Scribd
Upload
15 views2 pages

VL2021220701265 Ast03

The document outlines a procedure to estimate the chlorophyll content in leaves using solvent extraction and absorbance measurements at specific wavelengths. It provides mathematical formulas for calculating the amounts of chlorophyll a and b based on absorbance readings. The experiment involves crushing leaf samples, extracting chlorophyll with DMSO, and measuring absorbance to determine chlorophyll concentrations.

Uploaded by

lifeboring850
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
15 views2 pages

VL2021220701265 Ast03

The document outlines a procedure to estimate the chlorophyll content in leaves using solvent extraction and absorbance measurements at specific wavelengths. It provides mathematical formulas for calculating the amounts of chlorophyll a and b based on absorbance readings. The experiment involves crushing leaf samples, extracting chlorophyll with DMSO, and measuring absorbance to determine chlorophyll concentrations.

Uploaded by

lifeboring850
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

Estimation of Chlorophyll in Leaves

AIM:

To estimate the amount of chlorophyll present in particular type of leaf.

PRINCIPLE:

The principle of the experiment is based on solvent extraction in selective solvent in which
the chlorophyll will get completely leached when incubated. Chlorophyll a and b absorb
radiation in the visible region. Therefore, absorbance can be a measure of chlorophyll levels
in the leaf sample. Then the levels of chlorophyll a and b are calculated as per the
mathematical formulae given.

Chlorophyll a, b are calculated using the following mathematical expressions.

(12.7 x absorbance at 663nm)  (2.69 X absorbance at 645 nm)


Chlorophyll ' a'  xV
a x 1000 x W

(22.9 x absorbance at 645 nm)  (4.68 X absorbance at 663nm)


Chlorophyll ' b'  xV
a x 1000 x W

Where , a = Length of the height path in the cuvette (i.e. 1cm)


V = Volume of the extract in ml.
W = Weight of the sample in mg.
PROCEDURE:

Take 500mg of fresh leaves for different plants and crush to a fine pulp with the addition of
10mL of DMSO in water. Heat the solution on a water bath for 15-20 min and filter the
solution into a 50 mL std. flask. Again add 5 mL of DMSO to the leaves residue and heat on
the water bath for 15 min. and filter the solution into the same std. flask. A third extraction is
carried out by adding 3 mL of DMSO to the leaves residue and heating on a water bath for 10
min. and then filterimg the solution into the same standard flask. Make up the std. flask with
DMSO. Measure the absorbance at 645nm and 663nm for chlorophyll ‘a’ and ‘b’ respectively
against DMSO as a blank.
Then, prepare five dilutions of the extracted solution taking volumes ranging from 3 mL to 15
mL and making them up to mark in a 50 mL std. flask with DMSO. Prepare an unknown
concentration in a similar way. Measure the absorbance of these solutions at both the wave
lengths and draw a calibration plot between volumes of Chlorophyll solution and absorbance
at the two wavelengths. Report the unknown and Chlorophyll quantity in the result.

RESULT:

The estimated amount of


Chlorophyll ‘a’ = __________ μg /gram of leaves
Chlorophyll ‘b’ = __________ μg /gram of leaves

The amounts of Chlorophyll ‘a’ and Chlorophyll ‘b’ in the unknown sample are

______________ & ___________

You might also like