I'm purifying compounds from a RediSep Rf Strong Cation Exchange column on a Teledyne Isco EZ Prep system with a PurIon mass spectrometer system.
I dissolved methanolic extract of green tea in methanol containing acetic acid, and loaded it on the SCX column, washed the column with methanol, and ran the column according to the attached PDF. The first gradient (using water/ 5% acetic acid) washed off the xanthine alkaloids; I could see caffeine and theophylline in the mass spectrum.
The second gradient uses 5% ammonium hydroxide in water, and washes off stronger bases. During this second gradient, I recovered the sharp peak at ~47.4 column volumes (CV). The mass spectrum shows a peak at 549, suggesting possible masses of 548, 526 (if I'm looking at a [M+Na]+ at 549), or 516 (if my mass peak is an adduct of [M+MeOH+H]+ ). I'm using methanol as a carrier solvent.
It isn't a flavanoid or other phenolic compound since I'm using a cation exchange column. I haven't found a hit yet in the ACS journals, nor yet on http://www.ebi.ac.uk/chebi/
Any ideas what this could be? I guessing either an alkaloid or some small peptide?
Thanks in advance.
Maybe look at camellisin A! (Lei, C. et al., Chem. Pharm. Bull., 2010, 58, 939- 943); it could fit with [M+MeOH+H]+
@ Dr. Coulerie- interesting, I'll keep this in mind. Would you expect it to elute from an SCX (strong cation exchange) column? They appear acidic to me.
Another possibility is some sort of compound related to chlorophyll.
I have not much experience with SCX column; What retention did you observed with your compound? In other case, if you have UV or MSMS you can easily differenciate chlorophyll derivatives from a triterpene (and even by eyes)
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