Construct a chromatography setup (Fig. 8.1) Obtain a of chromatography paper, being careful to handle only by the sides Use scissors to cut one end of the to a point Use a pencil to a very faint dot in the middle of the strip 2 cm from the pointed tip Use a capillary tube to apply a small amount of chlorophyll extract on the dot. Do this by barely touching the end of the capillary tube to the paper and then immediately withdraw it. A small green dot should appear Wait until the green dot is dry completely and then repeat step 5 at least 15-20 times. At the end, the dot should be as small and as dark as possible Take the chromatography tube to the fume hood and pour enough chromatography solvent into it to fill the rounded portion Place the strip of chromatography paper (B) into the tube so that the point is in the solvent (D). Make sure only the tip is in the solvent. The green dot of chlorophyll should be above the line of solvent (C) Insert the rubber stopper (A) into the tube and use the test tube clamp (E) to hold it to the ring stand After 30 minutes, the solvent will have nearly traveled to the top of the strip of chromatography paper. Before it gets to the top, remove the strip of paper. Immediately use a pencil to mark the locations of the individual pigments on the paper strip. They will be more difficult to see if you wait until the solvent have dried completely. the strip to completely dry in the fume hood Return the chromatography solvent in the tube to the original container. Do not pour in the sink! Do not rinse the chromatography tubes with water. Allow to air dry You should see four of pigment corresponding to the following chlorophyll a - blue-green color, nearest original dot chlorophyll b yellow-green color, after chlorophyll a – xanthophylls light yellow, follow chlorophyll b – carotenes yellow-orange, furthest from original dot – Draw and label the approximate locations of these four pigments in Fig. 8.2 Fig. 8.1 Chromatography Setup Fig. 8.2 Chromotography of Chlorophyll
Construct a chromatography setup (Fig. 8.1) Obtain a of chromatography paper, being careful to handle only by the sides Use scissors to cut one end of the to a point Use a pencil to a very faint dot in the middle of the strip 2 cm from the pointed tip Use a capillary tube to apply a small amount of chlorophyll extract on the dot. Do this by barely touching the end of the capillary tube to the paper and then immediately withdraw it. A small green dot should appear Wait until the green dot is dry completely and then repeat step 5 at least 15-20 times. At the end, the dot should be as small and as dark as possible Take the chromatography tube to the fume hood and pour enough chromatography solvent into it to fill the rounded portion Place the strip of chromatography paper (B) into the tube so that the point is in the solvent (D). Make sure only the tip is in the solvent. The green dot of chlorophyll should be above the line of solvent (C) Insert the rubber stopper (A) into the tube and use the test tube clamp (E) to hold it to the ring stand After 30 minutes, the solvent will have nearly traveled to the top of the strip of chromatography paper. Before it gets to the top, remove the strip of paper. Immediately use a pencil to mark the locations of the individual pigments on the paper strip. They will be more difficult to see if you wait until the solvent have dried completely. the strip to completely dry in the fume hood Return the chromatography solvent in the tube to the original container. Do not pour in the sink! Do not rinse the chromatography tubes with water. Allow to air dry You should see four of pigment corresponding to the following chlorophyll a - blue-green color, nearest original dot chlorophyll b – yellow-green color, after chlorophyll a xanthophylls – light yellow, follow chlorophyll b carotenes – yellow-orange, furthest from original dot Draw and label the approximate locations of these four pigments in Fig. 8.2 Fig. 8.1 Chromatography Setup Fig. 8.2 Chromotography of Chlorophyll