Principle

to determine only this chloropphyll. The distribution of chlorophlls a, b an

Potosynthetic pigments present in the cells of green plants are the important apparatus
responsible for trapping the radiant energy to bring about photosynthetic reactions.
Chlorophyll aa is considered to be the most important phytopigment because od its
occurrence in all plants,. Therefore many workers prefer d c in some importaant algal
groups is as follows: Cyanophycease only a
Chlorophycia- a, b and c
Bacillariophycrae - a, c
Phaeopigments are the degration products of chlorophylls and they increase with the
age of the plants.
Chlorophylls being essentially involved with photosynthesis, their concentration is
considered to be a very good index of primary production.
ESTIMATION OF CHLOROPHYLL IN MACROPHYTES
Material and chemicals
Chemical Balance, mortar and pestle, centrifuge spectrophotometer, laboratory
glassware, magnesium carbonate and acetone 90%.
Method
Collect the macrophytes 1 gm of its fresh green part. Homogenize the material in
presence of 20 ml of 90% acetone and pinch of magnesium carbonate using a mortar and
pestle. Transfer the contents to a dark bottle, cap tightly and place it in refrigerator for 6
hrs (this period should be constant for all samples) to allow complete extraction of
chlorophyll. Centrifuge the contents at 3000 RPM for about 15 minutes and decant the
supernatant into a volumetric flask of 25 ml. Make up the volume of contents to 25 ml
by further adding 90% acetone and record the optical density (OD) of the extract on a
spectrophotometer at 565 nm and 645 nm.