[goldy]

Hello everybody,
I got one problem in Prep HPLC column. The single peak gets splitted into many. If anybody has idea about fixing this problem then please suggest me as soon as possible.

[cundi]

Use a buffered mobile phase!
Adjust the pH according your compund.

The sample preparation is also very important. Use the same mobile phase as solvent for your sample when possible. Don't employ more % of the higher elution force eluent.
i.e: mobile phase: 70% of KH2PO4 buffer pH=4, 30% ACN. Don't use more than 30% of ACN in your sample.

 Is your compound an amine?

[goldy]

Use a buffered mobile phase!
Adjust the pH according your compund.

The sample preparation is also very important. Use the same mobile phase as solvent for your sample when possible. Don't employ more % of the higher elution force eluent.
i.e: mobile phase: 70% of KH2PO4 buffer pH=4, 30% ACN. Don't use more than 30% of ACN in your sample.

 Is your compound an amine?

Actually the problem is not because of the type of the compound. It is rather than the column problem. Single peak in analytical column when injected to prep column shows the no. of splitting peaks. What should I do to improve the colum situation. Previously I separated the compound in prep columm (my sample was not dissolved in methanl so I used 10% chloroform in methanol to dissolve it and then injected.) Next day when the other samples was injected than I got such problem.
DO anybody has any idea to solve this problem.

Thank you so much.