CHROMATOGRAPHY AP BIO LAB #4A Image by Riedell http://www.valemount.com/joel/lightoptics/images/colour6.jpg

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 !!!!! Image by Riedell

ADD PLANT PIGMENTS Image from Biology Place: http://www.phschool.com/science/biology_place/labbench/lab4/deposit.html

PLACE IN JAR IMPORTANT: Make sure tip of paper is touching solvent Image by Riedell 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 Image by Riedell

Distance pigment migrated Distance solvent front migrated Rf = Image by Riedell

PLANT PIGMENTS Carotene is orange yellow Xanthophyll is lemon yellow Chlorophyll a is blue green Chlorophyll b is yellow green. Image by Riedell

Factors that affect movement of molecules during chromatography Solubility ___________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 (EX: Attraction of molecules to paper) Attraction to media 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 http://www1.istockphoto.com/file_thumbview_approve/581259/2/istockphoto_581259_big_time.jpg

CHLOROPHYLL FLUORESCENCE Image from Biology AP Edition by Campbell & Reece published by Pearson Benjamin Cummings