HPLC Assay of HCG

[Akthmps88]

Do you know what the predicted order of elution for the alpha and beta subunits of HCG would be on a C4 300A column? Gradient goes from high aqueous to high organic to elute the HCG.

I also see the 4 peaks you are observing. Two are fairly large, so I would think these are the alpha & beta subunits. I couldn't find a lot of information on the subunit structures. Do you think the alpha subunit is the first peak and the beta subunit is the second?

I am also going to evaluate an protein column from Phenomenex. It is 200A. Hoping this may also work and provide better resolution than the Water Symmetry 300A C4. The 4 peaks I see current don't show great resolution from each other.

[janoshik]

I don't think it matters for quantification - I believe (but am not 100% sure) that the beta subunit is eluting later.

This time I picked the second one, because the first one didn't get separated too well from the minor peaks, and used that for quantification.


200A will work for sure.


Chromforum is giving me issues and its a pain to answer here right now, if you want, you can let me know your linkedin and I can add you, in case you want to consult.