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Experiment # 3: Extraction and Chromatography

Estrada, Elle Nadine P. / Group 5

September 5, 2019
I. Introduction
A. Extraction
Extraction is a separation process where it separates the solution to a desired
substance. There are variety of aqueous solutions can be used to separate the organic acids
and basis to one another. It depends on its pH that affect the solubility of the organic
compound. Extraction gives several uses in the chemistry lab. Many organic compounds are
extracted from plants for medical purpose like morphine from the opium poppy, aspirin from
willow bark, digitalis from foxgloves and many more. Another example of extraction that we
do in our everyday life like washing laundry, adding bay leaf to the soup and tea bag for
drinking tea.
The experiment aims to know the effect of number of extractions to the distribution
coefficient and percentage yield.

B. Chromatography
Chromatography is one of the techniques in separating the solution to know the
components of the mixture. It is based on the distribution of the amounts of solutes within the
solid or liquid phase to the liquid or gas phase. Extraction or elution is involved in the
chromatography from extracting one material to another by using the solvent. In the
experiment, chromatography paper is used to know the small amount of analyte. As the liquid
phase is passing through the chromatography paper.
In the elution, the liquid phase moves up in the chromatography paper in the capillary
action. The separation of the mixture is contrasting depends on the relationship of the
chromatography paper, solvent, and solution. The components can either be absorbed or
carried along with the solvent. It may be caused by the solubility and absorption differences,
which may know the rate of separation of the components. This is affected by the molecular
structure of the substances along with the polarity differences. The separation of the
component is being served as a retention factor (Rf value) by dividing the measured distance
traveled by the solute to the solvent front.
The experiment aims to know the unknown compound by using the Rf values.
II. Materials and Methods
A. Extraction
a. Salting-out Effect
The material used are the micro test tubes and pipet. The reagents used are the
distilled water, aqueous crystal violet, sodium chloride and n-amyl alcohol. Fill the micro
test tube of 3mL of distilled water. In each tube, put 0.003M of aqueous crystal violet and
0.50mL of n-amyl alcohol. Shake each of the test tubes. Add sodium chloride (NaCl) to
one of the test tubes until the water is saturated. Compare the 2 test tubes through the
amount of the color in the water layer. Record the observations in the datasheet (Figure

Figure 1. Comparing the difference in the 2 test tube.

b. Determination of a Distribution Coefficient and Percentage Yield of Extraction
The materials used are Erlenmeyer flask, separator funnel, and beaker. The reagents
used are adipic acid, ether, sodium hydroxide, and phenolphthalein. In this experiment,
there should be no fire is present in the laboratory. Prepare the 250mL of the solution of
0.05M NaOH from the 2.50M NaOH. Put the 10Ml OF 10% adipic acid in the 50Ml
Erlenmeyer flask by using the pipet. A drop of phenolphthalein should be added in the
flask and titrate it with 0.05M NaOH (Figure 2) which is being prepared until the solution is
a light pink endpoint (Figure 3).

Figure 2. Adding NaOH in the solution.

Figure 3. The solution became light pink endpoint.

Record the volume of the titrate being used. Transfer the 10Ml of adipic acid in the
separator funnel. Extract the acid by using the 10mL of ether. During the extraction, shake
the mixture gently to avoid pressure build-up. The mixture should be able to stand for a
few minutes before the extraction to make the aqueous solution to settle down. A drop of
phenolphthalein should be added to the aqueous layer and titrate it using the 0.05M NaOH.
Record the volume the titrate is used and write the content of the separatory funnel after
it was being taken out in the separatory funnel and put it in the bottle with “Ether Extract”.
The acid should be extracted using the 5mL ether. Take out the aqueous layer in the clean
bottle. Empty the separatory funnel and place the solution in the bottle labeled “Ether
Extract”. Transfer the aqueous solution in the separatory funnel. The acid should be
extracted by using the 5mL of ether. Repeat the steps twice. Compute the distribution
coefficient and percentage yield in both the single and double extraction.
B. Chromatography

The materials used in this experiment are 600mL beaker, petri dish, and chromatography
paper. The reagents used in the experiments are methyl orange, fluorescein, methylene blue,
indigo carmine, ethanol, and unknown reagent. In the chromatography paper, draw a line of 1.5
cm at the top and in the bottom parallel. And through the line, place 6 dots which are equally
separated from one another. And through the dots, draw a horizontal line as an indicator of how
long will the solution will reach. In each dot, put one drop of each of the reagents (Figure 4).

Figure 4. 1 drop of each reagent in the chromatography paper.

Roll the chromatography paper and use the stapler to stick one end to have a cylindrical shape
paper. Put 5mL of 50% ethanol into the beaker. Put the paper into the beaker. Make sure that the
paper does not stick in the wall inside the beaker. Then cover the beaker by using the petri dish
(Figure 5).

Figure 5. The chromatography paper inside the beaker containing the ethanol.
Wait for 15 minutes or until the solution reaches the line in the upper part of the paper. Remove
the paper in the beaker and unfold it to put a line where each of the reagents can reach. Let the
paper dry in the petri dish (Figure 6).

Figure 6. Some of the reagent have reach to the solvent font.

Record the Rf by measuring the solution traveled in the horizontal line and divide it into the
solvent front. Identify the unknown by comparing the solutions of which one of them has the
nearest Rf to the unknown.
𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑙𝑒𝑑 𝑏𝑦 𝑑𝑦𝑒𝑠
𝑅𝑓 =
𝑆𝑜𝑙𝑣𝑒𝑛𝑡 𝑓𝑟𝑜𝑛𝑡 (6𝑐𝑚)

III. Results and Discussion

A. Extraction
a. Salting-out Effect
The micro test tube containing the distilled water, aqueous crystal violet and n-
amyl alcohol did not form any layer that separate the mixture. The second micro test
tube that contains NaCl (salt) have different color which is lighter than the first micro
test tube. It is because the NaCl formed 2 layer in the experiment through solubility of
nonelectrolyte in water that begins to decrease when the NaCl is added.

b. Determination of a Distribution Coefficient and Percentage Yield of Extraction

Distribution or partition coefficient is the ratio of how organic compounds have
difference in measure between the two phases. The ratio of the solubility of the
dissolved solute in the organic layer to the solubility of the material in the aqueous
layer. If the K is large then not all the solute can be found in the organic layer in the
single extraction. It need more extractions of the aqueous layer with the organic solvent
to be taken out to get the desired solution from the aqueous layer. Through the
experiment, it will show how much difference can be made through multiple extraction
to single extraction.

Through the comparison of the two values in the percentage yield and the
distribution coefficient of the extraction, the multiple extraction has greater percentage
yield and distribution coefficient than the single extraction. It means that the amount of
acid acquired is greater than extracting it once.
B. Chromatography
Through the chromatography paper inside the 50% ethanol in the beaker, the solvent
began to climb up to the solvent front. As the solvent climbs up, the samples are being
dissolved by the solvent through intermolecular attractions to the solvent. The samples
dissolve by the solvent, it also attracts in the liquid phase and began to move up to the
solvent front in the chromatography paper. The samples have different rate as some may
reach the solvent front and some did not. The unknown is identified by comparing the R f
of the given samples. The unknown sample was identified and it was methylene blue and
indigo carmine.
The Rf can be computed by dividing the measuring the solution traveled in the
horizontal line to the solvent front. Rf values may be depend on the polarity of the solvent,
solute and its liquid or solid phase. This can identify how high the solute and solvent are.
Ethanol has a strong polar solvent and it is strong absorbed. Low polarity or nonpolar of
the compound climb up very quickly and has a high value of Rf than the polar compound
which moves slowly and has low value of Rf. the methyl orange, fluorescein, and
erythrosine have high Rf so it is means that they are nonpolar while methylene blue, indigo
carmine, and unknown, which is methylene blue and indigo carmine, have low R f which
means that they are polar.

IV. Conclusions / Summary

A. Extraction
In the experiment, it concludes that double or multiple extraction is more efficient than
single extraction to get a higher percentage yield and distribution coefficient to get the
desired acid from the layer. There should be precautions in doing the experiment like
wearing face mask for the toxic compound like ether. Through the shaking of the
separatory funnel, it should be far away from the holder and its fellow neighbors. And lastly,
the stop cock must be open after being shake to release pressure.
B. Chromatography
In the experiment, it concludes Rf has an effect on the polarity of the compound. The
higher the Rf, then the compound has low polarity or nonpolar. While the lower the Rf then
the compound is polar. There should be precaution on doing the experiment like paper
should not stick in the wall inside the beaker, the chromatography chamber is sealed, the
sample spots must be separate from one another and the solvent front must not exceed
in the upper part of the chromatography paper.