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At least 217 records · Page 12Linked to original sources
PubMed1993

Software FIACRE: bioprocess monitoring on the basis of flow injection analysis using simultaneously a urea optode and a glucose luminescence sensor.

Various computer programs for large-scale bioprocess control and optimization have been developed as well as software for simple laboratory routine analysis. In comparison, software can hardly be found that works on laboratory scale and provides the control of complex flow injection analysis (FIA) systems, multisubstrate determination, data evaluation as well as minimal process control abilities. The sensors applied can be of different type (luminometric or other optical as well as electrochemical biosensors). The development of such a software may be very helpful for the transfer of FIA/biosensor systems from the state of development to industrial processes. Hence, each analysing system--even a well established biosensor--has to be individually adapted to the process, a task which is best done under laboratory conditions. Such a flexible, computer-controlled FIA system for research level based on the software FIACRE is presented. Five FIA/(bio)sensor system can be controlled simultaneously. Additionally, common temperature and pH recordings are possible. Determinations of substrate concentrations are performed by means of calibration curves which can be recorded at different times. This allows supervising the activities of the sensors during a cell cultivation and controlling the bioprocess, e.g. by adding substrate to a cell culture. The automated monitoring of the degradation of glucose and urea by two different optical sensing principles during a cell cultivation under the control of one microcomputer is presented for the first time. For this purpose, already well examined biosensors (a urease optode and a luminometric glucose sensor) were employed and their properties discussed under the aspect of working in real cultivation media. It will also be shown that substrates being of interest for bioprocess control can be detected by slight modifications of known reactions. For example, substrates of NADH-dependent enzymatic reactions can be detected by the luminol chemiluminescence system, and optodes can be employed for pH, penicillin and glucose determination.

Alcohol Dehydrogenase↗
PubMed1985

Flow-injection analysis of catecholamine secretion from bovine adrenal medulla cells on microbeads.

Bovine adrenal medullary cells have been cultured on microbeads which are placed in a low-volume flow system for measurements of stimulation-response parameters. Electronically controlled stream switching allows stimulation of cells with pulse lengths from 1 s to many minutes; pulses may be repeated indefinitely. Catecholamines secreted are detected by an electrochemical detector downstream from the cells. This flow-injection analysis technique provides a new level of sensitivity and precision for measurement of kinetic parameters of secretion. A manual injection valve allows stimulation by higher levels of stimulant in the presence of constant low levels of stimulant. Such experiments show interesting differences between the effects of K+ and acetylcholine on cells partially desensitized to acetylcholine.

Adrenal Medulla↗
PubMed1990

Determination of penicillin in pharmaceutical formulations by flow injection analysis using an optimised immobilised penicillinase reactor and iodometric detection.

An automated assay for the determination of penicillin in formulations suitable for use in pharmaceutical quality control is presented. The method is based on the classical iodometric penicillin assay which is incorporated in a flow injection analysis (FIA) system. The required hydrolysis is performed on-line by using an immobilised penicillinase reactor. Packed-bed and single-bead-string enzyme reactors are compared. It turns out that a packed-bed penicillinase reactor (10 cm x 1.5 mm i.d.) provides complete hydrolysis within short residence time, while only little back-pressure is generated. This enzyme reactor is stable for at least 9 months. Enzymatic hydrolysis of the beta-lactam ring results in the formation of the corresponding penicilloic acid, which consumes iodine. The iodine consumption is determined colorimetrically by measuring the decrease of the absorbance of the blue coloured iodine/starch complex. The optimum reactor length and flow rate for the colourimetrical detection reaction are determined. The optimised method is applied to the assay of penicillin in formulations and the results are compared with the "true" results obtained with a reference method: a mercurimetric titration. The reliability of the flow injection method is evaluated quantitatively by determining the maximum total error (MTE). The reliability is shown to be highest when measuring at a 0.3-mM level. Eight formulations including capsules, tablets and injectables containing penicillin G, amoxicillin or flucloxacillin are assayed. The MTE does not exceed the 6% level and the most probable MTE is between 1.5 and 3.5%.

Enzymes, Immobilized↗
PubMed1982

Direct determination of therapeutic concentrations of lithium in serum by flow-injection analysis with atomic absorption spectroscopic detection.

In this flow-injection system for direct determination of lithium in serum by atomic absorption spectroscopy, the 10-microL sample is manually injected into a continuously flowing non-segmented stream of de-ionized water, which is pumped, via a dispersion tube, to the spectrometer's nebulizer. Controlled dispersion of the sample zone, before it is introduced into the nebulizer, produces the required sample dilution. Effects of varying the length of the dispersion tube, the flow rate, and the sample size were studied. Analytical readout is obtained, in the form of transient peaks, 5 s after sample injection. It is necessary to include physiological concentrations of sodium and potassium in the standard because each of these cations enhances the lithium absorbance signal. Analytical recovery (98.5 to 101%) and CV (about 2%) are good, and results compare well with those obtained by aspiration of prediluted samples (n = 121, r = 0.99).

Chemistry, Clinical↗
PubMed1990

Flow injection analysis and biosensors: applications for biotechnology and environmental control.

Our experience in industrial bioprocess monitoring and environmental control let us develop a concept for biosensor research which distinguishes itself from other, more popular, approaches. Biosensors must improve and/or simplify existing state-of-the-art analysis systems. Only the parallel development of biosensors and their complementary metrology leads to industrially sound solutions. The combination of flow injection analysis with immobilized enzymes in the form of enzyme columns is already used today for the solution of on-line analytical problems in bioprocesses and environmental control.

Biosensing Techniques↗
PubMed2001

Flow-injection analysis of nitrate by reduction to nitrite and gas-phase molecular absorption spectrometry.

Two flow-injection manifolds have been investigated for the determination of nitrate. These manifolds are based on the reduction of nitrate to nitrite and determination of nitrite by gas-phase molecular absorption spectrophotometry. Nitrate sample solution (300 microL) which is injected to the flow line, is reduced to nitrite by reaction with hydrazine or passage through the on-line copperized cadmium (Cd-Cu) reduction column. The nitrite produced reacts with a stream of hydrochloric acid and the evolved gases are purged into the stream of O2 carrier gas. The gaseous phase is separated from the liquid phase using a gas-liquid separator and then swept into a flow-through cell which has been positioned in the cell compartment of an UV-visible spectrophotometer. The absorbance of the gaseous phase is measured at 204.7 nm. A linear relationship was obtained between the intensity of absorption signals and concentration of nitrate when Cd-Cu reduction method was used, but a logarithmic relationship was obtained when the hydrazine reduction method was used. By use of the Cd-Cu reduction method, up to 330 microg of nitrate was determined. The limit of detection was 2.97 microg nitrate and the relative standard deviations for the determination of 12.0, 30.0 and 150 microg nitrate were 3.32, 3.87 and 3.6%, respectively. Maximum sampling rate was approximately 30 samples per hour. The Cd-Cu reduction method was applied to the determination of nitrate and the simultaneous determination of nitrate and nitrite in meat products, vegetables, urine, and a water sample.

Absorption↗
PubMed1989

Automated determination of cholinesterase activity in plasma and erythrocytes by flow-injection analysis, and application to identify subjects sensitive to succinylcholine.

This automated spectrophotometric method for determination of cholinesterase activity in erythrocytes and plasma is based on measurement of the choline produced at 30 degrees C by the hydrolysis of acetyl-, butyryl-, or succinylcholine. Blanks, standards, and samples are prepared by a Gilson robotic unit. Use of flow-injection analysis for detection allows use of smaller volumes of reagent and sample. We applied this method to the study of 91 healthy subjects and members of two families with succinylcholine sensitivity. Results with use of the three different substrates for determination of activity in plasma correlated well (r greater than 0.94). Results for plasma and erythrocytes from healthy subjects are lower in women less than 50 years old than in women greater than 50 years or men. Values for plasma obtained with succinylcholine substrate (range: 31 to 100 U/L) allow detection of very sensitive subjects--AA phenotypes (less than 10 U/L)--but do not distinguish the UA from the UU phenotype.

Adult↗
PubMed2002

Quinoline-8-ol and its derivatives as preconcentration agents in flow injection analysis coupled to atomic and molecular spectrometric techniques.

Quinoline-8-ol, one of the most popular and versatile organic reagents, is known to react with several metals, because it has both a basic nitrogen and a phenolic group. Hence, quinoline-8-ol and its derivatives have been widely used as chelating and/or sorbent extraction agents in on-line flow injection analysis (FIA) along with a suitable detection technique. The state of the art for using quinoline-8-ol and its derivatives in FIA was studied in terms of the selectivity, sensitivity and precision. Emphasis was placed on validating these procedures by analyzing certified reference materials and applying these analytical methodologies to real samples for the determination of inorganics.

Journal Article↗
PubMed1990

Immobilization of glucose oxidase and peroxidase and their application in flow-injection analysis for glucose in serum.

Glucose oxidase (GOD) and Horseradish peroxidase (HRP) were covalently coupled to alkylamine controlled pore glass by means of glutaraldehyde. About 700-800 U/g of immobilized GOD and 300-400 U/g of immobilized HRP were obtained. Some factors of affecting enzyme immobilization were discussed. The immobilized enzymes were packed into a plastic tube and used in flow-injection analysis (FIA) for glucose in serum. A good linearity range was observed for this immobilized enzyme system at 20 mg/mL to 1000 mg/dL D-glucose, the recovery was 95.4-103.5%, the within-batch imprecision was 0.8-2.2%, and the between-batch imprecision was 2.2-4.2%. More than 100 samples were measured within an hour. One enzyme column with five units of immobilized GOD and HRP, applied for 50 assays/d, has been used for more than 2 mo.

Autoanalysis↗
PubMed2001

Fluorimetric determination of arsanilic acid by flow-injection analysis using on-line photo-oxidation.

A flow-injection-fluorimetric method for the determination of arsanilic acid is proposed. The assay is based on the on-line decomposition of arsanilic acid in the presence of peroxydisulfate on irradiation with UV light. The arsenate generated in the photochemical reaction was reacted with molybdate in dilute nitric acid to form arsenomolybdic acid, which oxidised thiamine to thiochrome. The thiochrome was monitored fluorimetrically at 440 nm with excitation at 375 nm. The calibration graph was linear in the range 0.10-10.8 microg mL(-1) with a correlation coefficient of 0.999. The detection limit was 0.01 microg mL(-1) and the sample throughput was 55 samples h(-1). The applicability of the method was demonstrated by determining arsanilic acid in animal foodstuffs and water.

Animal Feed↗
PubMed1993

Determination of minute amounts of ATP by flow injection analysis using enzyme amplification reactions and fluorescence detection.

A flow-injection system for assay of trace levels of ATP is described that incorporates a small column reactor containing co-immobilized hexokinase, pyruvate kinase and glucose-6-phosphate dehydrogenase. In the presence of appropriate cofactors, ATP is by the synergistic operation of the enzymes repeatedly recycled, resulting in substrate amplification. The ultimately generated NADH is measured fluorometrically. By this approach, where the enzymatic degradation step and the detection step are completely separated, it is possible to operate them individually under optimal conditions. The amplification factor is directly proportional to the residence time of the sample zone within the enzyme reactor, which time might be manipulated by altering the flow-rate and in the extreme by performing stopped-flow experiments. Amplification factors between 15 and 1000 were obtained, but it was found that increased amplifications did not lead to significantly lower detection limits; thus, it appears that a practical lower limit of detection is of the order of 1-5 nM. An investigation of this paradoxical feature, and a possible explanation for it, is given.

Adenosine Triphosphate↗
PubMed1992

Flow injection analysis with immobilized enzymes for process control of pullulan production by fermentation.

A flow injection system is described for the parallel determination of pullulan and glucose during a fermentation of the fungus Aureobasidium pullulans. The polysaccharide was hydrolyzed by pullulanase and amyloglucosidase, immobilized to controlled-pore glass (CPG). The glucose produced was oxidized by glucose dehydrogenase and the NADH formed determined photometrically. The pullulan concentration was calculated from the difference to the response obtained for free glucose. The calibration curves for monomer and polymer were both linear between 2 mg dm-3 and 20 mg dm-3. Analysis of one sample for the determination of glucose and pullulan took about 10 min.

Enzymes, Immobilized↗
PubMed2002

Electrochemical flow injection analysis study of ion partitioning at high surface area carbon fiber electrodes.

Charge-selective electrochemistry was previously shown to occur at high surface area carbon fibers that were produced by fracturing the outer periphery with anodic current or positive potential. The cyclic voltammetric behavior of electroactive species observed at these fibers exhibited a distinct pH dependence related to the protonation/deprotonation of oxygen-containing functional groups at the surface of the carbon fiber. In this paper, electrochemical flow injection analysis (EC-FIA) is used to probe ion partitioning in to and out of the interior microstructure of the treated carbon fiber, for both electroactive and electroinactive species. It was found that the extent of partitioning was the result of both ion charge and hydrated ionic radius, in addition to the level of fracture. It was further observed that the direction of movement for an injected ionic species could be controlled relative to the ion concentration, the pH of the carrier solution, or both. EC-FIA allowed the simultaneous observation of current due to ion movement and that due to electron transfer to a redox-active species. The results presented are consistent with a model in which fixed negatively charged sites in the interior of fractured fibers govern ion partitioning with positively charged ions in the carrier solution, with counterions located in the interior "free" volume.

Journal Article↗
PubMed1991

[Micro-determination of fluoride in biological samples by pyrohydrolysis and flow-injection analysis using a fluoride ion-selective electrode].

An apparatus has been developed for the isolation of fluoride in biological samples through pyrohydrolysis. With this apparatus, it is possible to determine both organic and inorganic fluorocompounds with a recovery close to 100% and precision within 5%. The high recovery rate can be expected even for highly heat-resistant compounds such as CaF2, without using WO3 as a catalyst. For determination of the isolated fluoride, a separate apparatus was developed in which flow-injection analysis was used in conjunction with a fluoride ion-selective electrode as a detector. With this apparatus, fluoride in a sample solution with a volume as small as 0.2 ml, and at a concentration as low as 0.5 microgram/l, can be determined within 3 minutes with a precision of several percent. Combined use of the two apparatuses makes it possible to determine fluoride in different biological samples within 10-15 minutes with a precision of several percent, free from external contamination. By selecting suitable conditions for analysis and using a 1 g sample, it is possible to determine fluoride at a concentration as low as 5 ng/g. By employing these apparatuses, the fluoride content in different biological samples has been determine and the effectiveness of their use confirmed.

Adult↗
PubMed1991

Flow-injection analysis with electrochemical detection of reduced nicotinamide adenine dinucleotide using 2,6-dichloroindophenol as a redox coupling agent.

The determination of reduced nicotinamide adenine dinucleotide (NADH) by electrochemical oxidation requires a more positive potential than is predicted by the formal reduction potential for the NAD+/NADH couple. This problem is alleviated by use of 2,6-dichloroindophenol (DCIP) as a redox coupling agent for NADH. The electrochemical characteristics of DCIP at the glassy carbon electrode are examined by cyclic voltammetry and hydrodynamic voltammetry. NADH is determined by reaction with DCIP to form NAD+ and DCIPH2. DCIPH2 is then quantitated by flow-injection analysis with electrochemical detection by oxidation at a detector potential of +0.25 V at pH 7. NADH is determined over a linear range of 0.5 to 200 microM and with a detection limit of 0.38 microM. The lower detection potential for DCIPH2 compared to NADH helps to minimize interference from oxidizable components in serum samples.

2,6-Dichloroindophenol↗
PubMed1995

Simultaneous determination of nitrate and nitrite in biological samples by multichannel flow injection analysis.

An automated method for the simultaneous determination of nitrite and nitrate in biological samples by using a multichannel flow injection analyzer has been developed. The method was based on the reaction of nitrite with Greiss reagent. The sample solution was injected and equally divided into two channels; channel one (1) represented total nitrite obtained by cadmium reduction of nitrate to nitrite while channel two (2) represented only nitrite. The absorbance of the color product was measured by photometric detectors with 540-nm filters. This method combines high reproducibility of sample introduction via flow injection and sensitivity of spectrophotometric detection. The detection limit is 25 nM for both nitrite and nitrate. The chemistry manifolds are constructed of Teflon tubing which, along with a low-pressure Flowfit connector system, provides for low maintenance, ease of use, and high sample throughput. We demonstrated that the system can be used for the determination of both nitrate and nitrite in a variety of biological samples as well as a comparison of the results from this system and the HPLC system.

Animals↗