Exercise 4C:

Testing for Fatty Acids

And Paper Chromatography
Acejo, Esguerra, Ordoñez, Buhayo
BIO 20 TF 1-4
Exercise 4C1

What are Fatty Acids?

Fatty acids are carboxylic acids with long

hydrocarbon chains.

They serve as precursors to other lipids.

They usually contain even numbers of carbon

atoms in straight chains (commonly C14 to C24),
and may be saturated or unsaturated
 Exercise 4C1

Objectives

01 To test or determine the presence of fatty acids in various seeds

02 To familiarize ourselves with the grease spot test, which shows the difference between
grease spot and wet spot
Exercise 4C1

Methodology

Rub a peanut seed on

one corner, coconut
Get a 10 x 10 cm filter Put a drop of vegetable
paper.
meat on another corner,
oil on the fourth corner.
corn oil, cocoa power
on another corner.

Hold the paper over a

In the center of the
flame for a few seconds Make the grease spot
paper, put a drop of
without burning the test.
water.
paper.
Exercise 4C1

Discussion
The grease-spot test is used to determine
the presence of lipids in a certain
substance.

Lipids that are derived from glycerol and

sphingosine will produce translucent spots
or stains on fabrics, while lipids that are
non-derivatives of glycerol and sphingosine
will not.
Exercise 4C1

Data and Results

 Exercise 4C1

What happened?
Grease and fat have high boiling points, enabling them to absorb more heat than water. As a result, it
takes much higher temperatures to evaporate grease and fat as compared to water, which has a much
lower boiling point.

When put over the open flame, the greasy and fatty spots remained, while the water absorbed the heat
and evaporated quickly.
 Exercise 4C2

Objectives

01 To be able to separate pigments by means of paper chromatography

02 To be able to compute for the Rf value of different pigments that appear in the
chromatography
Exercise 4C2

What is Chromatography?

A technique used to visually separate

mixtures into their individual
components

Can be used to isolate certain

components or to simply measure how
much of a substance there is in a
mixture.
 Exercise 4C2

Methodology

Place a leaf over

the filter paper
Keep rubbing the
Prepare and rub a coin
Measure about 1 coin until there is
chromatograph over the leaf.
cm from the a solid and
paper by cutting a Make sure that
bottom of the uniform green
strip of filter the pigment on
paper and mark. pigment on the
paper. the paper is
filter paper.
exactly on the
mark made.
 Exercise 4C2

Methodology

Place the filter paper with

In a beaker, combine 9
Using a dropper, place the the pigment in the test
parts lighter fluid or 9 parts
solvent in a test tube. tube, while making sure
hexane with 1 part
(Must be below 1 cm) that the solvent does not
acetone.
touch the pigment.

Wait for the solvent to go

Place the set-up in a dark
up along the filter paper
room, undisturbed.
until is stops.
Exercise 4C2

Getting Rf Values:

Retention factor (Rf) indicates how far a

particular compound will travel in a
particular solvent.

If Rf values are similar between two

unknown compounds, it is likely that
they are the same compound.
Exercise 4C1

Data and Results

Exercise 4C1

Data and Results

 Exercise 4C2

What happened?
The capillary action within the paper strip moved the solvent and the compounds along with it. Compounds such as
chlorophyll stayed near the bottom of the strip, while compounds like the carotenes moved all the way to the top near
the end of the solvent’s travel.
 Exercise 4C2

In Conclusion:
Paper chromatography operates on solubility. Compounds which are more soluble in the solvent (lighter fluid/hexane)
will go up a longer distance compared to compounds that are less soluble in the given solvent.

In this case, since lighter fluid is a non-polar substance, and based on the distances the compounds moved, we can
conclude that by order of non-polar to polar compounds it is carotene, xanthophyll, chlorophyll A, and chlorophyll B.
 Exercise 4C2

In Conclusion:
Although we can usually tell different compounds apart using their color, this is not always the case.

A more reliable way of telling compounds apart is by using their Rf values. A compound will always fall within a specific
range of Rf values.
Answers to Guide Questions

Common Drugs and Pharmaceutical Products with Lipids

1. Laxatives
• Main ingredient: Fiber Psyllium
• Effect: Loosens stool and increases frequency of bowel
movement
• Comes in oral and suppository forms
Answers to Guide Questions

Common Drugs and Pharmaceutical Products with Lipids

2. Castor Oil
• Main ingredient: Ricin oleic acid; Oil
• Also known as stimulant laxative
• Treats constipation; cleans out intestines before bowel
examination or surgery
Answers to Guide Questions

Common Drugs and Pharmaceutical Products with Lipids

3. Calamine Lotion
• Main ingredient: Zinc Oxide and Ferric Oxide
• Soothes and protects the skin following minor skin
irritations
4. Mint
• Main ingredient: mint
• For indigestion, stomach cramps, menstrual cramps and
can also be a appetite stimulant
Answers to Guide Questions

Why have plants been used as a source of many pharmaceutical products?

How is this related to photosynthesis?

• Plants have preventive characteristics regarding the

appearance of diseases.
• Plants produce compounds essential to both plants and
animals (humans).
• Plants are economically attainable.
Answers to Guide Questions

Why have plants been used as a source of many pharmaceutical products?

How is this related to photosynthesis?

• Photosynthesis is the process by which plants make their

own food.
• Carbohydrates, proteins and lipids are the primary products
of photosynthesis.
Answers to Guide Questions

Give the importance of using chemical test in determining plant constituents.

• Chemical tests are used to determine the pharmaceutical

and medicinal value of plants.
• Performing chemical tests incorrectly may result in
unwanted side effects in medicines produced, or may even
worsen the situation.
 References

Experiment 8: Grease spot test. Retrieved October 9, 2015, from

http://www.lungtp.com/biochem/e_bcdxb.html
Lipids. Retrieved October 9, 2015 from https://en.wikipedia.org/wiki/Lipid
Types of chromatography. Retrieved October 9, 2015 from http://www.rpi.edu/dept/
chem-eng/Biotech-Environ/CHROMO/chromtypes.html
Chromatograhy. Retrieved October 9, 2015 from
https://www.soinc.org/sites/default/files/uploaded_files/forensics/
For_Chromatography3.pdf

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