DETERMINATION OF CHLOROPHYLL IN LEAF EXTRACT

DETERMINATION OF CHLOROPHYLL IN LEAF EXTRACT

AIM:
To estimate chlorophyll content in a leaf sample.

APPARATUS:
Spectrophotometer, centrifuge, test tubes.

REAGENT:
80% acetone.

PRINCIPLE:
Chlorophyll is extracted and measured at specific absorbance values.

PROCEDURE:

• Grind 1g of leaf in 10ml acetone.

• Centrifuge and collect supernatant.

• Measure absorbance at 645 nm and 663 nm.

CALCULATION:
Use Arnon's formula:
Chlorophyll a = 12.7(A663) – 2.69(A645)
Chlorophyll b = 22.9(A645) – 4.68(A663)