Ketone Bodies Prac
MBARARA UNIVERSITY OF SCIENCE AND TECHNOLOGY
FACULTY OF MEDICINE
MEDICAL LABORATORY SCIENCE
CLINICAL CHEMISTRY DEPARTMENT
TOPIC; DETERMINATION OF KETONE BODIES IN URINE
STUDENT NAME; AKANYIJUKA KENNETH
REG NO. 2021/MLS/024/PS
ABSTRACT
We used the urine sample which was provided to determine the presence of ketone bodies in it,
We used two method that is the urine Dip stick method, Here we picked a Urine dip stick from a test
kit, then we dipped it in the provided urine sample, a colour change of the dip stick was matched
with the colour on the test kit and Rothera’s Test, here we added 1gm of Rothera’s powder mixture
with in a test tube which was containing 5ml of urine, we mixed and added 2ml of concentrated
Ammonium hydroxide to the mixture.
INTRODUCTION
Ketone bodies that is acetoacetic acid, Acetone, and Beta Hydroxybutyrate are products of Lipid
metabolism, they are present in urine when they are excessively produced in the body A condition
called Ketonuria, This can be seen in the following conditions; Diabetic Ketoacidosis, Persistent
Vomiting, High fat levels and low carbohydrate levels
Ketone bodies can be present in urine in the following percentages
Acetone 2% ( acetone is Volatile and can also be excreted in breath)
Acetoacetic acid 20%
Beta Hydroxybutyrate 78%
METHODOLOGY
URINE DIP STICK ROTHERA’S TEST
Urine sample Urine
Urine dip stick Test tube, Pipette,
Beaker Sodium Nitro prusside, Rothera’s Powder
paper 0.75 gm, Ammonium sulphate
Gloves Ammonium hydroxide
Gloves
PROCEDURE
URINE DIP STICK
We picked the Urine dip stick from the test kit and dipped it in urine, and 1 minute we noted the
colour change of on the dip stick, the matched the colour change of the trip with the colours on the
test kit
ROTHERA’S TEST
We added 5ml of urine in a clean test tube
We Transferred gm of Rothera’s powder mixture with in the test tube and we mixed well
We added 2ml of concentrated Ammoniun hydroxide to the mixture
RESULTS
URINE DIP STICK
COLOUR CONCLUSION
Purple colour change +++ 100(10) mg/dl (mmol/L) [ketone bodies
present]
ROTHERA’S TEST
Pink-Purple ring at the interface
EXPLANATION OF THE RESULTS
URINE DIP STICK
The kit contains dipsticks coated with chemicals that react with ketone bodies. The dipstick actually
measure many parameters including Glucose, Urobilinogen, Bilirubin, specific gravity, PH urine
protein, Urine Nitrites and Urine Leukocytes, all of these have different principles which finally lead
to a colour change.
For the case of Ketone bodies the principle behind is that Ketones react with Nitroprusside and
acetoacetic acid to produce a colour change ranging from light pink for negative results to a purple
colour for positive results.
ROTHERA’S TEST
Acetoacetic acid and acetone react with an alkaline solution of sodium nitroprusside to form a
purple coloured complex, this method can detect above 1-5mg/dl of acetoacetic and 10-20mg/dl of
acetone
Beta hydroxybutyrate is not detected by this test
CONCLUSION
