ABNORMAL CONSTITUENTS

OF URINE
 RULED

day
1. Test for Reducing Sugar: (Glucose)
(Ruled)
Test Observation Inference

Benedict’s test : Take 5ml Green or Reducing sugar

Benedict’s reagent in test yellowish confirmed Therefore
tube then add 8 drops of green or urine contains
urine sample and Boil it for reducing sugar
1-2 min and observe. orange or red
precipitate
obtained
• Benedict's solution: (a) Dissolve 1 gm of CuSO4.5H2O in 100 ml water.
Cool. (b) Dissolve 173 gm sodium citrate and 100gm anhydrous
sodium carbonate in 600ml water by heating. Cool and dilute to 850
ml. pour(a) into(b), stirring well and dilute at R. temp, to 1000ml.
• PRINICIPLE : In alkaline medium carbohydrates undergo
tautomerisation to form enediols which cause reduction of Cu++ to
Cu+ which form coloured complex.
• This test is done to know reducing property of sugars which is due to
presence of free aldehydes or keto group.
•Clinical Interpretation:
• Presence of glucose in urine is called
as glycosuria. Colour of the precipitate
indicates severity of glycosuria as
follows:
• Presence of glucose in urine is found
in,
• a. Diabetes mellitus.
• b. Hyperadrenalism.
• c. Renal glycosuria
2.KETONE BODIES
• The ketone bodies sometimes found in urine are: Acetoacetic acid,
beta hydroxy butyric acid and acetone. Their presence indicates
excessive metabolism of body fats. Which may occur in diabetic
acidosis, malnutrition or after anaesthesia and toxaemia of
pregnancy.
 (Ruled)
2.Test for ketone bodies
Test Observation Inference

ROTHERAS TEST: Saturate 5ml of Permanganate colour or Acetone and acetoacetic acid
urine by shaking it vigorously with an deep purple ring is present. Therefore urine contains
observed at the junction of ketone bodies.
excess of solid ammonium sulphate. two layers
Add (2) drops of freshly prepared
(2%) sodium nitroprusside and 1 mL
of NH3 FROM THE SIDES OF TEST
TUBE. allow to stand in the rack. A
permanganate colour indicates the
presence of acetone bodies.
• TEST FOR Ketone bodies :-
• Principle: Acetone in urine after saturation with ammonium sulphate
crystals dissociate into acetate ions which combines with sodium
nitroprusside in alkaline medium to from permanganate colour
sodium nitroprusside acetate ion complex.
• Clinical Interpretation
• Acetoacetic acid, β -hydroxy butyric acid & acetone are collectively
known as ketone bodies. Normal urine of 24hrs may contain ketone
bodies up to 1 mg. Excess of ketone bodies in blood is called as
ketonemia and higher excretion of ketone bodies in urine is known as
ketonuria and overall condition is known as ketosis. Ketone bodies are
observed in fasting, untreated diabetes mellitus, high fat, low
carbohydrate diet, prolonged starvation, pregnancy & lactation.
3.PROTEINS
• The excretion of plasma protein in urine is called "proteinuria".
Albumin and amounts of globulin are abnormal found in urine due to
renal damage. Physiological proteinuria occurs in heavy exercises, and
rich protein diet. Another kind of protein called Bence-Jones protein,
may be present in urine, but it is not a plasma protein. It originates in
the bone marrow and its presence in urine associated with a disease
of the bone marrow.
 (Ruled)
3.Test for proteins
Test Observation Inference

White Coagulum or Albumin is confirmed. Therefore

Heat coagulation turbidity is obtained which urine contains albumin
after the addition of acetic
test: Fill 3/4th of test acid gets intensified.
tube with urine sample
solution. Heat it at the
top and observe. Then
Add 1-2 drop of acetic
acid to it and again
observe.
.
Principle: Heating proteins causes them to denature, which changes
their structure. This process is irreversible and results in the formation
of an insoluble coagulum
Proteins which are insoluble in water or dilute salt solutions are known
as Fibrous proteins, while soluble proteins are collectively described as
globular proteins. Many globular proteins coagulate when their
solutions are heated and become permanently insoluble. Such proteins
are described as heat- globular proteins. Not all proteins are equally
denatured by heat, this fact can be used to distinguished between
plasma proteins and Bence- Jones protein by the heat-test.
• Clinical Interpretation
• Presence of detectable amount of protein in urine is known as
proteinuria. It is of two types, Physiological proteinuria : Less than 0.5
gm%. It occurs due to sever exercise, pregnancy, high protein diet etc.
• Pathological proteinuria: It occurs due to kidney damage
(Glomerulonephritis and Nephrotic syndrome). In some patients
suffering from multiple disease such as leukaemia lymphosarcoma.
Urine & blood contain specific type of protein called Bence Jones
protein. They are globulin & posses peculate solubility property i.e.
they coagulated at 40-60 0C & go in to the solution at 80 0C and
reappear after cooling
4.Bile salts
• Bile is a solution consists of water, bile salts, cholesterol, bile
pigments, lipids and electrolytes. Certain bile composition may
appear in excess in the urine of patient with:
• 1. Bile duct obstruction.
• 2. Liver disease.
• Bile salts test : Bile salts are formed in the liver and defined as
dehydrogenated bile acids where Na+ or K+ replaced by the lost
hydrogen atom.
 (Ruled)
4.Test for bile salts
Test Observation Inference

Hay’s Sulphur Sulphur powder Bile salts confirmed.

powder test: Take 3ml urine in a sinks down to Therefore urine contains
test tube and sprinkle bottom of the test bile salts.
a pinch of sulphur powder and tube.
observe.
Control:
Take 3ml water in a test tube and Sulphur powder f
sprinkle a pinch of sulphur floats.
powder and
observe
• Hay’s Sulphur powder test for bile salts
• Principle: Sulphur powder sinks down in test tube containing urine
sample because bile present in urine contain bile salts which acts as
emulsifying agent reduces the surface tension of urine sample.
 (Ruled)
5.Test for bile pigments
Test Observation Inference

Fouchets test Colour changes from Bile pigments confirmed.

add 3 mL of clear urine add 1 yellow to pista green Therefore urine contains
ml of BaCl2 and 1 ml MgSO4 bile pigments.
a white precipitate of barium
sulphate is formed. Mix well
and allow to stand till most of
the ppt. is settled down for 1-2
min Filter it and to the
precipitate on the filter paper
add few drops of Fouchet’s
reagent and observe.
• Principle: Bacl2 reacts with MgSo4 to for Baso4. Bile pigments are
absorbed on a precipitate of barium sulphate. The precipitate,
coloured with the type of the pigments, is filtered and then treated
on a filter paper with Fouchet's reagent (10%FeCl3 in a solution of
25%trichloroacentic acid (TCA). The bilirubin, which is yellow, is
oxidized to biliverdin (green) and cholecyanin (blue).
• Clinical Interpretation:
Bile salts are Na+ or K+ salts of glycocholates & taurocholates.
• These are found in urine in obstructive jaundice & amount of bile salt
is proportional to the degree of obstruction. These are found to some
extent in infective hepatitis due to partial obstruction.
• In haemolytic jaundice bile pigments are found in urine without bile
salts.