Thin Layer Chromatography

Thin Layer Chromatography

I. Abstract

The purpose of this lab was to determine the rate of flow values

(Rf=distance of component/distance of solvent) for certain pigments

found in chlorophyll and carotene. For the chlorophyll, we observed

the following five colors: yellow, orange, dark green, green, and

light green. For the cartenoid, we observed the following five colors:

yellow, auburn, purple, pink, and red. The solvent in both of our

experiments traveled a distance of 6.8cm. We obtained the following Rf

values for the chlorophyll. For yellow, Rf=0.89; for orange, Rf=0.78;

for dark green, Rf=0.51; for green, Rf=0.49; and for light green,

Rf=0.44. We obtained the following Rf values for the carotene. For

yellow, Rf=0.89; for auburn, Rf=0.81; for purple, Rf=0.69; for pink

Rf=0.51; and for red, Rf=0.49.

The second part of this lab dealt with statistical treatment of four

of our lowest Rf values of chlorophyll compared to the rest of the

class. Our personal S.D.M. value for the chlorophyll was 0.080. The

S.D.M. for the lowest level of the class' Rf value was 0.021. The

S.D.M. for the second level of the class' Rf value was 0.026. The

S.D.M. for the third level of the class' Rf value was 0.058. The

S.D.M. for the highest level of the class' Rf value was 0.068. The

mean value from lowest to highest of the class' Rf values are as

follows: .45, .48, .57, and .79.

Thin Layer Chromatography - Plant Pigments

II. Materials

a. Developer solvent - iso-octane-acetone.

b. 2 strips of chromatography paper.

c. Carotene pigment.

d. Chlorophyll pigment.

III. Apparatus

a. 2 Developing Chamber.

b. Chamber lid.

c. Capillary tube.

d. Ruler.

IV. Procedure

a. Gather the required apparatus items and materials.

b. Place 25 drops of your developer solvent into each one of the

developing chambers and place the lid on.

c. Mark each chromatography strip 1cm from each end with a pencil.

d. Using the capillary tube, apply three drops of the chlorophyll