Chromatography Forum

Hai Friends,

I am getting trouble while developing HPLC method for this molecule. There is no problem with elution. The problem is the resolution between the peaks after tailing. I am getting 3 peaks on tail of the principal peak. Because of the very very poor resolution, I could not able to quantify or even I unable to show the repeatability of the % of these impurities.

I am using triethylammonium phosphate buffer and acetonitrile and tried with C8 and C18 columns.
Also I tried with TBAHS one of the reagent in buffer.
Can anybody suggest how to resolve this problem

Regards,

Raman

Try methanol instead of acetonitrile!
Also, what is the pH?

Hai Uwe,

The pH of the buffer I am using is 3.5

Also I tried with Methanol. The tailing of principal peak is increased.

Raman

You can try to play with the pH. Maybe pH 2.5 gives you a slightly different result than pH 3.5. Or even better go to pH 7 and see what is happening. Then try methanol again.

This is not dissimilar to the standard method development approach in my book.