In my research I am trying to determinate nitroso- and amino- derivatives of 3,5-DNS. Currently, I am trying to obtain mobile phase suitable for their separation, but unfortunately, regardless whether in mixture or pure standard, I obtain single peak of retention time about 2 minutes - as if no interactions between the column or analyte occured.
I investigated C18 RP column, phases: MeOH:ACN (1:4, 1:1, 4:1), Water:ACN, ACN:Isopropanol (same pattern) and APS-1 amino column, phases: water-MeOH(4:1, 1:1, 1:4) or ACN:MeOH (4:1, 1:1, 1:4) . In every case, RT was very similar, no chance for any separation.
Do you have any suggestions?
If you are running an analytical system, try running a gradient from 5 to 100% organic in water and see if the reference elutes at the void. If not, you can flatten the gradient as desired. The amino column can act as normal phase or reverse phase depending on the compound and the solvent system, and so your compound may have eluted at the void. As an acidic compound is being run, a modifier such as acetic acid, formic acid, or TFA may be desirable.
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