Here I have got a HPLC trace with my desired pick at 25-26(t). However, the sharpness of my desired pick is pretty much distorted with an unfortunate side pick attached to it. I am seeking any practical approach to resolve this problem prior submitting my compound for Mass spectroscopy.
Dear Reza, I am no HPLC expert but more an end user. For me a 25min run to get the compound out is already too long, if no separation.
I think to get some valuable inputs you need to describe the type of compounds (no need to give details, but peptides, sugar type....) then please include your current method: column, solvents, buffers, isocratic or gradient, flow rate...
Here is some very valuable informations from another topic:
https://www.researchgate.net/post/HPLC_database_s
Dear Reza,
First of all, let us clearly know about the method you are following exactly. Because if you are following a gradient prog.. definitely will get a gradient peak corresponds to your programme. Check once with your blank by injecting the diluent. If i t is blank peak why you need to worry? Also try with temperature applications and with flow rate. All the best.
Hi Laurent,
Thank you for your input. Actually, I am working on Anthracycline Antibiotics. I take advantage of the C18 columns with H2O and Acetonitrile as my mobile phase. My flow rate is 3ml per min. Do you think TLC or size exclusion chromatography could be of any help?
Without knowing nature of your compounds, it's difficult to answer. However, C18 may not be the best phase to use. HILIC chromatography may be a better choice.
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