ADVANCING THE SEPARATION SCIENCES THROUGH THE DELIVERY OF NEW MATERIALS, TECHNOLOGY AND METHODOLOGY.

A thesis and collection of works submitted to Plymouth University in partial fulfilment for the degree of DOCTOR OF SCIENC

INVESTIGATION OF CHELATING DYE IMPREGNATED RESINS FOR THE ION CHROMATOGRAPHIC DETERMINATION OF TRACE METALS

A high-performance chelation ion chromatography (HPCIC) system using chelating dye impregnated resins for the determination of trace metals in various complex matrices is described. The parameters involved in the production of dye impregnated chelating columns have been identified and optimised, A range of chelating dyes have been impregnated successfully into high-performance substrates, producing a number of chelating columns capable of both metal preconcentration and separations. Each of the chelating columns produced exhibited unique chelating properties. These columns have been classified in terms of metal retention characteristics, separation capability and efficiency, dye loading and column capacities. A number of the above dye impregnated chelating columns have been employed successfully for the determination of trace metals in various sample types. This includes the determination of transition and heavy metals in seawater at low ug dm'^ levels, the determination of aluminium in seawater, the determination of alkaline earth metals in oil-well brines and the determination of barium and strontium in calcium containing matrices such as mineral waters and milk powder.Dionex (U.K) Lt

Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection

Capacitively coupled contactless conductivity detection (C4D) is presented as a novel and versatile means of visualising discrete zones of charged functional groups grafted onto polymer based monoliths. Monoliths were formed within 100 μm UV transparent fused silica capillaries and photografting methods were subsequently used to graft a charged functional monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) onto discrete regions of the “generic” monolith using a photomask. Post-modification monolith evaluation involves scanning the C4D detector along the length of the monolith to obtain a profile of the exact spatial location of grafted charged functionalities with millimetre accuracy. The methodology was extended to the visualisation of several zones of immobilised protein (bovine serum albumin) using photografted azlactone groups to enable covalent attachment of the protein to the monolith at precise locations along its length. In addition, the extent of non-specific binding of protein to the ungrafted regions of the monolith due to hydrophobic interactions could be monitored as an increase in background conductivity of the stationary phase. Finally, the technique was cross-validated using fluorescence microscopy by immobilising green fluorescent protein (GFP) in discrete zones and comparing the profiles obtained using both complementary techniques

AuNP-Agglomerated monoliths in pipette tips for lectin affinity extraction of glycoproteins

• To in situ fabricate ethylene dimethacrylate porous polymer monoliths within the confines of a commercial 20 μL polypropylene pipette tips. • To enhance the monolith surface area by immobilising AuNPs and then functionalise the AuNPs with ECL lectin for selective extraction of galactosylated proteins from complex media

Spiropyran modified microfluidic chip channels for photonically controlled sensor array detection of metal ions

Microfluidic chips are particularly attractive for analytical purposes because they provide a convenient small platform for rapid analysis and detection.1 Furthermore, spiropyrans dyes can be used as photonically controlled, self-indicating molecular recognition agents for the fabrication of sensors.2 Here, we show how through integrating the beneficial characteristics of microfluidic devices and spiropyrans dyes, a simple and very innovative chip for on-line metal ion sensor array can be realised. The chip (4x3cm) consists of four independent 180m depth, polydimethylsiloxane (PDMS) channels. 1’-(3-Carboxypropyl)-3,3’-dimethyl-6-nitrospiro-[2H-1]-benzopyran-2,2’-indoline is covalently immobilised on the ozone plasma activated PDMS microchannel surfaces. Upon exposure to UV light, the transparent PDMS channels change to light purple colour because the spiropyran molecules of the surface undergo a heterocyclic ring cleavage that result in the formation of the highly conjugated merocyanine form. When stock solutions of several ion metals (Ca2+,Zn2+,Hg2+,Cu2+) are pumped independently through the four channels, different optical responses were observed for each metal. 1-L.Basabe-Desmonts et al. Anal.Bioanal.Chem.(2008)390:307–315. 2-R.J.Byrne et al. J.Mat.Chem.(2006)16:1332-1337

Gold nano-particle modified silica monolithic micro-columns for selected chromatographic and biological applications.

Monolithic microcolumns and especially silica monoliths are showing several advantages compared to classical particle packed and organic polymeric monolithic columns: ease of production and functionalisation, excellent mechanical and thermal stability. Morphology of the monolithic columns can easily be tuned by simply changing the compositions of reaction mixtures. High porosity and interconnected flow-through pores ensure low back pressures at higher flow rates so increasing reaction speeds. High salt resistance allows use water based buffer solutions without any swelling of the stationary phase, large biomolecules can be utilised and conditions to prevent denaturation and comformation changes of these biomolecules can be maintained. Introduction of gold nano-particles on the surfaces of silica monoliths allows increase of the surface areas and alows creation of new, exotic surfaces. Gold shows strong affinity towards thiol groups, which can be found in different biomolecules so utilisation of this phenomena would allow production of micro-reactors and bioreactors in order to mimic biological reactions happening in living organisms and large biological systems. Silica monoliths were synthesised using classical sol-gel process. In order to immobilise gold nano-particles, surfaces of the silica monoliths were amminated using standard silanisation reaction with 3-aminopropyl-methyl-diethoxysilane. 20 nm citrate stabilised gold nano-particles were immobilised on the surfaces afterwards. Depending on the desired application, gold nano-oparticle modified silica monoliths were functionalised afterwards. Immobilisation of ionic species such as amino acids and small peptides would allow creation of stationary phase for ion chromatography, retention of enzymes and other biologically active molecules would allow to create micro-reactors. Leaving gold nano-particles unmodified would make ideal stationary phase for micro-extraction. These modified monoliths were characterised using microscopy techniques, such as scanning electron microscopy (SEM) and field emission SEM. They were used to characterise morphology of the monoliths as well as to evaluate the coverage of the surface with gold nano-particles. The fabricated stationary phases were used for selected biological and chromatographic applications (incorporanting classical chromatographic techniques in order to evaluate the performance of these new modified monolithic materials)

Effect of laser processing parameters and glass type on topology of micro-channels

Traditional processes to manufacture micro-fluidic devices include standard lithography, electron beam writing and photo-patterning. These techniques are well established but most are limited to surface micro-fabrication. Laser micro-machining provides an alternative for microfabrication of devices. This paper presents Design of Experiment models for the fabrication of micro-channel structures with four different types of glass, soda-lime, fused-silica, borosilicate and quartz. A 1.5kW CO2 laser with 90 μm spot size was used to fabricate micro-channels on the surface of glass sheets. Power, P, pulse repetition frequency, PRF, and translation speed, U, were set as control parameters. The resulting geometry of the channel (depth and width) and transmission capabilities were measured and analyzed. A comparison of the results of this experimental testing with the four glass types showed that quartz and fused-silica glasses would have better channel topologies for chemical sensing applications

Spiropyran modified PDMS micro-fluidic chip device for photonically controlled sensor array detection of metal ions

Micro‐fluidic chips are particularly attractive in biological and life sciences for analytical purposes because they provide a convenient small platform for rapid analysis and detection [1]. Using micro‐fluidic devices for the determination of ions emerges as a potential solution to some of the challenges not overtaken by conventional techniques e.g. atomic absorption, inductively‐coupled plasma‐optical emission, mass spectrometry and ion‐selective electrodes [2]. For example, these devices can integrate complex sample handling processes, calibration, and detection steps into a compact, portable system. Moreover they require small sample volumes (low μl or nl), consume little power, and are easily constructed for multi‐analyte detection, either through multiple parallel fluidic architectures or by using arrays of detection elements. Organic photochromic compounds like spiropyrans are particularly interesting targets for the development of new approaches to sensing since they offer new routes to multi‐functional materials that take advantage of their photo‐reversible interconversion between two thermodynamically stable states (a spiropyran (SP) form, and a merocyanine (MC) form), which have dramatically different charge, polarity and molecular conformations. Furthermore, they can be easily incorporated into membranes for improved robustness and ease of handling [3], but from our perspective, most interesting of all, they have metal ion‐binding and molecular recognition properties which are only manifested by the MC form. Based on the coordinationinduced photochromism characteristic of the MC form, spiropyrans have been employed as molecular probes for metal ions and organic molecules [4]. In this abstract, we show how through integrating the beneficial characteristics of micro‐fluidic devices and spiropyrans photoswitches, a simple and very innovative chip configured as an on‐line metal ion sensor array can be realised (Figure 1). The micro‐fluidic device consists of five independent 94 μm depth, 150 μm width channels fabricated in polydimethylsiloxane. The spiropyran 1’‐(3‐carboxypropyl)‐3,3’‐dimethyl‐6‐nitrospiro‐1‐benzopyran‐2,2’‐indoline (SP‐COOH) is immobilised by physical adsorption directly on ozone plasma activated PDMS micro‐channel walls. When the colourless, inactive, spiropyran coating absorbs UV light it switches to the highly coloured merocyanine form (MC‐COOH), which also has an active binding site for certain metal ions. Therefore metal ion uptake can be triggered using UV light and subsequently reversed on demand by shining white light on the coloured complex, which regenerates the inactive spiropyran form, and releases the metal ion. When stock solutions of several metal ions (Ca2+, Zn2+, Hg2+, Cu2+, Co2+) are pumped independently through the five channels, different optical responses were observed for each metal (Figure 2), (i.e. complex formation with metal ions is associated with characteristic shifts in the visible spectrum), and the platform can therefore be regarded as a micro‐structured device for online multi‐component monitoring of metal cations

In-process phase growth measurement technique in the fabrication of monolithic porous layer open tubular (monoPLOT) columns using capacitively coupled contactless conductivity

A technique for the in-process measurement of polymer stationary phase growth inside fused silica capillaries during the fabrication of monolithic porous layer open tubular (monoPLOT) columns is presented. In this work, capacitively coupled contactless conductivity detection (C4D) was applied as an online measurement tool for porous polymer layer growth within fused silica capillaries. The relationship between effective capillary diameter and C4D response was investigated for two polymers, butyl methacrylate–ethylene dimethacrylate (BuMA–EDMA) and polystyrene–divinylbenzene (PS–DVB) over a range of capillary diameters and layer thicknesses. The described technique can be used with both thermal and photo-initiated approaches for monoPLOT fabrication and provides an accurate, realtime measurement of the porous layer growth within the capillary, which should vastly improve columnto-column reproducibility. The technique was shown to be very precise, with a measured %RSD < 10%