High-speed cryo-focusing injection for gas chromatography: reduction of injection band broadening with concentration enrichment.

In order to maximize peak capacity and detection sensitivity of fast gas chromatography (GC) separations, it is necessary to minimize band broadening, and in particular due to injection since this is often a major contributor. A high-speed cryo-focusing injection (HSCFI) system was constructed to first cryogenically focus analyte compounds in a 6 cm long section of metal MXT column, and second, reinject the focused analytes by rapidly resistively heating the metal column via an in-house built electronic circuit. Since the cryogenically cooled section of column is small (∼750 nl) and the direct resistive heating is fast (∼6000 °C/s), HSCFI is demonstrated to produce an analyte peak with a 6.3 ms width at half height, w(1/2). This was achieved using a 1m long column with a 180 µm inner diameter (i.d.) operated at an absolute head pressure of 55 psi and an oven temperature of 60 °C, with a 10 V pulse applied to the metal column for 50 ms. HSCFI was also used to demonstrate the head space sampling and fast GC analysis of an aqueous solution containing six test analytes (acetone, methanol, ethanol, toluene, chlorobenzene, pentanol). Using Henry's law constants for each of the analytes, injected mass limits of detection (LODs) were typically in the low pg levels (e.g., 1.2 pg for acetone) for the high speed separation. Finally, to demonstrate the use of HSCFI with a complex sample, a gasoline was separated using a 20 m × 100 µm i.d. column and the stock GC oven for temperature programming, which provided a separation time of 200 s and an average peak width at the base of 440 ms resulting in a total peak capacity of 460 peaks (at unit resolution).

Diffusion coefficient measurement in a microfluidic analyzer using dual-beam microscale-refractive index gradient detection. Application to on-chip molecular size determination.

We report a microchip-based detection scheme to determine the diffusion coefficient and molecular mass (to the extent correlated to molecular size) of analytes of interest. The device works by simultaneously measuring the refractive index gradient (RIG) between adjacent laminar flows at two different positions along a microchannel. The device, referred to as a microscale molecular mass sensor (micro-MMS), takes advantage of laminar flow conditions where the mixing of two streams occurs essentially by diffusion across the boundary between the two streams. Two flows merge on the microchip, one containing solvent only, referred to as the mobile phase stream and one which contains the analyte(s) of interest in the solvent, i.e. the sample stream. As these two streams merge and flow parallel to each other down the microchannel a RIG is created by the concentration gradient. The RIG is further influenced by analyte diffusion from the sample stream into the mobile phase stream. Measuring the RIG at a position close to the merging point (upstream signal) and simultaneously a selected distance further down the microchannel (downstream signal) provides real-time data related to the extent a given analyte has diffused, which can be readily correlated to analyte molecular mass by taking the ratio of the downstream-to-upstream signals. For the dual-beam RIG measurements, a diode laser output is coupled to a single mode fiber optic splitter with two output fibers. Light from each fiber passes through a graded refractive index (GRIN) lens forming a collimated beam that then passes through the microchannel and then on to a position sensitive detector (PSD). The RIG at both detection positions deflects the two collimated probe beams. The deflection angle of each beam is then measured on two separate PSDs. The micro-MMS was evaluated using polyethylene glycols (PEGs), sugars, and as a detector for size-exclusion chromatography (SEC). Peak purity can be readily identified using the micro-MMS with SEC. The limit of detection was 0.9 ppm (PEG at 11 840 g/mol) at the upstream detection position corresponding to a RI limit of detection (LOD) (3sigma) of 7-10(-8) RI. The pathlength for the RIG measurement was 200 microm and the angular LOD was 0.23 micro(rad) with a detection volume of 8 nl at both positions. The average molecular mass resolution was 9% (relative standard deviation) for a series of PEGs ranging in molecular mass from 106 to 22 800 g/mol. With this excellent mass resolution, small molecules such as monosaccharides, disaccharides, and so on, are readily distinguished. The sensor is demonstrated to readily determine unknown diffusion coefficients.