High Performance Liquid Chromatography, popularly known as HPLC, had some common benefits that users looked forward to and these were such as high resolution, high sensitivity and the speed of analysis that it had to come with. As a matter of fact, most of these benefits were made a reality when UHPLC, Ultra High Performance Liquid Chromatography, came into the picture.
This is even looking at the fact that this column technology had proved the fact that column efficiency increased with a reduction in the particle sizes of stationary phases. Looking at the conventional analytical separations, the particle sizes that were used would often be within the ranges of 3 and 5 nanometer sizes. 2 nanometer was seen as a subpar barrier. As a matter of fact, in the later years after these assumptions and earlier years to this edition of article, this barrier was broken and there were actually demonstrated columns that had particles with 2 nanometer particle sizes. This is actually the development that saw the coming in of the UHPLC, Ultra High Performance Chromatography, in column separation in analytical separations.
As we seek to understand this better, we will be taking a look at some of the basic differences there are between UHPLC and the conventional HPLC.
As we look at these key differences, we start by looking at the column dimension differences. Generally speaking the UHPLC columns are often shorter and narrower as compared to the HPLC columns. Analytical columns are in most cases up to 4.6mm ID by 250mm length while the UHPLC columns would be 2.1mm ID by 50mm long.
Talking of the key differences that you can expect from these kinds of columns, we have to take a look at their operating pressures and pressure capabilities. When we look at the operating pressure factor, we see the fact that the UHPLC columns as well still outdo the HPLC columns, as well known as the analytical columns, where in we see the fact that the UHPLC columns can handle pressures of between 15000 and 18000 psi while the HPLC columns can only handle such pressures within a system between 5000 psi and 6000 psi as their maximum limits. You can trust the pumps to handle such extreme pressure limits and these will not have as much damage and effect on the components there are in them.
Not to forget mention the fact of better detection with the UHPLC columns. This can be explained by the fact that the UHPLC columns produce narrow peaks, which are often a few seconds or less, as compared to the conventional HPLC or analytical separations or columns that have such a slow response and as such generate some sort of compressed leaks as a result of lesser number of their data points making them somehow inaccurate in their detection in the end.