Multiple diastereomers and rotamers merging with reverse phase chromatography. UPLC with lower id and particle size columns helps certain level. I need well separated peaks for development and method validation. Please suggest some reverse phase columns and techniques.
Choice of column for proper separation would be a trial and error approach. There are examples of separation of multiple diastereoisomers of flavonolignans by simple RP 18 columns (look for papers on separation of silybin A &B,isosilybin A& B, silychristin, iosilychrstin, silydianin and isosilydianin by reverse phase hplc). Again it depends on the sample matrix complexity and the type of constituents that you are attempting to separate. Ideally, I would use a Reverse Phase HPTLC (c18, Merck) strips to get at an idea of mobile phase combinations and the best separation conditions before embarking on a reverse phase HPLC. Good luck in your try.
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