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PubMed2006

Determination of trace hydroxyl radicals by flow injection spectrofluorometry and its analytical application.

On the basis of the fluorescence increase of the reaction of ninhydrin with hydroxyl radicals, a new method for the determination of trace amounts of hydroxyl radicals by flow injection spectrofluorometry is presented. The introduction of flow injection analysis brought better reproducibility and avoided the effect of oxygen and other substances in the environment on the reaction of ninhydrin with hydroxyl radicals. Under optimum experimental conditions, the hydroxylated product of ninhydrin had excitation and emission maxima at 300 and 406 nm, respectively. The linear range was 2.60 x 10(-7) to 4.00 x 10(-5) M, and the limit of detection was 7.91 x 10(-8) M. A high analysis rate of 22 samples per hour was obtained. The proposed method has been applied successfully to the determination of scavenging effects of thiourea and vitamin C on hydroxyl radicals as well as to the evaluation of antioxidant capacities of some natural food.

Cobalt↗
PubMed1997

Chemiluminescence flow system for vanadium(v) with immobilized reagents.

A chemiluminescence (CL)-based system for vanadium(v) combined with flow-injection analysis is described. The analytical regents, luminol and hexacyanoferrate(II), were both immobilized on an anion-exchange resin column. When a volume of phosphoric acid was passed through the column, these two reagents were eluted from the resin and then mixed with a vanadium(v) stream under acidic conditions. By means of the fast oxidation reaction between vanadium(v) and hexacyanoferrate(II), vanadium(IV) and hexacyanoferrate (III) were generated, both of which catalyzed the oxidation of luminol by dissolved oxygen in aqueous alkaline solution to produce CL. The CL emission intensity was correlated with the standard vanadium (v) concentration in the range from 1.0 x 10(-2) to 10 micrograms cm-3, and the detection limit was 5.4 x 10(-3) micrograms cm-3 vanadium (v). Interfering metal ions co-existing in sample solutions could be effectively separated on-line by a cation-exchange column placed upstream. A complete analysis, including sampling and washing, could be performed in 1 min with a relative standard deviation of less than 5%. The system was stable for over 100 analyses and was applied successfully to the determination of vanadium in geochemical and human hair samples.

Flow Injection Analysis↗
PubMed1997

Methodological aspects of glucose monitoring with a slow continuous subcutaneous and intravenous ultrafiltration system in rats.

A method for the continuous ultrafiltration of venous blood or subcutaneous fluid is demonstrated with glucose monitoring in the living rat. Ultrafiltrate was withdrawn at a constant flow rate of approximately 100 nl/min. Glucose content of the ultrafiltrates was electrochemically determined with a flow injection analysis method and a bi-enzyme reactor. After glucose loading, the time course of glucose in the ultrafiltrate from the jugular vein was virtually identical, whereas that from the subcutaneous compartment was attenuated and the peaks blunted as compared to glucose levels in concomitantly assayed arterial blood. Our study demonstrates the potential of low rate ultrafiltration for monitoring metabolism with biosensor technology in vivo.

Animals↗
PubMed2005

In situ generation of Co(II) by use of a solid-phase reactor in an FIA assembly for the spectrophotometric determination of penicillamine.

A flow injection analysis (FIA) manifold for the determination of penicillamine in pharmaceutical preparations is proposed. The manifold includes a solid-phase reactor for the in situ production of the derivatizing reagent, Co(II) ion, which forms a coloured complex with penicillamine in an alkaline medium. The reactor is prepared by natural immobilization of cobalt carbonate on a polymer matrix, which endows it with a high mechanical and microbiological stability. The cobalt released by passage of a 5 x 10(-4) mol l(-1) sulphuric acid stream at a flow-rate of 2.3 ml min(-1) is merged with a volume of 314 microl of sample containing penicillamine in ammonium-ammonia buffer at pH 9.5 to measure the absorbance at 360 nm. Beer's law is obeyed over the penicillamine concentration range 5-60 mg l(-1). The limit of detection (LOD) of the method is 1 mg l(-1) and its throughput 70 samples h(-1).

Cobalt↗
PubMed2006

Synthesis of bilirubin imprinted polymer thin film for the continuous detection of bilirubin in an MIP/QCM/FIA system.

A bilirubin imprinted polymer (BIP) was coated on a thiol pretreated Au electrode on a quartz crystal microbalance (QCM) chip. The BIP thin film was synthesized using 4-vinylpyridine (4-Vpy) as the monomer, divinylbenzene (DVB) as the cross-linker, and benzophenone as the initiator. By using a photo-graft surface polymerization technique with irradiation by ultra-violet (UV) light, a thin BIP film was prepared, from which a biomimetic sensor for the detection of bilirubin was developed. The sensor was able to discriminate bilirubin in solution owing to the specific binding of the imprinted sites. The BIP/QCM chip has been repeatedly used for more than 7 months in many continuous experiments. The detection signal of bilirubin from the BIP thin film/QCM was compared with the non-BIP thin film/QCM. Biliverdin, an analogue of bilirubin, was used for comparison. The analogue comparison confirmed the binding specificity of the BIP film toward bilirubin. The selectivity can be as high as 31.2. The effect of pH on the detection of bilirubin is also discussed. With proper solvent for elution and recovery, flow injection analysis (FIA) could be applied to the system. The performance of the BIP/QCM chip was evaluated. A linear calibration of the bilirubin concentration with respect to the frequency shift was successfully obtained. The reproducibility of measurements from the same BIP/QCM chip was confirmed. In addition, repeatability of detection was also confirmed from different BIP/QCM chips. In conclusion, a combined BIP thin film/QCM/FIA method was successfully established for the detection of bilirubin concentration using a molecularly imprinted film.

Bilirubin↗
PubMed2000

Electrochemical sensor detecting free sulfhydryl groups: evaluation of milk heat treatment.

We describe a new and rapid method for the evaluation of reactive sulfhydryl groups in whey proteins obtained after precipitation of casein by acetic acid at pH 4.6. The procedure is based on the use of a wire tungsten electrode operating at -0.2 V versus saturated calomel electrode in flow injection analysis. The method was applied to raw milks and to commercial pasteurized and UHT milks. Results showed that the tungsten electrode constituted a robust amperometric sensor that could be used to differentiate milks that underwent different heat treatments. The decrease of thiol content in the whey proteins from samples was in agreement with the whey protein content found by HPLC. The procedure is suitable for on-line quality control of heat-treated milks.

Animals↗
PubMed2003

Evaluation and application of conducting polymer entrapment on quartz crystal microbalance in flow injection immunoassay.

An immunosensor employing conducting polymer entrapment (CPE) method to immobilize immuno-protein on the quartz crystal microbalance (QCM) for clinical flow-injection-analysis (FIA) purpose was exploited. By comparing the CPE approach and the conventional physical immobilization (PI) method, a frequency shift of the former was 47% higher than that of the latter when measuring at 0.5 mg/ml human serum albumin antibody concentration. This implied that CPE was a feasible approach for developing QCM-FIA process. An immunoassay of anti-pseudorabies virus antibody in mouse sera further exemplified its practical potential in diagnostic implication.

Animals↗
PubMedDate not supplied

Flow-injection with enhanced chemiluminescence detection of ofloxacin in human plasma.

A simple, rapid and sensitive method for the determination of ofloxacin in plasma has been developed based on flow-injection analysis with enhanced chemiluminescence (CL) detection. This method employs the CL reaction of cerium(IV) and sulphite sensitized by a Tb(III)-ofloxacin complex. Plasma samples were deproteinized with acetonitrile before analysis. Under optimal conditions, the method allowed the quantitative analysis of ofloxacin in plasma over the range of 22.5-900 ng/mL with a detection limit of 7.5 ng/mL (3sigma). The intra-day and inter-day precision was < 3.0% (n = 11) and < 7.0% (n = 3), respectively. The method was a convenient tool, rapid (20 s/analysis) and well suited for pharmacokinetics studies in which thousands of samples must be analysed daily. It was applied to pharmacokinetics studies of ofloxacin.

Acids↗
PubMed2003

Simultaneous determination of the monoamine neurotransmitters and glucose in rat brain by microdialysis sampling coupled with liquid chromatography-dual electrochemical detector.

A new type of chemically modified electrode based ring-disk electrode as the dual electrochemical detector (DECD) for high-performance liquid chromatography (HPLC) to simultaneously determine the monoamine neurotransmitters and glucose is described. The ring electrode was modified with an ion-exchange polymer-overoxidized polypyrrole (OPPy) and the disk electrode was modified with nano Au colloid and glucose oxidase (GOD). The electrochemical behaviors of dopamine (DA) and ascorbic acid (AA) at the OPPy chemically modified electrode (CME) were investigated by differential pulse voltammetry (DPV). It was found that the CME could permeate dopamine cations and repelled the ascorbate anions, which could be used to determine the monoamine neurotransmitters and avoid the interference of AA. The electrochemical behavior of glucose at the Nafion/GOD-Au colloid/GC CME was investigated by amperometry and flow injection analysis (FIA). It was found that the sensitivity of the CME increased apparently in determination of glucose. In order to obtain better separation and current responses of the analytes in HPLC-DECD, several operational parameters have been investigated. Under the optimum conditions, the method showed good stability and reproducibility. The application of this method coupled with microdialysis sampling for in vivo simultaneous determination of monoamine neurotransmitters and glucose in rat brain was satisfactory.

Animals↗
PubMed1992

Nonimmunological assay of urinary albumin based on laser-induced fluorescence.

We describe the first nonimmunological assay of albumin in urine with a detection limit of 1 mg/L. The method is simple, rapid, and accurate. It is based on the probe Albumin Blue 670, which becomes highly fluorescent on binding to albumin. An inexpensive diode laser was used as the light source for measurement of laser-induced fluorescence. The assay was coupled to a flow-injection analysis system capable of running 20 samples per hour. The working range was 1-100 mg/L, which covered albumin concentrations found in nonpathological urine and in urine with slightly increased albumin. This range makes prediction of nephropathy possible at an early stage. Other serum proteins and hemoglobin do not interfere. The coefficients of variation were < 4% and < 7% within one day and from day to day, respectively. A correlation coefficient of 0.990 (n = 100) was obtained for comparison with the Behring nephelometric assay.

Albuminuria↗
PubMedDate not supplied

A flow injection chemiluminescence method using Cr(III) as a catalyst for determining hydrogen peroxide. Application to H(2)O(2) determination in cultures of microalgae.

Flow injection analysis has been applied to the determination of hydrogen peroxide produced by some different species of microalgae. The method is based on the luminol-H(2)O(2) chemiluminescence reaction using Cr(III) as a catalyst. Optimum experimental conditions for the method have been studied and trace amounts of hydrogen peroxide determined with detection limits of 4 10(-8) mol/L. The method using Cr(III) was compared with that using horseradish peroxidase as the catalyst.

Catalysis↗
PubMed2002

On-line complexation/cloud point preconcentration for the sensitive determination of dysprosium in urine by flow injection inductively coupled plasma-optical emission spectrometry.

An on-line dysprosium preconcentration and determination system based on the hyphenation of cloud point extraction (CPE) to flow injection analysis (FIA) associated with ICP-OES was studied. For the preconcentration of dysprosium, a Dy(III)-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol complex was formed on-line at pH 9.22 in the presence of nonionic micelles of PONPE-7.5. The micellar system containing the complex was thermostated at 30 degrees C in order to promote phase separation, and the surfactant-rich phase was retained in a microcolumn packed with cotton at pH 9.2. The surfactant-rich phase was eluted with 4 mol L(-1) nitric acid at a flow rate of 1.5 mL min(-1), directly in the nebulizer of the plasma. An enhancement factor of 50 was obtained for the preconcentration of 50 mL of sample solution. The detection limit value for the preconcentration of 50 mL of aqueous solution of Dy was 0.03 microg L(-1). The precision for 10 replicate determinations at the 2.0 microg L(-1)Dy level was 2.2% relative standard deviation (RSD), calculated from the peak heights obtained. The calibration graph using the preconcentration system for dysprosium was linear with a correlation coefficient of 0.9994 at levels near the detection limits up to at least 100 microg L(-1). The method was successfully applied to the determination of dysprosium in urine.

Artifacts↗
PubMed2003

Flow-injection determination of ornidazole by chemiluminescence detection based on a luminol-ferricyanide reaction.

A flow-injection analysis (FIA) with a chemiluminescence detection method was developed for the determination of ornidazole based on the inhibition intensity of chemiluminescence from the luminol-ferricyanide system. Under the condition of 1.0 x 10(-3) mol/L luminol and 5.0 x 10(-6) mol/L potassium ferricyanide, the response to the concentration of omidazole is linear from 0.2 microg ml(-1) to 10 microg ml(-1), and a detection limit of 0.05 microg ml(-1) can be obtained. This method has been successfully applied to the determination of omidazole in pharmaceutical preparations.

Anti-Infective Agents↗
PubMed2006

Biosensor measurements of polar phenolics for the assessment of the bitterness and pungency of virgin olive oil.

Bitterness and pungency, sensory quality attributes of virgin olive oil, are related to the presence of phenolic compounds. Fast and reliable alternatives for the evaluation of sensory attributes and phenolic content are desirable, as sensory and traditional analytical methods are time-consuming and expensive. In this study, two amperometric enzyme-based biosensors (employing tyrosinase or peroxidase) for rapid measurement of polar phenolics of olive oil were tested. The biosensor was constructed using disposable screen-printed carbon electrodes with the enzyme as biorecognition element. The sensor was coupled with a simple extraction procedure and optimized for use in flow injection analysis. The performance of the biosensor was assessed by measuring a set of virgin olive oils and comparing the results with data obtained by the reference HPLC method and sensory scores. The correlations between the tyrosinase- and peroxidase-based biosensors and phenolic content in the samples were high (r = 0.82 and 0.87, respectively), which, together with a good repeatability (rsd = 6%), suggests that these biosensors may represent a promising tool in the analysis of the total content of phenolics in virgin olive oils. The correlation with sensory quality attributes of virgin olive oil was lower, which illustrates the complexity of sensory perception. The two biosensors possessed different specificities toward different groups of phenolics, affecting bitterness and pungency prediction. The peroxidase-based biosensor showed a significant correlation (r = 0.66) with pungency.

Biosensing Techniques↗
PubMed1999

Determination of organophosphate and carbamate pesticides in spiked samples of tap water and fruit juices by a biosensor with photothermal detection.

The determination of organophosphate (paraoxon, chlorpyrifos, diazinon) and carbamate (carbaryl, carbofuran) pesticides in spiked drinking water and fruit juices was carried out using a photothermal biosensor. The biosensor consists of a cartridge containing immobilised enzyme acetylcholinesterase (AChE) placed in a flow-injection analysis (FIA) manifold and a photothermal detector based on thermal lens spectrometry. With this approach, 0.2 ng/ml of paraoxon can be detected in less than 15 min. Limits of detection for other organophosphate (chlorpyrifos, diazinon) and carbamate (carbaryl, carbofuran) pesticides varied, depending on their antiacetylcholinesterase (AntiAChE) toxicity, from 1 ng/ml to 4 microg/ml. The biosensor was used for the direct detection of pesticides in spiked tap water and fruit juices without any pretreatment steps. In these cases, the LOD3sigma of 1.5, 2.8 and 4 ng/ml paraoxon in tap water, orange juice and apple juice were obtained, respectively.

Beverages↗
PubMed1993

Automated enzymatic determination of sodium in serum.

An automated method based on the principles of flow-injection analysis is proposed for the enzymatic determination of sodium ion in serum. The method relies on the activation of beta-galactosidase by the analyte. The features of the proposed method include linear range between 1 and 1700 mumol/L, a sampling rate of 50 samples/h, a sample volume of 50 microL, and the absence of interferences from species usually present in serum. The results obtained were consistent with those provided by widely used methods such as those based on flame spectrometry and direct potentiometry with ion-selective electrodes.

Flow Injection Analysis↗
PubMed2000

Clearance of increased serum advanced glycosylation end products in patients with end stage renal disease by hemodialysis.

AIM: To study the effect of removal of hemodialysis using acetate membrane on serum advanced glycosylation end products (AGEP) in 36 patients (59.1 +/- 1.6) with end stage renal disease (ESRD). METHODS: Serum AGEP levels were determined with quantitative fluorescence spectrometry, flow injection analysis (FIA), and competitive enzyme-linked immunosorbent assay (ELISA) using a polyclonal antibody directed against AGEP. RESULTS: The serum AGEP levels in patients with ESRD quantified by fluorescence spectrometry, FIA, and ELISA were higher than those in controls [(25 +/- 5) vs (7.5 +/- 1.5), (6.1 +/- 1.8) vs (1.4 +/- 0.5), and (37 +/- 20) vs (9 +/- 10) kU/L, respectively] and markedly reduced to [(22 +/- 6), (4.2 +/- 1.4), and (19 +/- 14) kU/L, respectively] after hemodialysis. CONCLUSION: Increased serum AGEP levels in the circulation of patients with ESRD were reduced effectively by hemodialysis.

Aged↗
PubMed1995

Evanescent fluorobiosensor for the detection of polyaromatic hydrocarbon based on DNA intercalation.

A flow-injection analysis (FIA) system coupled with an evanescent wave (EW) biosensor employing total internal reflection of fluorescence radiation (TIRF) for the detection of polyaromatic hydrocarbon that intercalates into DNA is reported. A highly fluorescent intercalator, "ethidium bromide," has been used as the reference compound for detection. The EW biosensor was developed according to the procedure described earlier (1,2). Data on the analysis of Naphthalene, 3-methylcholanthrene, 7,12-dimethylbenz(a)anthracene, 1,2-benzanthracene, and some standard reference materials supplied by the National Institute of Standards and Technology are reported. The relative ability of the polyaromatic hydrocarbon to displace ethidium bromide, based on the relative binding ratio, is found to be on the order of 7,12-dimethylbenz[a]anthracene > 3-methylcholanthrene > 1,2-benzanthracene > napthalene.

9,10-Dimethyl-1,2-benzanthracene↗