I'm looking at my HPLC chromatograms of Rubus leaves extracted twice by 20 and 10 ml of methanol respectively, and I can see a significant peak at 440 nm after 2.90 min elapsed time. Does anyone know a plant pigment refered to this peak ?
(I saw that chlorophyll c3 is oftenly attributed to this peak on micro-algae HPLC, but theorically not present in plants.)
Options: Collect it and re-analyze it to determine it's ID (or use: LC-MS. IR or other orthogonal techniques). Look up the plant's name and see what the literature reports about its chemical make up.
*Make sure you verify that your HPLC method is fit for purpose and selective for the compounds being analyzed. A "std" 4.6 x 250mm RP column run at 1 mL/min has a void volume of about 2.9 minutes (same as your peak) and any sample peak seen at that Rt would indicate the method used is invalid and non-selective.
It's probably one of the compounds mentioned in the attached reference, see 1st link. My guess is Quercetin see 2nd link.
http://www.mdpi.com/2076-3921/5/2/17/htm
http://www.sciencedirect.com/science/article/pii/S0102695X15000885
Retention time depends on solvents, column lenght, stationary phase, temperature, flow rate, etc., so RT only can give you an idea of what is in your sample if you compare with research done on a similar sample and in the same conditions. Furthermore, RT is not a identity confirmatory parameter. There are a lot of compounds that can gives a 440 nm peak. Did you use UV at 40 nm or diode array detection?
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