EXPERIMENT 13: URINE

I. Objectives

At the end of the experiment, the students should be able to:

a. explain the pathological constituents of urine;and

b. define the test that will detect the presence of different substances present in urine.

II. Materials

BEAKER GRADUATED CYLINDER TEST TUBES

BUNSEN BURNER WATER BATH DROPPER

URONIMETER
III. REAGENTS

Chemical Chemical Molecular Melting Boiling point Hazards

name formula weight point
Benzidine  
C12H12N2 184.24 120 deg cel 401 deg cel Cause eye
g/mol and skin
irritations
Hydrogen Solution N/A 108 degl cel -33 deg cel Corrosive,
Peroxide may cause
fire and
burns
Nitric acid HNO3 63.01 g/mol -42 deg cel 83 deg cel Sever skin
and burns
rapidly
Acetic acid C2H4O2 60.06 g/nol 17 deg cel 118 deg cel Skin
irrritations
Sodium C5H4FeN6Na2O3 297.95 N/A N/A Respiratory
nitroprusside g/mol irritations
Fehlings CuH2O4S 161.63 590 deg cel 650 deg cel Eye unjury
solution g/mol and burns
Ammonium NH4OH 35.04 g/mol -91.5 deg cel 24.7 deg cel Skin burn
hydroxide

IV. Procedures

I. Qualitative Tests for Constituents of Urine II. Pathological Urine

A. Physical Properties
1.Collect and measure total volume of urine
sample. Note the color.
2.Determine the transparency or cloudiness of
the sample.
3.Determine the specific gravity by using
urinometer.
4.pH –determine the approximate pH by dipping
a strip of litmus paper into the urine.
B. Chemical test for normal constituents of urine
shows:
a.Anions like chlorides, sulfates, and
phosphatesb.Cations like sodium, potassium,
ammonium, calcium, and magnesium
A. Sugar
1.Place 10 drops each of Fehling’s A and B in a
test tube then add 10 drops of urine sample.
2.Heat in boiling water for two minutes.
3.Observea color change.
B. Blood
1.To 1 mL of freshly prepared benzidine solution,
add 10 drops of urine sample.
2.Add 6 drops of 3% hydrogen peroxide. Take
note of the result.
 C. Chemical test for pathologic or abnormal
constituent of urine
The following constituents, protein, sugar, ketone
bodies, and bile pigments, are found in normal
urine in small quantities, too small to be detected
by ordinary laboratory procedures. They become
pathologic when excreted in abnormal
amounts. Blood, however, when present in
urine (except menstrual blood) is always
pathologically significant.
ORANGE – FIRST
RED - SECOND
C. Protein
1.Heat and Acetic Acid Test
a.Fill a test tube about ¾full of urine.
b.Heat the upper portion gently to boiling for 1-2
minutes being careful not to shake the tube.
c.Rotate the tube to prevent cracking.
Turbiditymay be due to albumin, phosphates or
carbonates.
d.Add 3 drops of acetic acid drop bydrop, boiling
between each drop. If turbidity disappears,it is
due to carbonates and phosphates. Afaint trace
of albumin may appear upon boiling and
increase upon addition of acetic acid.
2.Heller’s Nitric Acid Test
a.Place 1 mL urine sample in a test tube.
b.Hold the test tube in an inclined position and
carefully pour 10 drops of conc. Nitric Acid down
the side of the test tube.
c.Note the reaction at the zone of contact
between the solutions.
D. Ketone Bodies
1.To 1 mL urine sample, add a few dropsof freshly
prepared sodiumnitroprusside. Shake.
2.Add 1 mL of conc. ammonium
hydroxidecarefully down the side of the tube to
form a layer over urine.
3.Observe and note the color reaction.