I would like to estimate stevioside and rebaudiosid A content in dried leaf materials using HPLC. Could you reccomend me an efficient extraction protocol and appropriate HPLC conditions for that?
To estimate stevioside and rebaudiosid A content in dried leaf materials using HPLC use NH2 column, ACN:Water (80:20) mobile phase with ELSD detector.
For PDA at 210nm.
Dear Dr Robin, please guide that why have we used NH2 column insted of ordinary C18 or C8 column?
Thanks Dr ashrey. Basically sugars gives good separation in NH2 column instead of C-18. You can also use C-18 columns instead of NH2 column for steviosides but resolution, separation etc. are not very good for C-18 on ACN:Water as mobile phase. If you are restricted for PDA or C-18 then you have to standardise mobile phase, flow etc.
I have UV-dedector and C18 column. it seems HPLC conditions not available for this work. thank you...
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