and parathion-methyl) and carbamate insecticides (carbofuran and carbaryl) is presented. The detector is a simple pH electrode with a wall-jet entry. Variations in enzyme activity due to inhibition are measured from pH changes when the substrate (acetylcholine) is injected before and after the passage of the solution containing the insecticide. The percentage inhibition of enzyme activity is correlated to the insecticide concentration. Several parameters influencing the performance of the system are studied and discussed. The detection limits of the insecticides ranged from 0.5 to 275 ppb. The determination of these compounds was conducted in Hepes buffer and a synthetic sea water preparation. The enzyme reactor can be regenerated after inhibition with a dilute solution of 2-PAM and be reused for analysis. The immobilized enzyme did not lose any activity up to 12 weeks when stored at 4 degrees C.

Acetylcholine↗

PubMed1992

On-line determination of glucose and lactate concentrations in animal cell culture based on fibre optic detection of oxygen in flow-injection analysis.

A flow-injection analysis (FIA) system based on fibre optic detection of oxygen consumption using immobilized glucose oxidase (GOD) and lactate oxidase (LOD) is described for the on-line monitoring of glucose and lactate concentrations in animal cell cultures. The consumption of oxygen was determined via dynamic quenching by molecular oxygen of the fluorescence of an indicator. GOD and LOD were immobilized on controlled pore glass (CPG) in enzyme reactors which were directly linked to a specially designed fibre optic flow-through cell covering the oxygen optrode. The system is linear for 0-30 mM glucose, with an r.s.d. of 5% at 30 mM (five measurements) and for 0-30 mM lactate, with an r.s.d. of 5% at 30 mM (five measurements). The enzyme reactors used were stable for more than 4 weeks in continuous operation, and it was possible to analyse up to 20 samples per hour. The system has been successfully applied to the on-line monitoring of glucose and lactate concentrations of an animal cell culture designed for the production of recombinant human antithrombine III (AT-III). Results of the on-line measurement obtained by the FIA system were compared with the off-line results obtained by a glucose and lactate analyser from Yellow Springs Instrument Company (YSI).

Animals↗

PubMed1990

Kinetics of protease hydrolysis of extended peptide substrates: measurement by flow-injection analysis.

A flow-injection analysis (FIA) system was developed to study the enzyme-catalyzed hydrolysis of synthetic peptides, each of which contained one scissile bond. The concentrations of alpha-amino groups in reactions mixtures were determined by FIA with o-phthalaldehyde as a fluorescence reagent. The method allows a rapid, precise, and sensitive determination of kinetic constants for proteases acting on extended peptide substrates.

Chromatography, High Pressure Liquid↗

PubMed1991

On-line determination of intracellular beta-galactosidase activity in recombinant Escherichia coli using flow injection analysis (FIA).

A flow injection analysis (FIA) system was developed for the determination of cytoplasmic beta-galactosidase activity in recombinant Escherichia coli. The FIA system and its application for on-line monitoring of beta-galactosidase production during cultivation of recombinant E. coli in a 60-l airlift tower loop reactor is described. The results demonstrate that an FIA assay in conjunction with a cell disintegration step can be applied successfully for on-line monitoring of intracellular protein formation.

Colorimetry↗

PubMed1991

On-line determination of glucose concentration throughout animal cell cultures based on chemiluminescent detection of hydrogen peroxide coupled with flow-injection analysis.

A flow-injection analysis (FIA) system for the on-line determination of glucose in animal cell cultures is described. The system is based on immobilized glucose oxidase (GOD). The hydrogen peroxide generated in the enzyme reaction is determined via a highly sensitive chemiluminescent reaction with luminol. Based on the measurement of the maximum emitted light intensity, the system was able to analyse hydrogen peroxide over the concentration range of 10(-7) to 10(-2) M. For glucose determination, the system has a linear range of 10(-5) to 5 x 10(-2) M glucose, with an r.s.d. of 3% at the 1 mM level (5 measurements). The influence of luminol and buffer concentrations, pH and temperature on the chemiluminescent reaction were investigated. The enzyme reactor used was stable for more than 4 weeks in continuous operation, and it was possible to analyse up to 20 samples per h. The system has been successfully applied to on-line monitoring of glucose concentration during an animal cell culture, designed for the production of human antithrombin III factor. Results obtained with the FIA system were compared with off-line results, obtained with a Yellow Springs Instrument Company Model 27 (YSI).

Animals↗

PubMed1987

Selective determination of lithium in biological fluids using flow injection analysis.

The use of flow injection analysis to automated extraction methods for the determination of lithium ion utilizing crown ethers or cryptands is demonstrated. The ion-pair extraction of cryptand 211, lithium, and resazurin exhibits a linear range for lithium ion of 70 ppb to 2.1 ppm. This method could tolerate up to 1000 ppm sodium ion. The chromogenic crown ether, 1-(2-hydroxy-5-nitrobenzyl)-1-aza-4,7,10-trioxacyclododecane, exhibits a linear range for lithium ion of 0.3 to 2 ppm. A sodium ion concentration of 230 ppm can be tolerated. Both extraction systems were used in the automated determination of lithium in blood serum and urine. Both methods agreed well with the known and/or atomic absorption values.

Bridged Bicyclo Compounds↗

PubMed1990

Fluorometric determination of urinary kynurenic acid by flow injection analysis equipped with a "bypass line".

A flow injection analysis involving a photochemical reaction and fluorometric detection has been developed for the determination of urinary kynurenic acid. Kynurenic acid was found to fluoresce on irradiation with ultraviolet light at pH 7.2 in the presence of hydrogen peroxide. This method was applied to flow injection analysis using a new procedure involving a "bypass line" for the simultaneous determination of urinary kynurenic acid and background fluorescence. The calibration graph showed linearity over the range of 0.20 to 120 pmol. For pretreatment of urinary kynurenic acid, a PRE-SEP C18 cartridge was used. The mean recovery of kynurenic acid from urine was 94.5%. The content of urinary kynurenic acid was 13.0 +/- 2.68 mumol/day. There was good correlation (r = 0.9729) between values determined by flow injection analysis and high-performance liquid chromatography.

Chemistry Techniques, Analytical↗

PubMed1992

Spectrophotometric determination of promethazine by flow injection analysis and oxidation by CeIV.

A flow injection analysis (FIA) procedure is proposed for the determination of promethazine. The sample solution is directly injected into the carrier-reagent stream which comprises a solution of ceric ions in a sulphuric acid medium. The absorbance at 514 nm from the red colour developed by the oxidation of promethazine is measured. Effects of foreign substances have been investigated and the procedure has been applied to the determination of promethazine in a pharmaceutical formulation (tablets).

Cerium↗

PubMed1992

Solubilized substrates for the on-line measurement of lipases by flow injection analysis during chromatographic enzyme purification.

A flow injection analysis (FIA) system for the on-line measurement of lipases in chromatographic processes has been developed. The photometrically detectable substrates para-nitrophenylpalmitate, S,O,O'-tripropyryl-1-thioglycerol, and 1,2-O-dilauryl-rac-glycero-3-glutaric-resorufinester were investigated. Different detergents and qualities of assay emulsions were tested for optimal results in FIA applications. Emphasis was placed on increasing the stability of the assay emulsion. Lipases of different origin and specificity were detected. The linear detection range was adapted to the requirements of the chromatographic purification procedures. The connection of the FIA with a fast protein liquid chromatography system permitted the automatization of lipase purification by monitoring protein content, salinity, and enzyme activity of the effluent from column chromatography.

Animals↗

PubMed1989

Solid-phase electrochemical enzyme immunoassay with attomole detection limit by flow injection analysis.

A sandwich electrochemical enzyme immunoassay with flow injection analysis for the model antigen mouse IgG has been developed with alkaline phosphatase as the enzyme label. The enzyme substrate, 4-aminophenyl phosphate and its enzymatic reaction product, 4-aminophenol have been studied by cyclic and hydrodynamic voltammetry. The determination of 4-aminophenol by flow injection analysis with electrochemical detection (FIAEC) has a linear range of 5.0 x 10(-8) to 1.0 x 10(-5) M, a detection limit of 2.4 x 10(-8) M, and a sample throughput of 72 samples/h. The detection limit is set by a background capacitance response, which depends on the ionic strength difference between the sample and the mobile phase. The sandwich immunoassay has been characterized with respect to substrate concentration for the enzymatic reaction, detection limit, dynamic range and sources of error. Mouse IgG can be determined with a detection limit of 0.81 pg ml-1 by a 30-min substrate incubation time and a six orders of magnitude linear dynamic range.

Aminophenols↗

PubMed1979

Flow-injection analysis with chemiluminescence detection.

In "flow-injection analysis," the sample is inserted into a stream of reagent by use of a sample-injection valve. Mixing occurs downstream from the valve in a coil of tubing. With chemiluminescence detection this coil is positioned in front of a photomultiplier. We have evaluated the system for detection of hydrogen peroxide, using the luminol reaction with cupric ion as a catalyst. The effects of flow rate, sample volume, and reaction kinetics on the magnitude, duration, and repeatability of the chemiluminescent response have been evaluated. Precisions of 1 to 2% relative standard deviation on replicate measurements are readily achievable. A sample throughput of six samples per minute is possible with very little peak overlap. This detection system can be coupled to any process in which peroxide is generated.

Autoanalysis↗

PubMed1991

Flow injection analysis with immobilized reagents.

Immobilized reagent phase flow injection analysis can be configured as discrete reagent cells upstream of the sensor element or as an integral reagent/transduction system (flow injection analysis-biosensor). The former approach has attracted greater attention because several assays can be assembled with greater versatility in reagent column units employing a single sensor, than can be co-immobilized on the surface of a transducer.

Animals↗

PubMed1991

Quantitation of DNA and RNA in crude tissue extracts by flow injection analysis.

An automated two-dye flow injection analysis system to quantitate DNA and RNA in crude extracts of tissues is described. The method uses the fluorochrome dyes ethidium bromide and Hoechst 33258. DNA concentration is determined directly from its fluorescence in Hoechst dye. RNA is estimated from fluorescence in ethidium bromide after subtraction of the fluorescence due to DNA. This method has several advantages: a simple extraction procedure, a low detection limit (0.01 micrograms DNA and 0.10 micrograms RNA), automation, and a high sample throughput.

Animals↗