Subramanian and Nagarajan (1969). I'm using this protocol in this methanol extract fractionation, done by pet. ether, ethyl acetate and diethyl ether.
Can I use n-hexane instead of pet. ether for fatty substances removal, or should I use something else?
Dear Sunda, you should fractionated your crude methanol extract first by step wise gradient polarization solvent. n-hexane could be used for removal of fatty compound. In my experience, flavonoid compound mostly found in ethyl acetate fraction. For fractionation you can use liquid-liquid partition method. It is quite simple.
Most flavonoids are phenolic, so SAX (strong anion exchange) workson these compounds. After that, I've used LH-20 followed by C18
Here are some app notes & posters:
http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN85_Purification_Strategies_for_Flavones_and_Related_Compounds.pdf
http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN87_RediSep%20Rf_SAX_Column_Applications.pdf
http://www.isco.com/WebProductFiles/Applications/101/Poster_and_Paper_Reprints/Purification_of_Phenolic_Flavonoids.pdf
thanx jack & yhiya .if further i encounter any problem can i ask for help?
which protocol i follow in this DCM is not included ..can u elaborate what is the role of DCM in fractionation ? and diethyl ether is seapreting free flavonoids r n-butanol do the same? sorry for these bulk of ques but i wana clear understanding about these.
We have to extract the flavonoid with DCM from crude. Further, Chromatography is more useful tool to separate the products, if we get two or more spots in TLC.
Dear Sundas,
Try fractionating your methanol extract using dichloromethane, ethyl acetate and n-butanol to obtain their respective fractions having much better chances of bearing flavonoid compounds. Following threads may be useful for you:
http://academicjournals.org/article/article1390558568_Yusuf%20et%20al.pdf
http://www.academicjournals.org/article/article1391011777_Yakubu%20et%20al.pdf
Further you can use n-hexane instead of pet. ether for removal of fatty substances.
Good Luck.
Dear Sunda, you should fractionated your crude methanol extract first by step wise gradient polarization solvent. n-hexane could be used for removal of fatty compound. In my experience, flavonoid compound mostly found in ethyl acetate fraction. For fractionation you can use liquid-liquid partition method. It is quite simple.
thanx everyone.... i got all three layers of fatty substance, free flavonoids & conjugated flavonoids.... ;-)
Check this thread and link:
https://www.researchgate.net/post/How_to_fractionate_methanol_extract?_tpcectx=topic_detail&_trid=55409c80cf57d7241b8b4618_1
http://www.slideshare.net/rahulbs89/extraction-of-plant-contituents
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