Estimation of total chlorophyll by Aron's method
Purpose:- Estimation of total chlorophyll by Arnon's method. Requirement:- Spectrophotometer Buchner's funnel, filter paper test tube, Fresh leaves. measuring cylinder pestle & mortar 80% Acetone Formula:- 20.2 (A645) +802 A663) / 1000 x W Principle:- Chlorophyll is extracted in 80% acetone & the absorption at 660 nm& 645 nm are read in s spectrophotometer using the absorption co-efficient, the amount of chlorophyll is calculated Procedure:- 1) Weigh 1 gm of finely cut & well mixed representative sample of a leaf or fruit tissue into a clean mortar 2) Grind the tissue to a fine pulp with the addition of 20mil of 80% acetone 3) Centrifuge (5000 rpm for 5 min) & transfer the supernatant to a 100 ml volumetric flask 4) Repeat this procedure until the residue is colorless wash the mortar and pestle thoroughly with 80% acetone and collect the clear washing in the volumetric flask 5) Make up the volume to 100 ml with 80% acetone 6) ...