LABORATORY EXERCISE NO.

ROUTINE URINALYSIS

Urinalysis or the examination of the urine is probably the oldest clinical

laboratory practice. A commonly performed laboratory procedure both in the clinical
laboratory as well as the physician’s office.

It is an essential procedure for hospital admissions and physical examinations. It

is one of the most useful indicators of health and disease, and it is especially helpful
in the detection of renal or metabolic disorders.

The routine urinalysis procedure is comprised of the following components:

A. Specimen Evaluation

B. Physical Examination
1. Color
2. Appearance/Transparency
3. Reaction
4. Specific Gravity

C. Chemical Examination
1. Protein (Qualitative)
2. Reducing Substances
3. Glucose

D. Microscopic Examination of the Urine Sediment

 EXERCISE NO. I-A

PHYSICAL EXAMINATION OF THE URINE

The physical examination of the urine involves the determination of the various
properties of urine: color, odor, appearance, reaction and specific gravity. Results in
the physical examination provides preliminary information concerning renal diseases
and other metabolic or endocrine disorders. Furthermore, information gathered in this
examination may provide explanation or confirm significant findings in the chemical
microscopic examination of the urine.

INSTRUCTIONS:

1. Individually perform the physical examination of your own urine sample.

2. Agitate the urine container to mix well the sample. Examine and describe the urine
with regards to its color, appearance or transparency.

3. Record the data in the table provided.

A. APPEARANCE

1. Color
Record as straw, pale straw, yellow, amber, red, pink, black, etc….

The amber yellow color of urine is due largely to the pigment urochrome and
to small amounts of urobilin and uroerythrin.

The intensity of the normal amber color may be related directly to the
concentration or specific gravity of the urine.

Due to the presence of abnormal pigments, the color of urine changes in

many disease states.

APPEARANCE AND COLOR OF URINE

APPEARANCE CAUSE REMARKS

COLORLESS Very diluted urine Polyuria, diabetes insipidus
CLOUDY Phosphates, carbonates Soluble in dilute acetic acid
Urates, uric acid Dissolve at 60oC and in
Alkali
Leukocytes Insoluble in dilute acetic
acid
Red Cells (“smoky”) Lyse in dilute acetic acid
Bacteria, yeasts Insoluble in dilute acetic
acid
Spermatozoa Insoluble in dilute acetic
acid
Prostatic fluid
APPEARANCE CAUSE REMARKS
Mucin, mucous threads May be flocculent
Calculi, “gravel” Phosphates, oxulates
Clumps, Pus, Tissue
Fecal contamination Rectovaginal fistula
Radiographic dye In acid urine
MILKY Many neutrophils (Pyuria) Insoluble in dilute acetic
acid
Fat
Lipiduria, opalescent Nephrosis, crush injury -
soluble in ether
Chyluria, Milky Lymphatic obstruction -
soluble in ether
Emulsified paraffin Vaginal creams
YELLOW Acriflavin Green fluorescence
YELLOW-ORANGE Concentrated urine Dehydration, fever
Urobilin in excess No yellow foam
Bilirubin Yellow foam in sufficient
YELLOW-GREEN Bilirubin-biliverdin Yellow foam
YELLOW-BROWN Bilirubin-biliverdin “Beer” brown, yellow foam
RED Hemoglobin Positive - reagent strip for
blood
Erythrocytes Positive - reagent strip for
blood
Myoglobin Positive - reagent strip for
blood
Porphyrin May be colorless
Fuscin, aniline dye Foods, candy
Beets Yellow alkaline genetic
Menstrual contamination Clots, mucus
RED-PURPLE Porphyrins May be colorless
RED-BROWN Erythrocytes
Hemoglobin on standing
Methemoglobin Acid pH
Myoglobin Muscle injury
Bilifuscin (dipyrrole) Result of unstable
hemoglobin
BROWN-BLACK Methemoglobin Blood, acid pH
Homogentisic acid On standing, alkaline,
alkaptonuria
On standing, rare
BLUE-GREEN Indicans Small intestine, infections
Pseudomonas infections
Chlorophyll Mouth deodorants
2. Turbidity

Fresh urine is clear to slightly hazy. The appearance of cloudy urine provides
a warning of possible abnormality such as the presence of pus, red blood cells, or
bacteria. However, excretion of cloudy urine may not be abnormal since the
change in urine pH may cause precipitation within the bladder of normal urinary
constituents. Alkaline urine may appear cloudy because of the presence of
phosphates and acid urine may appear cloudy because of urates.

PROCEDURE:

Observe the appearance of a fresh urine sample.

Interfering Factors:
1. After ingestion of food, urates or phosphates may produce cloudiness
in normal urine
2. Vaginal contamination from female patients is a common cause of
turbidity.
3. “Greasy” cloudiness may be caused by large amounts of fat.
4. Many normal urines will develop haze or turbidity after refrigeration
or standing at room temperature.

B. REACTION (pH)

The pH of urine is a reflection of the ability of the kidney to maintain normal

hydrogen ion concentration in plasma and extracellular fluid.

Methods for measuring urine pH:

a. ) Potentiometric Method - accurate method but not practical for routine

screening.
b. Indicator paper strips:
1. Litmus paper - much too crude for laboratory use
2. Reagent strip - currently used in most laboratories; fast, inexpensive

Procedure
Litmus paper:
1. Drop a small square of red litmus paper into the sample.
2. Note for any change in color.
3. Drop a small square of blue litmus paper into the sample.
4. Note for any change in color.

Reagent strip:
1. Dip reagent strip onto sample.
2. Remove excess urine by running the edge of the strip against the rim
of the container. Do not blot.
3. Observe the pH reagent pad and compare color to the strip container
to read result.
C. SPECIFIC GRAVITY

One of the most important functions of the kidney (physiologic) is the

maintenance of fluid and osmolar balance. These are determined by the specific
gravity, osmolality and refractive index. These terms are related to each other
since they reflect the quantity of dissolved solids in the urine. However, these
terms are not identical.

METHODS FOR MEASURING SPECIFIC GRAVITY OF URINE:

1. In the measurement of specific gravity, the hydrometer (urinometer) is usually used.
The urinometer is a hydrometer adapted to measure the specific gravity of urine at
room temperature (variable). It should be checked prior to use by measuring the
specific gravity of distilled water, which has a specific gravity of 1.000. If the
urinometer does not give a reading of 1.000, an appropriate correction must be
applied to all readings taken with that urinometer (I.e., if it reads 1.001 subtract 0.001
from all subsequent readings at that temperature.)

Because temperature influence the specific gravity, urine should be allowed to come
to room temperature or a correction of 0.001 should be made for 3oC or 5oF, the
specimen is above the calibration temperature.

Example: A sample of urine was received in the laboratory. Its specific gravity was
found to be 1.018 at a temperature of 34oC. The urinometer used was calibrated at
77oF. What was the true specific gravity?

The temperature of calibration must be expressed in degrees centrigrade. To convert

from Fahrenheit to Centrigrade, the following formula is used:

5/9(F-32) = oC
5/9(77-32) = 25oC

The difference between the temperature of reading and the temperature of calibration
divided by 3 and multiplied by 0.001 gives the figure to be added to the specific
gravity reading to obtain the true specific gravity.

34 - 25
--------------- x 0.001 = 0.003
3

1.018 + 0.003 =1.021 - true specific gravity of the specimen

PROCEDURE:
1. Fill the cylinder 3/4 full of urine.
2. Place the urinometer in the cylinder and spin.
3. Read the bottom of the meniscus when the urinometer has stopped spinning and
does not adhere to the side of the cylinder.
4. The specific gravity is always reported in 4 figures (e.g. 1.025)
5. Compute for the true specific gravity and record data.
2. Another method for measuring specific gravity specially for small volumes and for
routine purposes uses the modified Goldberg refractometer (Model 10400, American
Optical Co.)

The refractometer is a popular instrument for estimating urine specific gravity. Only a
few drops of urine are required and the instrument is temperature compensated. The
method depends on the relationship between the REFRACTIVE INDEX of a solution
and its content of dissolved solids.

The Goldberg refractometer is calibrated to read directly the specific gravity of urine.
The instrument is temperature compensated between 60oF and 100oF. It is damaged
by heat above 150oF and by immersion of eyepiece and focusing ring in water. It
should read 1.000 with distilled water.

PROCEDURE:
1. Clean the surfaces of the cover and prism with a damp cloth and then dry.
2. Close the cover.
3. Apply a drop of urine at the notched bottom of the cover so that it flows over the
prism surface by the capillary action.
4. Point the instrument towards a light source at an angle that gives optimum contrast.
5. Rotate the eyepiece until the scale is in focus.
6. Read directly on the specific gravity sclae (left side), the sharp dividing line
between light and dark contrast.
7. Repeat the entire procedure with a second drop of urine form the same sample.