I am currently treating my crude extract with acid to obtain flavonoid C-glycoside. The conditions that I used for acid hydrolysis was the dried crude extract was reconstituted with 1 mL of water. 1 mL 12 M concentrated hydrochloric acid was added to the extracts in capped tube. The mixture was incubated in water bath at 85℃ for 2 hours (Tahir et al 2012). Flavonoid c-glycosides present in crude and acid hydrolysed extract were quantified using LCMS with reference standards (vitexin, isovitexin, orientin and isoorientin). The calibration curves for each standard were plotted. However, the acid hydrolysed extract showed low amount of flavonoid c-glyscoside in comparison with crude extract. May I know what are the possibilities that may cause for this results? Thank you
The European Pharmacopeia uses ~1M HCl to hydrolyse the Flavonol-O-glycosides in Ginkgo for a quantitative assay, 12M appears quite harsh to me.
I could imagine that flavon-C-glycosides like vitexin could eliminate water rather easily, so that a double-bond is formed in the glycosidic part that is in conjunction to the pi-electron-system of the flavon part.
I agree with Jandrik, 12 M HCl is quite strong, you have to find out the optimum concentration. This strong acid might cleavage the glycoside bond.
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