Development of a positive pressure driven micro-fabricated liquid chromatographic analyzer through rapid-prototyping with poly(dimethylsiloxane): Optimizing chromatographic efficiency with sub-nanoliter injections

Abstract

A rapid and low-cost means of developing a working prototype for a positive-displacement driven open tubular liquid chromatography (OTLC) analyzer is demonstrated. A novel flow programming and injection strategy was developed and implemented using soft lithography, and evaluated in terms of chromatographic band broadening and efficiency. A separation of two food dyes served as the model sample system. Sample and mobile phase flowed continuously by positive displacement through the OTLC analyzer. Rectangular channels, of dimensions 10 μm deep by 100 μm wide, were micro-fabricated in poly-dimethylsiloxane (PDMS), with the separation portion 6.6 cm long. Using a novel flow programming method, in contrast to electroosmotic flow, sample injection volumes from 0.5 to 10 nl were made in real-time. Band broadening increased substantially for injection volumes over 1 nl. Although underivatized PDMS proved to be a sub-optimal stationary phase, plate heights, H, of 12 μm were experimentally achieved for an unretained analyte with the rectangular channel resulting in a reduced plate height, h, of 1.2. Chromatographic efficiency of the unretained analyte followed the model of an OTLC system limited by mass-transfer in the mobile phase. Flow rates from 6 nl min −1 up to 200 nl min −1 were tested, and van Deemter plots confirmed plate heights were optimum at 6 nl min −1 over the tested flow rate range. Thus, the best separation efficiency, N of 5500 for the 6.6 cm length separation channel, was achieved at the minimum flow rate through the column of 6 nl min −1, or 3 ml year −1. This analyzer is a low-cost sampling and chemical analysis tool that is intended to complement micro-fabricated electrophoretic and related separation devices.