The entire HPLC system must be thoroughly cleaned before installing the column. Flush your system with a filtered, buffer-free eluent at a low flow rate (0.1 ml / min) for at least 30 minutes (preferably several hours or overnight) to remove traces of strongly retrained, aggressive substances such as acids, bases or salts. Older HPLC systems tend to release metal ions and should therefore be cleaned even more thoroughly.
The choice of tubings, capillaries and fittings has a significant impact on the dead volume of the system. This is especially important when using short columns and small inner diameters, where the dead volume should be reduced to a minimum.
It is recommended to use finger-tight male nuts with ferrules. Using the wrong ferrules may lead to dead volume, resulting in band broadening and leaks. Every manufacturer has its unique connection system. Please consult the relevant product information on nuts, ferrules and tubings, as well as the connections of the injection system carefully before installation.
Special care must be taken when connecting the tubings and the column to ensure there is no gap between the column head and the end of the tubing. Press the tubing as far as possible into the column head so that it will not slide back when screwing, i.e. tubing must be seated all the way down into the column end-fitting. This applies particularly to stainless steel capillaries.
It is recommended to test the column and the entire system immediately after receiving the products, e.g. with an RP test mix.