Abstract:

Miniaturization is an important trend in modern analytical instrument development, including miniaturized gas chromatography and liquid chromatography, as well as micro bore columns and capillary-to-microfluidics-based platforms. Apart from the miniaturization of the separation column, which is the core part of a chromatographic system, other parts of the system, including the sampler, pumping system, gradient generation, and detection systems, have been miniaturized. Miniaturized liquid chromatography significantly reduces solvent and sample consumption while providing comparable or even better separation efficiency. When liquid chromatography is coupled with mass spectroscopy, a low flow rate can increase the ionization efficiency, leading to enhanced sensitivity of the mass spectrometer. In contrast, normal-scale liquid chromatography suffers from its relatively high volumetric flow rate, which challenges the scanning frequency of the mass spectrometer. On the other hand because of the small sample size, other detection strategies such as spectrometric methods cannot provide sufficient sensitivity and limits of detection. In this sense, mass spectrometry has become the detection method of choice for micro-scale liquid-phase chromatography. Miniaturized liquid chromatography can diminish sample dilution efficiently when extremely small amounts of samples are used. The main driving force for this miniaturization trend, especially in liquid-phase separations, is the desperate need for microscale analyses of biological and clinical samples, given these samples are precious and the sample size is usually very small. At present, microscale liquid-phase chromatography is the only method of choice for such small, precious, and highly informative samples. The miniaturization of liquid chromatography systems, especially chromatographic columns, would be advantageous to the modularization and integration of liquid chromatography instrumental systems. Chip liquid chromatography is an integration of chromatography columns, liquid control systems, and detection methods on a single microfluidic chip. Chip liquid chromatography is an excellent format for the miniaturization of liquid chromatography systems, and it has already attracted significant attention from academia and industry. However, this attempt is challenging, and great effort is required on fundamental techniques, such as the substrate material of the microfluidic chip, structure of the micro-chromatography column, fluid control method, and detection methods, in order to make the chips suitable for liquid chromatography. Currently, the major problem in chip liquid chromatography is that the properties of the chip substrate materials cannot meet the requirements for further miniaturization and integration of chip liquid chromatography. The strength of the existing chip substrate materials is generally below 60 MPa, and the material properties limit further advances in the miniaturization and integration of chromatographic chips. Therefore, new chip substrate materials and the standard of chip channel design such as channel size and channel structure should be the key for further development of chip liquid chromatography. Mainstream instrumentation companies as well as new start-up innovation companies are now undertaking efforts toward the development of microchip liquid chromatographic products. Agilent, the first instrumentation company that introduced commercial microchip liquid chromatographic columns to the market, has led this field. Apart from microchip-based columns, Agilent introduced trap columns for different kinds of biological molecules as well as gradient generation systems for microchip-based liquid phase chromatography. Recently, another start-up company introduced microchip columns based on the in situ microfabrication of the column bed rather than packing the column with a particulate material. Such developments in microfabrication may further propel the advancement of micro-scale liquid-phase chromatography to an unprecedented level, which is beyond the conventional components and materials employed in normal-scale liquid chromatography. This review introduces the recent research progress in microchip liquid chromatography technologies, and briefly discusses the current state of commercialization of microchips for liquid chromatography by major instrumentation companies.

Key words: liquid chromatography (LC), microchip chromatography, microfluidics, miniaturization, review