Exercise 5
SEPARATION AND PURIFICATION OF ORGANIC COMPOUNDS
EXTRACTION
A frequently used method of withdrawing or separating a substance from a mixture is extraction by
the use of solvents. In applying this method, the mixture is dissolve in one of two immiscible solvents and
then shaken with the other to distribute the components between them. The amount of components in each of
the two solvents will be in direct proportion to their relative solubilities in each of them.
Water is usually one of the solvents in the extraction process and the other solvent is usually an
organic liquid of a non- polar character. Ethyl ether, one of the most important organic solvents, is used
extensively as an extracting solvent. It has a high solvent power for hydrocarbons and for oxygen containing
compounds, and is so highly volatile (b. p.34.6) that is easily removed from an extract at a low temperature
that even sensitive compounds are not liable to decompose. Although often preferred for research work
because of these properties, ether is avoided in oxidation to peroxide on long exposure to air which presents
the hazard of explosibility.
Ether is slightly soluble in water (7.5% by weight). However, its efficiency in use can be improved
considerably by the addition of a small amount of ionazable salt such as NaCI to the water layer. The
increase in polar property of the water solution will cause a decrease in the solubility of the non-polar
compounds. This is referred to as “salting-out effect.”. Hence, the bulk of the water present in a wet ether
extract with saturated sodium chloride solution and drawing of the lower layer. Drying is completed
simplified by filtering the ether solution with anhydrous sodium sulfate which abstract water by formation of
the pentahydrate.
Alternative water-immiscible solvents which are sometimes preferred even though they do not
match all the favorable properties of ether are: petroleum ether, ligroin, benzene, carbon tetra chloride,
chloroform, methylene chloride, ethylene dechloride and n-butanol.
Extraction with water-immiscible solvent is useful for isolation of natural products that occur in
animal and plant tissues having high water content. For exhaustive extraction of organic compounds from
dried leaves or seeds, the solid is packed into a filter water thimble place in Soxhlet extractor . In the soxhlet
extractor, solvent vapor rises in the tube and condensed solvent drops into the solids, leaches out the soluble
material and carries it to the boiling flask where the non-volatile extracted material accumulates. The same
solvent is used over and even substances of very slight solubility can be extracted by prolonging the
operation.
Test Compounds: Apparatus/Materials:
red gumamela flowers beakers
pechay leaves amber-colored bottles
other flowers separatory funnel
Reagents: filter paper
95% ethyl alcohol methanol knife
1 N-HCl distilled water funnel
1 N-NaOH anhydrous Na2SO4 vials
Petroluem ether Vaseline dessicator or wide mouthed jar with cover
Benzene
PROCEDURE:
A. Extraction of Colored Substance from Gumamela Flowers
1. Get the petals of two medium-sized red gumamela flowers and immerse them in 25 ml of 95%
alcohol in the beaker for one hour.
2. Decant and filter to remove any foreign solid substance. Collect the filtrate n an amber-colored
bottle.
3. Test the filtrate for its indicator property by adding 1-2 drops of it to 3 ml of 0.1 N-HCl
solution in a vial or test tube. Repeat using the 3 ml of 0.1 N-NaOH solution. Record the
colors observed in each case.
4. Repeat steps 1-3, uwing other flowers
Note: Step 1 and 2 are preferably done by one group only, and the resulting filtrate is
divided and distributed among the other groups for testing of the indicator property.
B. Partial Solubility in Immiscible Solvents Extraction of Chlorophyll (for demonstration)
1. Remove the white stems of pechay leaves and wash the green portion thoroughly to remove
dirt and soil adhering to it. Air-dry the leaves.
2. Slice the green leaves finely and weight 10 grams of it. Immerse in mixture of 45 ml of
petroleum ether, 5 ml of benzene and 15 ml of methanol. Let stand for 1 hour in a dark, cool
place.
3. Decant the liquid, then filter.
4. Place the filtrate in a separatory funnel and add distilled water to dissolve the methanol.
Collect the green extract in an evaporating dish.
NOTE: Avoid shaking vigorously.
5. Dry the extract over anhydrous sodium sulfate in a dessicator for 30 minutes. Describe its
color and consistency. Transfer to a vial, label property, and submit to your instructor.
NOTE: The same extraction process can be done with other natural products like
carrots and “atsuete.”
C. Keep the extracts from A and B for use in the following exercise.
QUESTIONS:
1. What are the indications that the desired substances have been extracted?
2. Which solvent extracted the colored substances from
a) Gumamela?
b) pechay?
3. What is the role of methanol in the extraction of chlorophyll?
4. What is “salting-out?” When is it used in the extraction process?
DATA AND ANSWER SHEET
Name ___________________________________ Date __________________________
Class Schedule ____________________________ Rating __________________________
Group No. _______________________________
Exercise 5
Separation and Purification of Compounds
Extraction
I. Results and Observations
A. 1. Extraction form Gumamela Flowers:
Solvent used: _______________________________________________________________
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Color of extract:
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2. Indicator Test Results
a. with acid: ________________________________________________________________
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b. with alkali: _______________________________________________________________
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3. Extraction of Chlorophyll:
Color of extract
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Other description
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Exercise 5
II. Answer to Questions
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