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Salkowskis test The presence of a double bond in onecholesterol rings is responsible for itsability to form color products in thepresence

of concentrated inorganic acids Sulfuric acid Results in dehydration of cholesterol moleculewith a formation of ared bicholestadiendisulphonate Bluish color between the 1st layer(chloroform) and 2ndlayer (H2SO4)

Liebermann Burchard test

The LiebermannBurchard or acetic anhydride test is used for the detection of cholesterol. The formation of a green or green-blue colour after a few minutes is positive. LiebermanBurchard is a reagent used in a colourimetric test to detect cholesterol, which gives a deep green colour. This colour begins as a purplish, pink colour and progresses through to a light green then very dark green colour. The colour is due to the hydroxyl group (-OH) of cholesterol reacting with the reagents and increasing the conjugation of the un-saturation in the adjacent fused ring.

The "Acrolein Test" is for the presence of glycerin or fats. A sample is heated with potassium bisulfate, and acrolein is released if the test is positive. When a fat is heated strongly in the presence of a dehydrating agent such as potassium bisulfate (KHSO4), the glycerol portion of the molecule is dehydrated to form the unsaturated aldehyde, acrolein (CH2=CHCHO), which has the odor peculiar to burnt cooking grease

The Benedict's test allows us to detect the presence of reducing sugars (sugars with a free aldehyde or ketone group). All monosaccharides are reducing sugars; they all have a free reactive carbonyl group. Some disaccharides have exposed carbonyl groups and are also reducing sugars. Other disaccharides such as sucrose are non-reducing sugars and will not react with Benedict's solution. Starches are also non-reducing sugars. The copper sulfate (CuSO4) present in Benedict's solution reacts with electrons from the aldehyde or ketone group of the reducing sugar to form cuprous oxide (Cu2O), a red-brown precipitate. CuSO4
++ -Cu + SO4

2 Cu++ + Reducing Sugar (electron donor) Cu+


Cu2O (precipitate)

The final color of the solution depends on how much of this precipitate was formed, and therefore the color gives an indication of how much reducing sugar was present.

Increasing amounts of reducing sugar

green orange red brown