Salkowski reaction is the reaction of cholesterol with concentrated sulphuric acid.

Concentrated
sulphuric acid is highly hygroscopic and it removes two molecules of water from two molecules of
cholesterol, it causes a connection at position 3, forming bi- cholestadien . Simultaneously the sulphuric
acid sulphonates the molecule of bi-cholestadien at positions 7,7’ of aromatic ring and, as a final
product, red colour bi-sulphonic acid of bi-cholestadiene is formed

This taste is named after Leopold Salkowski, a German physiological chemist. In this test, only sulfuric
acid was added in a chloroform solution of the sample. In the presence of the acid, dehydration occurs
forming a bisteroid which gives a red color interphase (Espino-Cabatit, 1988). The red interphase can
also be blue. This interphase is observed in the chloroform layer while in the acid layer a green
fluorescence. This test is done in the sample and the color of interphase was noted.

Sample Documetation Observation

 Cholesterol Dark red at the junction

Raizaalmirah. (n.d.). Salkowski reaction is the reaction of cholesterol with concentrated sulphuric:
Course Hero. Retrieved from https://www.coursehero.com/file/p359l0k/Salkowski-reaction-is-the-
reaction-of-cholesterol-with-concentrated-sulphuric/.

bianca5195. (n.d.). The second test performed is the Salkowski test This test is similar to: Course Hero.
Retrieved from https://www.coursehero.com/file/p77u2la/The-second-test-performed-is-the-
Salkowski-test-This-test-is-similar-to/.

Procedure:

1. A small pinch of cholesterol was placed in a test tube.

2. Chloroform was added then added drop-by-drop until it is dissolved.
3. 5 drops of Sulphuric acid was then slowly added at the side of the inclined test tube.
4. The students stood the test tube slowly without shaking and observed the color at the juncture
of the two liquids.

Small pinch The change

Chloroform 5 drops of
of of color at
was added Sulphuric
cholesterol the
added acid was
was placed junction
drop-by- then slowly
in a tes was
drop added
tube observed
Conclusion:

Based on the experiment, the students concluded that all the test results of the samples of cholesterol
and bile complements the results of the control hence, ensuring that the sample do all contain the
properties that were in specific tests. In addition, the students determined the chemical reactions
undergone by cholesterol such as the Leibermann-Burchard Test and Salkowski Test which yielded
positive results. The properties of bile was also analyzed such as pH value, emulsification value and
presence of bile salt through Petten Kofer’s Test, which all yielded positive results.