| Contributor: |
The Laboratory of William Heidcamp at Gustavus Adolphus University
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| This protocol describes how to determine the chlorophyll content of chloroplasts using a spectrophotometric assay. |
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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 Hint #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
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This bioProtocol does not use any solutions
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Acetone
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1. Use water as a blank to zero the spectrophotometer.
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