You can visit thesis of Asma ben farhat! she works on different methods of extraction of orange!

The easiest way for you to start your work soon is to buy hesperidin as authentic sample from known company like Sigma. Make the necessary modifications to the protocol and start. Good luck.

Look in the literature;

I would try using a strong anion exchange column followed by polyamide or Sephadex LH-20, then C18. This can all be done on flash chromatography. Dr. Fathy's suggestion is a good one to verify the procedure works!

See:

http://www.teledyneisco.com/en-uk/liquidChromatography/Chromatography%20Documents/Posters/Purification%20of%20Phenolic%20Flavonoids%20with%20FC%20Poster.pdf#search=phenolic

http://www.teledyneisco.com/en-us/liquidChromatography/Chromatography%20Documents/Application%20Notes/RediSep%20Rf%20Strong%20Anion%20Exchange%20Column%20App%20Note.pdf#search=phenolic

http://www.teledyneisco.com/en-us/liquidChromatography/Chromatography%20Documents/Application%20Notes/Hints%20for%20Strong%20Ion%20Exchange%20Resins%20App%20Note.pdf#search=exchange

Also see:

http://www.teledyneisco.com/en-us/liquidChromatography/Chromatography%20Documents/Application%20Notes/Purification%20Strategies%20for%20Flavones%20and%20Related%20Compounds%20App%20Note.pdf#search=flav%2A

You can isolate hesperidin in good yield from orange peels by using the following method:

One-liter of petroleum ether is added to 250 gm of dried powder of orange peel in 2-liter volumetric flask, the mixture is heated for 4 hours under reflux. The petroleum ether extract is discarded. And the peels were reextracted using 1 liter of methanol, and the extraction continued for 2 hours under reflux.

The methanolic extract was filtered and evaporated under vacuum until syrup consistency is reached. The residue is mixed with 50 mL of 6% acetic acid, to precipitate solid is the crude hesperidin. It is sucked off with a Buchner funnel, washed with 6% acetic acid and dried 60 °C until it is constant in weight.

The crude product was dissolved in dimethyl sulfoxide under stirring and heating to 60–80 °C, water is added slowly whilst stirring. When cooling to room temperature the hesperidin precipitates. It is sucked off, first washed with little warm water and then with isopropanol and dried in the desiccator until it is constant in weight to give 2.5 gm of white powder to give pure hesperidin