Practical 4 – Shedding Light on Biomolecules: Photosynthetic pigments

1). You scan your seaweed extract in the spectrophotometer and you notice that the top of one peak
is missing off the top of the screen. What does this mean and what would you do?

This probably means that your sample is too concentrated and the peak is off the top of the scale.
You could alter the maximum absorbance setting (see ''setting up the spectrophotometer'' in the
handbook) and scan again. However, this would decrease the sensitivity of your machine; you
would lose resolution and may not see other, smaller peaks. A better idea would be to dilute your
sample and run it again.

2). What extraction medium would you use to determine if your seaweed contained
Allophycocyanin?

a) Phosphate

b) Ethanol – Ethanol is a non-polar solvent and is used to extract chlorophyll pigments.

Allophycocyanin is a phycobilin and these are soluble only in polar solvents.

c) Acetone - Acetone is a non-polar solvent and is used to extract chlorophyll pigments.

Allophycocyanin is a phycobilin and these are soluble only in polar solvents.

Correct: Allophycocyanin is a phycobilin and these are soluble only in polar solvents such as
phosphate.

The colour of seaweed does not always tell you how it should be classified.

3). You are given 2 seaweed samples from a species of Rhodophyta, one is red but one is green as it
has a mutation that blocks the production of phycoerythrin. How could you demonstrate from
pigment analyses that the green sample belonged in the Rhodophyta and not Chlorophyta?

a) You would see absorption peaks at 457 and 645nm in an ethanol extract of the green sample
- these peaks are from chlorophyll b absorption and this pigment is not found in
Rhodophyta.

b) You would not see absorption peaks at 457 and 645nm in an ethanol extract of the green
sample.

c) You would not see any absorption peaks - seaweeds contain a number of pigments so you
would see other peaks even if phycoerythrin is not present.

Correct: these peaks are from chlorophyll b and so you would not see them in Rhodophyta
species.