Scaling-up ultrasound standing wave enhanced sedimentation filters.

Particle concentration and filtration is a key stage in a wide range of processing industries and also one that can be present challenges for high throughput, continuous operation. Here we demonstrate some features which increase the efficiency of ultrasound enhanced sedimentation and could enable the technology the potential to be scaled up. In this work, 20 mm piezoelectric plates were used to drive 100 mm high chambers formed from single structural elements. The coherent structural resonances were able to drive particles (yeast cells) in the water to nodes throughout the chamber. Ultrasound enhanced sedimentation was used to demonstrate the efficiency of the system (>99% particle clearance). Sub-wavelength pin protrusions were used for the contacts between the resonant chamber and other elements. The pins provided support and transferred power, replacing glue which is inefficient for power transfer. Filtration energies of ∼4 J/ml of suspension were measured. A calculation of thermal convection indicates that the circulation could disrupt cell alignment in ducts >35 mm high when a 1K temperature gradient is present; we predict higher efficiencies when this maximum height is observed. For the acoustic design, although modelling was minimal before construction, the very simple construction allowed us to form 3D models of the nodal patterns in the fluid and the duct structure. The models were compared with visual observations of particle movement, Chladni figures and scanning laser vibrometer mapping. This demonstrates that nodal planes in the fluid can be controlled by the position of clamping points and that the contacts could be positioned to increase the efficiency and reliability of particle manipulations in standing waves.

Separation of alkali metals using isotachophoresis with cryptand 222 as a leading electrolyte additive.

This work shows how the inclusion of cryptand 222 as a leading electrolyte additive in isotachophoresis affects the electrophoretic mobilities of alkali metal cations. Using isotachophoresis the separation of alkali metals can be difficult due to the similar electrophoretic mobilities of three of the ions: caesium, rubidium and potassium. However, the presence of cryptand 222 in the leading electrolyte retards the mobilities of the caesium, rubidium and potassium to a different extent allowing separations to be made. A novel electrolyte system was formulated which consisted of a leading electrolyte of 10 mM caesium hydroxide, 0.75 mM cryptand 222 buffered to pH 9.4 with glycylglycine and a terminating electrolyte of 10 mM tetrabutylammonium hydroxide. The use of this electrolyte system allowed good separations of mixtures of rubidium, potassium, sodium and lithium to be achieved. The method was also applied to the analysis of edible salt samples.

Miniaturised free flow isotachophoresis of bacteria using an injection moulded separation device.

A new design of miniaturised free flow electrophoresis device has been produced. The design contains a separation chamber that is 45 mm long by 31.7 mm wide with a depth of 50 µm and has nine inlet and nine outlet holes to allow for fraction collection. The devices were formed of polystyrene with carbon fibre loaded polystyrene drive electrodes and produced using injection moulding. This means that the devices are low cost and can potentially be mass produced. The devices were used for free flow isotachophoresis (FFITP), a technique that can be used for focussing and concentrating analytes contained within complex sample matrices. The operation of the devices was demonstrated by performing separations of dyes and bacterial samples. Analysis of the output from FFITP separations of samples containing the bacterium Erwinia herbicola, a biological pathogen, by cell culturing and counting showed that fractionation of the output was achieved.

Thiocyanate and nitrite analysis using miniaturised isotachophoresis on a planar polymer chip.

A new method has been developed to improve the determination of thiocyanate using isotachophoresis. This method uses complexation with copper(II) as a mechanism for improving the separation of thiocyanate from chlorate and perchlorate. By using a pH of 3.25 the method can also be used to analyse nitrite. Separations were carried out using a miniaturised poly(methyl methacrylate) (PMMA) separation device. Linearity was observed from 1.25 to 75 mg dm(-3) with a correlation coefficient of 0.998 for both thiocyanate and nitrite. Limits of detection for these two species were calculated to be 0.8 mg dm(-3) and 0.9 mg dm(-3) respectively. The method was successfully applied to the analysis of these anions in a range of samples including explosive residues.

Determination of the potassium content of explosive residues using miniaturised isotachophoresis.

A new method has been developed to allow the determination of potassium in post-explosion residues to be made using miniaturised isotachophoresis. The method is based on the use of a caesium leading ion with 4.5 mM 18-crown-6 ether added to retard the potassium to allow reliable determinations to be made. With the conditions selected no interference was noted from other small inorganic cations, such as ammonium, barium, calcium, magnesium, sodium or strontium. The method was successfully applied to the analysis of seven samples containing explosive residues obtained from the unconfined burning of several flash powders. The procedure was found to offer good linearity for potassium determinations over the concentration range of 1.25-150 µg/mL with a coefficient of determination of 0.999 achieved.

Isotachophoresis-based sample preparation of cellulases in sugarcane juice using bovine serum albumin as a model protein.

The foremost requirement of quantification of cellulases expressed in genetically modified sugarcane is an efficient sample clean-up. This work investigates the feasibility of isotachophoresis for this purpose. An electrolyte system comprising a leading electrolyte of 10mM formic acid at pH 9.0 and a terminating electrolyte of 10mM ß-alanine was devised and used to perform isotachophoresis of cellulases. The use of a simple front cutting method removed a majority of interfering species in the juice, thereby resulting in the formation of a distinct zone of desired proteins. In comparison to techniques such as ultrafiltration and liming, the analysis time and loss of desired proteins was lower when the sample was prepared by using isotachophoresis. Hence, isotachophoresis was an ideal choice for purification of the proteins in question from the remaining components in the juice.

A miniaturised isotachophoresis method for magnesium determination.

The use of malonic acid as a complexing agent has enabled a new method to be devised to allow the determination of magnesium to be made using miniaturised isotachophoresis. Using a leading electrolyte of 10 mmol L(-1) caesium hydroxide and 2 mmol L(-1) malonic acid at pH 5.1 gave the method a high specificity towards magnesium. Investigations using a poly(methyl methacrylate) chip device with an integrated conductivity detector showed that no interference from calcium, strontium, barium and sodium should occur. The method was found to be linear over the range of magnesium concentrations from 0.625 to 75 mg L(-1) and the limit of detection was calculated to be 0.45 mg L(-1). Separations were demonstrated with water samples but the procedure should also be applicable to more complex sample matrices such as inorganic explosive residues, blood or urine.

The simultaneous determination of chloride, nitrate and sulphate by isotachophoresis using bromide as a leading ion.

A new method has been devised to allow the determination of small inorganic anions using isotachophoresis. This method makes use of indium(III) as a counter ion to manipulate the effective mobilities of inorganic anion species by means of complexation reactions. This new procedure successfully allowed the simultaneous determination of nitrate, chloride and sulphate to be realised on a capillary scale instrument and in a chip-based separation device. The electrolyte system developed to allow the separation to be achieved employed a 10mM bromide-based leading electrolyte containing 1.25 mM indium(III) at pH 3.15 and a terminating electrolyte of cyanoacetic acid.

Determination of chlorine containing species in explosive residues using chip-based isotachophoresis.

A new method has been developed to allow the determination of the chlorate, chloride and perchlorate anions in inorganic explosive residues to be made using isotachophoresis (ITP). To enable a good separation of these species to be achieved the method involves the use of two complexing agents. Indium(III) is used to allow the determination of chloride whilst using nitrate as the leading ion and alpha-cyclodextrin is used to allow the separation of chlorate and perchlorate. Separations were carried out using a miniaturised poly(methyl methacrylate) (PMMA) separation device. The method was applied to analysing both model samples and actual inorganic explosive containing residue samples. Successful determinations of these samples were achieved with no interference from other anions typically found in inorganic explosive residues. Limits of detection (LOD) for the species of interest were calculated to be 0.80 mg l(-1) for chloride, 1.75 mg l(-1) for chlorate and 1.40 mg l(-1) for perchlorate.

Miniaturised isotachophoresis of DNA.

This paper presents the findings of a feasibility study investigating the behaviour of DNA under conditions of miniaturised isotachophoresis. An electrolyte system comprising a leading electrolyte of 5mM perchloric acid at pH 6.0 and a terminating electrolyte of 10mM gallic acid was devised and used to perform isotachophoresis of DNA containing samples on a miniaturised poly(methyl methacrylate) device. Under such conditions it was found that no separation of DNA fragments was observed with the substance migrating instead as a single isotachophoretic zone. Whilst such a result shows the method is unsuitable for analysis DNA it offers significant potential as a means of sample preparation for subsequent analysis using another method. This is because the single zone of DNA formed is preconcentrated to a constant concentration governed by the leading ion and is separated from all species with different effective electrophoretic mobilities.

Miniaturised isotachophoresis analysis.

The application of miniaturized total analysis systems (microTAS) has seen rapid development over the past few years. Isotachophoresis (ITP) has been transferred into microchip format for both electrophoretic separation and pretreatment purposes, due to its advantageous features including separation parameters controlled by electrolyte composition and high sample load capacity. The primary focus of this concise review is to summarize the basic features of microchip based ITP and its applications to the analysis and pretreatment of ionic compounds and biomolecules that have arisen since 1998.

Rapid chloride analysis using miniaturised isotachophoresis.

A new design of miniaturised separation device for performing isotachophoresis (ITP) has been produced. The device contains a simple arrangement of channels comprising a single separation channel with a 'double T' injection geometry. The device was produced in poly(methyl methacrylate) and incorporates an on-column conductivity detector. A new electrolyte system was developed to enable the rapid determination of chloride to be made. This electrolyte system uses a leading ion of 3.5 mM nitrate at pH 3.0 with 0.5 mM indium(III) added as a complexing agent. Use of this electrolyte system with the new separation device allowed chloride samples to be analysed in under 100 s, with a limit of detection (LOD) calculated to be 2.2 mg l(-1).

Analysis of chloride, bromide and iodide using miniaturised isotachophoresis on a planar polymer chip.

A new method has been developed to allow the determination of the halide anions chloride, bromide and iodide using isotachophoresis. This method employs a new electrolyte system which incorporates the novel application of indium(III) as a complexing agent. This electrolyte system was devised based on the findings of an investigation into the potential for using indium(III) as a complexing counter ion to selectively manipulate the effective mobilities of halide ions. A leading electrolyte incorporating 3.5 mmol dm(-3) of indium(III) allowed the simultaneous determination of chloride, bromide and iodide to be successfully achieved. The new procedure allows such separations to be made without interference from common inorganic anions such as sulfate and nitrate. Separations were performed using a miniaturised planar poly(methyl methacrylate) chip with integrated platinum wire conductivity detection electrodes. Using this instrumentation the limits of detection were calculated to be 0.7 mg dm(-3), 1.7 mg dm(-3) and 2.2 mg dm(-3) for chloride, bromide and iodide respectively.

The use of indium(III) as a complexing counter-ion to enable the separation of chloride, bromide, and iodide using isotachophoresis.

A new method has been devised to enable the determination of halide anions by isotachophoresis. This method uses an electrolyte system that employs indium(III) as a counter-ion to manipulate the effective mobilities of sample species by means of complexation reactions. This new procedure successfully enabled the simultaneous determination of the halide ions chloride, bromide, and iodide when a 12 mmol L(-1) nitrate-based leading electrolyte containing 3.5 mmol L(-1) indium(III) at pH 3.0 was used.

Analysis of amino acids by miniaturised isotachophoresis.

A method allowing the miniaturised isotachophoretic analysis of amino acids has been developed. To overcome the problems of carbonate contamination which occur when performing separations at alkaline pH levels glycolate was used as the leading ion. Addition of magnesium to the leading electrolyte as a counter species was found to improve the separations. The method has been used on a poly(methyl methacrylate) microdevice with integrated on-column conductivity detectors. The behaviour of a range of common amino acids was investigated and successful separations of up to seven amino acids were made. Good linearity was observed with calibration curves for aspartic acid and phenylalanine over the range 0.063-1.0 mM. Limits of detection for these two species were calculated to be 0.060 and 0.018 mM, respectively.

Use of miniaturised isotachophoresis on a planar polymer chip to analyse transition metal ions in solutions from metal processing plants.

An electrolyte system, using malic acid as a complexing agent, has been developed to allow the determination of transition metal cations using miniaturised isotachophoresis. The method allowed the simultaneous determination of Mn2+, Cr3+, Fe2+, Co2+, Zn2+ and Ni2+ to be made without interference from other common ions. Limits of detection were calculated to be in the range 0.5-1.0mg l(-1) for Mn2+, Cr3+ Co2+ and Zn2+ and 2.0 mg l(-1) for Fe2+ and 4.7 mg l(-1) for Ni2+. The successful analysis of five industrial samples, containing a range of these metal ions, obtained from metal processing plants were achieved in under 13 min. The separations were performed on a poly(methyl methacrylate) chip with integrated platinum wire conductivity detection electrodes.

Determination of the ascorbate content of photographic developer solutions using miniaturised isotachophoresis on a planar chip.

The use of miniaturised isotachophoresis, performed on a planar poly(methyl methacrylate) device with integrated platinum conductivity electrodes, for the analysis of the ascorbate content of photographic developer solutions has been investigated. An electrolyte system has been developed which enabled the analysis to be made without interference from any of the other components in the developer solution, a number of which were present at significantly higher concentrations than that of the ascorbate ions. Using this system, the ascorbate content of the developer solutions could be analysed in under 6 min. The limit of detection for ascorbate using miniaturised isotachophoresis was calculated to be 0.011 mmol dm(-3).

Determination of inorganic selenium species by miniaturised isotachophoresis on a planar polymer chip.

The use of miniaturised isotachophoresis to allow the simultaneous determination of two inorganic selenium species has been investigated using a poly(methyl methacrylate) chip with a 44-mm-long, 200-microm-wide, 300-microm-deep separation channel. The miniaturised device included an integrated on-column, dual-electrode conductivity detector and was used in conjunction with a hydrodynamic fluid transport system. A simple electrolyte system has been developed which allowed the separation of selenium(IV) and selenium(VI) species to be made in under 210 s. The limits of detection were calculated to be 0.52 mg L(-1) for selenium(IV) and 0.65 mg L(-1 )for selenium(VI). The method allowed the separation of the selenium species from a range of common anions including fluoride, nitrate, nitrite, phosphate, sulfate and sulfite.

Miniaturised isotachophoretic analysis of inorganic arsenic speciation using a planar polymer chip with integrated conductivity detection.

A new method allowing the analysis of inorganic arsenic species using isotachophoresis has been developed. This method has been shown to be suitable for use on both miniaturised planar polymer separation devices and capillary scale devices. A poly(methyl methacrylate) chip with integrated conductivity electrodes has been successfully used for the rapid analysis of inorganic arsenic species in under 600 s. Limits of detection of 1.8 mg l(-1) and 4.8 mg l(-1) for arsenic(V) and arsenic(II), respectively, have been achieved with the miniaturised device. The device has also been used to perform the simultaneous separation of arsenic(III), arsenic(V), antimony(III), molybdenum(VI) and tellurium(IV).

Bidirectional isotachophoresis on a planar chip with integrated conductivity detection.

The use of a miniaturised planar separation device with integrated conductivity detection for performing bidirectional isotachophoresis (ITP) is described. The chips were produced in poly(methyl methacrylate) (PMMA) using a milling procedure. To enable bidirectional ITP the devices were designed to inject samples into the centre of the section channel and incorporated two integrated on-column conductivity detectors, positioned at opposite ends of this channel. When used with a hydrodynamic sample transport system the devices were used for the analysis of a range of small ions: NH4+; Na+; Mg2+; Ca2+; Li+; NO3-; ClO4-; SO4(2-); F-. Results sucessfully achieved included the simultaneous separation of three anions and three cations.