Experiment 3(a): To

inks by ascending seperate

the coloured
components in a given mixture of red and blue
chromatography and find the Rf-values of the
components present.
Apparatus Required
Strip of Whatman filter paper No.1 2 Capillary tube
Glass rod. 4. Gas jar with lid.
Materials Required
Distilled water Alcohol
Procedure or Method (i) Mixture of blue and red ink.
Take a strip of Whatman filter
paper and draw a line about 4 cm above the lower end of the
with the help of a pencil. strip parallel to the width
(i) Now placea drop of the mixture of red and blue inks
the strip in air to get it dry. at the centre of this line with the help of a capillary tube, keep
Take a gas jar with the cork of suitable size which can be fitted
(v) Make a narrow hole in the cork and pass a bent tightly into its mouth.
glass rod through it. This rod can serve as a hook.
Suspend the paper strip into the gas jar with the help of a glass hook.
(vi) Add suitable amount of
developing solvent (50% alcohol+Distilled water) so that it occupies 4 cm of the jar,cork
the jar tightly and deep it undisturbed.
(vi) Allow to dip the strip in the solvent in such a
way that the marked line is 2 cm above the solvent.
Vii) Keep the strip in such position for 1/2 an hour.
ix) Sovent starts rising up with the mixture of red blue inks.
(x Two different spots of red and blue colour are seen in the strip after the solvent has
risen a certain height.
(xi) Take out the strip from the jar and mark the level of the solvent on the
paper with a pencil called solvent front
(xx) Dry the paper. Different spots will travel different
height.
 Chromatography or Chromatographic Analysis
(xii) Measure the distance of the two spots red and blue form the original point and also measure the solvent from form
the same starting point.
(xiv) Then determine the R-values of these two inks by applying the following formula:

Distance travelled by blue/red ink form the point of application.

R, Distance travelled by the solvent form the
original line.

Observations:
S. No. Name ofthe substance Distance travelled by components Distance travelled by solvent R
Red-Blue Ink A cm.(Red) X Cm. AWx
Red-Blue Ink B cm. blue
L cm. B/x
Result:-() R,for Red component=A/x
R, for Blue component=-B/x
Necessary Precautions:
Always make the use of a fine capillary tube.
Keep the jar undisturbed and covered during the experiment.
A spot should be small and rich in mixture.
4. Allow the spot to dry up completely before
putting the strip in the
Keep the strip erect. Donot let it to be curled.
jar.
3.
6. Always keep it away form the walls of the jar. It should not touch the walls of the jar.
7. Use only distilled water.
8. Do not allow the drop to dip in the solvent.
9. Watch and wait till the solvent has risen up about 10-15 cm above the original line.

Experiment 3(b): To separate the coloured components present in the given grass/flower
juice by Ascending paper chromatography and determine their Rf Value.
1. Apparatus/Requirements: (As in
expt. 3)
Theory/Method: (Same as in expt. No. 3)
Chromatography or Chromatographic Analysis
Observations:

S. No. Name of the substance Distance travelled by components Distance travelled by solvent R

Flower/Grass/Juice Mix. a cm.(Green Chlorophyll) A cm. alA

- do-
b cm.(Yellow Xanthophyll) AA cm. b/A
- do - Red
3. c cm. Carotene A cm c/A
Result:
R,of Green Component(chlorophyll)=a/A
RofYellow Component (Xanthophyl)=b/A
R,of Red Component (Carotene)=c/A
Necessary Precautions
Same as in Expt. No. 3.
Instructions for the Lab. Assistants
1. Take red and blue inks of good quality and mix them in equal quantties, dilute with water and supply.
2. Firstallowtocrush some flowers and grass and bring about the extraction of coloured components with the help
of dil. alcohol or acetone.