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Rothera’s test
Definition: Rothera’s test is a type of laboratory test which is performed for the qualitative detection of ketone bodies in urine. During the “ketosis” three ketones bodies or acetone bodies are found in the urine which are the products of fat metabolism, known as acetone (2%), acetoacetic acid (20%) and beta-hydroxybutyrate (78%). During starvation and uncontrolled diabetes, the Ketone bodies are synthesized within the liver to re-utilized energy. These are types of acid that can results in metabolic acidosis in uncontrolled diabetes. Ketosis may be correlated with diabetes mellitus termed Diabetic ketoacidosis, or it may be due to starvation, persistent vomiting and high fat and low carbohydrate diet.
Rothera’s Test Principle
Acetoacetic acid and acetone react with an alkaline solution of sodium nitroprusside to form a purple-colored complex. This method can detect above 1-5 mg/dl of acetoacetic acid and 10-20 mg/dl of acetone. Beta-hydroxybutyrate is not detected.
Rothera’s Test Procedure
1. Take a clean test tube and add 5 ml of urine to it.
2. Transfers 1 gm of Rothera’s powder mixture within the test tube and mix well.
3. Add 1-2 ml of concentrated ammonium hydroxide to the urine sample within the test tube. It will create a thin layer over the urine sample.
4. Observe the pink-purple ring at the interface.
Results
Positive Test: If a purple permanganate-colored ring is immediately formed at the interface, means Ketone bodies are present.
Negative Test: If no permanganate colored ring is formed at the interface, means ketone bodies are absent within the test sample.
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6 сен 2024
