I have methanol extract of a plant, Flavonoids are expected in it.
I run reverse phase HPLS, C18 Luna column, Acetonitrile and 0.1% Formic acid mobile phase.
I got some peaks, one of them identified as rutin.
I have to identify the compounds in the big peak on the right side of chromatogram (it contains more than one less polar compounds). Kindly have a look on the atached chromatogram and guide me.
Thansk in advance
Hi Abdul,
You should try many options :
1. Simplest is to increase flow rate in gradient method, it increases resolution usually
2. Change the column oven T', try lowering and increasing.
3. Increase percent of HCOOH, 0.15 or 0.2...or try acetic.. Acidify the sample
4.Most effective should be replace the column with best end-capped and base-deactivated, like that from ACE advanced technologies, Inertsil C18, Phenomenex C18 Luna2, Purosher C18e STAR, and many similar...
Hope that some of these will help
You can try also TFA or H2SO4.
You can tune the selectivity by xhanging AcCN to EtOH or MeOH. Addition od THF also could change the selectivity.
Regards
GB
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