Basic Concepts in Clinical Biochemistry-A Practical Guide.7z

30.2 Rothera’s Test for Acetoacetic Acid and Acetone

30.2.1 Principle

Alkaline nitroprusside reacts with keto-group of acetone and acetoacetic acid to form
a purple-colored complex.

30.2.2 Reagents

(a) Rothera’s reagent: dry mixture

Take ammonium sulfate and sodium nitroprusside in 100:1 ratio. Grind well to mix
powder of salts.

(b)Conc. ammoniain liquid form

36.5 Procedure

5 ml of urine is saturated with Rothera’s reagent in a test tube. Then 0.5–1.0 ml of
conc. ammonia is added through the sides of the tube in such a way that it layers on
top of the urine. Any change in color is observed within 30–60 s.

gluconeogenes

citric

acid

cycle

Ketone

bodies

Acetyl CoA

Acetyl CoA

Liver Blood Extrahepatic tissue

Glucose

FFA

excreted in urine

citric acid cycle

Acetyl CoA

ketone bodies

acetone in lungs

glucose utilisation

acetone and

β-hydroxy butyrate

exported as energy

source for heart,

skeletal muscle,

kidney and brain

Fig. 30.1 Utilization of ketone bodies

120 30 Qualitative Analysis of Ketone Bodies in Urine