Experiment 8: PHYSICAL PROPERTIES & CONSTITUENTS

OF URINE

OBJECTIVES

 To become familiar with the routine urinalysis

 To examine urine for the presence of normal and abnormal constituents
 To acquire an appreciation for the value of urine analysis as a diagnostic tool

PROCEDURE

A. Physical properties of urine

Color, Odor, and transparency

1. Examine the urine specimen for color, odor, and transparency

2. Record your observation

Specific gravity

1. Weigh a 100-mL graduated cylinder

2. Fill the graduated cylinder with 50mL of urine sample and weigh.

3. Get the weight of the urine by subtracting the weight of the empty graduated cylinder from the
weight of the graduated cylinder with the urine sample

4. Divide the weight of urine by volume. The value obtained will be your urine density. Assuming
that the density of water is 1 g/mL, the density of the urine is equal to its specific gravity.

pH

1. Using a pH paper or a pH meter, determine the pH of the urine

B. Inorganic constituents of urine

Chlorides

1. Place 5L of urine sample in a test tube and acidify with 2 drops of HNO 3 to prevent
precipitation of phosphorous.
2. Add 3 drops of 12% silver nitrate solution

3. Observe what is formed.

Phosphates

1. Place 5mL of urine in a test tube and add NH 4OH solution until the urine becomes alkaline to
precipitate the phosphate.

2. Record your observation

Sulfates

1. Pour 5mL of urine in a test tube and acidify with a drop of HCl

2. Add 3 drops of BaCl2 solution to precipitate the sulfates

3. Record your observation

C. Organic constituents

Urea

1. Mix 15mL and 5mL of Baryta mixture (1 volume of saturated solution of barium nitrate and 2
volumes of saturated solution of barium hydroxide)

2. Filter off the precipitate and evaporate the filtrate to thick syrup

3. Add an equal volume of 95% ethyl alcohol and filter

4. Allow the filtrate to crystallize in a cool place until next laboratory period.

Creatinine

1. To make 5mL of urine add 5 drops of nitroprusside

2. Make this solution alkaline with dilute NaOH

3. Record your observation

Pigments

1. Place 2mL of urine in a test tube and add 1mL of dilute ammonium hydroxide.

2. Let stand for a while and then filter

3. To the filtrate, add 2mL of zinc chloride solution

4. Observe the presence of a greenish fluorescence which indicates the presence of urobilin
D. Pathological constituents

Preparation of pathologic urine

1. Add a pinch of albumin, 10 drops of acetone; add 15 drops of 10% glucose solution to 15mL
of urine sample. This is your pathologic sample.

Helloer ring test for albumin

1. Place 3mL of normal urine and the prepared pathologic urine on separate test tubes

2. Slant each tube and carefully add 2mL of concentrated nitric acid down sides of each tube.
DO NOT MIX THE CONTENTS OF EACH TUBE.

3. Observe the formation of a white ring at the juncture of the lipids each sample

Lange’s test for acetone

1. In separate test tubes, place 5mL of normal urine and pathologic urine.

2. Add 5 drops of glacial acetic acid and 4 drops of sodium nitroprusside and mix.

3. Slant each test tubes and with a medicine dropper, carefully add about 20 drops of 28% of
NH4OH to cover the surface of the sample

4. Observe the formation of purple or purplish ring at the juncture of the liquid.

Benedict’s test

1. Fill two test tubes to about 1/3 with Benedict’s solution and add 8 drops of normal urine to
one tube, and 8 drops of pathologic urine to the other tube.

2. Place the tubes in boiling water bath for 5 minutes.

3. Observe the formation of brick red precipitate in the urine sample.

Experiment 8: PHYSICAL PROPERTIES & CONSTITUENTS
OF URINE

Score:

Last name Given name MI

Course/Year Date performed Date

submitted

Laboratory class schedule Laboratory Instructor/Professor

POST-LAB ACTIVITY
RESULTS

Table 1. Physical properties of urine sample [2 pts]

Properties Observation

Color

Odor

Transparency

Specific gravity

pH
Exploratory questions:

1. What is responsible for the color of urine? How do these substances formed? [3 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

2. What is the significance of knowing the specific gravity? What is the implication of the specific
gravity is high or low?

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

3. What is the pH of normal urine? What is/are the reason for the observed pH of urine? [3 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

Table 2. Test for the presence of inorganic and organic constituents of urine. [6 pts]

Reaction w/ Observation Inference

AgNO3 solution

NH4OH solution

BaCl2 solution

Baryta mixture

Sodium nitroprusside

Zinc chloride solution

Exploratory questions:

4. What is the normal value of chloride? How does chloride become a component of urine?
What does a value above normal for chloride would indicate? [4 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

5. What is the normal value of phosphate? How does a phosphate become a component of
urine? Would an increase in the value of phosphate above normal indicate a pathologic
condition? Why or why not?

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

6. What is the normal value of sulphate? How does sulphate become a component of urine?
What are the causes of an increase in sulphate excretion? Explain. [4 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

7. What is the normal value of urea? How does urea become a component of urine? What
conditions are indicated by an increase of urea? Why? [4 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________

8. What is the normal value of creatinine? How does creatinine become a component of urine?
Why is the value of creatinine used to diagnose renal failure? [4 pts]
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________

9. What are the pigments usually in urine? How are these pigments formed? [3 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________

Table 3. Test for pathologic urine [6 pts]

Tests Observation Inference

Helloer Ring

Lange

Benedict’s

Exploratory questions:

10. Why is albumin present in pathological urine? What does the presence of albumin
indicates? Is an increase albumin excretion observed only in pathologic condition? Why or why
not?

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

11. Why is acetone present in pathological urine? What does the presence of acetone
indicates? Explain. [4 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
_________________________________________________________________________

12. Why is glucose present in pathological urine? What does the presence of glucose indicates?
Explain. [4 pts]

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
__________________________________________________________________________

REFERENCES:

____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
____________________________________________________________________________
________________________________________________________________________