Experiment 5: CHROMATOGRAPHY

GROUP III

Obtain an 8 x 22 cm chromatogram
paper. Draw light pencil line parallel to
the longer side 2 cm from edge.

Dyes

MO

MO+MB

MB

F+MB

Order
1
2
3
4
5
Legend: MO Methyl orange E Erythrosine
F Fluorescein
MB Methylene blue

Starting 1 cm from the vertical

edge along the line put ten
equally-spaced dots (2 cm apart).
Using a capillary tube, spot the
chromatogram paper with dyes.

I+MB

E+I

Un

7
8
I Indigo carmine
Un Unknown

10

Roll the paper in such a way that

the spots are visible using a staple
wire by connecting the two shorter
edges. The edges should be 3
mm apart. Allow the spots to dry.

Fill up 600 mL beaker with 5 mL

50% ethanol. Place the rolled
chromatogram paper inside the
beaker; spots at the bottom. It
should not touch the sides.

Cover the beaker with Petri dish.

Allow the chromatogram to
develop for 15 mins or until the
solvent front is about 2 cm from
the upper edge of the paper.

Measure the distance travelled by

the dyes. (Distance from where
the dye was spotted to the center
of the colored zone or to the
center of two colored regions.)

Measure the distance travelled by

the solvent. (Distance from the
horizontal line draw up to the
solvent front.)

Remove and unroll the

chromatogram. Immediately pencil
out the solvent front. Dry in air.

Determine and record the Rf

values of the known and unknown
dyes.

Identify the unknown through the

Rf values calculated for the known
dyes.

Attach the chromatogram to the

data sheet and submit for
checking.