Hi Moustafa,

Here are some usefull links.

Cheers

https://www.researchgate.net/post/What_is_the_best_way_to_isolate_leaves_with_a_lot_of_chlorophyll

https://www.researchgate.net/post/Before_extraction_from_leaf_material_how_can_I_get_rid_of_pigments_like_chlorophyll

https://www.researchgate.net/post/How_can_I_remove_chlorophyll_from_leaf_extract_without_losing_other_secondary_metabolites

try ethanol

When you start your extraction (crude extract obtainment); avoid using Ethanol (since it has a high affinity for chlorophyll), Methanol is an excellent solvent to start with. If your  target is a bioactive compound, then you should obtain first your methanol crude extract;  dry it from methanol; then try to extract from this crude extract, the bioactve substance using other solvents with decreasing polarity. But you need to follow your obtained fractions with tests of their activity, using the methodology named: Bioassays guided fractionation; then you continue until you obtain the most active fraction.

N.B: you can refer to my first paper: publ. in J. Pharm. Pharmac., uploaded & visible in RG.

Chlorophyll forms micelles in ethanol if you add a little water.  They can be centrifuged out.

Buffer acetone 80% works fine for me, but requires cold grinding (better if you can use liquid nitrogen). Another option is DMF, but requires leaving overnight to one day.

Do you want to eliminate the chlorophyll before you analyze (HPLC?) another compound, in order to not introduce chlorophylls in the chromatograph? Do chlorophylls really interfere with the detection of your analyte?

In that case... is really a great problem to completely remove the chlorophyll (at trace levels)... so you should separate in the chromatography, and clean the chromatograph column (HPLC) before next analysis.... or, you must "purify" your extract, for example using a SPE column.

Check these interesting threads

https://www.researchgate.net/post/How_could_we_remove_chlorophyll_from_plant_extracts_during_purification_of_natural_compounds?tpr_view=cf963c49-6fdb-43d5-a01f-f1e47d31e8b6_1

https://www.researchgate.net/post/I_would_like_to_know_about_the_method_of_separating_chlorophyll_from_ethanolic_extracts_from_celtis_africana#557153915dbbbd835b8b4571

https://www.researchgate.net/post/How_can_I_remove_chlorophyll_from_plant_extract?tpr_view=12502d85-0415-4a8a-9419-c5db2a876cb3_2

Please check this article. The safe and effective method was described. Article A green and effective method using oils to remove chlorophyl...

Chlorophylls are very highly hydrophobic and so can be removed using organic solvents that are not miscible with water. For example cooking oils.

If you have an extract in an organic solvent such as acetone, ethanol or methanol they quickly come out of solution if you add water.

The best method for removing chlorophyll is SPE (10 g) for 1g extract or you can pass the extract through activated charcoal, repeatedly, 2 or 3 times.