Ketones

DISCUSSION

Ketone bodies are the intermediate products of fat metabolism. These include acetone,
acetoacetic acid and beta-hydroxybutyric acid. Measurable amounts of ketones do not
appear in the urine because of the complete metabolism of fats into carbon dioxide
and water. Ketone bodies apppear in the urine when there is an increase in fat
metabolism. This can be seen in the following conditions: inability to metabolize
carbohydrates, as occurs in Diabetes mellitus, increased loss of carbohydrate from
vomiting and inadequate intake of carbohydrate associated with starvation and weight
reduction.

Seventy-eight percent of ketone compounds are beta-hydroxybutyric acid, 20 percent

acetoacetic acid and 2 percent acetone.

Procedure:

TESTS FOR ACETONE AND DIACETIC ACID/ACETOACETIC ACID

1. Legal’s test
Principle: In a reaction with the –NO group, sodium
nitroprusside, acetone and acetoacetic acid form isonitroacetone which
remains trapped in the complex anion.

 To a few mL of urine, add enough NaOH or KOH solution to render

it slightly alkaline.
 Add a few drops of sodium nitroprusside solution.
 Add a few drops of concentrated acetic acid.
 Observe the result and record as positive or negative.

A purple or violet-red color indicates acetone. Alcohol or acetic

aldehyde and diacetic acid also appear red.

2. Gunning’s test
Principle: This test is based on the reaction between acetone bodies and
alcoholic iodine resulting in the production of iodoform crystals.
 Pour 5 mL of urine into a test tube.
 Add 5 drps of strong ammonia
 Add Lugol’s solution enough to produce a black cloud which does
not disappear immediately.
 Let stand for a few minutes.
 Take a small quantity of the sediment and examine
microscopically. Iodoform crystals appear as yellowish or six-
pointed stars or six-sided plates.
  Report as positive or negative.

3. Gerhardt’s test
 To a few mL of urine in a test tube, add 10% ferric chloride
solution drop by drop until no further precipitation occurs.
 If diacetic acid is present, the urine will turn a Bordeaux red. If
the diacetic acid is due to drugs, it does not disappear upon
heating, and does not reappear upon cooling.