Urinalysis

R.A. Villaraza, D. Young, D.F. Yu, *R.A Zafra

Department of Biological Sciences

College of Science, University of Santo Tomas
España, Manila Philippines

ABSTRACT

Urine is a waste product synthesized by the body and secreted by the kidneys
through filtration and subsequently excreted out from the urethra. Urinalysis is performed
commonly as diagnosis to find out a possible illness of a person, it is not only a single test
but rather numerous ones to ensure accuracy. The experiment’s objectives are to subject an
acquired sample of urine to various tests and to qualitatively examine or observe for the
existence of both normal and pathological organic constituents. Before thorough
experimentation the sample collection time, turbidity, color and pH level are recorded.
Qualitative exam of normal organic constituents includes the tests for urea, uric acid, and
indican. The tests resulted in normal findings. As for the qualitative examination for
pathologic organic constituents, Gunnings’s test, Benedict’s test, Exton’s test, Smith’s test
and Occult blood test were performed. The tests yielded optimal results indicating the
healthy well-being of the volunteer.

INTRODUCTION

Urine is a liquid, transparent solution whose color ranges from colorless to dark
yellow, even orange in some occurrences. Urine is composed of wastes such as urea,
dissolved salts and organic compounds which are byproducts of the kidneys. It is crucial in
maintaining the homeostasis of an organism.

Due to the pigment known as urochrome, urine may be observed in a wide array of
colors such as pale yellow, dark yellow, light orange, amber and any variations of these.
These colors may be utilized to indicate the urine concentration.

The presence of volatile acids causes the odor of a fresh urine sample. The analysis
of a freshly voided urine’s pH may indicate its acid-base balance.

Urinalysis provides an overview of a person’s health, as such it is employed as one

of the basic screening and diagnostic methods in the field of medicine. As a diagnostic tool,
it can help with the detection of cellular substances in the urine that may indicate disorders
affecting the kidneys or renal tract or ailments not directly related to these.

The experiment aims to subject urine sample to various tests and to examine it for the
presence of normal and pathological organic constituents in a qualitative manner.

METHODOLOGY

A. Initial Examination of Urine

Time collected, color, turbidity and pH level were recorded

B. Qualitative Examination for Normal Organic Constituents

1.) Test for Urea

0.5mL of 70% NaOH as well as 4 drops of bromine water were added to

1mL of the fresh urine sample. Evolution N2 gas was observed thereafter.

2.) Test for Uric Acid

5mL of 20% Na2CO3 was added to the sample followed by 5 drops of

phosphotungstic acid reagent and was subsequently mixed. Blue solution
formation was then observed.

3.) Indican Test

5mL Obermayer’s reagent was added to 5mL of the sample and then
thoroughly mixed. After, 3 mL chloroform was added, before being shaken and
allowed the chloroform layer to settle. A blue color in the lower layer was
observed.

4.) Test for Creatinine

2mL of urine sample was added with 1mL of alkaline picrate solution.
Formation of a solution of orange color was then recorded.
C. Qualitative Examination for Pathogenic Organic Constituents

1.) Gunning’s Test

5mL of the sample was basified with 5 drops of concentrated ammonium

hydroxide through the utilization of a red litmus paper. Lugol’s solution was then
added to the basic urine to produce a black cloud, however this does not disappear
immediately. It was left to stand for 5 minutes. It was done in a similar fashion on
positive control and then the results were compared.

2.) Benedict’s Test

To begin, 5mL of Benedict’s reagent was added to 8-10 drops of sampled in

a test tube and was thoroughly mixed. Subsequently, it was heated in a boiling water
bath for about 2-3 minutes then it was left to stand for cooling. Precipitate formation
was observed. Similar procedure was performed for the positive control and the
results obtained were compared.

3.) Exton’s Test

3mL of urine and 3mL of Exton’s reagent were put in a test tube and mixed.
The solution was then warmed until the appearance of a cloudlike phenomenon. For
the positive control, similar procedures were followed. Yielded results were
compared.

4.) Smith’s Test

5mL of the urine sample was placed inside a test tube. It was inclined and
overlaid with 3mL tincture of alcoholic iodine mixture. The same manner was
performed for the positive control. Yielded results were compared.

5.) Test for Occult Blood

5mL of 95% Ethanol was added to a half spatula guaiac powder inside a test
tube and was mixed thoroughly. 5mL hydrogen peroxide was then added to the
mixture. Finally, 5mL of the solution was added to 3mL of acidified urine. The
 procedure done was performed in the same manner for the positive control and the
results obtained were compared.

RESULT AND DISCUSSION

Ammonia is considered as a base of extreme toxicity and over accumulation in the

body proves fatal in most cases. However, the liver holds a system of molecules and enzymes
designed to hastily convert ammonia as well as carbon dioxide into urea, this process is
known as urea cycle. Though high urea concentrations cannot be tolerated by the bodies, it
is a fact that it is less harmful than its ammonia predecessor. The kidneys deal with the
excretion of urea.

Aside from urea, humans excrete another nitrogenous waste called uric acid. Uric
acid is slightly water soluble and has the tendency to precipitate out of a solution, forming
crystals of sodium urate. These in turn play a factor in the formation of kidney stones and the
gout pain.

Amino acid tryptophan undergoes conversion to indole by tryptophan side chain

bacteria. Subsequently, indole is further converted to 3- hydroxy indole in the liver where it
is then conjugated with potassium sulfate and/or glucuronic acid. Finally, it is transported
through the blood to the kidneys for excretion. High levels are deemed by physicians as a
sign of toxemia.

Creatinine is a protein synthesized by the muscle released in the blood. A normal

person’s production is relatively stable. Creatinine can be used for measuring kidney function
by observing the level of creatinine level of serum.

Ketones are chemicals the body produces when insulin supply is low in the blood and
requires fat breakdown for the synthesis of its energy. Though having an emergency function,
ketone bodies have the ability to kill body cells due to its poisonous property.

Carbohydrates present in dietary supplements become glucose in blood, Glucose

levels are measured to determine or diagnose diabetes or to see how diabetes is controlled.

Usually, urine must be void of proteins, this is due to the kidneys’ function in keeping
large molecules in the blood while filtering only small impurities. High protein count in urine
signifies an ailing kidney.

Breakdown of hemoglobin results in the production of bilirubin. Should there be a

detection of abnormal values of bilirubin, ailments such as anemia, cirrhosis, hepatitis and
the like may be assumed.

Blood in urine may be explained with many potential reasons, but most of the time,
urinary tract or kidney is the surmised problem.

Initial Examination of Urine Sample

Time collected Early

morning; 6:32
AM
Color Dark Yellow
Transparency Clear
pH Level 6

Qualitative Exam of Normal Organic Constituents

Test Result
UREA Gas formation
URIC ACID No blue color solution
INDICAN Slight blue color in lower layer
CREATININE Orange solution with yellow bottom

Qualitative Exam for Pathologic Constituents

Test Positive Control Urine Sample
GUNNING’S TEST Formation of Iodoform Formation of Iodoform
crystals crystals

BENEDICT’S TEST Turbid, dark yellow Clear blue solution with

precipitate yellow, cloudy formation

EXTON’S TEST Cloudy solution Clear solution

 SMITH’S TEST Emerald top layer No Emerald green color

OCCULT BLOOD Mossy green solution with Turbid, brown solution

TEST blue bottom layer

CONCLUSION

In conclusion, inferring from the tabulation of results above, the volunteer is

deemed healthy as no indication of any ailment can be seen based on the conducted set of
urinalysis tests.

V. References

Simerville, J. A., Maxted, W. C., & Pahira, J. J. (2005). Urinalysis: a comprehensive

review. American family physician, 71(6).
Chen, G. L., El-Gabry, E. A., & Bagley, D. H. (2000). SURVEILLANCE OF UPPER URINARY
TRACT TRANSITIONAL CELL CARCINOMA:: THE ROLE OF URETEROSCOPY,
RETROGRADE PYELOGRAPHY, CYTOLOGY AND URINALYSIS. The Journal of
urology, 164(6), 1901-1904.

Hoberman, A., Wald, E. R., Penchansky, L., Reynolds, E. A., & Young, S. (1993).
Enhanced urinalysis as a screening test for urinary tract infection. Pediatrics, 91(6), 1196-
1199.
Wallach, J. Interpretation of Diagnostic Tests. Philadelphia: Wolters Kluwer
Health/Lippincott Wiliams & Wilkins, 2007.
Urine – Retrieved May 7, 2018
http://en.wikipedia.org/wiki/Urine
Urinalysis – Retrieved May 7, 2018
http://en.wikipedia.org/wiki/Urinalysis