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Evaluation of stability of arsenic species in rice.

Although most edible vegetables do not accumulate As at a high rate, rice, carrots and certain others are exceptions. In addition to nutritional or toxicological considerations, the relatively high level and variety of As species present in rice make it a very suitable matrix for a candidate reference material representative of terrestrial biological samples.An analytical procedure was developed for As speciation in rice based on the use of a 1:1 methanol-water mixture for species extraction, an anion Hamilton PRPX-100 column (at pH 6, and phosphate mobile phase 10 mM), and a cation Hamilton PRP-X200 column (at pH 2.8 in pyridine formiate 4 mM) for species separation and final determination by HPLC-ICP-MS. The detection limits for dry flour rice expressed as As were 2 and 3 ng g(-1) for As(III) and AsB on the cation column and 3, 6 and 5 ng g(-1) for As(V), MMA and DMA, respectively, on the anion column. The methodology developed was applied to check the stability of As species in the water-methanol extract and also under different processing steps and storage time and temperature conditions. It was demonstrated that the As species in the water-methanol extracts stored at +4 degrees C remained stable for at least one month. Once the rice grains are ground, the MMA and As(V) species are not stable under any storage conditions probably due to microbiological activity. When ground rice is gamma-irradiated species remain stable although the AsB does not appear.

Arsenic↗

Chemiluminescence determination of melatonin and some of its derivatives using potassium permanganate and formaldehyde system.

It was found that melatonin and its derivatives, such as N-acetyl- 5-methoxytryptamine (MT), N-acetyl-5-hydroxytryptamine (NAS), 5-Methoxytrypt- amine (5-MT), 5-Methoxyindolyl acetic acid (5-MIAA) and N-acetyl-5-methoxy- 6-hydroxytryptamine (6-HMT) would give chemiluminescence in the acidic potassium permanganate solution, and formaldehyde would enhance this chemiluminescent reaction greatly. The optimum conditions for this chemiluminescent reaction were studied in detail by a flow injection system. A new simple rapid method has been developed under the optimum conditions for determination of melatonin. This method has the advantages of high sensitivity, wide range of linear response and low detection limit. On the basis of investigation of chemiluminescent, fluorescent and UV spectra of melatonin in acidic solution containing potassium permanganate and formaldehyde, a possible mechanism of this reaction was proposed.

Flow Injection Analysis↗

Screen-printed multienzyme arrays for use in amperometric batch and flow systems.

Screen-printing technology for electrode fabrication enables construction of amperometric devices suitable for combination of several enzyme electrodes. To develop a biosensor array for characterisation of wastewaters, tyrosinase and horseradish peroxidase (HRP) or cholinesterase-modified electrodes were combined on the same array. The behaviour of the tyrosinase-modified electrode in the presence of hydrogen peroxide (required co-substrate for the HRP-modified electrode) and acetylthiocholine chloride (required co-substrate for cholinesterase) was studied. Performance of bi-enzyme biosensor arrays in the batch mode and in the flow-injection system are discussed.

Acetylcholinesterase↗

Simple optical fibre biosensor based on immobilised enzyme for monitoring of trace heavy metal ions.

A simple optical fibre biosensor based on immobilised enzyme for monitoring of trace heavy metal ions has been developed. The biosensor recognition system was designed based on the inhibition of urease activity, where the urease is immobilised on ultrabind membrane. The studies of inhibition by the heavy metal ions Hg(II), Ag(I), Cu(II), Ni(II), Zn(II), Co(II) and Pb(II) were performed using a fibre-optic biosensor configuration, where the pH change resulting from the bio-catalytic hydrolysis of urea was monitored at the wavelength 615 nm spectroscopically, using commercial pH indicator strip before and after the exposure to the heavy metal ions. The immobilised urease was regenerated by l-cysteine. The linear response range between 1 x 10(-9)-1 x 10(-5) M and the limit of detection 1 x 10(-9 )M (0.2 microg/L) for Hg(II) ions was achieved by employing the flow method. The optimisation of experimental parameters, including flow method, is also discussed.

Biosensing Techniques↗

A study of the chemiluminescence behavior of myoglobin with luminol and its analytical applications.

A chemiluminescence signal at 425 nm was observed when ferric state myoglobin was mixed with luminol in alkaline medium. Because the signal was remarkably enhanced in the presence of Fe(CN)6(4-), analytical applications were investigated in a flow-injection system. The increase in chemiluminescence was linearly dependent on myoglobin concentration in the range 0.1 to 100 nmol L(-1), and the limit of detection was 0.04 nmol L(-1) with relative standard deviation 3.2% (3 sigma). It was also found that binding of Mb with the ligands CN-, SCN-, and F- significantly inhibited the chemiluminescence reaction. The linear dynamic ranges for the ligands were 1.0-300.0, 0.1-3.0, and 0.5-100.0 nmol L(-1), and the limits of detection (S/N=3) 0.4, 0.04, and 0.2 nmol L(-1), for F-, CN-, and SCN-, respectively. The relative standard deviations were 5.32%, 6.13%, and 3.38% for 0.1 nmol L(-1) CN-, 0.5 nmol L(-1) SCN-, and 1.0 nmol L(-1) F-, respectively. At a flow rate of 2.0 mL min(-1) the assay could be accomplished in 1 min, including sampling and washing. The method has been successfully applied to the determination of myoglobin in human urine and F- in water samples. A possible mechanism of chemiluminescence production by myoglobin and luminol is presented.

Female↗

Trace element determination in seawater by ICP-SFMS coupled with a microflow nebulization/desolvation system.

Direct and simultaneous determination of Al, Ag, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Pb, Sb, U, V and Zn in diluted (1:10 v:v) seawater from the Antarctic Ocean and the Venice Lagoon at the ng mL(-1) and pg mL(-1) level has been performed by using an inductively coupled plasma sector field mass spectrometer (ICP-SFMS). Samples were analysed by using a PFA microflow nebulizer coupled with a desolvation system or a PFA microflow nebulizer coupled with a Teflon spray chamber, respectively. Measurements were carried out at low (LR, m/Delta m=300), medium (MR, m/Delta m=3,000) and high (HR, m/Delta m=7,500) resolutions depending on the studied isotope. To avoid contamination, sample pre-treatment was carried out in a clean laboratory equipped with a Class 100 vertical laminar flow hood. Concentration ranges (minimum-maximum in ng mL(-1)) found in the Antarctic seawater samples (in depth profiles) were: Ag 0.0004-0.0018, As 0.69-1.32, Cd 0.031-0.096, Co 0.018-0.065, Cr 0.18-0.46, Cu 0.04-1.58, Fe 0.13-1.63, Mn 0.02-0.12, Mo 5.97-12.46, Pb 0.007-0.074, Sb 0.033-0.088, U 0.5-1.9, V 0.6-2.5 and Zn 0.16-0.80. Concentration ranges (min-max in ng mL(-1)) found in the Venice Lagoon water samples (temporal profile from a benthic chamber experiment) were: Al 0.24-0.61, Ag 0.007-0.031, As 1.42-2.27, Cd 0.050-0.182, Co 0.440-1.461, Cr 0.15-0.34, Cu 0.81-2.46, Fe 0.25-1.66, Mn 11.6-31.7, Mo 6.50-10.6, Pb 0.047-0.225, Sb 0.240-0.492, U 1.7-3.3, V 1.3-2.8 and Zn 5.20-21.5. The detection limits range between 0.06 pg mL(-1) for Ag and U to 15 pg mL(-1) for Fe. In order to check the accuracy of the analytical procedure, measurements of the trace elements in a certified reference material (coastal Atlantic seawater, CASS-4-NRCC) were compared with the certified values. In addition, the results from the Antarctic and Venice Lagoon samples were compared with those obtained by using different analytical techniques.

Antarctic Regions↗

Solid-phase reactor flow-injection on-line oxidizing spectrofluorimetry for determination and dissolution studies of folic acid.

A simple, sensitive and specific flow-injection spectrofluorimetric method has been developed for the determination of folic acid in pharmaceuticals. The method is based on use of a lead dioxide solid-phase reactor for on-line oxidation of folic acid into a strongly fluorescent compound with a maximum excitation wavelength of 281 nm and an emission wavelength of 450 nm. Under optimum conditions the fluorescence intensity of oxidation product is proportional to the concentration of folic acid over the range 0.008-2.5 microg mL(-1). The detection limit is 0.0001 microg mL(-1), the relative standard deviation is 0.85% for 11 replicate determinations of 0.05 microg mL(-1) folic acid, and the sample throughput is 20 h(-1). In combination with an on-line filter and dilution, an automated drug-dissolution system was established. The proposed method has been successfully applied to the determination of folic acid in pharmaceutical preparations and dissolution testing.

Flow Injection Analysis↗

Direct determination of selenium in urine samples by electrothermal atomic absorption spectrometry using a Zr plus Rh-treated graphite tube and co-injection of Rh as chemical modifier.

Different chemical modifiers for use with electrothermal atomic absorption spectrometry (ET AAS) were investigated in relation to determining the selenium in human urine samples. The samples were diluted in a solution containing 1% v/v HNO3 and 0.02% m/v cetyltrimethylammonium chloride (CTAC). Studying the modifiers showed that the use of either Ru or Ir as the permanent modifier gave low sensitivity to Se and the peak shape was very noisy, while Zr or Rh gave no peak at all. The same occurred when Zr was used in solution. For mixtures of permanent modifiers, Ir plus Rh or Zr plus Rh gave very low sensitivity, Zr plus Rh with co-injection of Ir in solution was also not efficient, Zr plus Rh in solution gave good sensitivity, but the best results were obtained with a mixture of Zr and Rh as the permanent modifier and co-injection of Rh in solution. Using this last modifier, the following dilutions with the HNO3 and CTAC were studied: 1:1, 1:2, 1:3 and 1:4. The best dilution was 1:1, which promoted good sensitivity and a more defined peak shape and made it possible to correct for the background using a deuterium arc lamp. Under these conditions, a characteristic mass of 26+/-0.2 pg was obtained for Se in aqueous solution. Six certified urine samples were analyzed using matrix matching calibration and the measured concentrations were in agreement with the certified values, according to a t-test at the 95% confidence level. Recovery tests were carried out and the recoveries were in the range 100-103%, with relative standard deviation better than 9%. The limit of detection (LOD, 3 sd, n = 10) was 3.0 microg L(-1) in the sample. The treated graphite tube could be used for at least 600 atomization cycles without significant alteration of the analytical signal.

Electrochemistry↗

Simultaneous determination of rifampicin and isoniazid by continuous-flow chemiluminescence with artificial neural network calibration.

In this paper a continuous-flow chemiluminescence (CL) system with artificial neural network calibration is proposed for simultaneous determination of rifampicin and isoniazid. This method is based on the different kinetic spectra of the analytes in their CL reaction with alkaline N-bromosuccinimide as oxidant. The CL intensity was measured and recorded every second from 1 to 300 s. The data obtained were processed chemometrically by use of an artificial neural network. The experimental calibration set was 20 sample solutions. The relative standard errors of prediction for both analytes were approximately 5%. The proposed method was successfully applied to the simultaneous determination of rifampicin and isoniazid in a combined pharmaceutical formulation.

Algorithms↗

Interfacing liquid chromatography with atmospheric pressure MALDI-MS.

Two different strategies for coupling liquid chromatography with atmospheric pressure matrix assisted laser desorption/ionization (AP MALDI) are presented. The first method is flow-injection liquid AP UV-MALDI. Compared with previous similar research, the detection limit was improved 10 times to 8.3 fmol using a solution of 50 nM peptide with 25 mM alpha-cyano-4-hydroxycinnamic acid. The applicability of this method to measure oligosaccharides, actinomycin antibiotics, antibiotics, phosphopeptides, and proteins is demonstrated. The upper mass limit achieved with the current instrumentation is 6,500 Da (doubly charged cytochrome c). The feasibility of a second strategy based on single-droplet IR AP MALDI is demonstrated here. Aqueous peptide solutions were successfully measured by this method.

Atmospheric Pressure↗

Flow injection chemiluminescence immunoassay for 17beta-estradiol using an immunoaffinity column.

A new flow injection chemiluminescent immunoassay was developed for the detection of 17beta-estradiol (E2). The method uses p-iodophenol (PIP) as enhancer and is based on a solid-phase immunoassay format in which an E(2)-OVA immobilized immunoaffinity column inserted in the flow system is used to trap unbound horseradish peroxidase (HRP)-labeled anti-E2 antibody after an off-line incubation of E2 with HRP-labeled anti-E2 antibody. The trapped enzyme conjugate was detected by injecting substrates to produce an enhanced chemiluminescence (CL) response. The linear range for E2 was 10.0-1,000.0 ng mL(-1) with a correlation coefficient of 0.996 and a detection limit of 3.0 ng mL(-1). The sampling and chemiluminescence detection time for one sample was 400 s after a pre-incubation procedure of 30 min. Serum samples detected by this method were in good agreement with the results obtained by EIA with E2-biotin.

Calibration↗

Flow-injection enhanced chemiluminescence method for determination of ciprofloxacin in pharmaceutical preparations and biological fluids.

A novel rapid and sensitive analytical method, enhanced chemiluminescence with flow-injection sampling, is described for determination of ciprofloxacin. The method is based on the chemiluminescence reaction of the potassium permanganate-sodium thiosulfate-ciprofloxacin system. An enhanced chemiluminescence reaction was developed, and optimum conditions for CL emission were investigated. The chemiluminescence intensity was linearly dependent on ciprofloxacin concentration in the range 1.0 x 10(-8)-1.0 x 10(-5) g mL(-1). The detection limit was 4 x 10(-9) g mL(-1). The relative standard deviation was 1.8% for eleven measurements of 2.0 x 10(-7) g mL(-1) ciprofloxacin standard solution. The new method enables simple, sensitive, and rapid determination of ciprofloxacin and has been successfully used for determination of ciprofloxacin in biological fluids and in ciprofloxacin hydrochloride tablet and injection.

Antioxidants↗

Flow-injection technique for determination of uranium and thorium isotopes in urine by inductively coupled plasma mass spectrometry.

A sensitive and efficient flow-injection (FI) preconcentration and matrix-separation technique coupled to sector field ICP-mass spectrometry (SF-ICP-MS) has been developed and validated for simultaneous determination of ultra-low levels of uranium (U) and thorium (Th) in human urine. The method is based on selective retention of U and Th from a urine matrix, after microwave digestion, on an extraction chromatographic TRU resin, as an alternative to U/TEVA resin, and their subsequent elution with ammonium oxalate. Using a 10 mL sample, the limits of detection achieved for 238U and 232Th were 0.02 and 0.03 ng L(-1), respectively. The accuracy of the method was checked by spike-recovery measurements. Levels of U and Th in human urine were found to be in the ranges 1.86-5.50 and 0.176-2.35 ng L(-1), respectively, well in agreement with levels considered normal for non-occupationally exposed persons. The precision obtained for five replicate measurements of a urine sample was 2 and 3% for U and Th, respectively. The method also enables on-line measurements of the 235U/238U isotope ratios in urine. Precision of 0.82-1.04% (RSD) was obtained for 235U/238U at low ng L(-1) levels, using the FI transient signal approach.

Flow Injection Analysis↗

Electrochemical reduction and flow detection of iodate on (Bu4N)2Mo6O19 self-assembled monolayer.

A stable monolayer of the inorganic-organic hybrid polyoxometalate (Bu4N)2Mo6O19, denoted as Mo6O19, was formed on a sodium-3-mercapto-1-propanesulfonate (MPPS)-covered gold electrode surface, interlaced with an anionic poly(dimethyldiallylammonium chloride) (PDDA) binding layer based on the electrostatic self-assembled (ESA) technique. Electrochemical characterization of the Mo6O19 self-assembled thin films on the solid surface by cyclic voltammetry and AC impedance spectroscopy revealed a stable and sensitive electrocatalytic response to the reduction of iodate. Iodate was determined amperometrically through a flow injection cell at the modified electrode in the concentration range of 1.0 x 10(-6) to 1.0 x 10(-1) M with a detection limit of 8 x 10(-8) M (signal-to-noise ratio = 3). Performance was improved to meet practical needs compared with previously reported analogues.

Electrochemistry↗

Resolution of an intense sweetener mixture by use of a flow injection sensor with on-line solid-phase extraction. Application to saccharin and aspartame in sweets and drinks.

An integrated solid-phase spectrophotometry-FIA method is proposed for simultaneous determination of the mixture of saccharin (1,2-benzisothiazol-3(2H)-one-1,1-dioxide; E-954) (SA) and aspartame (N-L-alpha-aspartyl-L-phenylalanine-1-methyl ester; E-951) (AS). The procedure is based on on-line preconcentration of AS on a C18 silica gel minicolumn and separation from SA, followed by measurement, at lambda = 210 nm, of the absorbance of SA which is transiently retained on the adsorbent Sephadex G-25 placed in the flow-through cell of a monochannel FIA setup using pH 3.0 orthophosphoric acid-dihydrogen phosphate buffer, 3.75x10(-3) mol L(-1), as carrier. Subsequent desorption of AS with methanol enables its determination at lambda = 205 nm. With a sampling frequency of 10 h(-1), the applicable concentration range, the detection limit, and the relative standard deviation were from 1.0 to 200.0 microg mL(-1), 0.30 microg mL(-1), and 1.0% (80 microg mL(-1), n = 10), respectively, for SA and from 10.0 to 200.0 microg mL(-1), 1.4 microg mL(-1), and 1.6% (100 microg mL(-1), n = 10) for AS. The method was used to determine the amounts of aspartame and saccharin in sweets and drinks. Recovery was always between 99 and 101%. The method enabled satisfactory determination of blends of SA and AS in low-calorie and dietary products and the results were compared with those from an HPLC reference method.

Aspartame↗

Total determination of metal-binding carbohydrates in plant extracts by using FIA with electrochemical detection.

FIA with pulsed amperometric detection (PAD) is used for the determination of metal-binding carbohydrates in plant extracts. Results of direct FIA measurements agree very well with those of corresponding HPLC-PAD analyses. The proposed method is used to determine total carbohydrates in isolated low-molecular-weight fractions of plant roots, which have been obtained after extraction at different pH values. The results are compared with those obtained by corresponding metal determinations (Ca, Mg, Zn, Mn) by AAS and also with constant potential amperometric detection at a copper working electrode. The latter detection mode is more sensitive, but less selective for carbohydrates.

Carbohydrates↗

Extraction of antimony and arsenic from fresh and freeze-dried plant samples as determined by HG-AAS.

Six extraction media (acetic acid, EDTA, tetrabutylammonium hydroxide, NaOH, MeOH/H2O, acetonitrile/H2O) were tested for their ability to extract antimony (Sb) and arsenic (As) from freeze-dried poplar leaves, pine shoots and spruce shoots, as well as from a peat matrix. Additionally, the extraction efficiency of Sb and As in fresh and freeze-dried elder leaves and poplar leaves was compared. Total concentrations of Sb and As of aliquots (approximately 220 mg) of the freeze-dried samples were analysed by flow injection hydride generation atomic absorption spectrometry (FI-HG-AAS) after open vessel digestion with adequate mixtures of nitric, sulfuric, hydrochloric, and perchloric acid. Three reference materials GBW 07602 Bush Branches and Leaves, GBW 07604 Poplar Leaves, and SRM 1575 Pine Needles were analysed with every batch of samples to ensure the accuracy and precision of the applied analytical procedures. The use of hydrofluoric acid in the digestion mixture leads to distinctly lower As values (down to 40%) than actual concentrations in the investigated plant materials. Extraction efficiencies were generally low and lower for Sb than for As. Solutions of 0.66 mol L(-1) NaOH liberated highest amounts of Sb with approximately 10% for poplar leaves, and approximately 19% each for pine shoots and spruce shoots. Distinctly higher concentrations of As in NaOH extracts of poplar leaves (22%), pine shoots (32%), and spruce shoots (36%) were quantified. Extraction experiments resulted in yields of 7-9% from fresh elder and poplar leaves, respectively, and 8-13% for freeze-dried samples for Sb. The corresponding values for As were 10-35% for the fresh material and 7-37% for the freeze-dried samples.

Antimony↗

Spectrophotometric flow injection determination of total phosphorus in beer using on-line UV/thermal induced digestion.

A flow injection system for the automatic determination of total phosphorus in beer is described. The developed manifold uses a two-stage photooxidation/thermal digestion procedure together with oxidizing and hydrolyzing reagents to convert all forms of phosphorus compounds to orthophosphate. Polyphosphates are hydrolyzed by acid and heat, and organo-phosphates are digested by UV-catalyzed peroxodisulfate oxidation. The orthophosphate formed is then spectrophotometrically determined by the phosphomolybdenum blue reaction, using stannous chloride as reducing agent. The results obtained for a set of 19 beer samples (with concentrations from 120 to 735 mg P/L) were in good agreement with the reference method, the maximum relative deviation found being 4.7%. Relative standard deviations for ten consecutive determinations were lower than 1.5%, and a detection limit of 1 mg P/L was achieved.

Beer↗