Ketone Bodies test procedure: Nitroprusside tube test or Rothera's test

Nitroprusside tube test or Rothera's test (Testing for Ketone)......

The Nitroprusside tube test or Rothera's test is performed with urine to determine the presence of acetone or acetoacetic acid in the urine and the symptoms of diabetes mellitus.

Nitroprusside dry reagent making procedure:

Ammonium sulphate ........................................ 100g
Anhydrous Sodium carbonate ........................ 50g
Sodium nitroprusside ...................................... 3g

Sodium nitroprusside should be well grained and taken in a clean container. Ammonium Sulphate and Sodium carbonate should be mixed well with nitroprusside. This reagent stays good for several months at room temperature.

Test method:

1. Take 0.5-1.0g Nitroprusside reagent in a test tube.
2. Then one drop of fresh urine should be given with Nitroprusside reagent of test tube.
3. After one minute it will look good, if the mixture is pale reddish purple then the result is positive, if the color of the mixture remains unchanged then the result is negative.

Results:

If the color of the mixture does not change, the result is Negative
If the color of the mixture is light purple the result is Positive (+)
If the color of the mixture is medium purple the result is Positive (++)
If the color of the mixture darkens the result is positive (+++)