Urinalysis

Lacaran, Jared Vincent T.*, & Matthew Eliseo A. Saligumba

a
Department of Chemistry, College of Science, Adamson University, 900 San Marcelino, Ermita, Manila
1000, Philippines

Abstract
Urine is a liquid waste product excreted from the urethra. Urinalysis is the chemical analysis of urine used
to determine the presence of diseases and infections in a person’s body. In this experiment, fresh urine
was obtained and analyzed for the presence of urea, uric acid, indicant, creatine, ketone bodies, glucose,
protein, bile pigments, and occult blood. The pH, turbidity, and color were also analyzed.

1.0 Background

Urine is a liquid waste product excreted from the body through the urethra. It is a solution that can

range in color from colorless to pale amber. Urine contains metabolic wastes such as urea, dissolved salts,

and organic compounds. Diuresis is the process of urine production. It is essential in maintaining the

body’s homeostasis. The compound urochrome is responsible for the yellowish color of urine. This color

is indicative of the urine concentration: dilute urine is pale yellow in color while concentrated urine is

deep amber.

Volatile acids are responsible for the distinctive odor of urine. Fresh urine has a balanced acid –

base composition. This is due to the kidney’s work in the maintenance of normal bodily pH in

maintaining homeostasis. The acid – base balance is maintained by the kidneys mainly through the

reabsorption of sodium, ammonium, and hydrogen ions from tubular secretions. The composition of urine

is indicative of kidney function and changes in blood. Urine analysis can yield important information

about the health of a person’s kidneys and body. Various diseases and infections causes abnormal

metabolic byproducts to appear in urine. The process of urine diagnosis and screening is known as

urinalysis.

The objective of the experiment is to subject a urine sample to several tests to qualitatively

examine the presence of some normal constituents and pathologic organic constituents.
2.0 Methodology

2.1 Materials, reagents, and instruments

The materials used were Pasteur pipet, watch glass, beaker, burner, dropper, buret, and test tube.

The reagents used were distilled water, freshly voided urine sample, 70% NaOH, bromine water,

phosphotungstic acid, Obermayer’s reagent, saturated picric acid – 10% NaOH, 20% Na 2CO3,

concentrated NH4OH, Lugol’s solution, Benedict’s reagent, Exton’s reagent, alcoholic iodine mixture,

guaiac powder, 95% ethanol, 3% H2O2, and HOAc,

The instrument used was analytical balance and hot plate.

2.2 Test for Urea

0.5 mL of 70% NaOH and 4 drops of bromine water were added to a 1 mL urine sample. The

evolution of N2 gas was observed.

2.3 Test for uric acid

5 mL of 20% NA2CO3 was added to 1 mL of urine sample and mixed. 5 mL of phosphotungstic

acid was added and mixed. The formation of a blue solution was observed.

2.4 Indican test

5 mL of Obermayer’s reagent was added to a 5 mL urine sample and mixed. 3 mL of chloroform

was added and shook to allow the chloroform layer to settle. The formation of a blue color in the layer

was observed.

2.5 Test for creatine

 1 mL of alkaline picrate solution was added to a 2 mL urine sample. The formation of an orange-

colored solution was observed.

2.6 Gunning’s test for ketone bodies

5 mL of urine sample was basified with 5 drops of concentrated ammonium hydroxide. Sufficient

Lugol’s solution was added to the basic urine to produce a black cloud which does not disappear

immediately. It was allowed to stand for 5 minutes. The formation of iodoform crystals was observed.

2.7 Benedict’s test for glucose

5 mL of Benedict’s reagent was added to 8-10 drops of urine sample in a test tube and mixed. It

was heated in a boiling water bath for 2 – 3 minutes and cooled. The formation of precipitate was

observed.

2.8 Exton’s test for albumin

3 mL of urine and 3 mL of Exton’s reagent were mixed in a test tube. Warm if there’s an

appearance of cloudiness. If cloudiness persisted or increased upon heating, albumin is present.

2.9 Heat and acetic acid test for protein

5 to 10 mL of clear urine was placed in a test tube. The sample was boiled in a water bath.

Development of turbidity was followed by addition of 1 – 2 drops of HOAc until the turbidity disappears.

Protein is present if turbidity disappears.

2.10 Smith’s test for bile pigments

5 mL of urine sample was placed in a test tube. The test tube was inclined and overlaid with 3 mL

of tincture of alcoholic iodine mixture. Formation of emerald green color at the point of contact is

observed.
2.11 Guaiacum test for occult blood

5 mL of 95% ethanol was added to a half spatula guaiac powder in a test tube and mixed. 5 mL of

hydrogen peroxide was added to this mixture. 5 mL of this solution was added to 3 mL of urine acidified

with acetic acid. Formation of blue ring is indicative of presence of blood.

2.12 Benzidine test for occult blood

A pinch of benzidine was placed in a test tube. 2-3 drops of 5% HOAc was added and mixed. 2

mL of hydrogen peroxide solution was added. The urine sample was transferred to a test tube. Formation

of a dark blue color is indicative of the presence of blood.

2.13 Kastle-Meyer test for hemoglobin

A cotton swab was moistened with water and slightly immersed in the urine sample. It was stood

for 3 minutes. 1-2 drops of 95% ethanol was added to the swab along with 1-2 drops of colorless

phenolphthalein solution, and hydrogen peroxide. Formation of pink color in the swab indicates presence

of blood.

3.0 Results and Discussion

3.1 Initial examination of urine

Sample Time Color Turbidity pH

Urine 2 pm Dark yellow Not turbid 6

The urine should be clear because cloudiness may be an indication of pus, bacteria, or red blood

cells. Abnormally colored urine may result from pathological conditions or medicine intake. The pH is

borderline neutral indicating that there is no acidemia or alkalemia.

3.2 Qualitative examination for normal organic constituents

 Qualitative test Observation Inference
Test for urea Bubbles formed Urea is present
Test for uric acid Solution turned blue Uric acid is present
Test for indican 2 layers formed, no blue layer Indican is not present
Test for creatine Solution turned dark orange Creatine is present

Urea is the end product of nitrogen metabolism in humans and is primarily excreted through the

kidneys in urine. Humans is the final product of purine metabolism and is present normally in urine.

Indicant is produced in the body by bacterial action on the amino acid tryptophan in the intestine.

However, it is normally excreted in the feces and is rarely present in urine. Low creatine levels in urine is

an indication of damaged kidneys.

3.3 Qualitative test for pathological organic constitutents

Qualitative test Observation Inference

Test for ketone bodies Crystal were formed at the bottom No ketone bodies
Test for glucose No precipitate formed No glucose
Test for protein No change in solution No proteins
Test for bile pigments 2 layers formed; no emerald green No bile pigments

color
Test for occult blood Solution became turbid No occult blood

Presence of ketones in urine is an indication of diabetes or altered carbohydrate metabolism. Glucose is

also commonly found in diabetic patients. There is normally no protein in urine and presence of it would

indicate kidney disease. Presence of bile and occult blood in urine can indicate hepatitis, cirrhosis, or

toxic liver damage.

4.0 Conclusion

It is concluded that the owner of the urine sample is relatively normal and can be assumed to be free

from disease though the slight acidity of his urine can indicate borderline acidosis.

5.0 References
 Nelson, D. L., & Cox, M. M. (2008). Lehninger Principles of Biochemistry (Fifth Edit). New

York: W. H. Freeman and Company.

 Shivananda Nayak B. Manipal Manual of Clinical Biochemistry. 2007. Jaypee Brothers

Medical Publication.

 Fischbach, F. and Dunning, M. B. III. A Manual of Laboratory and Diagnostic Tests.

Philadelphia: Williams & Wilkins, 2004.

 Lehmann, C. A. Saunder’s Manual of Clinical Laboratory Science. Philadelphia: W.B.

Saunders, 1998.