High throughput liquid chromatography with sub-2 μm particles at high pressure and high temperature

In this study, ultra performance liquid chromatography (UPLC) using pressures up to 1000 bar and columns packed with sub-2 μm particles has been combined with high temperature mobile phase conditions (up to 90 °C). By using high temperature ultra performance liquid chromatography (HT-UPLC), it is possible to drastically decrease the analysis time without loss in efficiency. The stability and chromatographic behavior of sub-2 μm particles were evaluated at high temperature and high pressure. The chromatographic support remained stable after 500 injections (equivalent to 7500 column volumes) and plate height curves demonstrated the capability of HT-UPLC to obtain fast separations. For example, a separation of nine doping agents was performed in less than 1 min with sub-2 μm particles at 90 °C. Furthermore, a shorter column (30 mm length) was used and allowed a separation of eight pharmaceutical compounds in only 40 s.