Broadband absorption spectroscopy for rapid pH measurement in small volumes using an integrated porous waveguide.

Broadband absorption spectroscopy integrated with microfluidics is widely used to measure the pH of samples available in small volumes. Single pass absorption spectroscopy, however, suffers from poor minimum detectable absorption coefficient because the optical path length is determined by the physical dimensions of microchannels. As a result, either large amounts of indicator dyes are added or instrumentally complex techniques are used to determine sample pH. This work addresses these limitations by the integration of metal-clad leaky waveguide (MCLW) with microfluidics to perform broadband absorption spectroscopy. The MCLW consisted of a porous agarose waveguide with an estimated thickness and swelling ratio values of 1.405 ± 0.009 µm and 20.96 ± 0.65% respectively coated onto a titanium layer 32.8 ± 0.43 nm thick on a glass slide. MCLW permits obtaining absorption spectra over most of the wavelength range of the visible region (i.e. 450 nm to 700 nm) with a speed of 12.5 spectra per second using 3.2 nl of sample. The minimum detectable decadic absorption coefficient measured using MCLW was 0.43 cm-1, which is intermediate between single pass absorption spectroscopy and instrumentally complex techniques such as broadband cavity enhanced absorption spectroscopy and whispering gallery mode resonators.

A novel leaky waveguide grating (LWG) device for evanescent wave broadband absorption spectroscopy in microfluidic flow cells.

Evanescent wave (EW) broadband absorption spectroscopy is commonly interfaced with a range of analytical systems, including microfluidic flow cells, for identification and quantitation of species. A miniaturised spectrometer integrated with a microfluidic flow cell is useful to permit on-site analysis of samples. This work reports a novel leaky waveguide grating (LWG) device, which is able to obtain an absorption spectrum of an analyte of interest (in this case, methylene blue) without an external spectrometer. At 600 nm, the spectral resolution and the minimum detectable absorption coefficient of the LWG device is 49.5 nm and 1.65 cm(-1), respectively. The LWG can be fabricated on birefringent substrates such as plastics because the performance of the device is independent of the polarisation of the excitation source. The LWG device was fabricated using microcontact printing and hence can be easily mass produced.

A polymeric waveguide resonant mirror (RM) device for detection in microfluidic flow cells.

A novel resonant mirror (RM) device, which consisted of silica sol-gel spacer and polystyrene waveguide layers on a standard microscope slide glass substrate, was developed in this work. The device was successfully used to measure the absorption spectrum of methylene blue with a limit of detection (LOD) of at most 20.8 µM at 635 nm and a minimum detectable absorption coefficient of 0.94 cm(-1). A RM device consisting of dye-doped polystyrene waveguide layer was then demonstrated to be suitable to monitor antibody-antigen (in this case, anti-IgG and IgG) binding and was shown to be capable of detecting at least 100 nM IgG. The sensitivity of the device was estimated to be 17.27° per refractive index unit (RIU), which corresponds to a resolution of 1.45 × 10(-4) RIU for the set-up used. The RM device developed in this work can be easily integrated with microfluidic devices to identify and quantify (bio) chemical species by either absorption spectroscopy or measurement of effective optical thickness or both. In addition, the device was fabricated using a simple and low cost fabrication technique, spin coating. Hence, it can be easily mass produced.

Using droplet-based microfluidic technology to study the precipitation of a poorly water-soluble weakly basic drug upon a pH-shift.

The purpose of this study is to develop a droplet-based microfluidic device capable of monitoring drug precipitation upon a shift from gastric pH (pH 1.5) to intestinal pH (pH 6.5-7.0). The extent of precipitation occurring in droplets over time was measured using a novel on-chip laser scattering technique specifically developed for this study. The precipitation of ketoconazole, a poorly water-soluble basic drug, was investigated under different concentrations and pH values. It has been shown that the drug precipitates rapidly under supersaturation. Two water-soluble aqueous polymers, namely, polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) have been evaluated as precipitation inhibitors. HPMC was shown to be the most potent precipitation inhibitor. It is envisaged that the microfluidic pH-shift method developed in this study would form a proof-of-concept study, towards the development of a high throughput method for screening pharmaceutical excipients/precipitation inhibitors.

Chemiluminescence detection flow cells for flow injection analysis and high-performance liquid chromatography.

We have examined a range of new and previously described flow cells for chemiluminescence detection. The reactions of acidic potassium permanganate with morphine and amoxicillin were used as model systems representing the many fast chemiluminescence reactions between oxidising agents and organic analytes, and the preliminary partial reduction of the reagent was exploited to further increase the rates of reaction. The comparison was then extended to high-performance liquid chromatography separations of α- and ß-adrenergic agonists, with permanganate chemiluminescence detection. Flow cells constructed by machining novel channel designs into white polymer materials (sealed with transparent films or plates) have enabled improvements in mixing efficiency and overall transmission of light to the photodetector.

Assessment of the feasibility of the use of conductive polymers in the fabrication of ion mobility spectrometers.

The development of an ion mobility spectrometer with an injection molded plastic drift tube made from carbon-loaded nylon and the cyclo-olefinpolymer Zeonex is described. Thermogravimetric assessment combined with headspace analysis by ion mobility spectrometry and gas chromatography-mass spectrometry indicated that Zeonex encapsulated carbon-loaded nylon could be used to fabricate a snap-together injection molded stacked ring drift tube, 4.25 cm long that could be substituted for a conventional wire-wound heated ceramic drift tube of the same length into a high temperature ion mobility spectrometer. Temperature stability experiments indicated that such a combination of polymers produced stable water-based reactant ion peaks [(H(2)O)(n)H](+) up to a temperature of approximately 50 °C. Above this temperature, ammonia appeared to outgas, resulting in the production of [(H(2)O)(n)(NH(4))(m)H](+) type species before, at higher temperatures, the release of oligomeric entities suppressed resolved ion responses. Surface charging effects were also observed, and over a period of continuous operation of 4 h, these caused suppression of the signal intensity (1.11-0.954 V) and an apparent mobility shift in the observed responses (K(0) = 1.86-1.90 cm(2) V(-1) s(-1)). Substituting nylon, a polymer with a significantly lower surface resistivity, for the Zeonex demonstrated how surface charging phenomena could be managed though control of surface resistivity in future polymer formulations. The device was challenged successfully with test atmospheres of hexan-1-ol (K(0) = 1.66 cm(2) V(-1) s(-1) (monomer) and 1.32 cm(2) V(-1) s(-1)(dimer)) and dimethylmethyl phosphonate (K(0) = 1.70 cm(2) V(-1) s(-1) (monomer) and 1.44 cm(2) V(-1) s(-1) (dimer)). The potential advantages of developing polymeric systems using more advanced polymer formulations are discussed.

Continuous two-phase flow miniaturised bioreactor for monitoring anaerobic biocatalysis by pentaerythritol tetranitrate reductase.

A novel continuous recirculating two-phase flow miniaturised bioreactor was developed for biocatalytic transformations with the enzyme pentaerythritol tetranitrate reductase using on-chip spectroscopic detection of the organic and aqueous phases. A phase separation technique is described that uses electrostatic attraction to force charged droplets to merge back into the aqueous phase and thus allow the monitoring of both reaction phases during enzymatic turnover. We report an increased rate of enzyme catalysed reduction of trans-2-(2-nitrovinyl)thiophene, which was used as a model system to demonstrate the principles of the bioreactor design, compared to conventional macroscale experiments. Additional data obtained with ketoisophorone, trans-cinnamaldehyde and 2-methylmaleimide support our findings and provide a basis for improving the chemistry of biocatalysis.

Optimisation and analysis of microreactor designs for microfluidic gradient generation using a purpose built optical detection system for entire chip imaging.

This paper presents and fully characterises a novel simplification approach for the development of microsystem based concentration gradient generators with significantly reduced microfluidic networks. Three microreactors are presented; a pair of two-inlet six-outlet (2-6) networks and a two-inlet eleven-outlet (2-11) network design. The mathematical approach has been validated experimentally using a purpose built optical detection system. The experimental results are shown to be in very good agreement with the theoretical predictions from the model. The developed networks are proven to deliver precise linear concentration gradients (R(2) = 0.9973 and 0.9991 for the (2-6) designs) and the simplified networks are shown to provide enhanced performance over conventional designs, overcoming some of the practical issues associated with traditional networks. The optical measurements were precise enough to validate the linearity in each level of the conventional (2-6) networks (R(2) ranged from 0.9999 to 0.9973) compared to R(2) = 1 for the theoretical model. CFD results show that there is an effective upper limit on the operating flow rate. The new simplified (2-11) design was able to maintain a linear outlet profile up to 0.8 microl/s per inlet (R(2) = 0.9992). The proposed approach is widely applicable for the production of linear and arbitrary concentration profiles, with the potential for high throughput applications that span a wide range of chemical and biological studies.

Precision milled flow-cells for chemiluminescence detection.

Novel flow-cells with integrated confluence points and reaction channels designed for efficient mixing of fast chemiluminescence systems were constructed by machining opposing sides of a polymer chip and sealing the channels with transparent epoxy-acetate films. A hole drilled through the chip provided the conduit from the confluence point on one side to the centre of the reaction zone on the other side, allowing rapid presentation of the reacting mixture to the photodetector. The effectiveness of each flow-cell was evaluated by comparing the chemiluminescence intensity using flow injection analysis methodology, and examining the distribution of light emanating from the reaction zone (captured by photography in a dark room) when the reactants were continuously merged. Although previously reported chemiluminescence detectors constructed by machining channels into polymers have almost exclusively been prepared using transparent materials, we obtained far greater emission intensities using an opaque white chip with a thin transparent seal, which minimised the loss of light through surfaces not exposed to the photomultiplier tube. Furthermore, this approach enabled the exploration of reactor designs that could not be incorporated in traditional coiled-tubing flow-cells.

Free flow isotachophoresis in an injection moulded miniaturised separation chamber with integrated electrodes.

An injection moulded free flow isotachophoresis (FFITP) microdevice with integrated carbon fibre loaded electrodes with a separation chamber of 36.4mm wide, 28.7 mm long and 100 microm deep is presented. The microdevice was completely fabricated by injection moulding in carbon fibre loaded polystyrene for the electrodes and crystal polystyrene for the remainder of the chip and was bonded together using ultrasonic welding. Two injection moulded electrode designs were compared, one with the electrode surface level with the separation chamber and one with a recessed electrode. Separations of two anionic dyes, 0.2mM each of amaranth and acid green and separations of 0.2mM each of amaranth, bromophenol blue and glutamate were performed on the microdevice. Flow rates of 1.25 ml min(-1) for the leading and terminating electrolytes were used and a flow rate of 0.63 ml min(-1) for the sample. Electric fields of up to 370 V cm(-1) were applied across the separation chamber. Joule heating was not found to be significant although out-gassing was observed at drive currents greater than 3 mA.

A microfluidic device for self-synchronised production of droplets.

The primary requirement for a mixing operation in droplet-based microfluidic devices is an accurate pairing of droplets of reaction fluids over an extended period of time. In this paper, a novel device for self-synchronous production of droplets has been demonstrated. The device uses a change in impedance across a pair of electrodes introduced due to the passage of a pre-formed droplet to generate a second droplet at a second pair of electrodes. The device was characterised using image analysis. Droplets with a volume of ~23.5 ± 3.1 nl (i.e.~93% of the volume of pre-formed droplets) were produced on applying a voltage of 500 V. The synchronisation efficiency of the device was 83%. As the device enables self-synchronised production of droplets, it has a potential to increase the reliability and robustness of mixing operations in droplet-based microfluidic devices.