Chromatography Forum

Hi

Sorry if the title of the post is a bit cryptic!
I'm in the process of developing a method that aims at separating 2 compounds by HPLC with UV-VIS detection. However neither of the 2 compounds displays profound UV activity, and sometimes I'm really sure whether the compounds have eluted or not.
Hence, I inject the reference sample with no column installed, and use that as a reference point with respect to the peak area, just to have a bearing with regards to the expected peak area.
But is there any pitfalls to this approach, and does any body else use this?

In advance thanks
Erik

Could you please delete this duplicate post of yours? Thank you.

You might be seeing effect of the injection, particularly if you are looking at lower wavelength. It might make sense to consider derivatization or an alternative detection technique (ELSD, MS, RI)