[2][3][4] This biochemical method got its name after German biochemist Ernst Leopold Salkowski,[2] who is known for development of multiple new chemical tests, that are used for detection of different kinds of molecules (besides cholesterol and other sterols also for creatinine, carbon monoxide, glucose and indoles).
Usually the solution of chloroform and the sample is prepared first and later treated with concentrated sulfuric (VI) acid.
[2][6] It is important to use only dried glassware, as dehydration reaction occurs during the test's procedure.
In such cases a sample is treated with nitric acid and 2% solution of potassium nitrite, with positive reaction being shown by presence of red colour.
[11] Treating a solution of a sample, containing sterols, with chloroform and highly hygroscopic sulfuric acid, leads to a dehydration reaction (two water molecules are removed from two cholesterol molecules) and formation of new double bonds.