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AP Lab 4: Plant Pigments and
Photosynthesis
Prelab Discussion!
What is chromatography?– A separation
technique that separates molecules by size and solubility
AP Lab 4 Part A: Pigment
Chromatography
AP Lab 4 Part A: Pigment Chromatography
What will our setup look like?
We will substitute a screw-top jar for a graduated cylinder!
Part A: Chromatography Remember to make a dark pigment
line! Allow pigment to dry between
applications. Do not allow solvent above pigment
line at start!
Be sure to mark solvent line at end for calculations!
solvent
AP Lab 4 Part A: Pigment Chromatography
What is Rf and how do you calculate it?– It is the relationship between the distance
moved by a pigment and the distance moved by the solvent.
AP Lab 4 Part B: Photosynthesis How could one measure photosynthetic
rates?– Amount of O2 produced
– Amount of glucose produced– Amount of ATP produced– Amount of NADPH created by the
reduction reaction: NADP+ + H+ NADPH
AP Lab 4 Part B: Photosynthesis
We will focus on the NADPH production.
We know that NADPH is the final electron and H receptor in the light dependent reactions.
We will be substituting the compound DPIP (a blue dye to be the final electron acceptor.
Therefore as photosynthesis progresses, the blue dye will become increasingly clear.
Light Dependent Reactions:sunlight
photolysis
THYLAKOID COMPARTMENT
second electron transport system
H2O
NADP+ NADPH
e–
ATP
ATP SYNTHASE
PHOTOSYSTEM IPHOTOSYSTEM II ADP + Pi
e–
first electron transport system
STROMA
Fig. 7.12a, p. 121
http://www.stolaf.edu/people/giannini/flashanimat/metabolism/photosynthesis.swf
DPIP!
Light Independent Rxns: Stroma
Light Dependent Rxns: Thylakoid Membrane
Sun = Energy Source
Chlorophyll absorbs solar energy
H2O = reactant
O2 waste
NADPH
ATP
CO2 reactant
CO
O
C
OO
NADPH drops off HH H
HH
Glucose made with ATP energy
NADP+
AP Lab 4 Part B: Photosynthesis
We will measure the % absorbance with probeware and a colorimeter.– What would a high % absorbance mean
reagrding photosynthetic rate?– A low % absorbance?
AP Lab 4: Plant Pigments and
Photosynthesis
AP Lab 4 Day 2: Take out your flowcharts and lab packets.
Light Dependent Reactions:sunlight
photolysis
THYLAKOID COMPARTMENT
second electron transport system
H2O
NADP+ NADPH
e–
ATP
ATP SYNTHASE
PHOTOSYSTEM IPHOTOSYSTEM II ADP + Pi
e–
first electron transport system
STROMA
Fig. 7.12a, p. 121
http://www.stolaf.edu/people/giannini/flashanimat/metabolism/photosynthesis.swf
DPIP!
Light Independent Rxns: Stroma
Light Dependent Rxns: Thylakoid Membrane
Sun = Energy Source
Chlorophyll absorbs solar energy
H2O = reactant
O2 waste
NADPH
ATP
CO2 reactant
CO
O
C
OO
NADPH drops off HH H
HH
Glucose made with ATP energy
NADP+
Part B: Photosynthesis Note setup!
Part B: Photosynthesis Troubleshooting Tips:
– Calibrate once! Two people can calibrate the sensor then
help out with the cuvette setup!
– Remember to mix each cuvette prior to taking an absorbance reading!
– Remember to handle cuvettes by the curved sides and load properly!