HPLC-UV method for nucleotides

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Hi all,

I need to analyse nucleotides using HPLC-UV. I have a supelco c18-T column. Does anyone have a hplc method I could use?
Hi HPLC chemist,

Yes I did and only one of the methods worked but i had baseline problems where the baseline drifted quite significantly affecting my chromatogram.

The method I tried:

Mobile phase A: 0.03M KH2PO4, 10mM TBAHS(tetrabutylammonium hydrogen sulphate) adjusted to ph 6 with NaOH.

Mobile phase B: Mobile phase A:methanol (80:20 v/v)

injection vol: 10 microlitre, flow rate: 0.7 ml/min, column temp: 35oC

When I run it without TBAHS, I have no problems with the baseline. However I need TBAHS to separate the nucleotides.
Sounds like your 'buffering/ion pairing' is not stable for your sample matrix. Increase the concentration of the TBAHS to 25 mM.
Also increase the concentration of the Phosphate buffer to 50 mM (not 0.03 mM).
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