FORMULAS

Butter No definite formula Lard C21H42O5

Olive oil C18H34O2 Coconut oil C4H8NNaO2

Castor oil C57H104O9 Sudan IV C24H20N4O

Ethanol C2H6O Diethyl ether (C2H5)2O

DATA AND OBSERVATIONS

Part A. Physical Properties of Lipid Samples

Solubility
SAMPLE STATE COLOR Odor
H2O Diethyl ether

1. Butter semi-solid yellow milky insoluble Soluble

2. Olive oil liquid light yellow rubber-like insoluble Soluble
3. Lard semi-solid white rubber-like insoluble Soluble
4. Coconut oil liquid light yellow rubber-like insoluble Soluble
5. Castor oil liquid light yellow rubber-like insoluble Soluble

All of the samples showed a similar manner: being miscible in diethyl ether and immiscible with water. Also,
butter and lard showed a similar characteristic of being semi-solid in room temperature and both came from animal
origin while the rest showed a similar contrasting characteristic of being liquid at room temperature and they came from
plant origin. The milky odor of the butter explains its source which is milk. Overall, all the samples showed a physical
properties of lipids.

Part B. Grease Spot Test

Table 1 Measurements of stain patches of the lipid samples

SAMPLE BEFORE DIETHYL ETHER AFTER DIETHY ETHER SIZE
Butter translucent spot present Translucent spot faded 0.15 cm
Olive oil translucent spot present Translucent spot faded 0.6 cm
Lard translucent spot present Translucent spot faded 0.2 cm
Coconut oil translucent spot present Translucent spot faded 0.5 cm
Castor oil translucent spot present Translucent spot faded 0.8 cm

Grease spot test is a test to indicate whether a food sample is a lipid. All of the sample showed a translucent
spot which implies they are lipids and after the addition of the organic solvent, diethyl ether, the translucent spot spread
out a little bit and faded. The disappearance of the translucent spot indicates that the sample were dissolved with the
organic solvent and are indeed, lipids.

Part C. Emulsion Test

Table 2 Results of emulsion test for lipid samples
SAMPLE APPEARANCE INDICATION
Butter Yellow layer above cloudy white solution +

Olive oil Yellowish white layer above cloudy solution +

Lard Cloudy solution +

Coconut oil Yellow layer above cloudy solution +

Castor oil Yellowish white layer above cloudy solution +

Legend: (+) – the sample is contains lipid, (-) – the sample does not contain lipids
This test is done to show the presence of lipids in a substance. The substance is first dissolved in ethanol. This
solution is then dissolved in water. If lipids are present in the mixture, it will precipitates and forms an emulsion. The
milky cloudy white emulsion is a positive result: lipid is present. If the mixture remains clear, there are no fats present
in the sample. All of the samples showed a positive indication which implies they contains lipids. Lipids are insoluble in
water and soluble in ethanol (an alcohol). After lipids have been dissolved in ethanol and then added to H2O, they will
form tiny dispersed droplets in the water. This is called an emulsion. These droplets scatter light as it passes through
the water so it appears white and cloudy. The yellowish color is a pigment of the oils except for lard, which is not a
yellow pigmented fat.

Part D. Sudan Test

Table 3 Results of Sudan Test
SAMPLE RESULT APPEARANCE
Butter Layers formed Yellow – red – orange - cloudy
Olive oil Layers formed Red – red orange - cloudy
Lard Layers formed Red – red orange - cloudy
Coconut oil Layers formed Red – whitish - clear
Castor oil Layers formed Red – red orange - clear

Sudan is a red fat-soluble dye that is utilized in the identification of the presence of lipids, triglycerides and
lipoproteins. Sudan III reacts with the lipids or triglycerides to stain red in color. The oil will stain red with Sudan III dye
since it is a lipid and contains triglycerides. However, since the oil is less dense than water and insoluble in water, the
oil will form a layer or globules above the water and appear as a red layer above the water in the test tube. A negative
result will show a diluted appearance of red color. All of the samples showed a red globules which is an indication that
the samples contain lipids.

QUESTIONS AND ANALYSIS

1. Why some lipids are solid or liquid in room temperature? And which one is preferred in the health industry?

2. In grease spot test it is uncommon to see a procedure involving addition of diethyl ether after translucent spot
is observed, what is the use of it? Also, what causes the spot look translucent?

REFERENCES

Brilliant Society. (2007). Brilliant-BioChem . Retrieved from Fats and Oils:

http://brilliantbiologystudent.weebly.com/ethanol-emulsion-test-for-lipids.html
Fearon, W. F. (2009). Introduction to Biochemistry. London: Heinmann.
Klein, D. (2013). Organic Chemistry: 2nd ed. Hoboken: Wiley.
M., B. J., L., T. J., & L., S. (2007). Biochemistry. New York: W. H. Freeman and Company.
Maton, A. (1993). Human Biology and Health. New Jersey: Prentice Hall.
Park, Y., Subar, A., Hollenbeck, A., & Schatzkin, A. (2011). Dietary fiber intake and mortality in the NIH-AARP Diet and
Health Study. Archives of Internal Medicine, 301-302.