CHROMATOGRAPHY
AP BIO LAB #4A
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PLANT PIGMENTS
.
http://members.aol.com/diaspore/vhouse/biology/projects/photosynthesis/woida/meaning_of_light.html#question_1
CHROMATOGRAPHY
USE PENCIL not pen, to
mark starting line !!!!!
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ADD PLANT PIGMENTS
Image from Biology Place: http://www.phschool.com/science/biology_place/labbench/lab4/deposit.html
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PLACE IN JAR
IMPORTANT:
Make sure tip of paper
is touching solvent
Make sure pigment line
is ABOVE solvent
Put lid on jar !!!!!!!!
Important
Don’t let solvent front
run off end of paper !
http://www.phschool.com/science/biology_place/labbench/lab4/pigsep.html
Marker Chromatography
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Distance pigment migrated
Rf =
Distance solvent front migrated
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PLANT PIGMENTS
Carotene is orange yellow
Xanthophyll is lemon yellow
Chlorophyll a is blue green
Chlorophyll b is yellow green.
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Factors that affect movement of molecules
during chromatography
Solubility
1. ___________of
molecule in solvent used
(better it dissolves… farther it will move)
SOLVENT used:
Chlorophyll:
9:1 Petroleum ether : acetone
Markers:
water
Factors that affect movement of molecules
during chromatography
Size
2. _________of
molecule
(bigger molecules
move slower)
http://elvis.phys.lsu.edu/svoboda/post/figs/SumoWrestler.BMP
Factors that affect movement of molecules
during chromatography
3. _______________________
used
Attraction to media
(EX: Attraction of molecules to paper)
Lots of different kinds of chromatography
http://courses.cm.utexas.edu/jrobertus/ch339k/overheads-1/ch5_mol-seive.jpg
http://www.cofc.edu/~kinard/Applets/GasChromatographFID.gif
Factors that affect movement of molecules
during chromatography
Time
4. __________allowed
to run
Longer time =
More separation
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CHLOROPHYLL FLUORESCENCE
Image from Biology AP Edition by Campbell & Reece published by Pearson Benjamin Cummings