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Multi-element analysis of saline matrices by inductively coupled plasma mass spectrometry following on-line preconcentration using a knotted reactor.

A versatile preconcentration system for trace element determination by ICPMS was developed. It is composed of a commercial flow injection analysis system (FIAS) retrofitted with a home-made control unit containing three solenoid valves and working concomitantly with the FIAS, permitting selection and segmentation of sample, reagent, washing solution and elution flow. The knotted reactor used had a length of 200 cm and was made from 0.05 cm i.d. PTFE tubing. The method applies ammonium 1-pyrrolidinedithiocarbamate (APDC) as complexant, allowing the preconcentration and quantitative multi-element determination of Cu, Ni, V, Co, Nb, Mo, In, Sb and Bi. Ethanol and 4-methyl-2-pentanone were tested as eluents. Recovery tests using complex matrices and spike concentrations of 200 ng L(-1) showed typical values in the range of 90% to 110%. Relative standard deviations were < 7% for elution with ethanol and < 5% with methyl isobutyl ketone. For simulated freshwater samples using 4-methyl-2-pentanone as an eluent, a sample loading rate of 5.0 ml min(-1), and a preconcentration time of 60 s, detection limits (ng L(-1)) were in the range of 0.02 (Bi) to 30 (Cu). Under these conditions, analytical frequency was about 15 samples per hour. The feasibility of the method was demonstrated by the succesful analysis of wastewater and seawater certified reference materials.

Journal Article↗

Hydrolysis of triphosphate from detergents in a rural waste water system.

The concentrations of detergent phosphates in raw sewage entering a small, predominantly domestic waste water treatment facility were determined using an ion chromatographic-flow injection analysis technique. Hourly loads of detergent phosphates were measured between 0600 and 2300 hrs (the major flow period in the plant) on days of both low and high phosphorus loads. The calculated loads of detergent phosphorus entering the plant on low and high load days were 260 g P/day and 350 g P/day, respectively. The half-life of detergent phosphates (triphosphate) in waste waters was measured to be 7.3 hours at 15 degrees C and 3.0 h at 20 degrees C. The major factor contributing to triphosphate degradation in waste water was shown to be biological in nature, with the most likely mechanism being enzymatic hydrolysis.

Calcium↗

Electroanalytical study of fluvoxamine.

Fluvoxamine (FVX) can be reduced at a mercury-drop electrode, with a maximum peak current intensity being obtained at a potential of -0.7 V vs. Ag/AgCl, in an aqueous electrolyte solution of pH 2. The compound was determined in a pharmaceutical product and in spiked human serum by square-wave adsorptive-stripping voltammetry (SWAdSV) after accumulation at the electrode surface, under batch conditions. Because the presence of dissolved oxygen did not interfere significantly with the analysis, it was also possible to determine FVX in the pharmaceutical product by use of a flow-injection analysis (FIA) system with SWAdSV detection. The methods developed were validated and successfully applied to the quantification of FVX in a pharmaceutical product. Recoveries between 76 and 89% were obtained in serum analysis. The FIA-SWAdSV method enabled analysis of up to 120 samples per hour at reduced cost, implying the possibility of competing with the chromatographic methods usually used for this analysis.

Electrochemistry↗

Free flow electrophoresis device for continuous on-line separation in analytical systems. An application in biochemical detection.

A free flow electrophoresis (FFE) device was developed for continuous electrophoretic separation of charged compounds and implemented in a continuous flow biochemical detection (BCD) system. These continuous separation characteristics make FFE well suitable for online implementation in a chromatographic or flow injection analysis system, in which an additional separation step of charged compounds is desired. In a heterogeneous biochemical flow assay for the determination of biotin, an analyte zone reacts with an excess of an affinity protein. Subsequently, the free binding sites of the affinity protein react with an excess of fluorescein-labeled ligand. Free and affinity protein-bound label are separated on the FFE device prior to fluorescence detection of the separated fractions. Biotin and streptavidin were chosen as, respectively, model ligand and affinity protein. Since all the compounds that are involved possess different electrophoretic properties, quantitative analysis is performed after completely separating the fluorescent affinity complex and labeled biotin in the FFE device within 2 min. Since the device is optically transparent, the separated zones can be detected in the separation compartment, using laser-induced fluorescence. The applicability of the BCD-FFE system in combination with a HPLC separation is demonstrated in the bioanalysis of biotin in human urine at the micromole per liter level.

Biotin↗

Comparison of different flow injection approaches to the automatic determination of enzymatic activity.

Three configurations based on the principles behind flow injection analysis (FIA) are proposed for the automatic determination of enzymatic activity. The proposed approaches are normal, stopped-flow and open-closed FIA. The comparative study of the methods developed from these approaches allows the establishment of the scope of the application of each, with inherent advantages and disadvantages.

Enzymes↗

Compensation voltage shifting in high-field asymmetric waveform ion mobility spectrometry-mass spectrometry.

The separation and ion focusing properties of High-Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS) depend on desolvated ions entering the device, leading to a compound-specific, reproducible compensation voltage (CV) for each ion. This study shows that the conditions identified for stable spray and satisfactory ion desolvation in normal electrospray ionization mass spectrometry (ESI-MS) operation might significantly differ from those required for FAIMS-MS. In a typical setup with high-flow electrospray conditions, ions could be incompletely desolvated, resulting in the formation of unidentified clusters with differing behavior in a FAIMS environment. This causes compound-specific shifts of as much as 10 V in CV values when the mobile phase composition and/or flow rate are varied. The shifts diminish and finally disappear when the flow rate of methanol, used as mobile phase, is reduced to 40 microL/min and that of acetonitrile to 20 microL/min. The reproducibility of the observed CV was determined by scanning the CV while infusing a five-component mixture into a 400 microL/min flow of methanol or 50:50 acetonitrile/water. The relative standard deviation (RSD) for these multiple scans ranged from 0.7% to 6%. Therefore, under a constant set of experimental parameters, the CV does not shift appreciably. These observations have an impact on method development strategies. High flow rates can be used with the FAIMS device, since the CV values are reproducible, but it is likely that clusters are forming. Therefore, CV scans should be performed under conditions which mimic the chromatographic elution or flow injection analysis conditions, including matrix composition, to minimize errors in CV determination. An alternative approach is to determine the liquid flow rate at which the CV becomes compound-specific and to split the mobile phase stream accordingly. These experimental results may be specific to the setup used for this study and may not be directly applicable to other instrument FAIMS devices.

Journal Article↗

Analysis of aprotinin on the mean arterial pressure, carotid artery blood flow, and hindlimb vascular resistance in the live rat, and pulmonary vascular resistance in the isolated perfused rat lung.

The effects of aprotinin on mean arterial pressure, carotid artery blood flow, pulmonary vascular resistance, and systemic vascular resistance have not been well documented. Therefore, the responses of aprotinin on the mean arterial pressure, carotid artery blood flow and the changes in pulmonary and hindlimb vascular resistances were investigated in the rat using ultrasonic flow probe analysis, and in two isolated vascular bed preparations. In studies on cardiac output using ultrasonic flow analysis, injections of aprotinin, in doses of 70-7000 KIU intravenously (iv), resulted in no changes in carotid artery ultrasonic flow, suggesting no changes in cardiac output. In isolated blood-perfused lung studies under conditions of controlled pulmonary blood flow, aprotinin, in doses of 7-7000 KIU intra-arterially (ia), caused no significant changes in pulmonary arterial perfusion pressure. Aprotinin, in doses of 7-240 KIU, was injected iv into the hindquarters perfusion circuit, and hindquarters arterial perfusion pressure did not change. Additionally, in the hindquarters perfusion preparation, aprotinin, in doses of 70-7000 KIU, ia resulted in no changes in mean arterial pressure. The present data demonstrate aprotinin has no significant response in MAP, carotid artery blood flow, pulmonary vascular resistance, or hindlimb vascular resistance in the rat.

Animals↗

Flow-through micro sensor using immobilized peroxidase with chemiluminometric FIA system for determining hydrogen peroxide.

A micromachined flow cell (overall size; 25 x 25 x 1 mm3) was designed for the fast determination of hydrogen peroxide, based on a luminol-H2O2 chemiluminescence reaction catalyzed by immobilized peroxidase (POD). The flow cell consisted of a sandwich of anisotropically etched silicon and glass chips and contained a spiral channel (20 turns, 50 cm long, 150 microm wide, 20 microm depth, channel volume 1.4 microl) and two holes (1 mm diameter). POD was covalently immobilized with 3-(trimethoxysilyl)propyldietylenetriamine and glutaraldehyde on the inner surface of the channel. The chip was placed in front of a window of a photomultiplier tube and used as a flow cell in a single-line flow-injection analysis system using a luminol solution as a carrier solution. The sample volume for one measurement was 0.2 microl. The maximal sampling rate was 315 h(-1) at a carrier solution flow rate of 10 microl min(-1). A calibration graph for H2O2 was linear for 5 nM - 5 microM; the detection limit (signal-to-noise = 3) was 1 nM (7 fg in 0.2 microl injection). The H2O2 concentration in rainwater was determined using this sensor system.

Calibration↗

A continuous film-recirculable drop gas-liquid equilibration device. Measurement of trace gaseous ammonia

A miniature gas-liquid equilibrator or a gas collector, intended as a low-volume interface between a soluble gaseous sample and a liquid phase analyzer or between a liquid phase sample and a detector designed for use with gas samples, is described. This paper addresses the application of the device for the measurement of trace atmospheric ammonia. Gas collection occurs solely by diffusive sampling such that aerosol particles are not collected. The device essentially consists of a tube surrounded externally by a jacket. Gas flows through the jacket and contacts a liquid film flowing on the surface of the tube. The flowing film forms a drop at the tube terminus and is aspirated off through the inner bore of the tube. The collected analyte can be (a) directly sent to an analysis system or (b) preconcentrated on a suitable stationary phase; the preconcentrator effluent can be recycled, if desired. With a fluorometric flow injection analysis system harnessed to measure ammonia with such a collector, the limit of detection (LOD, S/N = 3) for a sample drawn for 18 min at 200 mL/min was 4.5 parts per trillion by volume, with the linear range extending up to 30 parts per billion.

Journal Article↗

Determination of trace sodium in the water-steam system of power plants using an FIA/ISE method with an automatic penetration and alkalization apparatus.

A new method for the rapid determination of trace-level sodium ion based on flow-injection analysis (FIA) and an ion selective electrode (ISE) is proposed. Various effects on the sensitivity of the method such as the flow rate and alkalinity of the carrier, sampling volume, temperature, length of reaction coil, length and thickness of alkalization tube, concentration of the alkalizing reagent etc. were investigated. The optimum conditions were ascertained. The method showed good linearity in the concentration ranges of 0.5 - 10 microg L(-1) and 10 - 100 microg L(-1), and could deal with 40 - 50 samples per hour. The consumption of the sample is only 0.80 mL per time. The relative standard deviation was 0.55%, and the recovery range was 98 - 103%. By designing a hermetically sealed single-line FIA-ISE manifold, a problem removing the interference of sodium ion from air could be solved, and automatic alkalization of the sample was realized. This method has been used for successfully determining the trace-level sodium ions in the water-steam system at fossil power plants.

Journal Article↗

Selectivity of electrospray response in small polymer analysis by mass spectrometry.

The selectivity of electrospray was explored for a small poly(ethylene)glycol by comparing the oligomer response obtained from direct polymer introduction in flow injection analysis with the signal recorded in high-performance liquid chromatography/mass spectrometry (HPLC/MS). When the oligomer mixture was ionized, a suppression effect was measured for all but the more hydrophobic congeners for which the response was enhanced. This result would reflect the influence of electrospray droplet chemical composition on the equilibrium partitioning coefficient in Enke's model. On average, the electrospray selectivity observed for the studied poly(ethylene)glycol did not affect the molecular weight distribution parameters as response for the most concentrated oligomers was suppressed to the same extent while over-expressed largest congeners had a low contribution to the total polymer sample.

Journal Article↗

On-line determination of glucose in biotechnological processes: comparison between FIA and an in situ enzyme electrode.

Two different analysis techniques for on-line monitoring of glucose in biotechnological processes have been tested: an in situ enzyme electrode and a flow injection analysis system (FIA). The measuring ranges, detection limits, response times and the reliabilities of each system have been compared during monitoring of batch and continuous cultures of Saccharomyces cerevisiae.

Computers↗

An integrated silicon thermophile as biosensor for the thermal monitoring of glucose, urea and penicillin.

A new kind of calorimetric biosensor for the measurement of the heat (molar enthalpy change) of enzymatic reactions is presented. The device operates according to the Seebeck effect, the same principle on which thermocouples are based. The thermopile used in this work consists of an array of p-type silicon/aluminium strips integrated on a thin silicon membrane (5 microns). Its sensitivity is about 1 V output voltage per watt of heating power, corresponding to a temperature resolution in the order of 10(-5) K and a heating power resolution of some tenths of a mu W in the flow system used. Furthermore, this performance is obtained without any control of external temperature because of the high common-mode thermal noise rejection ratio of the thermopile. The universal technique of calorimetry combined with the specificity of biochemical reactions makes this biosensor very versatile, with a broad range of possible applications. Glucose oxidase together with catalase for the determination of glucose, urease and penicillinase for the monitoring of urea and penicillin G, respectively, were immobilized directly onto the back side of the thermopile. The sensor was operated in conjunction with flow injection analysis which, in addition to its traditional advantages, allows preconditioning of the samples. Thus, artefacts due to mixing effects were suppressed and interference caused by differences in ionic strength between sample and carrier was strongly decreased. Detection limits between 1 and 2 mM were reported in the flow injection conditions described.

Biosensing Techniques↗

Online concentration by field-amplified sample injection in acidic buffer for analysis of fangchinoline and tetrandrine in herbal medicine by flow injection-micellar electrokinetic capillary chromatography.

A novel, rapid, and continuous online concentration approach based on field-amplified sample injection for the analysis of fangchinoline and tetrandrine was developed in this paper by combination of flow injection-MEKC. The BGE used was a solution composed of 75 mM H3PO4-triethylamine-2.5% v/v polyoxyethylene sorbitan monolaurate-20% v/v methanol buffer (pH* 5.0). The analytes prepared in 50% v/v aqueous ethanol were used as the test analytes. Sample was injected electrokinetically between plugs of water. When the cations reached the boundary between the water plug and BGE, they slowed down and became concentrated. Thereafter, MEKC was initiated for the separation. This results in 6.8-8.9-fold improvement in concentration sensitivity relative to conventional CE methods. The separation could be achieved within 10 min and sample throughput rate can reach up to 50/h. The repeatability (defined as RSD) was 4.8, 4.4% with peak height evaluation and 3.6, 0.94% with peak area evaluation for TET and FAN, respectively.

Alkaloids↗

Solid-state PVC flow-through benzoate electrode.

A general construction procedure for conventional shape ion-selective electrodes based on a conductive support prepared with a mixture of a non-conductive epoxy and graphite was used in the preparation of a flow-through benzoate electrode to be used as potentiometric detector in flow injection analysis manifolds. The membranes were prepared from tetraoctylammonium benzoate in o-nitrophenyloctylether immobilized on PVC. The results of the assessment of the tubular electrodes behaviour in low dispersion manifolds against that of conventional electrodes with the same membrane are reported. Data obtained with a double-channel flow injection manifold incorporating these tubular detectors for benzoate determinations in several commercially available pharmaceutical preparations are also presented.

Benzoates↗

An analysis of avidin, biotin and their interaction at attomole levels by voltammetric and chromatographic techniques.

The electroanalytical determination of avidin in solution, in a carbon paste, and in a transgenic maize extract was performed in acidic medium at a carbon paste electrode (CPE). The oxidative voltammetric signal resulting from the presence of tyrosine and tryptophan in avidin was observed using square-wave voltammetry. The process could be used to determine avidin concentrations up to 3 fM (100 amol in 3 microl drop) in solution, 700 fM (174 fmol in 250 microl solution) in an avidin-modified electrode, and 174 nM in a maize seed extract. In the case of the avidin-modified CPE, several parameters were studied in order to optimize the measurements, such as electrode accumulation time, composition of the avidin-modified CPE, and the elution time of avidin. In addition, the avidin-modified electrode was used to detect biotin in solution (the detection limit was 7.6 pmol in a 6 mul drop) and to detect biotin in a pharmaceutical drug after various solvent extraction procedures. Comparable studies for the detection of biotin were developed using HPLC with diode array detection (HPLC-DAD) and flow injection analysis with electrochemical detection, which allowed biotin to be detected at levels as low as 614 pM and 6.6 nM, respectively. The effects of applied potential, acetonitrile content, and flow rate of the mobile phase on the FIA-ED signal were also studied.

Avidin↗

Sensitive determination of G-protein-coupled receptor binding ligands by solid phase extraction-electrospray ionization-mass spectrometry.

High affinity Histamine H2-receptor binding ligands were assayed by automated solid phase extraction (SPE) coupled via electrospray ionization with a Quadrupole-Time-of-Flight mass spectrometer (Q-ToF-MS). The mass spectrometric behavior of these analytes was tested in aqueous solutions with several (nine) volatile salts, in different pH, and with various methanol contents. Out of the high amount of available ligands, three fluorescent-labeled molecules (5706, 5707, and 5708) were studied in detail. The limits of detection (LODs) for all three compounds obtained in mass spectrometric detection was 1 fmol (absolute) in continuous flow and FIA (flow injection analysis) measurements. The results obtained with FIA-fluorescence detection gave LODs a factor 10-100 times higher. A systematic investigation of sample solving conditions, loading flow conditions, and elution flow conditions made the automated SPE-MS coupling efficient. Ideally, the ligands were dissolved in MeOH-25 mM phosphate buffer (30:70 v/v; pH 11), the SPE loading flow comprised MeOH-25 mM phosphate buffer (30:70 v/v; pH 11) and the SPE elution flow contained MeOH-100 mM ammonium formate solution (90:10 v/v; pH 3). Using this method on a C18-modified silica cartridge (C18, 5 microm, 100 A, 300 microm i.d. x 5 mm, LC Packings) assures high recovery and achieved LODs for all three compounds of 5 fmol (absolute). As an absolute amount of ligands specifically bound on H2-receptors in biochemical experiments is, as will be published elsewhere, between 10 and 100 fmol, the SPE-MS method for the basic compounds can be directly applied for these Histamine H2-receptors.

Buffers↗

Spectrophotometric bioanalytical flow-injection system for control of hemodialysis treatment.

A spectrophotometric flow-injection analysis (FIA) system for monitoring clinical hemodialysis is demonstrated. The role of a dialysate urea detector incorporated in this bioanalytical system is played by an optical flow-through biosensor based on Prussian Blue film with chemically linked urease forming a monomolecular layer of the enzyme. This pH-enzyme optode-FIA system is useful for the selective determination of post-dialysate urea in the range of concentration corresponding to its level in real clinical samples (2-16 mmol l(-1)). This bioanalytical system allows the analysis of about 15 samples of spent dialysate per hour. The operational and storage stabilities of the applied biosensor are longer than 2 weeks and 2 months, respectively. Clinical evaluation of the bioanalytical system was performed.

Biosensing Techniques↗