Actually I am currently working on saponin isolation but having very closed spot of four saponins in TLC and I have tried C18 column in preparative HPLC but it didn't get separated. After that we developed an HPLC method it’s got separated in used acid such as orthophosphoric acid, but the problem is that how can I extract acid from water (acid plus water) fractions, which come from preparative HPLC.
I have tried a new method and has managed to successfully isolated some kind of saponins from this method. I used soiid phase extraction method as it is usually used for polar and slightly polar compounds and is quite versatile. You can use water, acetonitrile or methanol for isolation. This method uses less solvents compared to column chromatography and is faster compared to HPLC. please try. Good luck.
I have tried a new method and has managed to successfully isolated some kind of saponins from this method. I used soiid phase extraction method as it is usually used for polar and slightly polar compounds and is quite versatile. You can use water, acetonitrile or methanol for isolation. This method uses less solvents compared to column chromatography and is faster compared to HPLC. please try. Good luck.
For saponin, extract your plant material with hexane followed by 70%methanol. Then concentrate your methanolic fraction and again fractionate with butanol for saponins. Then load into HP-20 resin followed by water and methanol. Then methanolic fraction again loaded in to C-18, Sephadex, prep HPLC for isolated of individual saponins.
Or you can follow our paper
Gudluck
:)
I agree with the Solid Phase Extraction suggested by Dr. Ali Mohd.
One can evaporate the organic solvent, adsorb the sample on a small C18 or CHP20 column, and elute with organic solvent. The reverse phase column could be hand-packed or a flash column.
I express my gratitude to Mustafa Ali mohdsir , robin sir, jack sir for your valuable suggestion but i have only prepartive C18 column,sephadex. In this both i have tried but couldn't get separate four saponin and Rt was very closed i have only option that used acid in my mobile phase then it get separate. but how can i exract acid from water as mobile phase because his acid may degrade my saponin in glycone and aglycone moiety during concentration.
Then try with prep TLC for separation of close saponins. I know saponins are difficult to isolate individually but you have to work hard.
Gudluck
:)
Before concentration, maybe add some base (ammonia, sodium hydroxide) to make the solution neutral, then you can concentrate it without the acid decomposing your compound. You can then extract or do SPE as described above.
You can use Sephadex for separation. Use chlorofom: methanol ratio to methanol water for isolation of individual saponins. It really works.
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