Saint Mary’s University

School of Health and Natural Sciences

BAYOMBONG, NUEVA VIZCAYA 3700
Nursing Department
Laboratory Activity No. 9: Urine

CARBONEL, Kyla Mae M.

BSN 1A

ABSTRACT

Urine is an unstable fluid, and changes to its composition commence to take vicinity as soon as it
is voided. As such, collection, storage, and handling are vital troubles in retaining the integrity of
this specimen. In the laboratory, urine can be characterized by bodily appearance, chemical
composition, and microscopically. Physical examination of urine consists of description of color,
odor, clarity, volume, and unique gravity. Chemical examination of urine consists of the
identification of protein, blood cells, glucose, pH, bilirubin, urobilinogen, ketone bodies, nitrites,
and leukocyte esterase. Finally, microscopic examination entails the detection of crystals, cells,
casts, and microorganisms.

Keywords: Characterized, Composition, Examination, Specimen, Vicinity

I. INTRODUCTION

Urine is the specimen most frequently submitted for culture. It also presents major problems
in terms of proper specimen collection, transport, culture techniques, and interpretation of results.
As with any other specimen submitted to the laboratory, the more information provided by the
submitting physician the more able is the laboratory to provide the best possible culture data. The
most common sites of urinary tract infection (UTI) are the urinary bladder (cystitis) and the urethra.
From these sites the infection may ascend into the ureters (ureteritis) and subsequently involve the
kidney (pyelonephritis). Females are more prone to infection of the urinary tract and also present
the greater problem in the proper collection of specimens. In both males and females, UTI may be
asymptomatic, acute, or chronic. Asymptomatic infection can be diagnosed by culture. Acute UTI
is more frequently seen in females of all ages; these patients are usually treated on an outpatient
basis and are rarely admitted to hospital. Chronic UTI in both males and females of all ages is
usually associated with an underlying disease (e.g., pyelonephritis, prostatic disease, or congenital
anomaly of the genitourinary tract) and these patients are most often hospitalized. Asymptomatic,
acute, and chronic UTI are three distinct entities and the laboratory results often require different
interpretation. Asymptomatic pyelonephritis in females may go undetected for some time, and is
often only diagnosed by carefully performed quantitative urine culture. Chronic prostatitis is
common and difficult to cure, and is often responsible for recurring UTI. In most UTI, irrespective

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 of type, enteric bacteria are the etiological agents, Escherichia coli being isolated far more
frequently than any other organism. In about 10% of patients with UTI, two organisms may be
present and both may contribute to the disease process. The presence of three or more different
organisms in a urine culture is strong presumptive evidence of improper collection or handling of
the urine specimen. However, multiple organisms are often seen in UTI in patients with indwelling
bladder catheters (digicollection.org)

II. OBJECTIVES

1. To test for the presence of some normal products of metabolism in the urine
2. To test for the presence of some pathological constituents of urine

III. MATERIALS

 Normal Urine
 5% sucrose
 Concentrated H2SO4
 Pathological urine sample (urine + 5% glucose, 5% albumin, acetone, bile, blood
solution)
 Concentrated NaOH
 Bromine Water
 Concentrated HCl
 10% NaOH
 (NH4)2SO4-NH3 Reagent
 Picric Acid
 5% H2O2
 Benizidine in glacial acetid acid
 Sodium Nitroprusside
 Benedict’s Reagent
 Concentrated HNO3
 2% Acetic Acid
 5% AgNO3

IV. PROCEDURES

A. Test for normal organic constituents

1. Urea

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 a. Place 10 drops of urine in a test tube, add 5 drops of 10% HCl, then add a few crystals
of NaNO3. Note the evolution of a gas.
2. Uric Acid
a. Place 5 mL of the urine in a beaker. Add 1 mL of Concentrated HCl, stir well and set aside
until the next laboratory period. Describe the appearance of the crystals that deposit on the
sides of the beaker.
b. Dissolve a few of the uric acid crystals in 2 mL of 5% sodium carbonate solution. Est its
reducing properly by adding 0.05% of CuSO4 (Benedict’s test). Boil in water bath for 15
minutes.
3. Creatinine. Jaffe’s Picric acid solution
a. Place 10 drops of urine in a test tube, then add 5 drops of saturated picric acid. Alkalinify
with 10% NaOH. A red color is produced which turns yellow when acidified.
4. Chlorides
a. To 3 mL of urine, add several drops of HNO3 and 5 drops of 5% AgNO3. Observe.
5. Phosphate
A. To 5 mL of urine.
a. Add dilute NH4OH. Observe the color of the solution.
b. Warm the mixture gently and filter. To the filtrate, add a few drops of magnesia mixture,
then warm gently.

B. Pathological Constituents.

Simulate 5 mL of the pathological urine sample by adding 5 mL each of 5% glucose, 5%

albumin, acetone, bile and blood solution to 225 mL of urine. (this will be shared by the whole
class) Test for the presence of the pathological constituents as follows:

1. Acetone: Rothera’s Nitroprusside Test

Place 5 mL of the pathological urine sample in a test tube, the add solid ammonium sulfate.
Shake until the urine is saturated. Now, add 2-4 drops of concentrated ammonium hydroxide
and 2-4 drops nitroprisside reagent. Shake and wait for the slow development of a purple tinge
that gradually deepens. This is a confimatory for the presence of acetoacetic acid.

2. Glucose: Benedict’s Test

Place 2-3 mL of Benedict’s reagent in a test tube, then add 5 mL of urine. Mix thoroughly.
Boil it in water bath for 10-15 minutes and allow to cool. Take note of the color of the
precipitate formed.

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 3. Albumin: Heller’s Ring Test

To 1.5 mL concentrated HNO3 in a test tube, deliver 1 ML urine down the side of the tube such
that it forms a separate layer. The presence of protein is shown by a pluffy zone at the urine-
acid interface.

4. Blood: Benzidine Test

Place about 3 mL of urine suspected of containing blood in an evaporating dish and add 5 drops
of benzidine. Mix, then add 2 drops of 3% H2Oo. Spread this solution over the surface of the
dish and describe the color that soon forms.

5. Bile Pigments: Gmelin’s Test

Place 1 mL concentrated HNO3 in a test tube. By means of a pipet, deliver down the side of the
tube 5 mL of urine. Do not shake. Note the color of the ring which appears at the interface of
the 2 liquids. Green, blue, or violet rings appear if bilirubin is present.

6. Bile Acid and Salts: Pettenkofer’s Test

Place 20 drops of urine in a test tube, then add 5 drops of 5% sucrose. Let 2 mL concentrated
H2SO4 slide down the side of the tube. Do not shake. A red ring develops at the point of contract
of the two solutions. Don’t mistake the brown color resulting from the charring of sucrose by
H2SO4 for the red ring. Stir the mixture and note the spreading of the red color throughout the
liquid.

C. Physiological Properties of Urine

Compare the characteristics listed below that are exhibited by normal urine and by pathological
urine.

V. QUESTIONS

1. Why must a 24-hour period sample of urine be used for examination if a detailed
composition is to be determined? For a simple routine qualitative analysis, why is anealy
morning sample of urine used for tests, and not a sample collected after meal?
2. What is meant by glucosuria? Albuminuria?

VI. RESULTS AND DISCUSSION

Table 1: Test for normal organic continents

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 Test Used Results
Urea A presence of moisture on the test tube
Uric Acid Crystal Formation
Creatinine Color turns to red when added 10% of NaOH and back to its normal
color when HNO3 and 5 drops of 5% AgNO3 was added
Chloride It has a white precipitation on the upper phase, light yellow on the
middle phase and transparent on the bottom phase
Phosphate A Yellowish color
Phosphate B Turned yellow solution with white precipitations at the bottom phase
In this table we could see the constituents that could be found in urine.

The urinary system’s ability to filter the blood resides in about 2 to 3 million tufts of
specialized capillaries—the glomeruli—distributed more or less equally between the two
kidneys. Because the glomeruli filter the blood based mostly on particle size, large elements like
blood cells, platelets, antibodies, and albumen are excluded. The glomerulus is the first part of
the nephron, which then continues as a highly specialized tubular structure responsible for
creating the final urine composition. All other solutes, such as ions, amino acids, vitamins, and
wastes, are filtered to create a filtrate composition very similar to plasma. The glomeruli create
about 200 liters (189 quarts) of this filtrate every day, yet you excrete less than two liters of
waste you call urine. Characteristics of the urine change, depending on influences such as water
intake, exercise, environmental temperature, nutrient intake, and other factors. Some of the
characteristics such as color and odor are rough descriptors of your state of hydration. For
example, if you exercise or work outside, and sweat a great deal, your urine will turn darker and
produce a slight odor, even if you drink plenty of water. Athletes are often advised to consume
water until their urine is clear. This is good advice; however, it takes time for the kidneys to
process body fluids and store it in the bladder. Another way of looking at this is that the quality
of the urine produced is an average over the time it takes to make that urine. Producing clear
urine may take only a few minutes if you are drinking a lot of water or several hours if you are
working outside and not drinking much. (opentextbc.ca)

Table 2: Pathological Constituents

Test Results
Acetone (Ketone Iodine) Purple tinge develops in the upper phase and white
precipitation
Glucose Blue solution turns to dark green with light yellow in the
upper phase and white precipitation
Albumin Light yellow solution in the upper phase, dark yellow ring
in the middle phase and transparent in the bottom phase
Bile Pigments Yellow in the upper phase, violet ring in the middle phase
and transparent solution in the bottom phase

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 Bile Acids and salts Red solutions in the upper phase, red ring develops in the
middle phase and transparent in the bottom phase and
when you stir the test solution it turns to red
Blood Turbid white yellow solution with precipitation in the
bottom phase
While in this table we could see the physical characteristics of a urine that could be test by
urinalysis in the laboratory.

1. 6 normal and 14 pathological urines have been analysed for inorganic ions, creatinine,
urea and protein. The results of these determinations are compared with the total salt
concentration, the total concentration of dissolved particles and the total solids. Na+, K+, Ca2+,
Mg2+ and NH+4 are shown normally to account for nearly all the cations present in urine. In one
pathological specimen containing large amounts of amino acids, a high concen- tration of
undetermined cations was shown to be present. Chloride, sulphate, phosphate and bicarbonate
are shown to account for about 80% of the anions present in urine. The urine of a schizophrenic
also suffering from a fractured femur, was shown to contain a concentration of undetermined
anions much higher than normal. By subtracting the vapour pressure due to urea, creatinine and
inorganic ions from the total vapour pressure, unanalysed constituents were found to make up
about 20% of the total in normal urine. Similar figures were obtained using the total solid
content. Abnormally high values for unanalysed constituents were found in urine from a patient
with the nephrotic syndrome, as well as in the two pathological urines described above with large
amounts of undetermined cations and anions. (H.J.Yardley, 1958)

Table 3: Physiological properties of urine

Normal urine Pathological Urine

Color Yellow Orange
Transparency Transparent Turbid
pH 7 neutral 6 acidic
Specific Gravity 1.4 3
Odor Aromatic fishy
This table indicates the characteristic of urine should be against the pathological urine.

Normal urine is actually a highly complex aqueous solution of organic and inorganic
substances. The majority of the constituents are either waste products of cellular metabolism or
products derived directly from certain foods that are eaten. The total amount of solids in a 24-hour
urine sample averages around 60 g. Of this total, 35 g are organic and 25 g are inorganic.The most
important organic substances are urea, uric acid and creatinine. Urea is a product formed by the

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liver from ammonia and carbon dioxide. Ninety-five percent of the nitrogen content of urine is in
the form of this substance. Uric acid is an end-product of the oxidation of purines in the body. By
weight, there is normally about 60 times as much urea as uric acid in urine. Creatinine is a hydrated
form of creatine. There may be twice as much creatinine as uric acid in the urine. The principle
inorganic constituents of urine are chlorides, phosphates, sulfates and ammonia. Sodium chloride
is the predominant chloride and makes up about half of the inorganic substances. Since ammonia
is toxic to the body and lacking in plasma, there is very little of it normally present in fresh urine.
The small amount that is present is probably secreted by nephron tubules. Urine that is allowed to
stand at room temperature for 24 hours or longer may give off an odor of ammonia due to the
breakdown or urea by bacterial action. Because of the efficient absorptive properties of renal tubule
cells there should be no appreciable amounts of glucose or amino acids in urine. About 0.3 to 1.0
g of glucose in a 24-hour urine sample would be considered normal excretion. Occasionally, higher
amounts may occur in individuals during emotional stress.(jmu.edu)

The first voided morning specimen is particularly valuable because it is more concentrated
and abnormalities are easier to detect. An early morning specimen is also relatively free of dietary
influences and changes due to physical activity. In collecting any urine specimen, it is always
important for the nurse to observe specific agency protocols, to check with the laboratory regarding
the need for refrigeration or preservation of specimens, and to follow universal precautions. Single
random specimens may be taken at any time of the day or night. Timed specimens range from
short-term 2-hour collections to 24-hour collections. A 24-hour urine specimen is an extremely
important diagnostic test because it reveals how the kidney adjusts to changing physiologic needs
over a long period. Substances excreted by the kidney are not excreted at the same rate or in the
same amounts during different periods of day and night; therefore, a random urine specimen does
not accurately represent the processes taking place over a 24-hour period. However, a 24-hour
urine specimen is useful only when all the patient's urine is collected for 24 hours. Even if just one
sample is discarded, the results will be inaccurate. The nurse must ensure that the patient and all
assistive personnel understand the importance of saving all the urine. To begin the 24-hour
collection, the person voids and discards the urine already in the bladder. All urine starting with
the next voiding is collected for the next 24 hours and put into a large collection bottle. To prevent
breakdown of urinary components, the collection has a preservative added to it or is refrigerated.(
rnceus.com)

Glucosuria, glucose in the urine, results from the glomerular filtration of more glucose than
the renal tubule can absorb. It occurs in all normal individuals in amounts up to 25 mg/dl.
Abnormally increased glucosuria [more than 25 mg/dl in random fresh urine, results from either
an elevated plasma glucose, an impaired renal glucose absorptive capacity, or both. The plasma
glucose concentration above which significant glucosuria occurs is called the renal threshold for
glucose. Its value is i, and deviations occur both above and below the commonly accepted "normal"

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 threshold of 180 mg/dl. In diabetic patients, the value is reported to vary from 54 to 300 mg/dl.
Although glucosuria greater than 25 mg/dl is considered pathologic, many commercial
semiquantitative urine tests for glucosuria that are available to patients fail to detect glucosuria
until it reaches a level of 50–250 mg/dl. (Cowart and Stachura, n.d)

Albumin is a type of protein that is normally found in the blood. Your body needs protein. It
is an important nutrient that helps build muscle, repair tissue, and fight infection. But it should be
in your blood, not your urine. When you have albumin (protein) in your urine, it is called
“albuminuria” or “proteinuria.” (kidney.org)

VII. GENERALIZATION

As a conclusion, I conclude that in this activity that the components that could see in the
urine could already indicate that you have a disease that could easily detect in a urinalysis test.
Also in the aroma from the urine which has a fishy smell that should be has an aromatic smell.

VIII. REFERENCE

Albuminuria, (n.d) Retrieved from https://www.kidney.org/atoz/content/albuminuria

ANATOMY AND PHYSIOLOGY: Physcal characteristics of Urine (n.d) Retrieved from

https://opentextbc.ca/anatomyandphysiology/chapter/25-1-physical-characteristics-of-urine/

Cowart and Stachura Glucosuria. (n.d) Retrieved from

https://www.ncbi.nlm.nih.gov/books/NBK245/

H.J.Yardley, The composition of normal and pathological urine with an estimate of the
concentration of unanalysed substances, (1958) Retrieved from
https://www.sciencedirect.com/science/article/pii/0009898158900901

Urinalysis (n.d) Retrieved from http://csmbio.csm.jmu.edu/biology/danie2jc/urinalysis.htm

Urine Collection (n.d) Retrieved from http://www.rnceus.com/ua/uacollect.html

World Health Organization, Basic Laboratory Procedures in Clinical Bacteriology (1991)

Retrieved from http://helid.digicollection.org/en/d/Jwho01e/4.3.1.html

IX. DOCUMENTATION

Procedure 3 shows the presence of No appearance of green or violet

protein in urine color so there’s no biliburin

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 Procedure results for the benedicts
test

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