seeking advice for poor HPLC separation

Record: 657

Category: Specifications
Product: C18
Sample:

Question
Hello. You have sold us a column a few months ago, and recently I have been using it but getting poor separation of the compounds I am interested in. I have included a chromatogram of a recent run using standards for several of the compounds of interest. I have been using rather simple eluent conditions – 50 mM sodium phosphate and 5% methanol. Can you advise me if there are other things I can try to get better separation? Maybe different mobile phase composition or run conditions? I would appreciate some help, because I do not know where to go from ehre.

Answer
You want to make your compounds non-polar in order to create reversed-phase retention. By using the acidic mobile phase may help do so. I’m wondering if following condition can be applied:http://shodex.net/index.php?seitenid=1&expd_prod=107&applic=1407

 
 
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