chemical examination of urine contain glycosuria, proteinuria, ketonuria, bile salt, bile pigment, urobilinogen.

1. Chemical examination of urine
Ms Ankita R Bhatiya
Assistant Professor
Shree P.M.Patel COLLEGE OF
PARAMEDICAL SCIENCE N
TECHNOLOGY

2. It include
 1.Glycosuria
 2.Proteinuria
 3.Ketonuria
 4.Bile salt
 5.Bile pigment
 6.Urobilonogen
 7.Occult blood test

3. 1.Glycosuria
 Present of sugar in urine in known as
glycosuria.
 Causes of Glycosuria
 Glycosuria with hyperglycaemia:
 Diabetes
 Cushing’s disease
 Hyperthyroidism
 Drugs like corticosteroids
 Glycosuria without hyperglycaemia:
 Renal tubular dysfunction

4. 3 method is available for detection of sugar :
1.Benedict ‘s Test
2.CliniTest
3.Reagent strip
1.Benedict’s Test:
 Requirement: Test tube, Test tube stand, Test tube holder
,burner.
Reagent: Copper sulphate reagent.
Principle: The copper sulphate present in the Benedict’s reagent
reacts with the reducing substances in the urine which convert
cupric sulphates to cuprous oxide in hot alkaline media.

5. Procedure:
Take 5 mL of Benedict’s qualitative reagent in a test tube.
 Add 8 drops (0.5 mL) of urine.
Boil the mixture for 5 min.
 Allow to cool ( under running tapwater).
Interpretation:
Blue clr: Negetive
Green clr with yellow ppts: +
Yellow green clr with yellow ppts:++
Yellow n orange ppts:+++
Orange n orange ppts: ++++

6. 2.CliniTest:
It is self heating test.
Principle: The copper sulphate present in the Benedict’s reagent
reacts with the reducing substances in the urine which convert
cupric sulphates to cuprous oxide in hot alkaline media.
Two types of method:
5 drop method
2 drop method
5 drop method:
Procedure:
1. Take 5 drop of urine.
2. 2.Add 10 drops of water n mix well.
3. 3.Drop 1 cliny test tube teblet.
4. 4. Compare reading with clr chart.
5. Note: 2 drop method is also availbale in which 2 drop of
urine is taken.

7. Procedure:
1. Take 2 drop of urine.
2. 2.Add 10 drops of water n mix well.
3. 3.Drop 1 cliny test tube teblet.
4. 4. Compare reading with clr chart.
Interpretation:
0.25 gm/dl: Trace
0.5 gm/dl: +
0.75gm/dl:++
1 gm/dl:+++
2 gm/dl: ++++
2 drop method:

8. Procedure:
1. Take 10 ml urine in test tube.
2. Dip reagent strip into urine.
3. Remove excess urine on tissue paper.
4. Now compare clr change with chart given
on bottle.
3.Reagent strip method:
Principle:
Glucose + 02-------------- Gluconic acid +H2O2
H2O2+ chromogenic----------- oxidised (pink clr) + h2O

9. 2.Proteinuria:
Present of protein in urine in known as Proteinuria.
Causes of Proteinuria
Glomerulonephritis
Fever
Emotional stress
Cold
Exercise
Drug therapy
Nephrotic syndrome
Kidney disease
High blood pressure
Malingancy

10. 3 method is available for detection of Protein :
1.Sulphosalisilic acid
2.Heat n acetic acid
3.Reagent strip
1.Suphosalic acid method:
 Requirement: Test tube, Test tube stand.
Reagent: Salphosasilic acid
Principle: It is based on principle that protein in the
presence of acid and when subjected to heat ,they get
denatured and as a result ppts are fomed.

11. Procedure:
Take 1.5 mL of sulphosalic acid in test tube.
 Add 0.5 mL or drops of urine sample.
Mix it well.
 Observe the turbidity.
Interpretation:
No turbidity= protien absent
Faint turbidity= protein present
Very faint turbidity= protein present.

12. 2.Heat n acetic acid test:
Pricniple: It is based on principle that protein in the
presence of acid and when subjected to heat ,they get
denatured and as a result ppts are fomed.
Requirement: Test tube,Test tube stand,test tube holder
Reagent: Acetic acid
Procedure:
1. Centrifuge 10 ml urine.
2 Take supernant in another test tube.
3. Boil urine only upper portion,lower portion acts as a
control.
4.Cloudiness that appears on boiling.
5.Add 3 to 5 drops od acetic acid n boil again.
6.Read the degree of cloudiness.

13. Interpretation:
No turbidity= protien absent
Faint turbidity= protein present
Very faint turbidity= protein present.
3.Reagent strip method:
Same as glucose test
4.Bence jones protein method:
1.Do SSA test,if positive than perform this test.
2.Centrifuge 10 ml urine.
3.Take supernant in another test tune n add few drops of 10
% of acetic acid.
4.Heat in boiling water bath at 56 Celsius for 15 min, ppts
appear protein is present.
5.Place tube in boling water bath for 3 min ppts will
disappear.
6.Upon cooling ppts will reappear n dislove again when
cooled below 40 celsius.

14. 2.Ketonuria:
Present of keton bodies in urine in known as
Ketonuria.
Fever
Starvation
Sever vomiting
Diarrhea
Prolong bleeding
Glycogen storage disease
Types of ketone bodies:
 Acetone
Acetoacetic acid
 β hydroxy butyric acid
They are products of fat metabolism

15. 2 method is available for detection of ketonbodies :
1.Rothra’s test
2.Reagent strip method
1.Rothra’s test:
Requirement: Test tube.test tube stand
Reagent: Ammonium sulphate ,sodium nitroprusside
Principal: Acetone and acetoacetic acid react with sodium
nitroprusside in the presence of alkali to produce purple
color.
Method:
Take 5ml of urine in a test tube and saturate it with
ammonium sulphate, then add one crystal of sodium
nitroprusside. then slowly run the liquor ammonia along the
sides of the test tube.

16. Interpretation:
. Formation of purple colored ring at junction
indicates positive test .
No purple clr test is negetive.
2. Reagent strip method:
Same as glucose test

17. 4.Bile salt: Hay’s surface tension test
Hemolytic jaundice
Early hepatitis
Hepatocellular jaundice
Causes of Bile salt
2 method is available for detection of Bile salt :
1.Hay’s sulphar test
2.Reagent strip method

18. 1.Hay’s sulphar test:
Requirement: Test tube
Reagent: Sulphar powder
Principle: Bile salt present when lower the surface tension of
urine, when sulphar powder is added on the surface of urine.
Procedure:
In 5ml of urine sprinkle a pinch of sulphur particles.
Observe the result.
Interpretation:
If bile salt is present sulphur particles will sink to the
bottom because bile salts lower the surface tension of
urine .

19. 5.Bile Pigment: Harrison’s spot test
Causes of Bile pigment
Hemolytic jaundice
Early hepatitis
Hepatocellular jaundice
2 method is available for detection of Bile Pigment :
1.Harrison spot test
2.Reagent strip method

20. 1.Harrison’s spot test:
Requirement: Test tube.funnel,filter paper
Reagent: barium sulphate, fouchet’s tragent
Principle: Bilirubin absorbs to Barium Chloride and results in
green color formation when fouchet’s reagent is added.
Procedure: In 5 ml of urine add 5 ml of 10% Barium chloride
and mix well .then filter to obtain the precipitate on a filter
paper .To precipitate add 1 drop of Fouchet’s reagent.
Developement of blue green color around the drop indicates
presence of bilirubin.

21. Interpretation:
If blue green clr is present bile pigment positive.
No blue green clr bile pigment negetive

22. 5.Urobilinogen: Ehrlich test
Causes of urobiliogen
Hemolytic jaundice
Early hepatitis
Hepatocellular jaundice
2 method is available for detection of Bile
Pigment :
1.Ehrlich teat
2.Reagent strip method

23. 1.Ehrlich’s test :
Requirement: Test tube
Reagent: Ehrlich’s reagent
Principle: P-diamethyaminibenzyldehde react with
urobilogen in urine n form pink clr.
Procedure:
In 5 ml of urine add 0.5 ml of Ehrlich’s reagent.
Mix well n not down reading.
Interpretation:
If pink urobilinogen present
No pink clr urobilonogen absent

24. 7.Occult blood test: hematuria
Presence of bloof in urine known as hematuria.
Causes of Hematuria
Disease of urinary tract
Glomerular disease :
Glomerulonephritis
Non glomerular disease: Calculus , tumor, infection,
tuberculosis, pyelonephritis, trauma, carcinoma of prostate
Hematological condition: Coagulation disorders ,sickle cell
disease
2 method is available for detection of
hematuria :
1.Benzidine test
2.Reagent strip method

25. 1.Benzidine test :
Requirement: Test tube
Reagent: glacial acetic acid,H2O2,Benzidine powder.
Principle : The peroxidase activity of haemoglobin decomposes
hydrogen peroxide releasing nascent oxygen ,which in turn oxidizes
benzidine to give blue colour.
Procedure: Mix 2ml of benzidine solution with 2ml of hydrogen
peroxide in a test tube .Take 2ml of urine and add 2ml of mixture .A
blue or green colour within 5 min indicates positive reaction.
Interpretation:
If blue green clr occult blood present
No blue green clr occule bloof negetive.