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Step 10Spotting and Developing
Carefully mark a line 1.5cm from the base of one plate.
Place as small a drop as possible on this line, centered as much as possible. Blow on it until it dries and repeat two or three more times.
Place a 1cm layer of 6:4 hexane:acetone in the bottom of the mason jar/developing chamber, put the lid on and set aside for a few minutes to allow the eluent to wet the filter paper. In the pictures I went in to work and used a beaker since the jar is too hard to see through.
Using tweezers gently place the plate in the jar such that the spot is just above the level of the hexane/acetone mixture, propped up against the filter paper.
Sit back and watch as the solvent line climbs up the plate and pulls the pigments along with it. The green is the chlorophylls and they should move further up the plate than the yellow lines which are the xanthophylls.
When the solvent is about 0.5cm from the top of the plate quickly remove it and mark the solvent line with a pencil.
To obtain the RF for any spot (or band) on the plate, divide the distance from the bottom pencil line to the spot by the distance between the two pencil lines. The above separation was adapted from Adams' Laboratory Experiments in Organic Chemistry 6th Ed., in which petroleum ether was used instead of hexane.
Place as small a drop as possible on this line, centered as much as possible. Blow on it until it dries and repeat two or three more times.
Place a 1cm layer of 6:4 hexane:acetone in the bottom of the mason jar/developing chamber, put the lid on and set aside for a few minutes to allow the eluent to wet the filter paper. In the pictures I went in to work and used a beaker since the jar is too hard to see through.
Using tweezers gently place the plate in the jar such that the spot is just above the level of the hexane/acetone mixture, propped up against the filter paper.
Sit back and watch as the solvent line climbs up the plate and pulls the pigments along with it. The green is the chlorophylls and they should move further up the plate than the yellow lines which are the xanthophylls.
When the solvent is about 0.5cm from the top of the plate quickly remove it and mark the solvent line with a pencil.
To obtain the RF for any spot (or band) on the plate, divide the distance from the bottom pencil line to the spot by the distance between the two pencil lines. The above separation was adapted from Adams' Laboratory Experiments in Organic Chemistry 6th Ed., in which petroleum ether was used instead of hexane.
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