This document summarizes common urine tests performed as part of a routine urine examination. It describes the three parts of a routine urine exam - physical, chemical, and microscopic. For the physical exam, it discusses evaluating the urine's volume, appearance, odor, specific gravity, and other physical properties. It then explains various chemical tests performed on urine to detect proteins, glucose, ketone bodies, and bile salts. These tests help diagnose and monitor various diseases.

1. URINE EXAMINATION
Dr sandeep singh
Pathologist…

2. Examination of urine is important for
diagnosis and assistance of various
diseases.
• Routine urine examination is divided into three
parts:
• Physical examination
• Chemical examination
• Microscopic examination

3. Physical examination
• Volume
• Appearance
• Odor
• Specific gravity

4. SPECIMEN COLLECTION
• For routine urine analysis, first
morning sample is best since it is
most concentrated . For
bacteriologic examination, mid-
stream sample is preferable.
• For 24 hours sample , collection
of urine is started in the morning at
8 AM till 8 AM on the next day

5. PHYSICAL EXAMINITION
• The following parameters are examined during
physical examination of urine.
• Volume
• Normal Volume of urine passed in adult is 600-
2000 ml in 24 hours and most of it is passed
during day time.

6. Polyuria :
• When excess of urine is passed in 24 hours more
than 2000 ml with low specific gravity. due to
excess water intake, may be pathological (e.g.in
diabetes insipidus, diabetes mellitus) or
medication like diuretics.
• Oliguria: When less than 400 ml of urine is
passed in 24 hours, it is termed as oliguria. It
can be due to less intake of water , dehydration,
renal ischemia.

7. Anuria :
• Means urinary output <100 ml/24 hours or
when there is almost complete suppression of
urine .It occurs in acute tubular necrosis (e.g. in
shock,hemolytic transfusion reaction) ,complete
urinary tract obstruction.

8. Colour
• Normally, urine is clear,pale or straw-coloured due to
presence of pigment urochrome.
• Colourless in diabetes mellitus, diabetes insipidus,
excess intake of water.
• Deep yellow- Jaundice.
• Red/ Pink-Due to presence of blood as in
hemoglobinuria, hematuria.
• Smoky or smoky brown-Blood along with protein.
• Milky- Due to presence of chyle, fat, pus.
• Green- Phenol poisioning.
• Turbid- Due to large number of pus cells or large
number of amorphous phosphate crystals.

10. Odour
• Normally urine has faint aromatic odour.
• Ammoniacal- If allowed to stand at room
temperature for some time.
• Putrid -Due to decomposition of protein in
cases of urinary tract infection
• Fruity -Due to ketone bodies.
• Mousy -Due to phenylketonuria

11. Reaction /pH
• Normal PH of Freshly voided urine
ranges from 4.6-8.0
• Methods for PH determination
by: Litmus paper test, PH meter,
Reagent strip test.
• Acidic urine is due to:
• High protein intake ( eg. Meat),UTI
• Respiratory and metabolic acidosis,
• Alkaline urine is due to:
• Chronic renal failure, Renal tubular
acidosis, Vegetables , Respiratory
and metabolic alkalosis ,UTI by
Proteus and Pseudomonas.

12. Specific Gravity
• This is the ratio of weight of 1ml volume of urine to
that of weight of 1 ml of distilled water. It depends
upon the concentration of various particles / solutes
in the urine.
• Urinometer
• Procedure: Fill urinometer container 3/4 th with
urine..Read the graduation on the arm of urinometer
at lower urinary meniscus. Add or substract 0.001
from final reading for each 3° C above or below the
calibration temperature respectively marked on the
urinometer.
• Reagent Strip Method
• Significance of specific Gravity: The normal is
1.003 to 1.030
• Low specific gravity of urine occurs in: Excess
water intake, Diabetes insipidus
• High specific gravity : Diabetes mellitus
(glycosuria), Nephrotic Syndrome
• Low and Fixed specific gravity (1.010) of urine
is seen in: Chronic renal failure

13. CHEMICAL EXAMINATION OF URINE
• Chemical constituents frequently tested
in urine are:
• Protein,
• glucose,
• ketone bodies,
• bile derivatives
• blood.

14. Tests For Protein In Urine:
• Normally Kidney excrete scant amount of
protein in urine( up to 150 mg/24 hours).
• Proteinuria refers to protein excretion in urine
greater than 150 mg/ 24 hours.
• If urine is not clear, it should be filtered or
centrifuged before testing. Urine may be tested
for proteinuria by qualitative tests and
quantitative method.

15. Test for protein estimation
• Qualitative Tests for Proteinuria
• Heat and acetic acid test
• Sulphosalicylic acid test
• Heller’s test
• Reagent strip method

16. HEAT AND ACETIC ACID TEST
• Aim: To detect presence of protein in given urine sample.
• Principle: Heat causes coagulation of proteins, and that can be
seen as change in turbidity, Cloudiness or flocculation in urine.
• Material and Method: Urine Sample, dropper, Test tube , glacial
acetic acid.
• Procedure:
• I. Take a 15 ml test tube. Fill 2/3 rd test tube with urine sample.
• Boil upper portion of test tube for 2 minutes lower part of test tube
acts as control . If precipitation or turbidity appears, add a few drop
of 3% acetic acid. If turbidity persists it shows the presence of
protein. If turbidity disappears it may be due to presence of
phosphate crystal in urine.
• Result/Interpretation: If turbidity or precipitation disappeares
on addition of acetic acid , it is due to phosphates; if persists after
addition of acetic acid , then it is due to proteins.
• Precaution:
• 1. Avoid heating of entire test tube as lower portion of tube acts as a
control

17. SULPHOSALICYLIC ACID TEST
• This is a very reliable test. Specimen should be
centrifuged and a clear supernatant used. Add 3
ml of 3% sulphosalicylic acid in 3 ml of
supernatant urine. Invert tube to mix properly.
Let stand exactly for 10 minutes then invert
twice again. Observe the turbidity and/or
precipitation and graded as +/++/+++/++++.
• Quantitative Estimation of Proteins in
urine:
• Diagnosis of Nephrotic syndrome
• Detection of microalbuminuria
• There are two methods :
• 1. Esbach’s albuminometer method
• 2. Turbidimetric Method
•

18. Esbach’s albuminometer method
• .
• I. Fill the albuminometer with
urine up to mark U.
• ii. Add Esbach’s reagent (picric
acid+ citric acid ) up to mark R.
• iii.Stopper the tube, mix it and
let it stand for 24 hours.
• iv. Take the reading from the
level of precipitation in the
albuminometer tube and divide
it by 10 to get the percentage of
protein (albumin)

19. Causes of proteinuria:
• Heavy Proteinuria (>4 gm/day)
▫ Nephrotic Syndrome.Diabetes mellitus. SLE
• Moderate Proteinuria (1-4 gm/day)
▫ Nephrosclerosis ,Multiple Myeloma(Bence Jones)
• Minimal Proteinuria (<1 gm/day)
▫ Polycystic kidney, Chronic pyelonephritis ,UTI
• Microalbuminuria: is excretion of albumin
30-300 mg/day. It is considered as the earliest
sign of renal damage in diabetes mellitus

20. Bence jones proteinuria:
• Bence jones protein are Monoclonal
immunoglobulin light chains. Excess production
occurs in plasma cell dyscrasias like multiple
myeloma. Because of their low molecular weight
they are excreted in urine. When heated Bence
Jones proteins precipitate at temperatures between
40ᵒC to 60ᵒ C and precipitate disappears on boiling
at 85ᵒC -100ᵒC . When cooled to around 60ᵒC Bence
Jones protein reappeares. Electrophoresis and
Immunofixation electrophoresis (IFE) methods are
the best detection and quantification methods.

21. TESTS FOR SUGAR IN URINE
• Normally a very small amount of glucose is
excreted in urine (< 500 mg/24 hours) that
cannot be detected by the routine tests. Presence
of detectable amount of glucose in urine is called
as glucosuria/glycosuria.
• Tests for glucosuria may be qualitative or
quantitative.
• Qualitative Tests
• These are as under:
• Benedict’s test 2. Reagent strip

22. BENEDICT’S TEST:
• Principle: In this test cupric ion is reduced by
glucose to cuprous oxide and a coloured
precipitate is formed.
• Material and Methods: Urine sample,
Benedict’s reagent, Dropper, spirit lamp,
• Procedure :
• i.Take 5 ml of benedict’s reagent in a test tube ,
Add 8 drops (or 0.5 ml) of urine sample. Heat to
boiling for 2 minutes.look for colour change and
precipitation.
• Precaution:
• Benedicts test is positive for all reducing sugars
(glucose,fructose,maltose,lactose but not for
sucrose which is a non reducing sugar) and other
reducing substances ( ascorbic acid, salicylates)

23. Result/Interpretation:
Appearance Grading Interpretation
No change / blue colour _ Negative
Greenish colour Trace <0.5g/dl
Green with ppt + 0.5-1 g/dl
Yellow ppt ++ 1.0-1.5 g/dl
Orange ppt +++ 1.5-2.0 g/dl
Brick red ppt ++++ >2.0g/dl

24. REAGENT STRIP TEST:
• These strips are coated with glucose oxidase and the
test is based on enzymatic reaction.
• This test is specific for glucose.
• Causes of Glucosuria
• Glucosuria with hyperglycemia
• Diabetes mellitus.
• Hyperthyroidism, Cushing’s syndromely
• Administration of corticosteroids.
• Pregnancy induced diabetes.
• Alimentary glucosuria. (Transient)
•
• Glucosuria without hyperglycemia
• Renal Glucosuria

25. TEST FOR KETONE BODIES IN URINE
• Excretion of ketone bodies (acetoacetic acid,β
hydroxybutyric acid and acetone) in urine is
called as ketonuria.
• Following are the tests for ketone bodies:
• i.Rothera’s Test
• ii. Gerhardt’s Test
• iii. Reagent strip

26. ROTHERA’S TEST:
• Aim: To detect the presence of ketone bodies in given urine
sample.
• Principle: Ketone bodies ( acetone and acetoacetic acid β
hydroxy butyric acid) when combine with alkaline solution
of sodium nitroprusside forming purple complex at the
junction of solution.
• Material and Methods:, Ammonium Sulphate, Sodium
Nitroprusside crystal, Liquor ammonia.
• Procedure:
• i.Take 5 ml of urine in a test tube ,.Saturate it with solid
ammonium sulphate salt.iii. Till no more salt can be
dissolved in sample.
• iv. Add a few crystals of sodium nitroprusside and shake.
• v. Add liquor ammonia from the side of test tube.
• Result:
• Formation of purple coloured ring at the junction

27. INTERPRETATION:
Formation of purple coloured ring at the junction
indicates presence of ketone bodies.

28. TEST FOR BILE SALT IN URINE
• Bile salts are salts of four different types of bile acids: cholic ,
deoxycholic, and chenodeoxycholic , lithocholic. These bile
acids combine with glycine or taurine to form complex salts.
Bile salts appear in urine in liver diseases only.
• HAY’S TEST:
• Aim: To detect the presence of bile salts in given urine sample.
• Principle : Presence of Bile salts lowers the surface tension of urine
hence the sulphur Powder sprinkled over urine will sink to the bottom
• Material and Method:
• i.Fill a 50 or 100 ml beaker with 2/3rd to 3/4 th with urine.
• ii. Sprinkle finely powdered sulphur powder over it.
• Result: Sulphur powder will sink to the bottom.
• Interpretation: If bile salts are present in the urine then sulphur powder
sink, otherwise it floats.

29. TESTS FOR BILE PIGMENT IN URINE
• Bilirubin ( a breakdown product of
haemoglobin) Bile pigment appear in
urine in liver diseases. Presence of
bilirubin in urine is called as bilirubinuria.
• Test method for bilirubin are :
• Fouchet’s test, Foam test, Gmelin’s test,
Lugol iodine test, Reagent strips test

30. FOUCHET’S TEST:•
• Aim: To detect the presence of bile pigment
• Principle: Ferric chloride present in fouchet’s reagent oxidizes bilirubin to
green biliverdin compound.
• Material and Method: Urine sample, Test tube , 10% Barium chloride
solution, Filter paper, Fouchet’s reagent.
• Procedure:
• i.Take 10 ml of urine in a test tube.
• ii.Add 5 ml of 10% barium chloride and mix well .
• iii. Filter through filter paper.
• iv.To the precipitate on filter paper, add a few drops of Fouchet’s reagent (
10 ml of 10%ferric chloride+ 25 gm trichloroacetic acid+distilled water
100ml).
• Result: Blue-Green colour develops around the drop.
• Interpretation:
• Development of blue-green colour indicates presence of bile pigment
(bilirubin) in urine sample.
• Presence of bilirubin indicates conjugated hyperbilirubinemia( obstructive
or hepatocellular jaundice).This is because only conjugated bilirubin is
water soluble. Bilirubin in urine is absent in haemolytic jaundice.This is
because unconjugated bilirubin is water insoluble.

31. TEST FOR UROBILINOGEN IN URINE
• Normally small amount( 0.5-4 mg/24 hours)
• EHRLICH’S TEST:Principle: Urobilinogen in urine combines with
Ehrlich’s aldehyde reagent to give a red purple coloured compound.
• Material and methods: Urine sample , Test tube , Ehrlich’s aldehyde
reagent, filter paper.
• Procedure: Take 5ml of fresh urine in a test tube. Add 0.5ml of Ehrlich
aldehyde reagent. Allow to stand at room temperature for 5 minutes
development of pink colour indicates normal amount of urobilinogen. Dark
red purple colour means increased amount of urobilinogen.
• Presence of bilirubin interferes with the reaction, and therefore if present
should be removed. For this equal volume of urine and 10% barium
chloride are mixed and then filtered. Test for urobilinogen is carried out on
the filterate.
• Result: red purple colour/ Rose colour develops.
• Interpretation : Development of red purple colour indicates presence of
urobilinogen.
• Ehrlich’s aldehyde gives positive result with both urobilinogen and
porphobilinogen, this can be differentiated by adding sodium acetate.
• Normally about small amount of urobilinogen is excreted in urine in 24
hours. Urobilinogen is increased in the case of haemolytic jaundice. It is
absent from urine in obstructive jaundice.

32. TEST FOR BLOOD IN URINE
• The presence of abnormal number of intact red blood cells in urine is called
as hematuria.
• Test for detection of blood in urine are as under:
• Chemical test: These detect both intracellular and extracellular
haemoglobin(intact and lysed RBCs) as well as myoglobin.
• 1. Benzidine Test , 2. OrthotoluidineTest
• 1. Benzidine Test:
• Aim: To detect the occult blood in given urine sample.
• Principle: peroxidase activity of haemoglobin decomposes hydrogen
peroxide releasing nascent oxygen which in turn oxidizes benzidine to give
blue colour .
• Procedure:.Take 2 ml of urine in a test tube. Add 2ml of saturated
solution of benizidine with glacial acetic acid. Add 1 ml of hydrogen
peroxide to it.
• Interpretation: Appearance of blue colour indicates presence of blood.
• Precaution:
• i.Avoid touching of Benzidine powder with finger as it’s a potent
carcinogenic agent.
• ii. Use freshly prepared H2 O2 solution.