Silica monolithic columns suitable for implementation on microchips have been evaluated by ion-exchange capillary electrochromatography. Two different silica monoliths were created from the alkyl silane, tetramethyl orthosilicate (TMOS), by introducing a water-soluble organic polymer, poly(ethylene oxide) (PEO), with varying molecular weights into the prehydrolyzed sol. Silica monoliths created using 10 kDa PEO were found to have a much more closed gel structure with a smaller percentage of pores in the microm size range than gels created using 100 kDa PEO. Additionally, the size of the mesopores in the 100 kDa PEO monolith was 5 nm, while those in the 10 kDa PEO gel were only 3 nm. This resulted in a strong dependence of the electroosmotic flow (EOF) on the ionic strength of the background electrolyte, with substantial pore flow through the nm size pores observed in the 10 kDa PEO gel. The chromatographic performance of the monolithic columns was evaluated by ion-exchange electrochromatography, with ion-exchange sites introduced via dynamic coating with the cationic polymer, poly(diallyldimethylammonium chloride) (PDDAC). Separating a mixture of inorganic anions, the 10 kDa PEO monolithic columns showed a higher effective capacity than the 100 kDa PEO column.
The potential for using polyelectrolyte multilayers (PEMs) to provide chromatographic functionality on continuous silica networks created from sol-gel chemistry has been evaluated by capillary electrochromatography (CEC). Construction of the PEM was achieved by flushing the column with polyelectrolytes of alternative charge, with variation of the properties of the exposed polyelectrolyte providing a unique means to vary the chromatographic surface. Variation of the exposed polyelectrolyte from poly(diallyldimethylammonium chloride) (PDDAC) to dextran sulfate (DS) allowed the direction of the electroosmotic flow (EOF) to be changed and also provided a means to vary the chromatographic capacity. Variation of negative polymer from DS to poly(styrene sulfonate) (PSS) significantly altered the EOF and the migration of peptides, with both the reversed-phase and ion-exchange capacities increasing. An alternative method for changing the column capacity was to change the thickness of the PEM, which was evaluated by anion-exchange CEC. A 70-80% increase in retention was observed for all anions without any increase in EOF suggesting significant penetration of the analytes through the PEM and interaction with buried charges within the PEM.
W ith a focus on low-cost and low-power consumption, a miniature laser-induced fluorescence (LIF) detection system was assembled using a 635 nm red diode laser as the excitation source and a photodiode element coupled with an operational amplifier for signal collection. The primary elements of the miniature system, namely the laser and the detection system, cost a combined $70 and required only 270 mW of power for operation. When compared to conventional systems assembled using an argon-ion laser source and a photomultiplier tube, this represents a 98% decrease in the cost, and greater than 5000-fold decrease in power consumption. Limits of detection (LOD) and quantitation (LOQ) of the miniature system, evaluated on a microfluidic device for Nile Blue dye diluted in ethanol, were approximately 15 and 40 nM, respectively. Detection of l-phage DNA on a microfluidic device using the miniature system was performed after mixing with an intercalating dye, TO-PRO 3. The LOD and LOQ of l-phage DNA after TO-PRO 3 intercalation were approximately 1 and 4 ng/mL, respectively. Quantitation of DNA on microdevices using the miniature LIF detection system was also performed with an error of less than 15%. This detection system is a step in the direction of commercializing microfluidic instrumentation by reducing the cost and power required for operation. ( JALA 2006;11:254-9)
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