Is it a good practice to incubate the organic solvent-algal mass mixture in a water bath of ~50 oC for lets say 20-30 minutes in order to enhance the extraction of pigments?

After that is it redundant to place the mixture in the freeze at 4 oC for 24 h or not?

Is it necessary to wash the centrifugation sediment of marine algal mass with ammonium formate in order to remove salts prior to the introduction of the solvent? I think it is unnecessary as the solvent will act on the algal pigments only irrespectively of the salt presence. If not please correct me.

Final question. Is it necessary to wash the marine algal sediment after ammonium formate treatment (with or no a.f. treatment) with distilled water? If yes then what about possible rupture of the cells due to d.w.? Such a thing is good or bad for the spectrophotometric measurements of the pigments?