It is a chromatogram of phenolics compounds (standard compounds) and with a red arrow I point the place where a high peak (Peak 1) appears when I inject a methanolic extract from black mulberry after acid hydrolisis...I tried to identify the peak adding each standard to the sample but always obtain two peaks (the one in the sample (peak one) and the standard one).....could it be 2,4,6-trihydroxybenzoic acid (acetylphloroglucinol) I did not run this because i do not have it?...Thanks!
I use a C18 column, with acidified methanol linear gradient...detected a 280 nm.
Hi Gonzalo,
Before anything, you should check if this is not an artefact from the extraction or any other product which could be stuck on your column.
If these options are OK, the literature should guide you for an answer. If you don't find anything in the literature, UV detection may be not sufficient (e.g. sensitivity and selectivity) for identification if you do not have a standard. If you are using a DAD detector with your HPLC, you could look at the UV spectra of the unknown peak to gather more information about the compound. If you work with a VWD detector you will not have enough information to know what's your compound is.
Then you should do at least HRMS for identification if the targeted compound is known.
I do not see much more information to help, I hope this will be a first step,
Cheers.
Nicolas.
As you are running UV, look at the UV spectrum and see if it potentially matches some of the compounds listed in Dr. Shannon's posting.
Also, try to collect the peak and submit it for ESI MS. Test it in both positive and negative ESI, since I did have one compound that gave a nice response in positive ESI, but not negative, but most of these compounds seem to show a mass in both positive and negative mode, which is a good confirmation of the molecular weight. This can be used to limit the possible compounds.
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