Retention Time Trajectory Matching for Peak Identification in Chromatographic Analysis.

Retention time drift caused by fluctuations in physical factors such as temperature ramping rate and carrier gas flow rate is ubiquitous in chromatographic measurements. Proper peak matching and identification across different chromatograms is critical prior to any subsequent analysis but is challenging without using mass spectrometry. The purpose of this work was to describe and validate a peak matching and identification method called retention time trajectory (RTT) matching that can be used in targeted analyses free of mass spectrometry. This method uses chromatographic retention times as the only input and identifies peaks associated with any subset of a predefined set of target compounds. An RTT is a two-dimensional (2D) curve formed uniquely by the retention times of the chromatographic peaks. The RTTs obtained from the chromatogram of a sample under test and those pre-installed in a library are matched and statistically compared. The best matched pair implies identification. Unlike most existing peak-alignment methods, no mathematical warping or transformation is involved. Based on the experimentally characterized RTT, an RTT hybridization method was also developed to rapidly generate more RTTs and expand the library without performing actual time-consuming chromatographic measurements, which enables successful peak matching even for chromatograms with severe retention time drifts. Additionally, 3.15 × 105 tests using experimentally obtained gas chromatograms and 2 × 1012 tests using two publicly available fruit metabolomics datasets validated the proposed method, demonstrating real-time peak/interferent identification.

Portable comprehensive two-dimensional micro-gas chromatography using an integrated flow-restricted pneumatic modulator.

Two-dimensional (2D) gas chromatography (GC) provides enhanced vapor separation capabilities in contrast to conventional one-dimensional GC and is useful for the analysis of highly complex chemical samples. We developed a microfabricated flow-restricted pneumatic modulator (FRPM) for portable comprehensive 2D micro-GC (µGC), which enables rapid 2D injection and separation without compromising the 1D separation speed and eluent peak profiles. 2D injection characteristics such as injection peak width and peak height were fully characterized by using flow-through micro-photoionization detectors (µPIDs) at the FRPM inlet and outlet. A 2D injection peak width of ~25 ms could be achieved with a 2D/1D flow rate ratio over 10. The FRPM was further integrated with a 0.5-m long 2D µcolumn on the same chip, and its performance was characterized. Finally, we developed an automated portable comprehensive 2D µGC consisting of a 10 m OV-1 1D µcolumn, an integrated FRPM with a built-in 0.5 m polyethylene glycol 2D µcolumn, and two µPIDs. Rapid separation of 40 volatile organic compounds in ~5 min was demonstrated. A hybrid 2D contour plot was constructed by using both 1D and 2D chromatograms obtained with the two µPIDs at the end of the 1D and 2D µcolumns, which was enabled by the presence of the flow resistor in the FRPM.

A Microcolumn DC Graphene Sensor for Rapid, Sensitive, and Universal Chemical Vapor Detection.

Nearly all existing direct current (DC) chemical vapor sensing methodologies are based on charge transfer between sensor and adsorbed molecules. However, the high binding energy at the charge-trapped sites, which is critical for high sensitivity, significantly slows sensors' responses and makes the detection of nonpolar molecules difficult. Herein, by exploiting the incomplete screening effect of graphene, we demonstrate a DC graphene electronic sensor for rapid (subsecond) and sensitive (ppb) detection of a broad range of vapor analytes, including polar, nonpolar, organic, and inorganic molecules. Molecular adsorption induced capacitance change in the graphene transistor is revealed to be the main sensing mechanism. A novel sensor design, which integrates a centimeter-scale graphene transistor and a microfabricated flow column, is pioneered to enhance the fringing capacitive gating effect. Our work provides an avenue for a broad spectrum real-time gas sensing technology and serves as an ideal testbed for probing molecular physisorption on graphene.

Microfabricated ionic liquid column for separations in dry air.

Micro gas chromatography (µGC) is a technique developed for rapid, in situ analysis of volatile organic compounds (VOCs) for environmental protection, industrial monitoring, and toxicology. While reduced µGC size and power requirements allow for increased portability, the low moisture and oxygen resilience of current microcolumn technology result in increased peak broadening and tailing for humid samples, which necessitates the use of bulky helium or nitrogen carrier gas cartridges. Developing a microcolumn to address these deficiencies is desirable to improve µGC field performance and further reduce µGC system size. This paper reports the development and characterization of a microfabricated phosphonium ionic liquid (µIL) column and demonstrates separation of both polar and nonpolar compounds using this column via analyses of alcohols, chloroalkanes, aromatics, aldehydes, fatty acid methyl esters, and alkanes. The µIL column achieved operation at temperatures up to 345 °C for fatty acid methyl ester and alkane separation. Notably, all separations in this study used dry air as the carrier gas, showing that analysis of a diverse range of compounds was possible in the presence of oxygen. After exposure to dry air for 48 h at temperatures up to 220 °C, the µIL column's peak capacity was only degraded by 8.92%, which validated its long-term robustness against oxygen. The column's separation performance was not degraded by high moisture concentrations or long-term moisture exposure, also manifesting its robustness to moisture. The high temperature, moisture, and oxygen resilience of the µIL column enable more rapid separations in varying field environments without requiring additional µGC accessories (e.g., humidity filters and carrier gas cartridges). The µIL column is therefore expected to be useful for integration into future µGC devices.

Fast and Reproducible ELISA Laser Platform for Ultrasensitive Protein Quantification.

Optofluidic lasers are currently of high interest for sensitive intracavity biochemical analysis. In comparison with conventional methods such as fluorescence and colorimetric detection, optofluidic lasers provide a method for amplifying small concentration differences in the gain medium, thus achieving high sensitivity. Here, we report the development of an on-chip ELISA (enzyme-linked immunosorbent assay) laser platform that is able to complete an assay in a short amount of time with small sample/reagent volumes, large dynamic range, and high sensitivity. The arrayed microscale reaction wells in the ELISA lasers can be microfabricated directly on dielectric mirrors, thus significantly improving the quality of the reaction wells and detection reproducibility. The details of the fabrication and characterization of those reaction wells on the mirror are described and the ELISA laser assay protocols are developed. Finally, we applied the ELISA laser to detecting IL-6, showing that a detection limit of about 0.1 pg/mL can be achieved in 1.5 h with 15 µL of sample/reagents per well. This work pushes the ELISA laser a step closer to solving problems in real-world biochemical analysis.

Peak focusing based on stationary phase thickness gradient.

This paper reports the development of a stationary phase thickness gradient gas chromatography (GC) column that enables analyte peak focusing and improves separation resolution. Theoretical analysis and simulation demonstrate focusing via a positive thickness gradient, i.e., the stationary phase thickness increases along the column. This effect was experimentally verified by coating a 5 m long capillary column with a film thickness varying from 34 nm at the column inlet to 241 nm at the column outlet. The column was analyzed in forward (thin to thick) and backward (thick to thin) modes and compared to a uniform thickness column with a thickness of 131 nm, using alkanes ranging from C5 to C16 and aromatics. Comparison of resolutions between forward mode and the uniform thickness column demonstrated an overall focusing rate (i.e., improvement in peak capacity) of 11.7% on alkanes and 28.2% on aromatics. The focusing effect was also demonstrated for isothermal room temperature separation of highly volatile compounds and temperature programmed separation with different ramping rates. In all cases, peak capacities from forward mode separations are higher than those from other modes, indicating the ability of a positive thickness gradient to focus analyte peaks. This thickness gradient technique can therefore be broadly applied to various stationary phases and column types as a general method for improving GC separation performance.

Microfabricated porous layer open tubular (PLOT) column.

Development of micro gas chromatography (µGC) is aimed at rapid and in situ analysis of volatile organic compounds (VOCs) for environmental protection, industrial monitoring, and toxicology. However, due to the lack of appropriate microcolumns and associated stationary phases, current µGC is unable to separate highly volatile chemicals such as methane, methanol, and formaldehyde, which are of great interest for their high toxicity and carcinogenicity. This inability has significantly limited µGC field applicability. To address this deficiency, this paper reports the development and characterization of a microfabricated porous layer open tubular (µPLOT) column with a divinylbenzene-based stationary phase. The separation capabilities of the µPLOT column are demonstrated by three distinct analyses of light alkanes, formaldehyde solution, and organic solvents, exhibiting its general utility for a wide range of highly volatile compounds. Further characterization shows the robust performance of the µPLOT column in the presence of high moisture and at high temperatures (up to 300 °C). The small footprint and the ability to separate highly volatile chemicals make the µPLOT column highly suitable for integration into µGC systems, thus significantly broadening µGC's applicability to rapid, field analysis of VOCs.