A ketones in urine test measures ketone levels in your urine. Normally, the cells in your body use glucose (sugar) from your blood for energy. If your cells can't get enough glucose, your body breaks down fat for energy instead. This produces an acid called ketones, which can build up in your blood and urine.

1. SCHOOL OF PARAMEDICS AND ALLIED
HEALTH SCIENCES, BBSR
Presented By: ARPITA MOHAPATRA
Programme: Master of Science
Branch: Applied & Clinical Microbiology
Guided By: Asst. Prof. Sonalika Mahapatra

2. CONTENTS
 Urine and its Composition
 Ketone bodies in urine
 Ketone bodies: Types
 Terminologies related to Ketone bodies
 Clinical significance of Ketone Bodies
 Detection of Ketone Bodies- Rothera’s Test
2024 C LINIC AL PATHO L OG Y 2

3. URINE AND ITS COMPOSITION
 Urine: the waste product of kidney, a blood
filtered waste product, compositing of 96%
water, 2% urea, 2% other ions (Ammonia,
sodium, Potassium, Chloride, etc.)
 Ph- Slightly acidic about 6.2
 The normal constituents excreted in urine are
urea, chloride, sodium, potassium, creatinine,
dissolved ions, organic and inorganic
compounds.
 Abnormally, are the excretion of detectable
amount of glucose, ketone bodies, bile salt, etc.
2024 C LINIC AL PATHO L OG Y 3

4. KETONE BODIES IN URINE
 Abnormal constituent of urine- Ketone bodies
 When food is digested, the body turns carbohydrates in glucose that enters the blood
stream. The body needs glucose as a fuel to perform activities.
 However, glucose has to be delivered, it does not automatically route itself to body site
requiring fuel insulin, a hormone secreted by the pancreas carry out the task, delivering
glucose to the cells throughout body cells and tissues.
 When there is not enough insulin in body is unable to use sugar for energy and body
start burn fat for energy, Ketone bodies are the byproduct for this process.
2024 C LINIC AL PATHO L OG Y 4

5. 2024 C LINIC AL PATHO L OG Y 5
Fig; 1 Fig; 2

6. KETONE BODIES: TYPES
Acetone
 Present in 2%
 True ketone body
Acetoacetic acid
 Present in 20%
 True ketone body
Betahydroxybutyrate
Present in 78%
2024 C LINIC AL PATHO L OG Y 6
Ketone bodies consists chemically of three substances:

7. TERMINOLOGIES
2024 C LINIC AL PATHO L OG Y 7
 Normal Plasma level of ketone bodies- <<1mg/dl
 Normal urinary level of ketone bodies- <<3mg/dl
 Ketonemia- The presence of an abnormally high concentration of ketone bodies in
blood.
 Ketonuria- A condition in which abnormally high amounts of ketone bodies are
present in urine.
 Ketonuria also known as ketoaciduria.
 Ketoacidosis- serious diabetes complications, where body produces excess blood acids
(ketones) i.e., Diabetic ketoacidosis, diabetic acidosis.

8. CLINICAL SIGNIFICANCE OF KETONE BODIES
The ketones bodies are seen in case of:
 Diabetes mellitus
 Starvation
 Fasting
 Fever
 Low carbohydrate diet
 Increased protein diet
 Prolonged vomiting
 Pregnancy
 Anorexia
2024 C LINIC AL PATHO L OG Y 8

9. DETECTION OF KETONE BODIES IN URINE
ROTHERA’S TEST is performed to detect the presence of ketone bodies in urine.
Principle: Acetoacetic acid and acetone reacts with sodium nitroprusside in alkaline solution to
form a purple colored complex.
 Rothera’s test is sensitive to 5mg/dl of acetoacetic acid and 10mg/dl of acetone in urine.
 Either we can use commercially made Rothera’s powder compositing of 0.75gms of Sodium
Nitroprusside and 20gms of Ammonium sulphate.
 Else single compounds could also be taken prior to use.
2024 C LINIC AL PATHO L OG Y 9

10. 2024 C LINIC AL PATHO L OG Y 10

11. 2024 C LINIC AL PATHO L OG Y 11
Requirements: Test tubes, Sodium Nitroprusside, Ammonium sulphate, Liquor Ammonia, Dropper.
Procedure:
1. Transfer about 5 ml of urine to a test tube.
2. To it ammonium sulphate was added in pinch for saturation.
3. The saturation process tend to bring up all the ketone bodies to the surface of the urine
4. Saturate till deposition comes on the bottom of the test tube
5. To it a crystal of Sodium Nitroprusside was added and mixed.
6. To the mixture then Liquor ammonia was added (8 drops) alongside of the wall.
Result and Interpretation:
 Immediate formation of purple permanganate coloured ring at the interface- Ketone bodies
“+ve”
 The intensity of the colour formed was graded +, ++, +++.

12. 2024 C LINIC AL PATHO L OG Y 12

13. QUANTITATIVE DETECTION
 Quantitative detection: Blood test
 Detection of ketones in blood.
2024 C LINIC AL PATHO L OG Y 13

14. UPDATE ON MEASURING KETONES
 Recently technology has announced for measuring of beta-hydroxybutyrate in interstitial
fluid.
 Understanding the potential mechanism of SGLT 2 which may be involved in the promotion
of ketosis and ketoacidosis in Patients.
 Understanding the mechanism of T1D & T2D patients.
 Differentiation on the terminologies- Ketogenesis, Ketosis, Ketoacidosis and Diabetic
Ketoacidosis
2024 C LINIC AL PATHO L OG Y 14

15. 2024 C LINIC AL PATHO L OG Y 15
REFERENCES
Giri, D. (2018, November 5). Rothera's Test for Ketone Bodies: Principle, Procedure and Clinical
significance Retrieved from LABORATORY TESTS.org. : https://laboratorytests.org/rotheras-test/
Godkar, P. B., & Godkar, D. P. (2018). DETERMINATION OF KETONES. In P. B. Godkar, & D. P.
Godkar, TEXTBOOK OF MEDICAL LABORATORY TECHNOLOGY (3 ed., Vol. 2, pp. 1436, 1437).
Mumbai: Bhalani Publishing Hose. ISBN 9789381496190
Jingtong, H., Yeung, A., & Bergenstal, R. (1.02.2023). Update on Measuring Ketones. San Francisco: UC
San Francisco Previously Published work. doi:10.1177/19322968231152236

16. 2024 C LINIC AL PATHO L OG Y 16
THANK YOU