My lab is interested in conducting an insect bioassay on phytochemicals from an uncommonly studied plant species, Silphium integrifolium. We are looking for a simple protocol we can use to extract secondary metabolites from leaf tissue samples, and just clean up the sample enough such that any nutritive compounds remaining (proteins, carbohydrates, etc.) are negligible when the extraction is mixed into an artificial diet. Does anyone know of an existing methodology to do this?
It depends on the objective of the extraction.
If you aimed at the maximal extraction yield, I suggest a mixture of ethanol-water. you could vary the percentage of ethanol from 25 to 100% and study the variation of the yield and several phytochemicals, and also their biological activities.
If you aimed at the isolation of the bioactive compounds, you could use the bio-guided method including extraction with different solvents or solvent mixtures following by fractionation and isolation.
Please look at the following below links which may help you in your analysis:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218439/pdf/AJT0801-0001.pdf
https://applications.emro.who.int/imemrf/pak_j_pharm_sci/pak_j_pharm_sci_2009_22_3_317_322.pdf
https://ro.uow.edu.au/cgi/viewcontent.cgi?article=5083&context=theses
Thanks
The common extraction protocol is:
1- collection 2- cleaning ( washing with tap water 3- drying at shade 4- grinding
5-extraction with different solvent ( different polarity )...start from non-polar
solvent ( hexane or petroleum ether ), ethyl acetate, methanol and finally 70% methanol-water. 6- evaporate the solvents under vacuum. 6- separation using
chromatography ( if you need single pure compounds) 7- identification using
spectroscopic methods ( if you need) 8- biological activities.
If you don't need single pure compounds, ignore step 6 and 7.
Extract the plant/ leaves in pure methanol or 70% ethanol water system. Dry the extract and then suspend the dry extract in water and extract with hexane or pet.ether and the rest of extract you may use for your experiments.
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