The use of a polymer inclusion membrane for separation and preconcentration of orthophosphate in flow analysis

A highly sensitive flow analysis system has been developed for the trace determination of reactive phosphate in natural waters, which uses a polymer inclusion membrane (PIM) with Aliquat 336 as the carrier for on-line analyte separation and preconcentration. The system operates under flow injection (FI) and continuous flow (CF) conditions. Under optimal FI conditions the system is characterised by a linear concentration range between 0.5 and 1000 μg L⁻¹ P, a sampling rate of 10 h⁻¹, a limit of detection of 0.5 μg L⁻¹ P and RSDs of 3.2% (n = 10, 100 μg L⁻¹) and 7.7% (n = 10, 10 μg L⁻¹). Under CF conditions with 10 min stop-flow time and sample solution flow rate of 1.32 mL min⁻¹ the flow system offers a limit of detection of 0.04 μg L⁻¹ P, a sampling rate of 5 h⁻¹ and an RSD of 3.4% (n = 5, 2.0 μg L⁻¹). Interference studies revealed that anions commonly found in natural waters did not interfere when in excess of at least one order of magnitude. The flow system, operating under CF conditions, was successfully applied to the analysis of natural water samples containing concentrations of phosphate in the low μg L⁻¹ P range, using the multipoint standard addition method.     

Transport extraction for trace element separation and preconcentration

Summary A new method of sorbent supported transport extraction based on the use of open-cell polyurethane foam sorbents in solvent sublation separation has been developed. The method has been shown to be effective for the separation and preconcentration of radioiodine and/or stable iodine from 41 of deionized, drinking (tap) and surface (river) waters using N-cetylpyridinium chloride as the cationic surfactant and N₂ carrier gas as the bubbling medium. A transport extraction efficiency of about 94% was obtained in 60 to 90 min; a 300-fold preconcentration was found for radioiodine as measured by direct radioactivity counting. Both radioiodine and stable iodine can be back-extracted from the polyurethane foam support (e.g. into acetone with an efficiency of almost 95%).Part II: This journal (1993) 346:905On leave from the Institute of Radioecology and Applied Nuclear Techniques, P.O. Box A-41, 04061 Koice, Slovakia     

Two-dimensional electrophoresis of membrane proteins: separation of myelin proteins

A method for two-dimensional electrophoretic separation of myelin proteins is presented. The first dimension consists of isoelectric focusing of lyophilized and delipidated membrane proteins, solubilized in a mixture of the nonionic detergent Triton X-100, the zwitterionic detergent CHAPS, 9 M urea and carrier ampholytes, and incorporated into a slab gel before separation. Subsequent discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis was performed by moulding the isoelectric focusing slab gel with its supporting glass plate into the stacking gel. This method proved to give highly reproducible results since mechanical forces and thus the risk of stretching, folding or rupture of the isoelectric focusing slab gel is minimized. Furthermore, by immunoblotting, the positions of myelin-associated glycoprotein and 2',3'-cyclic nucleotide 3'-phosphodiesterase were established with specific antibodies.     

Transport extraction for trace element separation and preconcentration

Summary Transport extraction based on solvent sublation has been proposed for separation and preconcentration of trace analytes from aqueous solutions. The experimental capabilities of this method were investigated using a complex of molecular iodine with ionic surfactants. A comparison of transport extraction with conventional solvent extraction was also made. The theoretical principles of transport extraction have been revealed and the basic difference between these two extractive separations has been explained. The results showed that the non-equilibrium nature of transport extraction gives it some distinctive advantages over equilibrium solvent extraction.On leave from the Institute of Radioecology and Applied Nuclear Techniques, P.O.Box A-41, CS-04061 Kosice, Czechoslovakia     

Emulsion membrane systems for preconcentration of uranium

Various double emulsion systems with liquid membranes containing tri-n-octylphosphine oxide /TOPO/, tri-n-butylphosphate /TBP/, KELEX 100^® and di-2-ethylhexylphosphoric acid /DEHPA/, as carriers for uranium/VI/ pertraction, were studied. The conditions were found at which the systems are most efficient, comparing with solvent extraction. The use of DEHPA as a membrane carrier with solutions of H₂SO₄ and H₃PO₄ encapsulated in the membrane was substanfiated.     

In-line coupling SPE and CE for DNA preconcentration and separation

An in-line SPE method coupled to CE was developed for the analysis of DNA. The amino silica monolith was prepared in situ by polymerization of tetraethoxysilane and N-(β-aminoethyl)-γ-aminopropyltriethoxysilane in ethanol aqueous solution at the inlet end of a 100?μm id fused-silica capillary, and the remaining part of the capillary was used as separation channel. The procedure for this in-line SPE-CE method was constructed on the basis of investigation on operational conditions such as the introduction mode of sieving matrix, the composition of elution solvent and the elution time. Twenty millimolar ammonium hydroxide was demonstrated to be effective for DNA desorption from the monolith, and linear poly(N-isopropylacrylamide) was used as the separation matrix. The proposed method could achieve limits of detection of 0.065-0.123?ng/mL for six DNA fragments ranging 100-2000?bp. Compared with conventional CE, preconcentration factors of over 100 times were obtained. The applicability of the in-line SPE-CE method was further demonstrated by analyzing plasmid DNA from Escherichia coli crude lysate.     

Isoelectric focusing of membrane proteins: high resolution separation of myelin proteins

A mixture of the nonionic detergent Triton X-100, the zwitterionic detergent 3-[(cholamidopropyl)dimethylammonio]-1-propanesulphonate (CHAPS), 9M urea and carrier ampholytes was found comparable to media containing sodium dodecyl sulfate in the capacity for solubilization of myelin proteins, including the highly hydrophobic proteolipid protein. The solubilized sample was incorporated into the polymerization mixture before moulding an ultrathin gel, with heat convection characteristics allowing a high wattage to be applied, thus allowing fast separation with high resolving power. Since the most basic protein in myelin focuses at a pH greater than 10, fast separation is essential in order to minimize decay of the cathodic end of the pH gradient.     

Model experiments to test the use of a liquid membrane for separation and preconcentration of copper from natural water

The transfer and separation of Cu(II) ions across a bulk liquid membrane (BLM) containing pyridine-2-acetaldehyde benzoylhydrazone (2-APBH) as the mobile carrier dissolved in toluene has been investigated and optimised. The system was applied to the preconcentration of copper from natural waters prior to analysis by flame atomic absorption spectroscopy. The flux of copper across the membrane has been studied, and characterised as a function of analytical variables such as the carrier concentration, volume of organic phase, pH of feed and strip receiving solutions, stirring rate and temperature of solutions. The preconcentration yield at optimum conditions was 100.54±0.94%, even with a high saline matrix (30 g l⁻¹ NaCl), with good precision (1.49%). A preconcentration factor of approximately 18 times could be obtained. The detection limit of a blank sample was 0.24 μg l⁻¹ of Cu.The method was validated using a certified reference material (TMDA-62) and was applied successfully to the analysis of copper in two samples of seawater collected from the coast of Huelva (Spain). The relative errors were 2.42% for CRM and 0.48 and 3.66%, for seawaters (obtained between the results of the proposed and DPASV methods), respectively.     

Bioanalytical separation and preconcentration using ionic liquids

Ionic liquids (ILs) are novel solvents that display a number of unique properties, such as negligible vapor pressure, thermal stability (even at high temperatures), favorable viscosity, and miscibility with water and organic solvents. These properties make them attractive alternatives to environmentally unfriendly solvents that produce volatile organic compounds. In this article, a critical review of state-of-the-art developments in the use of ILs for the separation and preconcentration of bioanalytes in biological samples is presented. Special attention is paid to the determination of various organic and inorganic analytes—including contaminants (e.g., pesticides, nicotine, opioids, gold, arsenic, lead, etc.) and functional biomolecules (e.g., testosterone, vitamin B₁₂, hemoglobin)—in urine, blood, saliva, hair, and nail samples. A brief introduction to modern microextraction techniques based on ILs, such as dispersive liquid–liquid microextraction (DLLME) and single-drop microextraction (SDME), is provided. A comparison of IL-based methods in terms of their limits of detection and environmental compatibilities is also made. Finally, critical issues and challenges that have arisen from the use of ILs in separation and preconcentration techniques are also discussed.

Hollow fiber membrane concentrator for on-line preconcentration

Solvent removal by membrane permeation is presented as a method for on-line preconcentration. Experiments were carried out to develop a one-step, on-line, concentration process using a microporous composite hydrophobic membrane, or a polar solvent-permeable Nafion membrane depending on the solvent. Both polar and non-polar hollow fiber membranes were found to be effective in concentrating trace analytes. A large increase in analyte enrichment factors was found for both concentrator modules. Enrichment factors as high as 18.9 were observed. Residence time and operating temperature were found to be important parameters. Several different model compounds were preconcentrated. Further, in a Nafion membrane (polar solvent-permeable), analyte interaction with membrane bound sulfonic acid residues resulted in the loss of reactive analytes such as atrazine (ATZ). All analytes were successfully concentrated and detected using a polypropylene-siloxane composite membrane system when hexane was used as the solvent.     

A novel solid-phase extraction method for separation and preconcentration of zirconium

This work assesses the use of modified natural clinoptilolite as an adsorptive material for separation and preconcentration of trace amounts of zirconium ions. A simple, rapid and economical method was developed for the preconcentration of trace amounts of zirconium in aqueous medium using 1-(2-pyridylazo)-2-naphthol as a complexing agent. Effect of sample pH, flow rate of sample and elution solutions, breakthrough volume and interference of several ions were studied. Determination of zirconium was made by ICP-AES technique. The sorption was quantitative in the pH range from 3.0 to 4.0, whereas quantitative desorption occurred instantaneously with 2 mol L^?1 hydrochloric acid. Linearity was maintained between 0.05 and 9.0 μg mL^?1. Relative standard deviations range from ±0.9% to ±2.3% (n?=?5). The detection limit was 0.1 ng mL^?1. Because of good recovery (>97%), this method is suitable for preconcentration and determination of zirconium in effluents containing trace amount of zirconium.     

Chelating polymers and related supports for separation and preconcentration of trace metals

This review is concerned mainly with the applications of chelating polymeric resins for the separation and concentration of trace metals from oceans, rivers, streams and other natural systems. Commercially available resins, specially prepared polymers and a selection of other sorbents are described and their uses outlined. Special emphasis is placed on the preconcentration of uranium from sea-water.     

On-line separation and preconcentration in flow injection analysis

Various improvements in flow-injection systems involving on-line separation and preconcentration by gas diffusion, ion-exchange and liquid—liquid extraction are reviewed and their merits are discussed. On-line preconcentration systems based on gas diffusion are described for the spectrophotometric determination of cyanide, fluoride and ammonia. Different gas/liquid separation devices for hydride-generation and cold-vapour atomic absorption spectrometry (a.a.s.) are compared. Systems are outlined for on-line column preconcentration of cobalt, nickel, vanadium, beryllium and cerium for inductively-coupled plasma/atomic emission spectrometry, and for combinations of column preconcentration with hydride-generation and cold-vapour a.a.s for the determination of ultratrace selenium and mercury. An on-line liquid—liquid extraction/flame a.a.s. system capable of achieving 60-fold enhancement for lead is reported; the limit of detection is 0.02 μg ml^?.     

RS-01微柱分离富集装置的研制

研制了一台自动化微柱分离富集装置。该装置由两台两通道蠕动泵、三支旋转阀、控制电路和微型分离富集柱组成。时序电路控制旋转阀切换。自动完成柱平衡、进样、淋洗、洗脱、收集、再生过程。采用三阀六时段方式,能够有效地截取待测组分的高浓度洗脱液。配备相应的分离柱,该装置能在30min内完成Nd2O3基体中微量Tb、Dy、Ho元素的分离;与电感耦合等离子体质谱仪联用,分离测定的RSD为2．5％～3．6％,加标回收率范围为95．5％～109．0％。用以分离一测定高纯Eu2O3、Tb4O7、Nd2O3中痕量稀土杂质。     

Different strategies for the preconcentration and separation of parabens by capillary electrophoresis

Several strategies, namely, large volume sample stacking (LVSS), field‐amplified sample injection (FASI), sweeping, and in‐line SPE‐CE, were investigated for the simultaneous separation and preconcentration of a group of parabens. A BGE consisting of 20 mM sodium dihydrogenphosphate (pH 2.28) and 150 mM SDS with 15% ACN was used for the separation and preconcentration of the compounds by sweeping, and a BGE consisting of 30 mM sodium borate (pH 9.5) was used for the separation and preconcentration of the compounds by LVSS, FASI, and in‐line SPE‐CE. Several factors affecting the preconcentration process were investigated in order to obtain the maximum enhancement of sensitivity. The LODs obtained for parabens were in the range of 18–27, 3–4, 2, and 0.01–0.02 ng/mL, and the sensitivity evaluated in terms of LODs was improved up to 29‐, 77‐, 120‐, and 18 400‐fold for sweeping, LVSS, FASI, and in‐line SPE‐CE, respectively. These preconcentration techniques showed potential as good strategies for focusing parabens. The four methods were validated with standard samples to show the potential of these techniques for future applications in real samples, such as biological and environmental samples.     

Efficient separation and analysis of peroxisomal membrane proteins using free-flow isoelectric focusing

We have elaborated a protocol for the fractionation of both hydrophilic and hydrophobic proteins using as a model the matrix and membrane compartments of highly purified rat liver peroxisomes because of their distinct proteomes and characteristic composition with a high quota of basic proteins. To keep highly hydrophobic proteins in solution, an urea/thiourea/detergent mixture, as used in traditional gel-based isoelectric focusing (IEF), was added to the electrophoresis buffer. Electrophoresis was conducted in the ProTeam free-flow electrophoresis (FFE) apparatus of TECAN separating proteins into 96 fractions on a pH 3-12 gradient. Consecutive sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated that both matrix and the integral membrane proteins of peroxisomes could be successfully fractionated and then identified by mass spectrometry. This is documented by the detection of PMP22, which is the most hydrophobic and basic protein of the peroxisomal membrane with a pI > 10. The identification of 96 prominent spots corresponding to polypeptides with different physical and chemical properties, e.g., the most abundant integral membrane polypeptides of peroxisomes and specific ones of the mitochondrial and microsomal membrane, reflects the fractionation potential of free-flow (FF)-IEF, accentuating its value in proteomic research as an alternative perhaps superior to gel-based IEF.     

Pillar-structured microchannels for on-chip liquid chromatography: evaluation of the permeability and separation performance

The chromatographic characteristics were determined for a set of microfabricated separation channels structured with cylindrical and diamond-shaped pillars with a characteristic size of 5 microm. Channels with different structures and porosities were etched in a silicon wafer using lithographic techniques. The permeability for flow of the channels was shown to increase strongly with the overall porosity. Diamond-shaped pillars appeared to yield a slightly higher permeability than cylindrical pillars at the same channel porosity. Compared to packed columns, permeabilities were higher by a factor of up to 5. Band dispersion in the channels was measured with an unretained fluorescent probe compound using a fluorescence microscope. A relatively large variation in the observed plate heights between channels was found, which was mainly attributed to the inaccurate geometry of the structure close to the side walls. Reduced plate heights between 0.2 and 1.0 were obtained. The lowest plate heights were found for channels with low porosity. The chromatographic impedances were calculated and compared to the values for the traditional chromatographic systems. For one of the structured microchannels the impedance was found to be more than ten times lower than for a column packed with nonporous spherical particles. With the data collected, predictions are given on the possibilities in terms of efficiency and speed offered by structured microchannels for pressure-driven separations, taking practical constraints into account.     

Coupled affinity-hydrophobic monolithic column for on-line removal of immunoglobulin G, preconcentration of low abundance proteins and separation by capillary zone electrophoresis

A butyl methacrylate-co-ethylene dimethacrylate (BuMA-co-EDMA) monolith was synthesized by UV initiated polymerization at the inlet end of a 75 microm I.D. fused silica capillary that had been previously coated with a protein compatible polymer, poly(vinyl)alcohol. The monolith was used for on-line preconcentration of proteins followed by capillary electrophoresis (CE) separation. For the analysis of standard proteins (cytochrome c, lysozyme and trypsinogen A) this system proved reproducible. The run-to-run %RSD values for migration time and corrected peak area were less than 5%, which is typical of CE. As measured by frontal analysis using lysozyme as solute, saturation of a 1cm monolith was reached after loading 48 ng of protein. Finally, the BuMA-co-EDMA monolithic preconcentrator was coupled to a protein G monolithic column via a zero dead volume union. The coupled system was used for on-line removal of IgG, preconcentration of standard proteins and CE separation. This system could be a valuable sample preparation tool for the analysis of low abundance proteins in complex samples such as human serum, in which high abundance proteins, e.g., human serum albumin (HSA) and immunoglobulin G (IgG), hinder identification and quantification of low abundance proteins.