Zarah Walsh

Polystyrene bead-based system for optical sensing using spiropyran photoswitches

Spiropyran derivatives have been immobilised on the surface of polystyrene microbeads using different immobilisation strategies. These functionalised polymeric beads can be reversibly switched between the colourless inactive spiropyran (SP) and highly coloured (purple) active merocyanine (MC) forms using low power light sources, such as light-emitting diodes (LEDs). A UV LED (375 nm) is used for the SP → MC conversion, and a white LED (430–760 nm) for the reverse MC → SP conversion. The photochromic behaviour of the coated beads has been characterised using different LEDs and reflection spectroscopy, employing optic fibres and an in-house-designed holder. Investigations into the metal-ion binding behaviour of the spiropyran-modified microbeads have shown that Cu2+ ions cause an appreciable colour and spectral change when brought into contact with the beads in the MC form, suggesting that a significant interaction is occurring. However, the Cu2+ ions can be completely expelled by photonic-conversion of the beads into the inactive SP form using a white LED. This sequence has been successfully repeated six times, suggesting that it is possible to cycle through activation of the functionalised beads from a non-binding to a binding form (SP → MC) using a UV LED, allow binding with Cu2+ ions to occur, and subsequently, expel the bound ions and regenerate the passive SP surface using a white LED. Other metals, such as calcium, do not cause any appreciable colour or spectral change over the same concentration range and in the presence of the same anion (final concentration 7.1 × 10−4 M nitrate salt in ethanol). The system is therefore self-indicating in terms of whether the active MC or inactive SP forms are present, and whether Cu2+ ions are bound to the MC form. In principle, therefore, these functionalised beads could form the basis of a photoswitchable stationary phase for metal ion binding and detection: irradiation of the stationary phase with UV LEDs causes retention of guest species due to the presence of the MC form, while subsequent exposure to white LEDs causes release of guest species into the mobile phase.

Visible light initiated polymerization of styrenic monolithic stationary phases using 470 nm light emitting diode arrays

Poly(styrene-co-divinylbenzene) monolithic stationary phases have been synthesized for the first time by photoinitiated polymerization. An initiator composed of (+)-(S)-camphorquinone/ethyl-4-dimethylaminobenzoate/N-methoxy-4-phenylpyridinium tetrafluoroborate was activated using a 470 nm light emitting diode array as the light source. Spatially controlled polymerization of styrenic monoliths has been achieved within specific sections of a 100 microm id polytetrafluoroethylene-coated fused-silica capillary using simple photo masking. The sharpness of the edges was confirmed by optical microscopy, while SEM was used to verify a typical porous, globular morphology. Flow resistance data were used to assess the permeability of the monoliths and they were found to have good flow through properties with a flow resistance of 0.725 MPa/cm at 1 microL/min (water, 20 degrees C). Conductivity profiling along the length of the capillary was used to assess their lateral homogeneity. Monoliths which were axially rotated during polymerization were found to be homogeneous along the whole length of the capillary. The monolithic stationary phases were applied to the RP gradient separation of a mixture of proteins. Column fabrication showed excellent reproducibility with the retention factor (k) having a RSD value of 2.6% for the batch and less than 1.73% on individual columns.

Polymerisation and surface modification of methacrylate monoliths in polyimide channels and polyimide coated capillaries using 660 nm light emitting diodes

An investigation into the preparation of monolithic separation media utilising a cyanine dye sensitiser/triphenylbutylborate/N-methoxy-4-phenylpyridinium tetrafluoroborate initiating system activated by 660 nm light emitting diodes is reported. The work demonstrates multiple uses of red-light initiated polymerisation in the preparation of monolithic stationary phases within polyimide and polyimide coated channels and the modification of monolithic materials with molecules which absorb strongly in the UV region. This initiator complex was used to synthesise poly(butyl methacrylate-co-ethylene dimethacrylate) and poly(methyl methacrylate-co-ethylene dimethacrylate) monolithic stationary phases in polyimide coated fused silica capillaries of varying internal diameters, as well as within polyimide micro-fluidic chips. The repeatability of the preparation procedure and resultant monolithic structure was demonstrated with a batch of poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths in 100 μm i.d. polyimide coated fused silica capillary, which were applied to the separation of a model protein mixture (ribonuclease A, cytochrome C, myoglobin and ovalbumin). Taking an average from 12 chromatograms originating from each batch, the maximum relative standard deviation of the retention factor (k) for the protein separations was recorded as 0.53%, the maximum variance for the selectivity factor (α) was 0.40% while the maximum relative standard deviation in peak resolution was 8.72%. All maxima were recorded for the Ribonuclease A/Cytochrome C peaks. Scanning electron microscopy confirmed the success of experiments in which poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths were prepared using the same initiation approach in capillary and micro-fluidic chips, respectively. The initiating system was also applied to the photo-initiated grafting of a chromophoric monomer onto poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths within poly(tetrafluoroethylene) coated fused silica capillaries.

Incorporation of Acrylate Based Spiropyran Monoliths in Micro-Fluidic Devices for Photo-Controlled Electroosmotic Flow

Spiropyran photochromic compounds can be switched using light exposure between a non-polar spiro form (SP) and a zwitterionic merocyanine form (MC) that is subject to protonation (MC-H+). It has recently been demonstrated by Walsh et al. that, under acidic conditions, electroosmotic flow (EOF) generated in vinyl based spiropyran monoliths can be modulated using light irradiation [1]. In this paper, we report a spiropyran-modified acrylate based monolith which is particularly sensitive to protonation in the MC form, producing a positively charged surface that converts to the unpolar SP form by exposure to white light. When the MC-H+ form is dominant, it produces a charged surface which enables a relatively high flow rate (up to 1.6 μl/min) to be generated under electroosmotic conditions. Upon exposure to white light, the concentration of MC-H+ decreases due to the photo-conversion to the uncharged SP form, with up to 20% reduction of the EOF. The process is reversible, and removal of the light source results in a flow increase back to the original rate. The ability to alter flow rates in micro-fluidic channels using light has very significant implications, as it could dramatically simplify the manner in which micro-flow systems are controlled.