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Determination of lead and nickel in environmental samples by flame atomic absorption spectrometry after column solid-phase extraction on Ambersorb-572 with EDTA.

Lead and nickel were preconcentrated as their ethylenediaminetetraacedic acid (EDTA) complexes from aqueous sample solutions using a column containing Ambersorb-572 and determined by flame atomic absorption spectrometry (FAAS). pH values, amount of solid phase, elution solution and flow rate of sample solution have been optimized in order to obtain quantitative recovery of the analytes. The effect of interfering ions on the recovery of the analytes has also been investigated. The recoveries of Pb and Ni under the optimum conditions were 99 +/- 2 and 97 +/- 3%, respectively, at 95% confidence level. Seventy-five-fold (using 750 mL of sample solution and 10 mL of eluent) and 50-fold (using 500 mL of sample solution and 10 mL of eluent) preconcentration was obtained for Pb and Ni, respectively. Time of analysis is about 4.5 h (for obtaining enrichment factor of 75). By applying these enrichment factors, the analytical detection limits of Pb and Ni were found as 3.65 and 1.42 ng mL(-1), respectively. The capacity of the sorbent was found as 0.17 and 0.21 mmol g(-1) for Pb and Ni, respectively. The interferences of some cations, such as Mn2+, Co2+, Fe3+, Al3+, Zn2+, Cd2+, Ca2+, Mg2+, K+ and Na+ usually present in water samples were also studied. This procedure was applied to the determination of lead and nickel in parsley, green onion, sea water and waste water samples. The accuracy of the procedure was checked by determining Pb and Ni in standard reference tea leaves sample (GBW-07605). The results demonstrated good agreement with the certified values.

Adsorption↗

Simultaneous determination of local anesthetics including ester-type anesthetics in human plasma and urine by gas chromatography-mass spectrometry with solid-phase extraction.

The present study describes the simultaneous determination of seven different kinds of local anesthetics and one metabolite by GC-MS with solid-state extraction: Mepivacaine, propitocaine, lidocaine, procaine (an ester-type local anesthetics), cocaine, tetracaine (an ester-type local anesthetics), dibucaine (Dib) and monoethylglycinexylidide (a metabolite of lidocaine) were clearly separated from each other and simultaneously determined by GC-MS using a DB-1 open tubular column. Their recoveries ranged from 73-95% at the target concentrations of 1.00, 10.0 and 100 microg/ml in plasma, urine and water. Coefficients of variation of the recoveries ranged from 2.3-13.1% at these concentrations. The quantitation limits of the method were approximately 100 ng/ml for monoethylglycinexylidide, propitocaine, procaine, cocaine, tetracaine and dibucaine, and 50 ng/ml for lidocaine and mepivacaine. This method was applied to specimens of patients who had been treated with drip infusion of lidocaine, and revealed that simultaneous determination of lidocaine and monoethylglycinexylidide in the blood and urine was possible.

Anesthetics, Local↗

Formation of artefacts during air analysis of volatile amines by solid-phase micro extraction.

Solid-phase micro extraction (SPME) is a promising technique for fast and low cost trace analysis. However, some limitations of the technique were encountered when using a PDMS (polydimethylsiloxane)/Carboxen fibre for sampling a mixture of volatile aliphatic amines in air. On the GC chromatogram, two supplementary peaks were noticed in addition to the analyte peaks, thus limiting qualitative and quantitative analysis in this particular case. This paper presents the investigations to identify the artefacts and determine the origin of their formation. First, GC-MS identification, by both electron impact and chemical ionisation modes, demonstrated that the two artefacts were unsaturated amines assumed to be formed by a dehydrogenation reaction of the target amines. This reaction was found to occur during thermal desorption of analytes in the GC injection port and to be catalysed by temperature and by metals consisting of the inox (stainless-steel) needle of the SPME device. It was also demonstrated that artefact formation was not significant when using PDMS or PDMS/divinylbenzene fibres. This difference with PDMS/Carboxen fibre can be explained by the high desorption temperature required for this fibre. Moreover, the microporosity of Carboxen induces a longer desorption time which increases the contact between analytes and inox and thereby enhances artefact formation.

Air Pollutants↗

High-performance liquid chromatographic determination of phenylephrine and its conjugates in human plasma using solid-phase extraction and electrochemical detection.

An HPLC method for the determination of phenylephrine and its conjugates in human plasma was developed and validated. The method for quantitation involved extraction of diluted plasma (subject to hydrolysis with beta-glucuronidase for 30 min with 500 units of enzyme per 0.1 ml of plasma at 37 degrees C for the conjugates) on solid-phase weak cation-exchange cartridges followed by elution of the analyte and the internal standard (ethylnorphenylephrine) with 5% triethylamine in methanol. Analysis was carried out on a 15 cm ODS stationary phase using ion-pair reversed-phase chromatography. An electrochemical detector operated at + 1.15 V vs. Ag/AgCl was employed for detection. The standard curves were linear in the range 1.0-50.0 ng ml-1 for phenylephrine and 25.0-500.0 ng ml-1 for phenylephrine obtained from its conjugates. The limit of quantitation was 2.0 ng ml-1 (RSD = 17%) and 25.0 ng ml-1 (RSD = 18%), respectively. Acceptable accuracy and precision were obtained during intra- and inter-batch analyses for both the assays.

Adrenergic alpha-Agonists↗

A study of ion suppression effects in electrospray ionization from mobile phase additives and solid-phase extracts.

Since the wide adoption of liquid chromatography/tandem mass spectrometry (LC/MS/MS), the ion suppression/enhancement phenomenon is the latest barrier to high-throughput analysis. This consequence of a nonoptimized analytical method can lead to adverse effects during quantitation (i.e. poor accuracy and precision). Previous papers have reported that ion suppression is a direct result of endogenous material present in biological samples. However, in the case of a solid-phase liquid chromatography/tandem mass spectrometry (SPE/LC/MS/MS) system, the measured result is the combination of several operating conditions and parameters. Little has been done to effectively monitor and/or choose optimized conditions for the complete sequence of extraction, clean up, separation and analysis. This paper describes a simple setup for quantification of ion suppression/enhancement. Several mobile phase additives, ion-pairing agents and SPE extracts were measured and compared against a standard reference. The results demonstrated that a clean up of plasma extracts based on ion exchange leads to minimal ion suppression/enhancement for the compounds that were investigated.

Artifacts↗

Solid-phase extraction of pesticides from water: possible interferences from dissolved organic material.

A multiresidue analysis for trifluralin, simazine, atrazine, propazine, diazinon, parathion-methyl, alachlor, malathion, parathion, chlorpyrifos, pendimethalin, methidathion, and DEF in water that utilizes liquid-solid extraction (LSE) with octadecyl-bonded silica cartridges (C18BSCs) followed by gas chromatography/mass spectrometric analysis was developed. Recoveries of most pesticides were greater than 80% with C18BSCs from fortified water at concentration levels from about 1 to 500 ppb. Recoveries with C18BSCs, from an optically adjusted humic acid solution (10 ppm dissolved organic carbon) made to simulate a natural water with a high dissolved organic content, ranged from 29 to 153% and in general were lower than recoveries obtained from pure water. 14C-Labeled diazinon and parathion were recovered from the humic acid solution at levels of 57 and 68%, respectively, with C18BSCs; the remainder of the labeled pesticides was found in the cartridge eluents. Partition coefficients with human acid were calculated based on recovery of 14C-labeled pesticides from the C18BSCs.

Indicators and Reagents↗

Determination of a renin inhibitor in plasma by solid-phase extraction using acetone as protein binding displacer followed by on-line high-performance liquid chromatography.

H 218/54 is a potent inhibitor of human renin activity (pIC50 = 8.3 at pH 6) and is therefore a potential agent for blood pressure reduction. This lipophilic compound is highly bound to plasma proteins, e.g. 99.7% in rats and 99.6% in humans. For pharmacokinetic studies, a quantitative assay for 3H-H 218/54 in plasma has been developed. On top of an AASP phenyl solid-phase cartridge 70 microliters of rat plasma or 1 ml of cynomolgus plasma was mixed with 200 microliters of water containing 20% acetone. The acetone displaced the substance from plasma proteins without precipitation of the sample and clogging of the extraction column. The mixture was passed through the cartridge, which adsorbed 3H-H 218/54. The cartridge was placed in an AASP autosampler connected to a reversed-phase LC system, with a Vydac C-18 column and CH3CN-H2O-TFA (60:40:0.1, v/v/v) as mobile phase. The effluent from the separation column was collected in fractions for radioactivity counting. Recovery, as measured after adding various amounts of tritium-labelled H 218/54 to blank plasma followed by repeated analysis of the samples, was close to 100% with relative standard deviations between 1.4 and 3.0%. At the lowest level tested, 200 dpm per sample, the recovery was 120% with a relative standard deviation of only 10%. The sensitivity of the method will depend on the specific radioactivity of the dose given.

Acetone↗

Solid-phase extraction and ion-pair reversed-phase HPLC of isometamidium in bovine serum and tissues.

An analytical method has been developed for the determination of isometamidium in bovine serum and tissues. Samples were enzymatically hydrolysed and cleaned up on a solid-phase system (C8 Bond Elut column). The drug was chromatographed by an ion-pair reversed-phase technique using heptane sulphonate as a pairing-ion and triethylamine as a counter-ion reagent. Detection was by fluorescence at 593 nm (excitation = 380 nm). The method is more sensitive and specific than existing methods and it is currently being used in evaluating the pharmacokinetics of isometamidium in cattle.

Animals↗

Determination of the antihypertensive drug cilazapril and its active metabolite cilazaprilat in pharmaceuticals and urine by solid-phase extraction and high-performance liquid chromatography with photometric detection.

A liquid chromatographic method with photometric detection for the determination of cilazapril and its active metabolite and degradation product cilazaprilat in urine and pharmaceuticals has been developed. The chromatographic method consisted of a microBondapak C18 column maintained at 30+/-0.2 degrees C, using a mixture of methanol-10 mM phosphoric acid (50:50 v/v) as mobile phase at a flow-rate of 1.0 ml/min. Enalapril maleate was used as internal standard. The detection was performed at a wavelength of 206 nm. A study of the retention of cilazapril and cilazaprilat using solid-liquid extraction has been carried out in order to optimise the clean-up procedure for urine samples, which consisted of a solid-liquid extraction using C(R) cartridges. Recoveries greater than 85% are obtained for both compounds. The method was sensitive, precise and accurate enough to be applied to the determination of urine samples obtained from three hypertensive patients up to 24 h after intake of a therapeutic dose (detection limit of 70 ng/ml for cilazapril and cilazaprilat in urine). A comparison of the method developed using photometric and amperometric detection has been carried out.

Antihypertensive Agents↗

Comparative study of solid phase extraction techniques for isolation of leukotrienes from plasma.

We have compared five commercially available absorbent materials (i.e. C18 Sep-Pak, C18 J.T. Baker, Amberlites XAD-7, XAD-2 and XAD-4) for their applicability as effective tools for extraction of leukotrienes from plasma samples. Leukotriene C4 (LTC4) and B4 (LTB4) were selected as representatives of peptidic and non-peptidic leukotrienes, respectively. These leukotrienes were added to 1 ml of plasma and passed through columns containing the above described adsorbent materials. The recovery was determined for each material using different combinations of solvents. XAD-4 gave the highest recovery for LTB4 (90%), whereas XAD-4 and XAD-2 gave identical recoveries for LTC4 (90%) when an eluting solvent mixture of pyridine-water--dimethylformamide (50:45:5) was used. The efficiency of the other three solid adsorbent materials for leukotriene extraction were in order of decreasing magnitude, C18 J.T. Baker greater than XAD-7 greater than C18 Sep-Pak. XAD-7 was shown to be more efficient for LTB4 than for LTC4, whereas the octadecylsilane C18 materials gave approximately similar recoveries for both of the leukotrienes. In addition to very good extraction properties of XAD-4 and XAD-2 as compared to octadecylsilane silica, these solid adsorbent materials retained less plasma impurities than the C18 materials, giving cleaner chromatograms for leukotrienes extracted from plasma. Therefore, XAD-4 or XAD-2 are the best overall choice for extraction of leukotrienes from plasma for reversed-phase high-performance liquid chromatographic analysis.

Chromatography, High Pressure Liquid↗

Thin-film solid-phase extraction to measure fugacities of organic chemicals with low volatility in biological samples.

To investigate the environmental fate, food chain bioaccumulation, and toxicity of organic chemicals, it is often preferable to measure the chemical's fugacity rather than its concentration. However, simple methods to do this are rare. This paper presents a novel yet simple method to measure fugacities of a range of poorly volatile hydrophobic organic chemicals ranging in octanol-air partition coefficients from 10(5.6) to 10(9.2). Thin films of ethylene vinyl acetate coated on glass surfaces are used as solid-phase samplers of contaminated biological tissues. The technique is applied to fish tissue samples and spiked fish diets to determine method feasibility, equilibration times, reproducibility, and property characteristics of the thin films. It is concluded that the method provides an attractive technique to measure chemical fugacities in biological tissues without requiring solvent extractions and cleanup. The method is further expected to be applicable to investigate the fugacity of semivolatile and poorly volatile organic chemicals in air, water, sediments, and soil.

Chemistry Techniques, Analytical↗

On-line solid phase extraction with polyurethane foam: trace level spectrophotometric determination of iron in natural waters and biological materials.

This paper reports the development of a simple and accurate on-line procedure for preconcentration and determination of dissolved iron in waters and biological materials using unloaded polyether-type polyurethane foam as solid extractor. In the developed flow injection system, the analyte was preconcentrated from acidic aqueous medium as iron-thiocyanate complex with post-elution with ascorbic acid solution and spectrophotometric measurement with 1,10-phenanthroline as colorimetric reagent. In order to improve the performance of the system several chemical and flow variables were investigated as well as the effect caused by the presence of possible interferents. The method was validated by the analysis of two certified reference materials. Application of the methodology was carried out by the determination of dissolved iron content in eight natural water samples with different characteristics. The results were compared with those obtained by electrothermal atomic absorption spectrometry (ETAAS) and no statistical difference was observed. The detection limit was 0.75 microgram l-1 and the RSD was 1.2% for 2 min preconcentration time. At this condition, a productivity of 20 samples h-1 was achieved. Increasing the preconcentration time up to 3 min, a detection limit of 0.45 microgram l-1, an RSD of 1.5% and an analytical throughput of 15 h-1 were verified.

Automation↗

Determination of salbutamol using on-line solid-phase extraction and sequential injection analysis. Comparison of chemiluminescence and fluorescence detection.

Determination of salbutamol using sequential injection analysis (SIA) with chemiluminescence and fluorescence detection has been devised. The chemiluminescence signal was emitted during the oxidation of salbutamol by potassium permanganate in sulfuric acid medium. Sodium polyphosphate was used as chemiluminescence enhancer. The fluorescence signal (excitation wavelength 230 nm) was also measured in sulfuric acid medium. Both detection techniques were compared with respect to the application of the methods to the determination of salbutamol in biological materials. The sample pre-treatment takes place directly in the SIA system, when salbutamol is adsorbed on the solid-phase (Baker-carboxylic acid) microcolumn integrated into the system. Sulfuric acid serves both as the reagent and the eluent. The lab-made SIA system consisted of a 2.5-mL Cavro syringe pump, ten-port Vici Valco selection valve and Spectra-Physics FS 970 fluorescence detector, which was lab-modified for chemiluminescence detection. The system was controlled by a PC using originally compiled LabVIEW-supported software. Concentrations, volumes of reagents and flow rates were optimised by a simplex method. Salbutamol was determined in the linear range 0.05-10 microg mL(-1) (RSD 1.53%), with the detection limit (3 sigma) 0.03 microg mL(-1) and sample throughput of 42 samples per hour with chemiluminescence detection in standard solutions. The fluorescence detection enabled the determination of salbutamol in standard solutions in the linear range 0.5-100 microg mL(-1) (RSD 2.69%), with the detection limit 0.2 microg mL(-1) and sample throughput of 24 h(-1). The proposed methods were applied to the determination of salbutamol in human serum and urine. However, serum is a very complicated matrix and the SIA-SPE analysis did not provide satisfactory results. It was possible to determine salbutamol in human urine using this technique. Better recovery was achieved with fluorescence detection.

Adrenergic beta-Agonists↗

Solid phase extraction method for selective determination of Pb(II) in water samples using 4-(4-methoxybenzylidenimine) thiophenole.

A sensitive and selective extractive preconcentration procedure for the determination of traces of lead in water samples has been developed. An alumina-sodium dodecyl sulfate (SDS) coated modified with 4-(4-methoxybenzylidenimine) thiophenole (MBITP) was used for preconcentration and determination of Pb(II) by flame atomic absorption spectrometry. Lead was adsorbed quantitatively on modified column due to its complexation with MBITP and quantitatively eluted using 5 mL 1 mol L(-1) nitric acid in acetone. The effects of parameters such as pH, amount of solid phase, amount of MBITP, flow rate, type and concentration of eluting agent were examined. The effect of interfering ions on the determination of Pb(II) was also investigated. The response of proposed method is linear in the concentration range 0.05-1.2 microg mL(-1) of Pb(II). The limit of detections (3S.D.(b)/m, n=4) and relative standard deviations (n=11) are 1.6 ng mL(-1) and 0.9%, respectively. The presented procedure was successfully applied for determination of lead content in real samples such as river, spring, waste and drinking water.

Benzene Derivatives↗

Determination of eprinomectin in plasma by high-performance liquid chromatography with automated solid phase extraction and fluorescence detection.

A method is described for the determination of eprinomectin in plasma using high-performance liquid chromatography with fluorescence detection (excitation and emission wavelengths 355 and 465 nm, respectively). The fluorescent derivative was obtained by a condensation reaction with trifluoroacetic anhydride and N-methylimidazole. The method employs 1 ml plasma samples and gives linear calibration graphs (r = 0.999) over the concentration range studied (0.5-50 ng ml-1). Solid extraction using the benchmate procedure was used for sample preparation. This method permits the determination of eprinomectin at levels as low as 0.1 ng ml-1 and is suitable for the pharmacokinetic study of eprinomectin in animals.

Animals↗

Simultaneous determination of Cd(II), Cu(II) and Pb(II) in surface waters by solid phase extraction and flow injection analysis with spectrophotometric detection.

A method for heavy metal monitoring using spectrophotometric detection is presented. Traces of Cu(II), Pb(II) and Cd(II) at the low microg l(-1) level can be determined simultaneously after both selective removal of metal interferences and preconcentration using 'extraction chromatographic resins'. Lewatit TP807'84, which contains di(2,4,4-trimethylpentyl)phosphinic acid as active component, was used as solid adsorbent. Two minicolumns containing this resin were used: one at pH 3.2 for the removal of interferences, such as Zn(II) and Fe(III), and the other at pH 5.5 for the selective preconcentration of the target analytes. Spectrophotometric determination used FIA methodology with sulfarsazene as chromogenic reagent and partial least-squares multivariate calibration. The method was successfully applied to the analysis of surface waters from the Llobregat river and ground water samples from wells in the Guadiamar basin. Accuracy, expressed in terms of recoveries, was in the range 80-120% and relative standard deviations were below 10%.

Cadmium↗

Solid-phase extraction and high-performance liquid chromatography applied to the determination of quinapril and its metabolite quinaprilat in urine.

Quinapril is an antihypertensive drug that belongs to the family of angiotensin-converting enzyme inhibitors. It is metabolized to quinaprilat, which is the compound that is really responsible for the therapeutic action. In this study, a rapid and simple liquid chromatographic method with photometric detection is described and applied to the determination of quinapril and quinaprilat in urine. The cleanup procedure for the urine samples consists of a solid-liquid extraction using C8 cartridges. Under these conditions, both compounds and the internal standard (enalapril maleate) are separated in less than 9 min. Recoveries for quinapril and quinaprilat are greater than 80%. The method is sensitive enough (detection limit of 60 ng/mL for quinapril and 50 ng/mL for quinaprilat) to be applied for the determination of quinapril and quinaprilat in urine samples obtained from four hypertensive patients after the intake of a therapeutic dose.

Antihypertensive Agents↗

A high-throughput solid-phase extraction assay capable of measuring diverse polyprenyl phosphate: sugar-1-phosphate transferases as exemplified by the WecA, MraY, and MurG proteins.

The bacterial proteins WecA and MraY are members of the polyprenyl phosphate:N-acetylhexosamine-1-phosphate transferase family, each of which catalyzes the transfer of a specific hexosamine 1-P from a soluble UDP-hexosamine substrate to a bactoprenyl phosphate carrier at the membrane surface. Currently, assays designed to quantitate the activity of these enzymes rely on paper chromatography or liquid-liquid extractions or are specialized to a few members of the family. We describe a generalizable, high-throughput, one-pot assay for these activities that uses a solid-liquid bead-based separation system to selectively adsorb the highly hydrophobic products of reaction. By judicious choice of radiolabeled UDP-hexosamine precursor, the same format can be used to quantitate not only diverse members of this transferase family, but also enzymes that catalyze the further modification of these transferase products. This possibility is exemplified by the MurG protein of bacterial cell wall synthesis, which catalyzes the addition of an N-acetylglucosamine residue to the product of the MraY reaction. Thus, the use of this flexible assay tool will allow a critical biochemical and enzymologic analysis of many such membrane-bound transferases in a similar setting.

Bacterial Outer Membrane Proteins↗