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. Interestingly, these actually became a reality with the coming in of the UHPLC, Ultra High Performance Liquid Chromatography.
Talking of this, bear in mind the fact that over the years, this column technology had actually proved or demonstrated that column efficiency had an inverse relationship with the particle size of the stationary phase, increasing with a reduction in the size of the particle of stationary phases. By and large, in the conventional analytical separations, the particle sizes used would often be between the ranges of 3 and 5 nanometers. 2 nanometer was seen as a subpar barrier. But this was broken in years that followed and columns packed with the 2 nanometer particle size were shown. By and large, it is with this particular development that we actually saw the final coming in of the Ultra High Performance Liquid Chromatography, UHPLC, in the whole chemistry of analytical separation.
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. The UHPLC columns have actually been found to be much shorter and a lot narrower as compared to what would be with the HPLC columns in a general sense. By and large, talking of their dimensions, height and length, the analytical columns would have such measurements or dimensions as high as 4.6mm ID by 250mm length while the UHPLC columns would feature such dimensions as 2.1mm ID by 50mm lengthwise.
The other notable difference there is between these two is their operating 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. The pumps will be able to withstand such high pressure conditions with the least of wear and tear on their parts.
Not to forget mention the fact of better detection with the UHPLC columns. The reason for this is looking at the fact that the UHPLC columns produce rather fewer peaks as compared to the HPLC columns which have lesser number of data points as such being rather poor in detection.