A novel microfluidic flow-injection analysis device with fluorescence detection for cation sensing. Application to potassium

A microfabricated device has been developed for fluorimetric detection of potassium ions without previous separation. It is based on use of a fluorescent molecular sensor, calix–bodipy, specially designed to be sensitive to and selective for the target ion. The device is essentially made of a Y-shape microchannel moulded in PDMS fixed on a glass substrate. A passive mixer is used for mixing the reactant and the analyte. The optical detection arrangement uses two optical fibres, one for excitation by a light-emitting diode, the other for collection of the fluorescence. This system enabled the flow-injection analysis of the concentration of potassium ions in aqueous solutions with a detection limit of 0.5 mmol L⁻¹ and without interference with sodium ions. A calibration plot was constructed using potassium standard solutions in the range 0–16 mmol L⁻¹, and was used for the determination of the potassium content of a pharmaceutical pill. Figure Photography of the microfluidic channel showing the ridges in the PDMS substrate at the top of the channel     

Speciation analysis of inorganic arsenic by a multisyringe flow injection system with hydride generation-atomic fluorescence spectrometric detection

In this study, a new technique by hydride generation-atomic fluorescence spectrometry (HG-AFS) for determination and speciation of inorganic arsenic using multisyringe flow injection analysis (MSFIA) is reported. The hydride (arsine) was generated by injecting precise known volumes of sample, a reducing sodium tetrahydroborate solution (0.2%), hydrochloric acid (6 M) and a pre-reducing solution (potassium iodide 10% and ascorbic acid 0.2%) to the system using a multisyringe burette coupled with one multi-port selection valve. This solution is used to pre-reduce As(V) to As(III), when the task is to speciate As(III) and As(V). As(V) is determined by the difference between total inorganic arsenic and As(III). The reagents are dispensed into a gas-liquid separation cell. An argon flow delivers the arsine into the flame of an atomic fluorescence spectrometer. A hydrogen flow has been used to support the flame. Nitrogen has been employed as a drier gas (Fig. 1).Several variables such as sample and reagents volumes, flow rates and reagent concentrations were investigated in detail. A linear calibration graph was obtained for arsenic determination between 0.1 and 3 μg l⁻¹. The detection limit of the proposed technique (3σ_b/S) was 0.05 μg l⁻¹. The relative standard deviation (R.S.D.) of As at 1 μg l⁻¹ was 4.4 % (n = 15). A sample throughput of 10 samples per hour was achieved. This technique was validated by means of reference solid and water materials with good agreement with the certified values. Satisfactory results for speciation of As(III) and As(V) by means of the developed technique were obtained.     

Determination of aluminum in beverages by automated non-segmented continuous flow analysis with fluorescent detection of the lumogallion complex

A method for the automated determination of aluminum in a variety of beverages is described. The method utilizes lumogallion as a complexing agent in a buffer solution. The system is very similar to flow-injection analysis (FIA), however, the tubing id is larger than that typically used in FIA. Therefore, the system is best described as non-segmented continuous flow analysis using fluorescence spectroscopy detection. The method is extremely simple, requiring virtually no sample preparation and only one reagent. The instrument detection limit for aluminum is 0.012 microgram ml-1 and calibration is linear to 3 micrograms ml-1. Results from a variety of beverage matrices are discussed and compared with the frequently used 8-hydroxyquinolone method utilizing a chloroform extraction and fluorescence spectroscopy detection.     

The kinetic determination of clinically significant enzymes in an automated flow-injection system with fluorescence detection

An automated flow-injection manifold is described for the kinetic determination of enzyme activities by a stopped-flow procedure with fluorescence detection. The linear calibration range for alkaline phosphatase is 0/2-250 U l^?1 with a precision of 2%; sample thoughput is 35/2-40 h^?1. Linear responses were also obtained for lipase (0/2-100 U l^?1), acethylcholinesterase (0/2-500 U l^?1) and chymotrypsin (0/2-200 U l^?1). The advantages of this approach to the determination of plasma enzyme activities include sensitivity and the small sample and reagent volumes needed.     

Indirect determination of iron in a flow-injection system with amperometric detection

The change in peak current resulting from the reaction of Fe(II) with nitroso-R salt in a flow-injection system is used to quantify Fe(II) with either single- or dual-electrode amperometric detectors. The current change varies linearly with Fe(II) concentration from 0 to 200 mg 1^?1. The relative standard deviation was about 5% with the single-electrode detector and about 10% with the dual-electrode detector. The method is evaluated for the determination of iron in dietary supplements.     

Laser-induced dispersed fluorescence detection of polycyclic aromatic compounds in soil extracts separated by capillary electrochromatography

Polycyclic aromatic hydrocarbons (PAHs) and nitrogen containing aromatic compounds (NCACs) are characterized in soil extracts and laboratory standards by capillary electrochromatography (CEC) with laser-induced dispersed fluorescence (LIDF) detection using a liquid-nitrogen cooled charge-coupled device detector. The LIDF detection technique provides information on compound identity and, when coupled with the high separation efficiencies of the CEC technique, proves useful in the analysis of complex mixtures. Differences in fluorescence spectra also provide a means of identifying co-eluting compounds by using deconvolution algorithms. Detection limits range from 0.5 to 96x10(-10) M for selected PAHs and 0.9-3.7x10(-10) M for selected NCACs. Soil extracts are also injected onto the CEC column to evaluate chromatographic method performance with respect to complex samples and the ability to withstand exposure to environmental samples.     

Determination of nitrate and nitrite by continuous liquid-liquid extraction with a flow-injection atomic-absorption detection system

Summary A flow injection system coupled on-line with a continuous liquid-liquid extractor is used for the indirect determination of nitrate and nitrite with an atomic-absorption detection system. These anions form ion-pairs with the copper(I)-neocuproine chelate which are extracted into methyl-isobutyl-ketone, the atomic-absorption signal of copper from the organic phase being proportional to the nitrate or nitrite concentration. The methods proposed herein are suitable for determining nitrate or nitrite at the g ml^–1 level with a sampling frequency of 35 ± 5 h^–1. The methods compare favourably with their batch counterparts with regard to sensitivity, selectivity, sample volume and sampling frequency.

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Bestimmung von nitrat und nitrit durch kontinuierliche flüssig-flüssig-extraction mit detektion durch ein fließinjektions-AAS-system

Zusammenfassung Ein Fließinjektionssytem, das on-line mit einem Flüssig-Flüssig-Extraktor gekoppelt ist, wird zur indirekten AAS-Bestimmung von Nitrat und Nitrit benutzt. Diese Anionen bilden mit Kupfer(I)-neocuproinchelat Ionenpaare, die mit Methylisobutylketon extrahiert werden. Das AAS-Signal des Kupfers aus der organischen Phase ist der Nitrat- bzw. Nitritkonzentration proportinal. Die vorgeschlagenen Verfahren eignen sich zur Nitrat- oder Nitritbestimmung im g/ml-Bereich mit einer Probenfrequenz von 35 ± 5 je Stunde. Diese Methoden sind den Batch-Verfahren in bezug auf Empfindlichkeit, Selektivität sowie Probevolumen und -frequenz überlegen.

     

Optically specific detection of D- and L-lactic acids by a flow-injection dual biosensor system with on-line microdialysis sampling.

A flow-injection dual biosensor system with microdialysis sampling is proposed for the simultaneous determination of D-lactic and L-lactic acids. The dialysate from the microdialysis tube is delivered to a sample loop of the six-way autoinjector and then automatically injected into the flow-injection line with a dual enzyme electrode arranged in perpendicular to the flow direction. The dual enzyme electrode is constructed by hybridizing a poly(1,2-diaminobenzene) film into two sensing parts which respond selectively to D-lactic and L-lactic acids, respectively, without any cross-reactivity. The proposed flow-injection analysis method can be successfully applied to the simultaneous determination of D,L-lactic acids in alcoholic beverages.     

Determination of phenylpropanolamine and methoxamine using flow-injection with fluorimetric detection

A new flow-injection procedure for the determination of phenylpropanolamine and methoxamine is proposed. The method is based on the derivatization reaction of the primary amine group with o-phthalaldehyde in the presence of 2-mercaptoethanol using fluorimetric detection. The calibration graphs based on peak areas were linear in the ranges 5-200 ng ml(-1) for phenylpropanolamine and 0.2-6 ng ml(-1) for methoxamine. The detection limits were 3.8 and 0.13 ng ml(-1), respectively. The methods were applied to the determination of the drugs in commercial pharmaceutical preparations.     

Molecularly imprinted polymer for the determination of trace ractopamine in pork using SPE followed by HPLC with fluorescence detection

In this study, a molecularly imprinted functionalized polymer for the selective separation of ractopamine (RAC) was prepared by combining a surface molecular imprinting technique with a sol–gel method process. The polymer was evaluated by static, kinetic adsorption, and selective experiments. Results indicated that the molecularly imprinted polymer had high adsorption capacity, selective ability, and fast mass transfer rate. The polymer was applied for the determination of trace RAC through online SPE‐HPLC. With a sample loading flow rate of 2 mL/min, the enhancement factor of 516.26 and the LOD (S/N = 3) of 4.6 ng/L were achieved, respectively, and the linear range of the calibration curve was 0.04–18 μg/L with r² >0.99. The RAC in pork was determined at three spiked levels (0.5, 1, and 2 ng/g) with recoveries ranging from 55.86 to 67.28%.     

Determination of aluminum in serum by capillary zone electrophoresis with laser-induced fluorescence detection

Summary A novel method of separating and detecting trace aluminum by capillary zone electrophoresis is described. Aluminum is reacted with lumogallion [4-chloro-3-(2,4-dihydroxyphenylazo)-2-hydroxybenzen-1-sulphonic acid] so that the complex can be selectively and sensitively detected by a laser-induced fluorescence detector after capillary electrophoretic separation. Using the proposed method, limits of detection in the sub parts per billion range are achieved. The technique is applied to the determination of aluminum in human serum.     

Determination of water in organic solvents by flow-injection analysis with Karl Fischer reagent and a biamperometric detection system

A flow-injection system with a biamperometric flow-through detector provided with two platinum plate electrodes was tested for the determination of water with a two-component pyridine-free Karl Fischer reagent. The response was shown to be linear in the concentration range 0.03-0.11% water in methanol, ethanol or 2-propanol, with methanol as the carrier solvent. The maximum sampling frequency was about 150 samples per hr. It appeared to be possible to introduce a membrane separation step, thus allowing for the determination of water in fouled process streams. To avoid direct contact between the Karl Fischer solution and the pumping tubes, and thus extend the lifetime of the tubes, an indirect delivery system, based on replacement of the solution by pumped silicone oil, was also applied.     

Cellphone-based detection platform for rbST biomarker analysis in milk extracts using a microsphere fluorescence immunoassay

Current contaminant and residue monitoring throughout the food chain is based on sampling, transport, administration, and analysis in specialized control laboratories. This is a highly inefficient and costly process since typically more than 99 % of the samples are found to be compliant. On-site simplified prescreening may provide a scenario in which only samples that are suspect are transported and further processed. Such a prescreening can be performed using a small attachment on a cellphone. To this end, a cellphone-based imaging platform for a microsphere fluorescence immunoassay that detects the presence of anti-recombinant bovine somatotropin (rbST) antibodies in milk extracts was developed. RbST administration to cows increases their milk production, but is illegal in the EU and a public health concern in the USA. The cellphone monitors the presence of anti-rbST antibodies (rbST biomarker), which are endogenously produced upon administration of rbST and excreted in milk. The rbST biomarker present in milk extracts was captured by rbST covalently coupled to paramagnetic microspheres and labeled by quantum dot (QD)-coupled detection antibodies. The emitted fluorescence light from these captured QDs was then imaged using the cellphone camera. Additionally, a dark-field image was taken in which all microspheres present were visible. The fluorescence and dark-field microimages were analyzed using a custom-developed Android application running on the same cellphone. With this setup, the microsphere fluorescence immunoassay and cellphone-based detection were successfully applied to milk sample extracts from rbST-treated and untreated cows. An 80 % true-positive rate and 95 % true-negative rate were achieved using this setup. Next, the cellphone-based detection platform was benchmarked against a newly developed planar imaging array alternative and found to be equally performing versus the much more sophisticated alternative. Using cellphone-based on-site analysis in future residue monitoring can limit the number of samples for laboratory analysis already at an early stage. Therewith, the entire monitoring process can become much more efficient and economical. Figure

Cellphone-based detection platform for rbST biomarker analysis in milk extracts using a microsphere fluorescence immunoassay     

Determination of nitrate with a flow-injection system combining square-wave polarographic detection with on-line deaeration

The combination of a flow-injection system with square-wave polarography and on-line deaeration is applied to the determination of nitrate, utilizing the catalytic reaction between nitrate and uranyl ion. The method is simple, rapid (60 samples h^?1), sensitive and accurate, with a detection limit of 2 μM nitrate. The method has been applied to the determination of nitrate in drinking and river waters.     

Speciation of glyphosate, phosphate and aminomethylphosphonic acid in soil extracts by ion chromatography with inductively coupled plasma mass spectrometry with an octopole reaction system

Ion-pairing chromatography coupled with inductively coupled plasma mass spectrometry (ICP-MS) used for the speciation of phosphorus is limited as the mobile phase containing organic solvents changes in detection sensitivity and the carbon precipitates on torch and cones. To address this issue, anion-exchange chromatography with ICP-MS has been used for the speciation of glyphosate, phosphate and aminomethylphosphonic acid in soil extracts. The separation of the targets on a new column was achieved within 5 min using an eluent containing 20 mM NH₄NO₃ at pH 5.1. Furthermore, since the polyatomic ions such as ¹⁴N¹⁶O¹H⁺ and ¹⁵N¹⁶O⁺ from a nitrogen-based ion-pairing reagent interfered with ICP-MS detection of ³¹P, an octopole reaction system was investigated to determine whether the polyatomic interferences could be reduced. The results show that addition of He to the cell can benefit analyses by reducing such interferences, but at the expense of reduced sensitivity. The detection limits in the range of 1.0-1.5 μg L⁻¹ (expressed as P) was achieved when 50 μL was injected using He as the collusion gas.     

Chromatographic behaviour of phenols on thin layers of cation and anion exchangers. II. Dowex 50-X4 and Rexyn 102.

The chromatographic characteristics of 58 phenols on Rexyn 102 and Dowex 50-X4 thin layers in both the acidic and sodium salt forms have been studied, using elution with water, water-alcohol mixtures and aqueous salt solutions at different pH values. The influence of the percentage of alcohol, the pH and the ionic strength on the chromatographic behaviour of these phenols was investigated. The validity of the relationships among the RF values, the pH of the eluent and the pKa of the phenol has been verified on Dowex 50-X4 (Na+) thin layers. It has been shown that from the RFac, RFalk and pKa values of the different phenols, it is possible to predict their behaviour over the whole pH range and therefore to select the best conditions for their chromatographic separation.     

Electrochemiluminescent determination of L-cysteine with a flow-injection analysis system.

A flow-injection electrochemiluminescent method for L-cysteine determination has been developed based on its enhancement of the electrochemiluminecence of luminol at a glassy carbon electrode. This method is simple and sensitive for cysteine determination. Under the selected experimental parameters, the linear range for cysteine concentration was 1.0 x 10(-6) - 5.0 x 10(-5) mol/l, and the detection limit was 0.67 micromol/l (S/N = 3). The relative standard deviation for 11 measurements of 1.0 x 10(-5) mol/l cysteine was 4.5%. The proposed method has been applied to the detection of cysteine in pharmaceutical injections with satisfactory results.     

Speciation of arsenic in coarse and fine urban aerosols using sequential extraction combined with liquid chromatography and atomic fluorescence detection

An analytical procedure for speciation of As in urban aerosol samples was developed. The aerosols were collected by sequential filtration through membrane filters. Part of each filter was investigated by INAA for the total amount of As. Another part of the filters was treated by a sequential extraction procedure to differentiate between water-extractable, phosphate-extractable and refractory chemical forms. Water-extractable forms were further differentiated into anionic As species by HPLC-HGAFS. Extractability of As into water exhibited a clear dependency on the aerosol size fraction (12% in coarse fraction and 50% in fine fraction). Dependency of the phosphate extractable As on the aerosol size fraction seems not to be significant (10–15% in both size fractions). The remaining amount, i.e., about 78% of the coarse As and about 40% of the fine As was considered to be refractory or environmentally immobile As. Water-extractable As forms could only be attributed to arsenate.     

Determination of low concentrations of chlorite and chlorate ions by using a flow-injection system

A flow-injection method is reported for the determination of chlorite ion and chlorite and chlorate ions in mixtures at the submilligram per liter level in drinking water. The chlorite ion concentration is selectively determined by using its reaction with iodide ion at pH 2, which liberates iodine. Both species react with iodide ion in6 M HCl to produce iodine, the concentration of which is measured spectrophotmetricaly at 370 nm. The individual species are determined using multiplel regression. The method exhibits a linear range from 2 to 150 μM (0.1–10.1 mg l^-1) for chlorite ion and from 2 to μM (0.1–8.3 mg l^-1 for chlorate ion, with relative standard deviations of 0.4 and 1.2%, respectively.     

Speciation of arsenic in coarse and fine urban aerosols using sequential extraction combined with liquid chromatography and atomic fluorescence detection

An analytical procedure for speciation of As in urban aerosol samples was developed. The aerosols were collected by sequential filtration through membrane filters. Part of each filter was investigated by INAA for the total amount of As. Another part of the filters was treated by a sequential extraction procedure to differentiate between water-extractable, phosphate-extractable and refractory chemical forms. Water-extractable forms were further differentiated into anionic As species by HPLC-HGAFS. Extractability of As into water exhibited a clear dependency on the aerosol size fraction (12% in coarse fraction and 50% in fine fraction). Dependency of the phosphate extractable As on the aerosol size fraction seems not to be significant (10-15% in both size fractions). The remaining amount, i.e., about 78% of the coarse As and about 40% of the fine As was considered to be refractory or environmentally immobile As. Water-extractable As forms could only be attributed to arsenate.