Method of Analyzing Chlorophyll

Add approx. 30mg Chlorella powder into a small amount of 85% acetone (1) to grind it down for dissolution.
Then, add 10ml of 85% acetone and mix well for pigment extraction.
Then, separate it centrifugally and remove the skimming into an evaporator flask.
Evaporate it completely under reduced pressure by a rotary evaporator.
Then, dissolve it in 1ml of anhydrous acetone, and streak it onto the plate (2).
Develop (3) it at the room tempertaure for approx. 30 minutes.
Chlorophyll b band (yellow green) can be observed to lie beneath Chlorophyll a band (blue green).
Rake out this Chlorophyll b band, and add 4ml to 5ml of anhydrous ether on to it and mix them well.
Centrifugalize it and take the skimming out.
Measure its absorption spectrum, which is to be synchronized with that of the chlorophyll b.

(1)Add H2O to 85ml acetone to make 100ml.
(2)TLC plate Cellulose precoated (without fluorescence indicator) Art. 5 716 Merck. 20 x 20 cm Wako No.506-00971
(3)Developing solvent: petroleum benzine (bp50-80): acetone: n-propanol