We have made a ginkgo extract by drying 60g of fresh ginkgo leaves. From there, the leaves were crushed and soaked in a 50/50 mixture of distilled water and ethanol. Then, the solution was filtered via gravity and vacuum filtration. Now that we have a clear sample free of particulates, we need to know how to separate them. We are trying to separate rutin, quercetin, and kaempferol individually from the solution, so that we can test each flavonoid on its own rather than the entire extract.
Hello, Ashton,
I suggest that you optimize or search for a replicable protocol on flavonoid separation using Reversed-Phase HPLC. If a HPLC unit is available in your lab, the extraction process will be fast and easy.
I found this online material which might help you: http://www.waters.com/webassets/cms/library/docs/720004839en.pdf
Best,
Arizaldo
Hi Ashton,
Have you checked percent yeild?
1. Solvent play a big role so either dries the extract first or make a calibration curve of standard in same ratio of solvents.
2. Calculate yield directly in percentage yield of curcumin using formula
Wt of ginkgo /Wt of raw material X 100.
3. So, some spectroscopic tests have to be carried out to figure out the concentration of a particular set of compounds in the extract. For example, Folin-Ciocalteu's test provides an estimate of 'Total Polyphenol' concentration. AlCl3 test would give an estimate of 'Total Flavonoid' concentration. For carrying out both these tests, the (dry powder of the) extract has to be dissolved in a suitable solvent or in the reagents meant for these tests. Therefore, evaporation of entire solvent by rotavap does not necessarily mean that the concentration is 100%. For separation of flavonoid HPLC is better option.
Best...
Manju
I think. that HPLC chromatography is the best variant. Try "water-acetonitril" gradient eluation
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