Chromatography with higher pressure, smaller particles and higher temperature: a bioanalytical perspective

Recent years have seen widespread use of ultra-high-pressure liquid chromatography (UHPLC) in biological fluids. Most commonly the emphasis is on developing high throughput assay methods to reduce the analysis time and cost. Particle size and temperature are chromatographic parameters that can be changed to improve efficiency and obtain rapid separations. UHPLC and high-temperature liquid chromatography (HTLC) are two techniques that reduce the analysis time by decreasing the size of the column packing material and increasing the column temperature, respectively. Both of these techniques have advantages and limitations. In this article we have summarized the history, theoretical background of UHPLC and HTLC and the various advantages and limitations of sample preparation techniques and the detection systems (mass spectrometry and ultraviolet) used for the bioanalytical assays. In addition, selected bioanalytical applications of the two techniques have been reviewed and tabulated.