KETONE BODY ANALYSIS By: Dr. Beenish Zaki

LEARNING OBJECTIVES  SKILL: At the end of the practical session, the student shall be able to detect the presence of ketones by qualitative analysis.  KNOWLEDGE At the end of the session the student shall be able to explain and interpret the principle of Ketone body analysis and its clinical application

CAUSES OF KETONURIA  Ketonuria occurs in association with  Fever  Anorexia  Starvation  Prolonged vomiting  Fasting  Diabetic ketoacidosis  Pregnancy  Strenuous exercise

INTERFERING FACTORS  Carbohydrate free diet will cause ketonuria.  Certain drugs like :  Isopropyl Alcohol  Insulin  Metformin  Levodopa

METHOD Rothera’s nitroprusside test

PROCEDURE Experiment Saturate 5 ml of urine with solid ammonium sulphate. Add 0.5 ml of sodium nitroprusside. Mix and add 0.5 ml of concentrated ammonia slowly through the side of the test tube. Do not mix

METHOD and PRINCIPLE  Method: Rothera’s nitroprusside test  Principle :  Ketone bodies react with sodium nitroprusside to produce purple coloring at the junction of the two layers  Ammonium sulphate removes the impurities  Ammonia provides the alkaline medium

PROCEDURE ExperimentObservationInference Saturate 5 ml of urine with solid ammonium Sulphate. Add 0.5 ml of sodium nitroprusside. Mix and add 0.5 ml of concentrated ammonia slowly through the side of the test tube. Do not mix Purple colored ring is observed Presence of Ketone bodies acetoacetic acid and acetone