Chlorophyll & Carotenoids Contents Protocol:

Arnon (1949) METHOD is considered while measuring the chlorophyll contents of plants along with chlorophyll a and chlorophyll b ratio and total chlorophyll contents. 0.5g of the leaf will be ground in 10ml of 80% methanol.  The extract will be centrifuged at 10,000rpm for 10 minutes. The supernatant will be stored in small bottles in a cool place. The absorbance will be taken at 663, 645, and 480 nm respectively using a spectrophotometer. The absorbances will be subjected to Yoshida (1976) for the calculation of chlorophyll a and chlorophyll b and Davis (1976) was consulted for the measurement of carotenoids.

Chl. a (mg/g) = [12.7(OD663)-2.69(OD645)] ×V/1000×W

Chl. b (mg/g) = [22.9 (OD645) -4.68(OD663)] ×V/1000×W

Total Chl. (mg/g) = [20.2 (OD645) +8.02 (OD663)] ×V/1000×W

Carotenoid (g ml-1) = {A Car/ Em×100} Where Em×100 = 2500,

A Car = [(OD480) +0.114(OD663) – 0.638(OD 645)]/2500

Where

V= volume of the methanol used in extract (ml)

W=Weight of fresh leaf tissue (g)

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