Hi everyone
I have been trying to resolve adenosine and Inosine on HPLC column. The details are as follows:
1) Column dimension: 150mmX4.6mm, with 5 micron bead size.
2) Mobile phase: 20mM phosphate buffer with methanol
3) gradient is: 7% methanol for 3 min then,
30% methanol for 3 min
finally, 7% methanol for 6 min
4) Run time : 15 min
5) UV dtection: 235nm and 260nm
However, I have not been able to resolve the peaks. Could anybody please suggest on this matter?
Thank You
Hi Ravindra,
you need a very flat gradient because your two compounds are structurally very similar. I guess your gradient ist to steep. Maybe this link will help you:
https://pdfs.semanticscholar.org/e63c/25c4807af5a2e5d5169cd66734076af24be7.pdf
Regards
Markus
You can also study the following articles:
https://www.sciencedirect.com/science/article/pii/S0003269796902468
https://onlinelibrary.wiley.com/doi/abs/10.1111/j.2042-7158.1988.tb05201.x
Thank you very much for the suggestions, I will try the methods given in the articles.
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