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Chromatography

The document discusses chromatography, a technique for separating components in a sample using a stationary and a mobile phase. It outlines the historical background, types of chromatography, and provides a detailed procedure for conducting experiments with leaf extracts and dye separation using paper chromatography. Additionally, it includes questions for discussion regarding the nature of the extracts and solvents used.

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4 views

Chromatography

The document discusses chromatography, a technique for separating components in a sample using a stationary and a mobile phase. It outlines the historical background, types of chromatography, and provides a detailed procedure for conducting experiments with leaf extracts and dye separation using paper chromatography. Additionally, it includes questions for discussion regarding the nature of the extracts and solvents used.

Uploaded by

venusabigail.gutierrez
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Experiment No.

Chromatography

Discussion:

In 1906 the Russian scientist Tswett reported separating different colored constituents
of leaves by passing an extract of the leaves through a column of calcium carbonate, alumina
and sucrose. He coined the termed chromatography from the Greek words meaning “color” and
“to write”. Chromatography is taken now to refer generally to the separation of components in
a sample by distribution between two phases - one that is stationary and one that moves,
usually but not necessarily in a column.

According to the distribution system employed, there are two types of chromatographic
procedures: adsorption and partition chromatography. In adsorption chromatography, the
stationary phase is a solid material while the mobile phase is a liquid. In partition
chromatography, the stationary phase is a liquid while the mobile phase maybe either a liquid
(liquid-liquid chromatography) or a gas (gas-liquid chromatography).

In all the different types of chromatography, the principle is basically the same. As the
mobile phase passes over, in and around the stationary phase, individual components of the
mixture maybe absorbed or desorbed by reversible distribution between the two phases.
Different components may have different attractions for either the stationary or mobile phase
and so may move at different rates along the direction of the solvent flow.

In paper chromatography, the stationary phase is a special filter-type of paper. Actually,


the water which is normally held by the cellulose (about 2H2O per glucose unit) known as bound
water is the functioning part of the stationary phase. The mobile phase is the solvent or mixture
of solvents.

The mixture of organic compounds which is applied as s spot on the paper maybe
considered as the solute which to be distributed between the two phases. Distribution of the
solute between the “bound” water on the paper and the developing solvent results in the
movement of the solute through the paper.

Procedure:

A. Separation of Leaves Extract


Cut up a handful of San Francisco leaves (or other leafy vegetable) into small pieces
with scissors. Grind the pieces in a mortar with enough ethyl alcohol to produce 2 to 3 ml of the
liquid. Decant the green liquid into a watch glass. The extract must be as concentrated as
possible.

Using a micropipette (a capillary tube will do), place a drop of the extract on for corners
of the filter paper resting across the mouth of an empty beaker. Allow the drops to dry then
place another drop on the same spots and dry. Continue until a total of 3 to 4 drops has been
added on each spot. Be sure to dry between drops.
Using another clean micropipette, add the solvent (toluene on the first spot, acetone on
the second, ethyl alcohol on the third and mixture of acetone and ethyl alcohol on the fourth)
on the spots made. As in making the spots, add the solvents drop by drop, drying between
drops by blowing until a total of about 15 drops has been added on each spot.

B. Paper Strip Chromatogram


Cut a strip 22 cm long of What man no. 1 filter paper 2 cm wide at one end tapering to
1 cm at the other end. Draw a pencil line 2 cm from, and parallel to, the narrow end of the
strip.

By means of capillary tip, apply a spot of a dye (to be prepared by your instructor) to the
paper at the center of the pencil line. The spot should be 3 to 5 mm in diameter. Dry the spot.

Prepare a mixture of 3 ml of water and 3 ml of ethyl alcohol in a hard glass tube. Have
ready a cork which fits the mouth of the tube.

Suspend the larger end of the filter paper to the tapered end of the cork by means of a
paper clip inserted through a cut in the middle of the cork. Adjust the length of the filter paper
such that when inserted in the tube, the narrow end of the filter paper is immersed in the
solution to a depth of 1 cm leaving a space between the surface of the solution and the dye
spot. Now suspend the strip in the test tube and fit the cork in to the tube. The paper should
hang straight and should not touch the sides of the tube. A staple placed at the bottom of the
paper will help to keep it straight.

Allow the tube to stand, undisturbed, until a separation of the mixture occurs or until the
solvent ascends about 5 mm from the broad end of the paper. Removed the paper and
observed.

Questions for Discussion:

A. Is the extract a substance or a mixture of substances? Explain.

Which solvent gives the best separation of components?

B. Is the dye a substance or a mixture of substances?


Organic Chemistry Individual Report Group No. ______

Name: _________________________ Date: ___________

Section: _______________________ Rating: __________

Experiment No. ___

Title

___________________________________

I. Discussion

II. Objective/s

III. Materials/Reagents

IV. Drawings/Set-up

V. Observation/Discussion of Results

VI. Conclusions/Generalizations

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