Well, it depends on how complex is the mixture and how much material it contains. If you have substantial amount and the mixture is of reasonable complexity, you can try column chromatography to seperate them. Hiwever, if you dont have much, then I would recommend doing preparative HPLC to purify the componenets. After purifying, you can employ exact mass (HRMS) and NMR, IR to determine the structures for each component. Good luck with your research.
Hi, in addition you can also use also TLC preparative (normal phase and/or reverse phase (RP) phase) , after this you use HR MS or NMR, good luck
Hi, you can use different solvent system such as tetra-system of EtoAc:CHCl3:MeOH:Water having composition of 15:8:4:1 and 13:7:4:1 with isocratic program, try firstly these solvent system on TLC, and then go for column chromatography. I hope you will be able to isolate secondary metabolites from butanol fraction.
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