I would like to quantify some of phenolic and flavonoid compounds by HPLC. How can I extract these compound from plant and what is the solvent of HPLC with what program and percent of solvent?
I have had success with an extraction solution of 70% methanol and 30% water. This yielded a wide variety of flavonoid compounds (e.g. naringenin, tremulacin, etc.) from poplar leaf and stem tissue. You need to decide whether or not to lyophilize your tissue prior to extraction, as this can affect the results somewhat. For some general guidelines, check out the following publication:
Rispail, N., P. Morris, and K.J. Webb. 2008. Phenolic compounds: Extraction and analysis. In Lotus japonicus Handbook. Springer.
Acetonitrile is a good choice as a solvent for the extraction of phenolic and flaonoid compounds from plant..u can try some combination of ACN with water..i ve done with 80% ACN in my study for the extraction gallic acid and catechins in tea...ACN was proven to be a better solvent in comparison to water and methanol..
Gud luk
(1) I am using 50mg dried sample + 1ml Methanol, sonication for 60 minutes at 50'C, filter with 0.45um membrane filter- 10 ul injcet to HPLC
(2) For the solvent : A=0.1 % Formic acid B=Acetonitrile
Time Flow (ml/min) %A %B
0 1.0 85 15
12 1.0 75 25
22 1.0 75 25
25 1.0 85 15
35 1.0 85 15
PDA detector
Gud luck
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