Determination of Chlorophyll Content in Chloroplasts

Friday, November 21, 2003

Description
This protocol describes how to determine the chlorophyll content of chloroplasts using a spectrophotometric assay.

Procedure
1. Isolate Chloroplasts from spinach leaves (See protocol for preparation of chloroplasts from spinach leaves).

2. Pipet 1 ml of a chloroplast suspension into a 15 ml centrifuge tube and add 8 ml of Acetone. Mix thoroughly.

3. Add 1 ml of ddH2O, mix again and centrifuge at 1,000 X g for 3 to 5 minutes.

4. Measure the absorbance of the sample from Step 3 on a spectrophotometer at 652 nm (See #1).

5. Calculate the amount of total chlorophyll in 1 ml of the Chloroplast suspension. The amount of chlorophyll is given by the equation:

mg chlorophyll per ml = Absorbance / 34.5

Recipes

Supplies

Tips
Use water as a blank to zero the spectrophotometer.

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Determination of Chlorophyll Content in Chloroplasts