CM134-1L: ORGANIC CHEMISTRY 2 LABORATORY
1ST Quarter SY 2019-2020
EXTRACTION AND CHROMATOGRAPHY
ORTEGA, MARY ALYSSA T. – GROUP 3
SEPTEMBER 5, 2019
Introduction
I. Extraction
Extraction is a process in which one selectively dissolves an organic compound in an acceptable
solvent. Organic acids and bases can be separated from each other by using a variety of aqueous solutions.
The solutions have varying levels of pH which in turn effects the solubility of the organic compound. We have
seen that recrystallization is used to purify solids that are contaminated by relatively small amounts of
impurities. On the contrary, the techniques of extraction are a more coarse or preliminary technique, in that
it can be used to separate one compound from another or from large amounts of impurities. Most uncharged
organic molecules are more soluble in organic solvents than in water. If the organic solvent itself is not very
soluble in water, then when the solvent or one of its solutions is mixed with water, two layers will form, and
the solvents are said to be immiscible. The layers are called the organic layer and the aqueous layer. If the
two layers are shaken together, small amounts of the components of the organic layer will dissolve the organic
layer, establishing equilibrium between the two layers. This experiment aims to know the effect of number of
extractions to the distribution coefficient and percentage yield.
II. Paper Chromatography
Separating mixtures of substances into their respective components can be done using
chromatography. There are various forms of chromatography, but they entirely work on a similar principle.
These forms all possess a stationary phase and a mobile phase. The stationary phase involves most likely a
solid form, or a liquid sustained on a solid. The mobile phase on the other hand involves a gas form or a
liquid form. Along the stationary phase, flows the mobile phase which then conveys the mixture components
with it. Each component of a mixture has varying travel rates. When it comes to paper chromatography, a
precisely even permeable paper is considered as the stationary phase while an appropriate solvent mixture
or simply a liquid in form is considered as the mobile phase. Paper chromatography has been proven to be
actually effective in examining chemical compounds. In this process, the solvent mixture is dropped to a piece
of chromatogram paper. The side of the paper is immersed in a specific solvent. By capillary action, the
solvent is eventually absorbed by the paper as it moves up. With changing degrees, the solvent carries the
different mixture components, subject on the fondness of the compound to undergo adsorption on the paper
against being conceded along the solvent. The paper is specialized to be composed of cellulose. This
cellulose is in which polar molecules coming from the water undergo adsorption. The solvent, being less
polar, is typically comprising of a mixture of an organic compound, liquid in form, and water. The fraction of
the distance travelled by a component to the distance travelled by the solvent front, is called the “Resolution
front (Rf)” value or the “Retention value” for that component. Both distances are measured from the point of
the application of the mixture.
Materials and Methods
I. Extraction
There are two parts for this experiment: Salting-out effect and the Determination of a
Distribution coefficient and percent yield of extract using a just amount of Ether. For the first
part of the experiment, the Salting-out effect, two samples having different composition were
carefully observed. Two micro test tubes were both filled with 3 ml of Distilled water. For each
test tube, a drop of 0.003 M aqueous crystal violet and 0.5 ml of n-anyl alcohol were added to
1
it. The test tube was mixed well to produce a solution. Sodium Chloride (NaCl) was added
only to one test tube until the water layer is saturated. The observations were recorded based
on the change of appearance of each micro test tube.
For the second part of this experiment, the determination of a distribution coefficient
and percent yield of extract, utilized reagents such as: Adipic Acid, Sodium Hydroxide (NaOH),
and Ether. There were three samples that were prepared for titration. In the first sample which
is the preparation of 1% Adipic Acid as the original sample. A 10 ml of 10% Adipic Acid was
transferred into a 100 ml Erlenmeyer flask using a pipette. A drop of phenolphthalein was
added to the sample, then was proceeded to titration until it shows a light pink end point. The
second sample to test used Ether. A 10 ml of Adipic Acid was transferred to quick fit separatory
funnel. To this, 10 ml of Ether was poured to extract the acid classifying it as a single
extraction. The mixture was mixed gently. A drop of phenolphthalein was added to the
aqueous layer and proceeded to titration with 0.05 M NaOH. For the last preparation of the
sample which is the double extraction, 10 ml and 5 ml of Ether was transferred in the
separatory funnel using a pipette. The aqueous layer of the mixture is drawn off from the
separatory funnel into a clean bottle leaving a certain layer of Ether. The Ether was then drawn
off to the “Ether Extract” bottle. The previously drawn aqueous layer was transferred into the
separatory funnel then was extracted using 5 ml of Ether. A drop of phenolphthalein was
added to the mixture and then proceeded to titration.
II. Chromatography
An 8 x 22 chromatography paper was used for this experiment. First, a rectangular
piece of chromatogram paper was obtained. Using a pencil, a parallel line, 1.5 cm from its
longer side, was drawn. It was then marked with six dots, having equal space of 2cm from
each other. These dots were spotted with different dyes using a capillary tube. The paper was
rolled, with its two edges connected but not touching through staple wire. The 600mL beaker
was filled with 5mL of 50% ethanol. Inside the beaker, the rolled paper was placed carefully,
not touching its sides. The beaker was then covered with large petri dish that is enough to
cover the beaker until the dyes reached the end of the line. The distance traveled by the dyes
were marked. The paper was then dried. Other necessary measurements were also done.
The Rf values of the dyes were calculated. The unknown was identified by its Rf value.
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Results and Discussion
I. Extraction
Table 1. Salting-out Effect
OBSERVATIONS
The change of appearance was observed with the addition of the NaCl. The n-amyl alcohol
was completely soluble with water and aqueous crystal violet. However, on the other test tube
with a composition of NaCl, the solution gradually separated into two phases or layers. The
sodium chloride interrupted with the interaction of aqueous crystal violet and n-amyl alcohol.
NaCl increased the solution’s ionic strength through the water molecules and the ions. This
resulted in the decreases of the solubility of n-amyl alcohol to aqueous crystal violet.
The observation above having the change of appearance from a test tube containing NaCL
was because the solubility of a nonelectrolyte in water is decreased when an electrolyte, like salt
is added.
Table 2. Determination of a Distribution Coefficient and Percent Yield
Volume of NaOH solution used for Titration (mL)
Original 1% sample 71
Solution extracted once 59.3
Solution extracted twice 58.6
Weight of Adipic Acid (g)
Original Solution 0.25915
2
Solution, extracted once 0.21720
Solution, extracted twice 0.21390
Distribution Coefficient
Single Extraction 0.193
Double Extraction 0.212
Percentage Yield (%)
Single Extraction 83.82
Double Extraction 82.54
A quantitative measure of the how an organic compound will distribute between aqueous and
organic phases is called the distribution coefficient. It is the ratio, K, of the solubility of solute
dissolved in the organic layer to the solubility of material dissolved in the aqueous layer.
II. Paper Chromatography
Table 3. Rf Values for each sample
Dye Sample Rf Values
Methyl Orange 0.6825
Methylene Blue 0.5417
Fluorescein 0.6667
Indigo Carmine 0.5833
Erythrosine 0.6667
Unknown 0.6872
Name of the Unknown Methyl Blue & Fluorescein
The Rf value is defined as the ratio of the distance moved by the solute and the distance
moved by the solvent along the paper. Based on the data above, Methyl Orange having 0.6875
as its Retention Factor was the fastest to reach the solvent front. MO has also the largest R f
among the samples that would correspond to the largest distance traveled by this sample. Also,
MO having the same Rf values with the unknown, this can be said that the unknown sample is
Methyl Orange.
Conclusions
I. Extraction
Extraction is a process in which one selectively dissolves an organic compound in an
acceptable solvent. Organic acids and bases can be separated from each other by using a variety of
aqueous solutions. The solutions have varying levels of pH which in turn effects the solubility of the
organic compound. The organic compounds were separated with the addition of an organic solvent,
Ether. There were present errors in this experiment resulting to different data, but it was corrected
and analyzed. At the end of the experiment, it was determined that double or multiple extraction is
more coefficient and can attain as much of the desired product from the layer.
II. Paper Chromatography
In general, the adsorptivity of compounds increases with increased polarity (i.e. the more polar
the compound then the stronger it binds to the adsorbent). The eluting power of solvents increases
with polarity. Therefore, low polarity compounds can be eluted with low polarity solvents, while higher
polarity compounds require solvents of higher polarity. The stronger a compound is bound to the
adsorbent, the slower it moves. Non-polar compounds move up most rapidly (higher Rf value),
whereas polar substances travel up the plate slowly or not at all (lower Rf value). Based on the data
gathered, Methyl Orange resulting to the same value as the unknown is an indication that this dye
sample can be considered as the unknown.
