Simple Laboratory Test For Urine-Final

Simple Laboratory Test for Urine
CHAPTER I:
THE PROBLEM AND ITS SCOPE
Background of the Study
Over the last decade, cases such as urinary tract infection, cirrhosis, diabetes,
protein in urine (proteinuria), and kidney inflammation (glomerulonephritis) have risen,
not by the hundreds but by the thousands (Fareed MD, 2013). These are just some of the
diseases and illnesses that have become rampant in the world today, where almost all the
lifestyles of the people include unhealthy diets and such. The age of life expectancy has
now become 69 years old, from what once was 80 years (Life Expectancy Statistics, 2019).
With people having unhealthy lifestyles, mortality rates will most likely increase.
Especially now, where people only go to doctors when it’s already to late. Diseases and
illnesses, as stated above, are some of which can be prevented and monitored, yet many
people still lose their lives to it.
Urinary tract infections (UTIs) are considered to be the most common bacterial
infection. An infection anywhere in the urinary tract is called a urinary tract infection
(UTI). That infection may be in the urethra (urethritis), bladder (cystitis), or kidneys
(pyelonephritis) which may cause irritation when urinating (B. Foxman, 2002). According
to the 2007 Department of Health survey, UTI accounted for nearly 7 million office visits
and 1 million emergency department visits, resulting in 100,000 hospitalizations. The risk
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of UTI increases with increasing duration of catheterization. In non-institutionalized
elderly populations, UTIs are the second most common form of infection, accounting for
nearly 25% of all infections (B. Foxman, 2002).
In connection to other illnesses, the next on the list is cirrhosis. Cirrhosis is a late
stage of scarring (fibrosis) of the liver caused by many forms of liver diseases and
conditions, such as hepatitis and chronic alcoholism ( Cirrhosis , 2018). Each time your
liver is injured — whether by disease, excessive alcohol consumption or another cause —
it tries to repair itself. In the process, scar tissue forms. As cirrhosis progresses, more and
more scar tissue forms, making it difficult for the liver to function (decompensated
cirrhosis). Advanced cirrhosis is life-threatening. The liver damage done by cirrhosis
generally can't be undone. But if liver cirrhosis is diagnosed early and the cause is treated,
further damage can be limited and, rarely, reversed (S. Starr, 2011).
In addition, diseases like diabetes are also on the rise. Diabetes is one of the most
common metabolic disorders in the world, and the prevalence of diabetes in adults has been
increasing in the last decades. Over time diabetes can damage the heart, blood vessels,
eyes, kidneys and nerves. Approximately 12 adults die of a diabetes-associated illness per
1,000 cases in 2011 in low- and middle-income countries: more than double the mortality
rate of high-income countries. Mortality rates are much lower in high-income countries
with the greater healthcare recourses (Guariguata L. et al., 2011). According to the
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International Diabetes Federation or IDF, over 3,721,900 people have been reported to
have diabetes in the Philippines as of 2017, and most likely these numbers are still rapidly
increasing. Philippines is also considered one of the diabetes’ “hot spots” in the Western
Pacific region, where the disease is already reaching epidemic proportions. Our
government knows this too well, and the increased taxes on sugary drinks is just one of the
steps being taken to stem the tide (Castilo, R. 2018).
Furthermore, protein in urine also poses as a threat to the health of many today.
Protein in urine — known as proteinuria — is excess protein found in a urine sample
(Proteinuria, 2018). Kidneys filter waste products from your blood while retaining what
your body needs — including proteins. However, some diseases and conditions allow
proteins to pass through the filters of kidneys, causing protein in urine. People
with proteinuria have urine containing an abnormal amount of protein. The condition is
often a sign of kidney disease (Khatri MD, 2018).
In addition, other rampant diseases include kidney inflammation
(glomerulonephritis). Glomerulonephritis is the inflammation of the tiny filters in kidneys
(glomeruli). Glomeruli remove excess fluid, electrolytes and waste from your bloodstream
and pass them into your urine. Glomerulonephritis can come on suddenly (acute) or
gradually (chronic) either of which still has the same level of danger (Glomerulonephritis,
2018). Glomerulonephritis occurs on its own or as part of another disease, such as lupus or
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diabetes. Severe or prolonged inflammation associated with glomerulonephritis can
damage your kidneys. Treatment depends on the type of glomerulonephritis you have
(Murrel MD, 2018).
Despite the rising cases of these diseases, many organizations are finding ways to
help alleviate the devastation it brings. Many are innovating ways to help find a solution,
a fix, and a cure. The efforts differ from small talks about these diseases to lab researchers
finding a new form of drug. But, we the researchers would want to help detect and prevent
these diseases from becoming more and more rampant. As the famous Desiderius Erasmus
once said, “Prevention is better than cure”. Knowing how to detect and prevent these
diseases costs less than actually treating it.
In the world today, people from around the globe share the same sentiments. People
want the things in life, to be fast and simple. Thus, in view of these factors, the researchers
recasted this study which aims to show a simple way of detecting these disease in people,
taking into the consideration of the Philippines economy, its people and the betterment of
the future world. We the researchers, have opted out to make a simple laboratory test using
urine to test for the presence of different kinds of substances that would indicate different
kinds of diseases.
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STATEMENT OF THE PROBLEM
Health is wealth, as the great Ralph Waldo Emerson once said that means
no matter how wealthy you are, if you are not healthy, you are still going to be poor. This
leads us to thinking that it is better to have health than wealth.
The Philippines being a 3rd world country struggles with providing good-
quality healthcare to each and every one of the Filipinos (Medilo 2018). These diseases
like diabetes, being one of the Philippines leading causes of death, is also one of the most
easily prevented but because of circumstances like the lack of knowledge, people’s
lifestyle, and financial stability, these diseases remain a problem in the Philippines and in
the world (D.O.H. 2015).
Having stated that the Philippines is a 3rd world country, that means that it is an
underdeveloped country or in simple terms there are less investments on education, health
care, etcetera. This means that most of the Filipinos don’t have the luxury of money to pay
for the tests needed in order to see if they have these diseases because full laboratory tests
cost a lot of money (Medilo 2018). Thus, making 3 rd class Filipinos become more likely to
be struck by such diseases and illnesses, and eventually die because of it.
This all boils down to one thing and that is the welfare of each Filipino. These
diseases don’t come knocking on one’s door, but instead they come slow and undetected.
They remain a problem in the Philippines until today, and these problems lead to many
more problems if left undiscovered and untreated.
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The researchers want to find a way to help diminish the devastation brought by
these diseases, and in lieu of this the researchers will conduct a simple laboratory test for
urine, which may give indicators if one has these specific diseases. The researchers aim to
make a better and simpler way of testing urine that is not only simple but also readily
available at any laboratory or facility. The researchers also want the people to be educated
on understanding the indicators of urinary tract infection, cirrhosis, diabetes, protein in
urine, and kidney inflammation.
Hence this research tries to find answers as to how the people in the 21st century
can use the modern day technology to test urine in a laboratory in the Philippines. More
specifically, it aims to find the answers of the following questions:
1. What are the tests that will be utilized by the researchers?
2. What result/s will indicate UTI, cirrhosis, diabetes, protein in urine, and
kidney inflammation?
3. What is the significance of the said laboratory tests?
4. How will this test affect the field of medicine in the future?
The researchers aspire that by answering these questions; it will bring a new light
in the field of simple laboratory test for urine and hopefully lead to a better development
on the field of medicine.
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HYPOTHESIS
In the world today, there are only 3 ways to get your urine tested, namely these are:
by visual examinations, by chemical examinations, and by microscopic examination
(Urinalysis, 2017). Under chemical examinations, the researchers hypothesize that a
simple laboratory test for urine can be done through the different reagents and chemical
reactions that will be utilized such as: the ammoniacal zinc chloride test for detection of
pigments, the adding of acetic acid for detection of sulphates, the silver nitrate test for
chlorides, the ammonium hydroxide test for phosphates, coagulation test for albumin,
Heller’s ring test for proteins, and the Benedicts reagent test for glucose. These specific
tests will be used to determine the presence of different kinds of compounds found in urine.
These are some of the oldest yet accurate tests when utilizing urine. Procedures like tests
for the detection of sugar, sulphates, chlorides, and phosphates in urine are among the
oldest tests known in clinical chemistry (Alfred H. et al 1957).
Through experimentation, the researchers will find out whether or not these
reagents can be used for such tests, and if the positives show, then the researchers will find
out what they indicate to the health factor of our body.
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CONCEPTUAL FRAMEWORK
Laboratory Tests for Urine
Different types of Laboratory

The tests the researchers will use
Tests for Urine
Visual Examination Ammoniacal zinc chloride Test
Chemical Examination Determination of sulphates
Microscopic Examination Silver Nitrate test for chlorides
Determination of phosphates
Coagulation Test for albumin
Heller’s Ring Test for proteins
Benedict’s Reagent Test for glucose.
- used as a simple test for urine.

Multiple reagents are used to
detect different kinds of
components of urine.
Visual Examination and
Microscopic Examination Simple Laboratory Test for Urine
Very costly and time consuming, Simple, specific, sensitive, and speedy
thus leading to less frequent test
testing
In alignment with the world’s different kinds of urinalysis,
the researchers decided to use different reagents as listed
above. These tests are simple, specific, sensitive, and Dependent Variable/s:
The outcome of the research

speedy, contrasting the visual and microscopic
Independent Variable/s:
examinations which are costly and time consuming to
The different reagents, materials, and urine
many. The dependent variables found in the test is the
outcome of the tests itself. On the other hand, the
independent variable found are: the different reagents, the

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materials used, and the urine.
SIGNIFICANCE OF STUDY
The findings of this research will redound to the benefit of those people in the
lower-middle class society, considering that UTI, cirrhosis, diabetes, proteinuria, and
glomerulonephritis are spreading fast and that people nowadays want something simple
and time efficient. The higher the cases of these diseases that will occur, justifies the
significance of these tests. Thus, people who will read this study may discover a simple
way of testing their urine.
The findings of the research will also be beneficiary to pharmacies and hospitals,
for they can use this study to further enhance their medical capacity, because this is cheap,
affordable and accurate. The diseases stated above are no doubt one of the many medical
illness that are prevalent here in the Philippines, thus utilizing this research will not only
help people with those kinds of illnesses, but it will also help the country in an economical
view.
In addition, this research will also benefit future researchers who are also in the
search for simple laboratory tests for urine. They may utilize this research and further
incorporate modern technology, thus creating more advanced medicinal approaches to
modern day urinalysis. Furthermore, this research shines a new light, to using not so used
reagents, to test urine.
To sum it all up, the study will help readers uncover alternative ways to determine
different kinds of components in urine that may lead to diseases if not looked into. It will
also be beneficial to the Filipinos who are in poverty, in rural areas, in the field of business,
and people who have little to no time for getting tested in clinical labs.
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LIMITATIONS OF THE STUDY
In view of the external factors of this research, the researchers have gathered the
limitations of the study. First and foremost, being student researchers we are not given
privileges to fully use all the equipment found in the laboratory, because we are still
underage. In addition, urine, being a waste product of humans, is strictly prohibited to be
used in laboratories found in school because they are not clinically ready for urine testing,
thus, forcing the researchers to make their very own simulated urine. Second, the
researchers have limited time, when in fact a large span of time is needed to accurately
complete the research because not only should there be multiple tests, to test for accuracy,
there should also be a span of 2-3 weeks observation which the researchers didn’t have.
Third, the researchers have no access to all the specific chemicals they need as to which
they need to purchase necessary equipment and chemicals for the study and our resources
are insufficient. This was because the laboratory used by the researchers does not have
complete sets of reagents needed for all the tests for urine. Fourth, the researchers over-all
resources were not sufficient to back up the study.
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DEFINITION OF TERMS
In lieu of this paper, the researchers have found 23 highfalutin words of
which definitions are given below.
Benedict’s Reagent - is used as a simple test for reducing sugars.
Cirrhosis- is late stage of scarring (fibrosis) of the liver caused by many forms of liver diseases
Conceptual framework- is an analytical tool with several variations and contexts. It is
used to make conceptual distinctions and organize ideas.
Devastation- great destruction or damage.
Diabetes- is a disease in which your blood glucose, or blood sugar, levels are too high.
Diminish- make or become less.
Epidemic- The occurrence of more cases of a disease than would be expected in a
community or region during a given time period.
Epidemic proportions- an outbreak of disease that spreads quickly and affects many
individuals at the same time.
Factors - matters affecting other medical conditions diagnosed when attitudes or
behaviors have a negative effect on a medical disorder that the person has
Glucose- it is a type of sugar you get from foods you eat, and your body uses it for
energy.
Healthcare- the organized provision of medical care to individuals or a community.
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Highfalutin- expressed in or marked by the use of language that is elaborated or heightened by
artificial or empty means.
Hypotheses- represent specific restatements of the purpose of the study, which can be
directional or non-directional.
Insulin- a hormone produced in the pancreas which regulates the amount of glucose in
the blood.
Lieu- on behalf of a whole
Lifestyle- the way in which a person or group lives
Metabolic disorder-inherited single gene anomaly, most of which are autosomal
recessive
Non-communicable - a medical condition or disease that is by definition non-infectious
and non-transmissible among people Mortality- the number of deaths in a given time or
place
Prevalent- widespread in a particular area or at a particular time.
Recast- write (something) again to alter or improve it.
Risk- refers to a possibility that harm may occur.
Third “3rd” world country- underdeveloped or developing countries
Undiagnosed- not diagnosed or having been subject to diagnosis.
UTI- infections in any part of the urinary tract
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CHAPTER II:
REVIEW OF RELATED LITERATURE
Preamble
UTI, cirrhosis, diabetes, proteinuria, and kidney inflammation are world-wide
public health problems. These are the leading causes of death and disability in the world.
The diagnosis and management of these diseases require tight monitoring. The challenge
of providing such tight and reliable control remains the subject of enormous amount of
researches (Wang, 2000). The presence of these diseases approximately doubles the risk of
a wide range of vascular diseases. Evidence is also emerging that these diseases is
associated with nonvascular conditions, including positive associations with certain
cancers (e.g., liver cancer) and negative associations with other cancers (e.g., prostate
cancer) (Diabetes and Kidney Diseases, 2011).
The research design that was used is the experimental approach, paired by the data
collection technique of tallying observation. This chapter aims to show the readers the
different related literature found in this study. In this chapter a theoretical framework will
be shown for the reader to further understand the related literature.
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Theoretical Framework
SIMPLE LABORATORY TEST FOR URINE
Ammoniacal Zinc Cirrhosis

Chloride test
Urobilin
Sulphate in Kidney
Detection of
Sulphates Urine Inflammation
Chlorides in Kidney
Detection of
Chloride Urine Inflammation
Detection of Phosphates
Phosphates in Urine UTI
Detection of Albumin in Proteinuria

Albumin Urine
Protein in
Heller’s ring test Urine UTI
Benedict’s Glucose in
Diabetes
reagent test Urine
Above is the theoretical framework. The researchers hypothesize that by studying
the causes of cirrhosis, kidney inflammation, UTI, proteinuria, and Diabetes, the
researchers will derive from it the indicators of which the tests will be based from. These
indicators are: urobilin, sulphate in urine, chloride in urine, phosphate in urine, albumin in
urine, proteins in urine, and glucose in urine. After knowing the indicators, the researchers
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studied the following test that are co-related to the indicators needed to be present for the
disease. The following tests were studied: Ammoniacal zinc chloride test for the detection
of urobilin, determination of sulphates, silver nitrate test for chlorides, determination of
phosphates, coagulation test for detection of albumin, Heller’s ring test for detection of
proteins, Benedict’s Reagent Test for detection of glucose, all of which will be used in
urine.
In view of this, the researchers have studied different kinds of literature in order to
obtain the results the researchers wanted. In the hopes of creating a simple laboratory test
for urine, the researchers have enlisted in this chapter the different related literature that
was utilized for this study to happen.
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Review of Related Literature
Diabetes mellitus
The Egyptians made the first mention of diabetes around 1500 BC. The Greek
physician Aretaeus (130–200 CE) noted a disease with symptoms of constant thirst,
excessive urination and loss of weight, and named the condition ‘diabetes’, meaning
‘flowing through’. The first clear reference to diabetes was made by an Arab physician,
Avicenna (980–1037 CE), who accurately described in detail the clinical features and
complications of the disease and its progress. During mediaeval times, an attempt was
made to identifyvarious diseases by examining urine samples for appearance, color,
sediment and often taste. It was not until the early 19th century that glucose was identified
as the sugar present (Clarke and Foster, 2012).
Diabetes mellitus is a principal cause of morbidity and mortality in human
populations. It prevents your body from properly using the energy from the food you eat.
It is a chronic disease that leads to complications including heart disease, stroke, kidney
failure, blindness and nerve damage (Steppan, 2000). There are two main types of diabetes.
Type 1 diabetes occurs because the insulin-producing cells of the pancreas (beta cells) are
damaged. People with type 1 diabetes must use insulin injections to control their blood
glucose. Type 1 is the most common form of diabetes in people who are under age 30, but
it can occur at any age. Ten percent of people with diabetes are diagnosed with type 1. In
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type 2 diabetes, the pancreas makes insulin, but it either doesn't produce enough, or the
insulin does not work properly. Nine out of 10 people with diabetes have type 2. This type
occurs most often in people who are over 40 years old but can occur even in childhood if
there are risk factors present. Type 2 diabetes may sometimes be controlled with a
combination of diet, weight management and exercise. However, treatment also may
include oral glucose-lowering medications (taken by mouth) or insulin injections (shots).
Aside from cancer and vascular disease, diabetes (vs. no diabetes) was also associated with
death from renal disease, liver disease, pneumonia and other infectious diseases, mental
disorders, no hepatic digestive diseases, external causes, intentional self-harm, nervous-
system disorders, and chronic obstructive pulmonary disease. Since diabetes is a
multisystem disorder, there is a need for adequately powered, standardized assessment of
associations of diabetes with the risk of death from a broad range of causes.
Glucose
Glucose is a simple sugar that is mainly made by plants and most algae during
photosynthesis from water and carbon dioxide, using energy from sunlight. It is the most
abundant monosaccharide, a subcategory of carbohydrates. In energy metabolism, glucose
is the most important source of energy in all organisms (Abraham, Kost and David, 1998).
Glucose is the primary energy source for the body's cells and the only energy source for
the brain and nervous system. It is a type of sugar that the body requires and uses for energy.
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The body converts the carbohydrates intake into glucose. Having too much glucose in the
body can be a sign of a health problem. If unable to receive the right treatment and glucose
levels in the body remain high, it might develop more serious complications. The most
common cause of elevated glucose levels is diabetes, a condition that affects your body’s
ability to manage glucose levels.
Urinalysis and Glucose Test in Urine
Urinalysis is a high-volume procedure that currently requires significant labor to
examine microscopic sediment (Ben-Ezra, Bork and McPhenson, 1998). Urinalysis is also
known as routine and microscopy (R&M). It is one of the most commonly ordered clinical
tests. This frequency is partly due to the ease of urine collection and testing. Urine testing
has been part of medicine for many centuries, with Hippocrates having written about urine
examination as early as 400 BC (Patel, 2006). A complete urinalysis includes physical,
chemical, and microscopic examinations. Cloudy urine often is a result of precipitated
phosphate crystals in alkaline urine, but pyuria also can be the cause. A strong odor may
be the result of a concentrated specimen rather than a urinary tract infection (Simerville,
Maxted and Pahira, 2005). Advances in Chemistry allowed significant progress in urine
testing during the nineteenth century, and the modern era of reagent strip (dipstick) testing
began in 1956. Urine testing can be used to screen for a number of disorders (Patel, 2006).
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Dipstick urinalysis is convenient, but false-positive and false-negative results can occur.
Specific gravity provides a reliable assessment of the patient’s hydration status (Simerville,
Maxted and Pahira, 2005). Urine glucose test is a quick and simple way to check for
abnormally high levels of glucose in your urine. People with diabetes could use the urine
glucose test as a way of monitoring the degree of sugar control, or efficacy of treatments.
Urine tests were once the main type of testing used to measure glucose levels for people
who potentially had diabetes.
Urobilinogen
An urobilin in urine test measures the amount of urobilinogen in a urine sample.
Urobilinogen is formed from the reduction of bilirubin. Bilirubin is a yellowish substance
found in the liver that helps break down red blood cells. Normal urine contains some
urobilinogen. If there is little or no urobilinogen in urine, it can mean that the liver isn’t
working correctly. Too much urobilinogen in urine can indicate a liver disease such as
hepatitis or cirrhosis (Hinkle, Cheever, Brunner and Suddarth, 2014). An urobilinogen test
may be part of a urinalysis, a test that measures different cells, chemicals, and other
substances in the urine. The normal urobilinogen range is 17 umol/L (<1 mg/dl). However,
it’s still considered normal to have values in the range of 0-8 mg/dl (Marchione, 2019).
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Detection of Pathological Constituents
Pathological urine constituents are substances which are not usually present in urine
such as glucose, protein, ketones, RBCs, Hb, bilirubin etc.
Abnormal urine constituents include protein urea, glucose urea, keto urea, haemato urea
and hemoglobinuria.
Protein urea is the presence of abnormal amount of protein in urine. The urine of a
healthy individual contains no protein because in normal physiology, small M.wt. protein
get to be reabsorbed by kidney tubules or proximal tubule while large M.wt. of protein
can’t pass from the kidney tubule to urine unless the kidney tubule has damage.
Glucose urea refers to the abnormal concentration of glucose in urine. Normally, glucose
is reabsorbed by active transport in proximal tubule and therefore doesn’t appear in urine
but if the blood glucose level exceeds the reabsorption capacity of kidney tubules (renal
threshold), glucose will appear in urine.
Keto urea is the presence of abnormal amount of ketone bodies in urine. Normally,
ketone bodies are removed by the liver. Elevated levels of ketone bodies in blood and urine
cause acidosis which leads to coma and death.
Haemato urea is the presence of red blood cells (RBCs) in the urine.
Hemoglobinuria is the presence of hemoglobin in urine due to rupturing of RBCs.
This may occur in malaria, typhoid, yellow fever, hemolytic jaundice and other diseases.
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Albumin
Albumin is a protein found in the blood. A healthy kidney doesn’t let albumin pass
from the blood into the urine meanwhile a damaged kidney lets some albumin pass into the
urine. The fewer albumins in your urine are better. Albuminuria is a sign of kidney disease
and it means you have too much albumin in your urine. Measuring urine albumin in
important for diagnosing kidney disease and monitoring the progression of any kidney
disease. People who have diabetes, high blood pressure, heart disease or a family history
of kidney failure are at risk for kidney disease (Albuminuria: Albumin in the Urine, para.1).
Coagulation Test
In many parts of the developing world, the urine protein heat coagulation test is
routinely used to screen for proteinuria in pregnancy. Coagulation is highly conserved
throughout biology; in all mammals, coagulation involves both a cellular (platelet) and a
protein (coagulation factor) component (Michelson, 2006). The system in humans has been
the most extensively researched and is the best understood. Standardized heat coagulation
test reliably detects significant proteinuria.
Urinary Tract Infections (UTI)
Urinary Tract Infections are among the most common bacterial infections acquired
in the community and in hospitals (Foxman, 2010). This is an infection that occurs in any
part of urinary system- kidneys, ureters, bladder and urethra. UTIs typically occur when
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bacteria enters the urinary tract through the urethra and begin to multiply in the bladder.
Women are at greater risk of developing a UTI than men. There are risk factors specific to
women for UTIs; these include female anatomy, sexual activity, certain types of birth
control, menopause, urinary tract abnormalities, and blockages in the urinary tract and
suppressed immune system.
Cirrhosis
Cirrhosis is a late stage of scarring (fibrosis) of the liver caused by many forms of
liver diseases and conditions, such as hepatitis and chronic alcoholism. Each time the liver
is injured- whether by disease, excessive alcohol consumption or another cause- it tries to
repair itself. As cirrhosis progresses more and more tissue forms making it difficult for the
liver to function. Cirrhosis often has no signs or symptoms until liver damage is extensive.
A wide range of diseases and conditions can damage the liver and lead to cirrhosis.
Kidney Inflammation
Nephritis is a condition in which the nephrons, the functional units of the kidneys,
become inflamed. This inflammation, which is also known as glomerulonephritis, can
adversely affect kidney function. There are many types of nephritis with a range of causes.
While some types occur suddenly, others develop as part of a chronic condition and require
ongoing management. It can be acute or chronic. Early symptoms may include changes in
the color of the urine and swelling of the hands and feet. Laboratory tests can confirm or
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rule out the presence of an infection. These tests include urinalysis, which tests for the
presence of blood, bacteria, and white blood cells (WBCs). A significant presence of any
of these may indicate an infection. Kidney failure occurs when one or both kidneys stop
working for a short time or permanently.
Benedict’s reagent
Benedict's Solution, or one of the many variants that evolved over the years, was
used as the reagent of choice for measuring sugar content for more than 50 years (R.
Simoni, 2002).is a chemical reagent named after American chemist Stanley Rossiter
Benedict.
It is an equal-volume mixture of aqueous solution of Copper Sulphate (CuSO4) and
sodium citrate added with sodium carbonate. Here sodium citrate is the complexing agent.
Benedict’s solution is a deep-blue alkaline solution used to test for the presence of the
aldehyde functional group- CHO (Whitson MD, 2011).
Formation of red colored copper (I) oxide indicates the formation of a precipitate.
This precipitate is insoluble in water. As Benedict’s test continues, the concentration of
reducing sugar increases. Following this condition, high amount of brick-red color
precipitate will be formed at the end of the test tube. Sometimes you will find small
amounts of copper oxide along with brick-red precipitate. Sodium carbonate of Benedict’s
reagent facilitates the alkaline conditions which are required for the redox reaction.
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Another compound Sodium citrate complexes with the copper (II) ions to avoid
degradation into copper(I) ions during storage(“Benedict’s test”, 2013).
Generally, Benedict's test detects the presence of aldehydes and alpha-hydroxy-
ketones, also by hemiacetal, including those that occur in certain ketoses. Thus, although
the ketose fructose is not strictly a reducing sugar, it is an alpha-hydroxy-ketone, and gives
a positive test because it is converted to the aldoses glucose and mannose by the base in
the reagent.
The principle of Benedict's test is that when reducing sugars are heated in the
presence of an alkali they are converted to powerful reducing species known as enediols.
Enediols reduce the cupric compounds (Cu2+) present in the Benedict's reagent to cuprous
compounds (Cu+) which are precipitated as insoluble red copper(I) oxide(Cu2O).
The color of the obtained precipitate gives an idea about the quantity of sugar
present in the solution, hence the test is semi-quantitative. A greenish precipitate indicates
about 0.5 g% concentration; yellow precipitate indicates 1 g% concentration; orange
indicates 1.5 g% and red indicates 2 g% or higher concentration.
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Chemical Formula:
Standard Outcomes from Benedict’s Test:
Conclusion
In conclusion, the literature studied were about the diseases that the researchers
want to tackle, the indicators they need, and the specific tests that will find for those
indicators.
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CHAPTER III
METHODOLOGY
Preamble
Countless of people die from diseases such as urinary tract infections, cirrhosis,
diabetes, protein in urine (proteinuria), and kidney inflammation (glomerulonephritis),
continue to pose a threat to the Filipinos and all of mankind. The purpose of this study is
to help detect these kinds of diseases by creating a simple laboratory test for urine. This
chapter aims to answer the research questions, such as, the tests that will be utilized and
how the researchers did this tests in complete detail.
The nature of this research solely lies on the different kinds of tests that will be
used by the researchers, in the hope of creating a simple laboratory tests for urine to find
indicators of the specific diseases stated above.
This chapter is divided into several sections in which addresses the research design
and method, the data collection technique, the instrumentation, the ethical consideration,
and the conclusion. For the research design and method, the researchers used the clinical
research approach, specifically for the trends of diagnostic products. For the data collection
technique the researchers used the tallying observation technique. These were used because
they were best fitted for this experimental kind of research.
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Research Design and Method
The researchers in this study have used the clinical research approach, which is a
branch of medical science that determines the safety and effectiveness of medications,
devices, diagnostic products, and treatments intended for human use. These may be used
for prevention, treatment, diagnosis or for relief of symptoms in a disease (J. Lind, 2001).
The clinical research approach was used in this study because, it can evaluate diagnostic
products and it can cater to the need of the researchers to make a product used for the
prevention of a disease, for this case, UTI, cirrhosis, diabetes, protein in urine, and kidney
inflammation (Gates, 2014).
The researchers will be doing different kinds of tests in the hopes of creating a
simple laboratory test for urine. The researchers will use the following reagents for the
different kinds of test, namely: Ammoniacal zinc chloride test for the detection of urobilin,
determination of sulphates, silver nitrate test for chlorides, determination of phosphates,
coagulation test for detection of albumin, Heller’s ring test for detection of proteins,
Benedict’s Reagent Test for detection of glucose, all of which will be used in urine. The
urine sample that will be used is a simulated urine consisting mountain dew a carbonated
drink plus egg whites.
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Data Collection Technique
In this study, the researchers used the tallying observation technique as the main
way to get data. This technique was used because it was best fitted for the study in terms
of subjects and in terms of gathering data, because the researchers were conducting
experiments. This was done by observing the reactions that would take place in each set
of tests, and after observing, the researchers would write down all the noticeable changes,
and look for the unseen changes in its chemical composition.
Instrumentation
The researchers used the CIC – Cebu Laboratory for all the tests and experiments
done. For the simulated urine, the researchers had to buy mountain dew and an egg. For
the chemicals and apparatuses, the laboratory of CIC- Cebu has provided for all the needed
chemicals and apparatuses that were used in order to complete the tests. Following this,
are the short descriptions, and the detailed experimentations done by the researchers. This
includes, the chemical equations, the materials and apparatuses used, and the steps done
in order to successfully do the experimentation.
28 | P a g e
Making the Simulated Urine

Mountain dew was used as the simulated urine for three reasons: it’s color, it’s
glucose content, and the presence of elements such as sodium citrate, chlorides,
phosphoric acid, and sodium carbonate, which are all found in urine. The egg white was
added for the researchers to add the presence of proteins. Because naturally the urine of
humans have proteins. But, since mountain dew does not have proteins, the egg white of
1 egg was added to the mountain dew. Both of these are a relatively close representation
of urine.
Equation:
30 ml mountain dew + 15ml of egg white Simulated urine
Materials Quantity
Mountain Dew 30ml
Egg white 15ml
Stirring Rod 1
Spatula 1
Beaker 2
Steps used for the experiment
1. Add 30ml of mountain dew into a beaker .
2. In another beaker, separate the egg yolk and egg white.
3. Mix 15ml of egg white to the 30ml of mountain dew and stir.
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Detection of Pigments
The reagent used for the detection of pigments is the ammoniacal zinc chloride test. Urine
added with ammonium hydroxide (NH 4OH) plus zinc chloride will then react to make a greenish
fluorescence that will indicate the presence of Urobilin. The equation, materials and their
respective quantity are stated below.
Equation:
2ml Urine + 1ml NH4OH + 1ml Zinc Chloride Light yellow fluorescence
Materials Quantity
Simulated Urine 2ml
Ammonium Hydroxide (NH4OH) 1ml
Zinc Chloride 1ml
Test tube 1 piece
Test tube rack 1
Dropper 2 for each chemical placed
1. Add 2ml of simulated urine into a test tube and slowly drop 1ml of ammonium
hydroxide and 1ml of zinc chloride
2. Observe how the urine reacts with ammonium hydroxide and zinc chloride.
30 | P a g e
Detection of Inorganic Physiological Constituents
For the detection of inorganic physiological constituents, the researchers have utilized
three different tests, namely; the detection of sulphates, the detection of chlorides, and the
detection of phosphates, respectively.
Detection of Sulphates
For the detection of sulphates, the researchers used 5ml of urine added with 5 drops of
diluted acetic acid solution. A white precipitate will form indicating bariun sulphate.
The equation, materials and their respective quantity are stated below.
Equation:
Urine + 5 drops of acetic acid & barium chloride solution white precipitate is formed
Materials Quantity
Simulated Urine 5ml
Acetic Acid (CH3COOH) 5 drops
Test tube 1 piece
Test tube rack 1
Dropper 1
1. Add 5ml of simulated urine into a test tube and slowly drop 5 drops of acetic acid and
barium chloride solution
2. Observe how the urine reacts with acetic acid.
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Detection of Chlorides
For the detection of chlorides, the researchers used 5ml of urine added with 2 drops of
diluted HNO3, 2 drops of AgNO3, and 2 drops of NH4OH. A white precipitate will form indicating
the presence of chlorides.
Equation:
5ml Urine + 5 drops of diluted HNO 3 + white precipitate is formed

2 drops of AgNO3 + 2 drops of NH4OH
Materials Quantity
Simulated Urine 5ml
Diluted Nitric Acid (HNO 3) 2 drops
Silver Nitrate (AgNO3) 2 drops
Ammonium Hydroxide (NH4OH) 2 drops
Test tube 1 piece
Test tube rack 1
1. Add 5ml of simulated urine into a test tube and slowly drop 2 drops of diluted nitric
acid, 2 drops of silver nitrate, and 2 drops of ammonium hydroxide
2. Observe how the urine reacts with nitric acid, silver nitrate, and ammonium
hydroxide.
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For the detection of phosphates, the researchers added 5ml of urine and ammonium
hydroxide, then after the solution is heated by a hot water bath. After heating there will be a white
precipitate that will be formed, the researchers filtered this out thus indicating the presence of
phosphates.
Equation:
heating and filter
5ml Urine + 5 ml of NH4OH white precipitate is formed
Materials Quantity
Simulated Urine 5ml
Ammonium Hydroxide (NH4OH) 5ml
Test tube 1 piece
Test tube rack 1
Beaker 1
Hot water 100ml
Filter Paper 1 piece
Funnel 1
1. Add 5ml of simulated urine into a test tube and add 5ml of ammonium hydroxide
2. Heat the solution in a hot water bath of 100ml
3. Observe and filter out the precipitate formed then add the magnesium chloride.
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Detection of Pathologic Constituents
For the detection of physiological constituents, the researchers have utilized three different
tests, namely; the detection of albumin, the detection of protein, and the detection of glucose,
respectively.
Detection of Albumin
The coagulation test will be used for the detection of albumin in urine. This is done by
utilizing 5ml urine boiled then added with 3 to 4 drops of diluted CH3COOH then heat by a
water bath. Then a flocculent precipitate indicates the albumin.
Equation:
5ml Urine + 5 drops of diluted acetic acid flocculent precipitate is formed
Materials Quantity
Simulated Urine 5ml
Acetic Acid (CH3COOH) 3 drops
Test tube 1 piece
Test tube rack 1
Dropper 1
Bunsen Burner 1
Test tube holder 1
1. Add 5ml of simulated urine into a test tube and slowly drop 3 drops of acetic acid,
then heat.
2. Observe how the urine reacts with acetic acid and filter out the ppt. formed.
34 | P a g e
Detection of Protein
For the detection of protein, the researchers used the Heller’s ring test. To do this test, one
must prepare a protein containing solution then add to it concentrated HNO 3. Afterwards, the
urine will float in the nitric acid, observe for 5-10 minutes. After observing, a white ring will be
formed indicating the presence of protein.
Equation:
5ml Urine + 5 ml HNO3 white ring is formed
Materials Quantity
Simulated Urine 5ml
Nitric Acid ( HNO3) 5ml
Test tube 1 piece
Test tube rack 1
Dropper 1
1. Add 5ml of simulated urine into a test tube and 5ml of nitric acid.
2. Wait for 5 to 10 minutes and observe how the urine reacts with nitric acid.
35 | P a g e
Detection of Glucose
For the detection of glucose, the researchers used the Benedict’s reagent test.
Benedict’s reagent contains copper sulphate, sodium carbonate, sodium citrate. The Benedict’s
will be added to 5ml of urine, after adding the reagent, the solution will be boiled, the color will
change from the spectrum of yellow to red depending on the level of glucose present in urine.
Equation:
5ml Urine + 5ml Benedict’s reagent Color changes from yellow to red
Materials Quantity
Simulated Urine 5ml
Benedict’s Reagent 5ml
Test tube 1 piece
Test tube rack 1
Dropper 1
Bunsen Burner 1
Test tube holder 1
1. Add 5ml of simulated urine into a test tube and 5ml of Benedict’s reagent.
2. Heat until there is a change of color occurs.
3. Observe the reaction that takes place.
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Chemical Formulas
Simulated Urine
For the simulated urine, the composition contains mountain dew and egg whites, to
produce a solution relative to urine.
CO 2 + H 2O + C 12H 24O 12 + H 3PO 4 + C 6H 8O 7 + Cl - + Na 2CO 3 + C 6H 8N 2O 4

( Carbon dioxide) (Water) (Glucose) (Phosphoric Acid) (Citric Acid) (Chloride) (Sodium Carbonate) (Egg white)
+(NH4)2SO4
(ammonium sulphate)
CO 2 + H 2O + C 12H 24O 12 + H 3PO 4 + CON 2H 4 + C 5H 4N 4O 3

( Carbon dioxide) ( Water) ( Glucose) ( Phosphoric Acid ) ( Urea) (Uric Acid )
+ C 6H 8O 7 + NaCl - + Na3C6H 5O 7 + C 6H 8N 2O 4 +(NH4)2SO4

(Citric Acid) (Sodium Chloride) (Sodium Citrate) ( Proteins) (ammonium sulphate)
Ammoniacal Zinc Chloride Test for Determination of Urobilin

Urine added with ammonium hydroxide and zinc chloride to form urobilin.
Urine + NH4OH + ZnCl2 H2O + CON2H4 + C5H4N4O3 + C33H42N4O6

(Ammonium OH) (zinc chloride) (Water) (Urea) (Uric Acid) (Urobilin)
Urine added with acetic acid & barium chloride form a white precipitate or barium sulphate.
Urine + CH3COOH + BaCl2 H2O + CON2H4 + C5H4N4O3 + BaSO4

(Acetic Acid Solution) (Barium Chloride) (Water) (Urea) (Uric Acid) (Barium Sulphate)
Urine added with nitric acid, silver nitrate, and ammonium hydroxide to make a white
precipitate that is an indicator of chlorides.
Urine + HNO3 + AgNO3 + NH4OH H2O + CON2H4 + C5H4N4O3 + Cl- + AgNO3

(Nitric Acid) (Silver Nitrate) (Ammonium OH) (Water) (Urea) (Uric Acid) (Chloride) (Silver Nitrate)
37 | P a g e
Urine added with ammonium hydroxide then heated to make a white precipitate that is an
indicator of phosphates.
Urine + NH4OH heated H2O + CON2H4 + C5H4N4O3 + (NH4)3PO4

(Ammonium OH) (Water) (Urea) (Uric Acid) (Ammonium Phosphate)
Urine added with acetic acid solution then boiled to form a flocculent precipitate that indicates
albumin.
boil
Urine + CH3COOH H2O + CON2H4 + C5H4N4O3 + C123H193N35O37
(Acetic Acid Solution) (Water) (Urea) (Uric Acid) (Albumin)
Urine added with nitric acid to make a white ring around the solution that is an indicator of
simple proteins.
Urine + HNO3 H2O + CON2H4 + C5H4N4O3 + C6H8N2O4

(Nitric Acid) (Water) (Urea) (Uric Acid) (Proteins)
Urine added with Benedict’s reagent to form a precipitate that ranges from the spectrum of
yellow to red depending on the glucose level of the solution.
Urine + Cu3C 12H10O14 H2O + CON2H4 + C 5H4N4O3 + Cu2 O

(Benedicts Reagent) (Water) (Urea) (Uric Acid) (Oxidized Copper)
38 | P a g e
Sample
The samples used by the researchers were just the simulated urine, consisting of the
mountain dew and egg white. The simulated urine was divided into 7 test tubes each of
which would have a different test done on them for detection of the constituents of urine.
This sample size was enough because the researchers wanted to produce seemingly
simple, and doable tests for urine, because the researchers only aimed to observe seven
constituents that can be found in urine, and because only the seven are most likely to cause
diseases.
Ethical Considerations
Before experimenting the researchers booked the laboratory with consent from their
adviser, and the laboratory safe-keeper. The experimentation was conducted with full
supervision of Engineer Jocelyn Gonzales. It was insured that there will be minimal risks
to the researchers as well as the assistants of the researchers. While doing the experiments,
the laboratory assistants were fully aware of all the things the researchers were doing,
because they were told beforehand about what will happen, and what to expect. The
researchers guarantee 100% safety before, on, and after the experiment has been done.
39 | P a g e
Conclusion
In conclusion, the researchers has done 7 different tests, which are the ammoniacal
zinc chloride test for the detection of urobilin, determination of sulphates, test silver nitrate
test for chlorides, determination of phosphates, the coagulation test for detection of
albumin, Heller’s ring test for detection of proteins, Benedict’s Reagent Test for detection
of glucose, all of which will be used in urine.
The researchers used the tallying observation technique as the main way to get
data. This was done by observing the reactions that would take place in each set of tests,
and after observing the researchers would write down all the noticeable changes, and look
for the unseen changes in its chemical composition.
In addition, the researchers used the CIC – Cebu Laboratory for all the tests and
experiments done. All the materials and apparatuses used were from the laboratory, except
for the mountain dew and egg which were bought by the researchers.
The experimentation was done with thorough care and deliberation, and was
supervised all throughout by Engineer Jocelyn Gonzales. The researchers guarantee that
the experiments were 100% safe and doable.
40 | P a g e
CHAPTER IV:
DATA ANALYSIS
Preamble
In the years to come, cases such as urinary tract infection, cirrhosis, diabetes,
protein in urine (proteinuria), and kidney inflammation (glomerulonephritis), will continue
to rise, especially with the new lifestyle that is being incorporated by each and every human
being today. This study aims to find a way to help diminish the devastation brought by
these diseases, thus the researchers made a simple laboratory test for urine, that will detect
these kinds of diseases.
This researchers aim to objectify the following: the tests that will be utilized by the
researchers, the result/s that will indicate UTI, cirrhosis, diabetes, protein in urine, and
kidney inflammation, the significance of the said laboratory tests, and this research’s effect
on the field of medicine. The researchers hypothesize that by using the following tests they
can indicate the diseases said above.
The tests that will be utilized are the following: the ammoniacal zinc chloride test
for detection of pigments, the adding of acetic acid for detection of sulphates, the silver
nitrate test for chlorides, the ammonium hydroxide test for phosphates, coagulation test for
41 | P a g e
albumin, Heller’s ring test for proteins, and the benedicts reagent test for glucose. These
specific tests will be used to determine the presence of different kinds of compounds found
in urine.
The researchers used an experimental design for the research. The data collection
used was the tallying observation technique, for this was most fit for experiments.
Presentation and Analysis of Data
The researcher have seven different kinds of experiment each of which will be
analyzed below. The experiments include a test for detection of pigments, detection of
sulphates, detection of chlorides, detection of phosphates, detection of albumin, detection
of proteins, and detection of glucose.
Additionally, all the materials and apparatuses used were from the CIC-Cebu
laboratory. For each experiment, there will be a picture of actual experiment that was
conducted, an analysis of the tallied observation, and a detailed explanation.
42 | P a g e
Making of the Simulated Urine
The simulated urine was accomplished by mixing mountain dew plus egg whites.
The ratio of the mountain dew and egg white is 2:1, specifically 30ml for mountain dew,
and 15ml for the egg white.
Visualization
Above is the visualization, of the simulated
urine. This was used for all the tests. Mountain dew
was used because it’s color, it’s glucose content,
and the presence of elements such as sodium
citrate, citric acid, chlorides, phosphoric acid, and
sodium carbonate, which are all found in urine.
The egg white was added to add the presence of
proteins.
The simulated urine was separated into 7 different tubes, for each of the following
tests: Ammoniacal zinc chloride test for the detection of urobilin, determination of sulphates, silver
nitrate test for chlorides, determination of phosphates, coagulation test for detection of albumin,
Heller’s ring test for detection of proteins, Benedict’s Reagent Test for detection of glucose, all of
which will be used in urine.
43 | P a g e
Detection of Pigments
The reagent used for the detection of pigments is the ammoniacal zinc chloride test.
Urine added with ammonium hydroxide (NH4OH) plus zinc chloride will then react to
make a greenish fluorescence that will indicate the presence of Urobilin.
Visualization
The adding of simulated urine to the test tubes
The adding of ammonium hydroxide and zinc chloride to the simulated urine
44 | P a g e
The final solution
Analysis
After adding the ammonium hydroxide and zinc chloride, the solution turned to a
light yellow thus indicating the presence of minimal of urobilin. Urobilin is the substance
that is formed from the reduction of bilirubin. Bilirubin is a yellowish substance found in
your liver that helps break down red blood cells. Normal urine contains some urobilinogen.
If there is little or no urobilinogen in urine, it can mean your liver isn't working correctly.
Too much urobilinogen in urine can indicate a liver disease such as hepatitis or cirrhosis
(J. Hinkle, 2014). But since the simulated urine is not exactly the same as urine, the
presence of urobilin is not clearly seen, because urobilin is only synthesized inside the liver
where enzymes breakdown bilirubin to form urobilin.
Thus, the researchers can say that using ammonium hydroxide and zinc chloride
can be used for the detection of pigments in urine.
45 | P a g e
Inorganic Physiological Constituents
For the detection of sulphates, the researchers used 5ml of urine added with 5 drops
of diluted acetic acid and barium chloride solution.
Visualization
7th
6th 5th 4th 3rd 2nd 1st
The adding of simulated urine to the test tubes – the second test tube
7th
The adding of acetic acid and barium chloride to the simulated urine –
the second test tube
46 | P a g e
3rd 2nd 1st
The final solution – the second test tube
Analysis
The researchers have observed that after adding the acetic acid and barium chloride
solution, there were white precipitate that formed, indicating the presence of barium
sulphate. Though very minimal, there were white precipitate floating in the solution
indicating the presence of barium sulphate.
barium sulphate is a kind of sulphate that is formed when acetic acid is added to
urine. The acetic acid breaks down the bonds of the ammonium sulphate, thus creating free
radicals such as, the ammonia and the sulphate group, and by adding barium chloride the
sulphates react with the barium, creating barium chloride.Urinary sulfate excretion is
higher in patients who have kidney stones than in individuals who do not form stones, and
higher for diabetes (W. Tschope, 1985).

Thus, the researchers conclude that one can test for the presence of sulphates by
adding the acetic acid.
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For the detection of chlorides, the researchers used 5ml of urine added with 2 drops
of diluted HNO3, 2 drops of AgNO3, and 2 drops of NH4OH.
Visualization
The preparation of test tube 3
7th
3rd
The adding of nitric acid, silver nitrate, and ammonium hydroxide
48 | P a g e
3rd
After one week observation
Analysis
The researchers have observed that after adding nitric acid, silver nitrate, and
ammonium hydroxide, there was a white precipitate that was formed. This white precipitate
indicates the presence of chlorides, after one week the researchers have observed that the
excess silver nitrate has deteriorated, thus turning the white precipitate into dark brown.
An increased level of urine chloride usually indicates dehydration, but can also
occur with other problems that cause high urine chloride, such as kidney diseases.
Hypochloremia also occurs when too much base is lost from the body or when a person
hyperventilates (Battle D., 2013).
Decreased urine chloride level may be due to fluid loss, decreased salt intake,
sodium retention, diarrhea, vomiting, and/or gastric suction ( Tolwani AJ, 2017)
Thus, the researchers conclude a positive for this test.
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For the detection of phosphates, the researchers added 5ml of urine and ammonium
hydroxide, then after the solution is heated by a hot water bath.
Visualization
Preparation of hot water bath
4th
The adding of ammonium hydroxide to the 4th test tube
50 | P a g e
The heating of the test tube through hot water bath
Analysis
The researchers, after adding ammonium hydroxide and after heating through hot
water bath, lumps of white precipitate formed indicating phosphates. The phosphates that
was formed was ammonium phosphate.
The ammonium hydroxide reacts with the urine, and when heated the ammonium
hydroxide changes in enthalpy, thus creating ammonia gas, and causes the ammonium to
react with the phosphoric acid found in urine, which in return creates ammonium
phosphate.
Phosphates in urine would most likely indicate kidney stones and cirrhosis (Khatri
MD, 2017). Thus, in lieu of this reaction, the researchers conclude that using ammonium
hydroxide for urine and then heating it can be used for detection of phosphates.
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Detection of Pathological Constituents
The coagulation test will be used for the detection of albumin in urine. This is done
by utilizing 5ml urine that is boiled. after which is added with 3 to 4 drops of diluted acetic
acid solution, then heated again by a water bath.
Visualization
The boiling of the simulated urine then adding of acetic acid
The re-heating of solution
52 | P a g e
Filtering of the white precipitate that was formed

after re-heating the solution
Residue
The researcher have
observed that when they boiled and
then added acetic acid and then re-
heated it again, there was a white
precipitate that was formed.
The white precipitate formed indicates the presence of albumin. Albumin is a
type of protein found in large amounts in the blood. Because it is a small molecule in
size, it is one of the first proteins able to pass through the kidneys into the urine when
there are kidney problems. This presence of small amounts of albumin in the urine is
the condition called microalbuminuria. As kidney damage progresses and the amount
of albumin in the urine increases, the name of the condition changes from
microalbuminuria to albuminuria or proteinuria (M. Afkarian, 2016).
With the results, the researchers concluded that one can use this test for the
detection of albumin.
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Detection of Proteins
For the detection of protein, the researchers used the Heller’s ring test. One must
prepare a protein containing solution then add to it concentrated HNO3.
Visualization
The adding of nitric acid to the simulated urine
The reaction is taking place
54 | P a g e
6th
After one week observation
The researchers have observed that when nitric acid is added to the solution, a white
precipitate is formed, but more seemingly, there was a ring that was formed after 2-3
minutes. The ring indicates the presence of proteins.
Similar to the detection of albumin, the Heller’s test also tests for proteins. The
difference is that the proteins tested for the Heller’s test is for all the types of proteins,
while albumin is just a kind of protein that is specifically from the kidney. Total protein
measurements can reflect nutritional status and may be used to screen for and help
diagnose kidney disease or liver disease, for example. Sometimes conditions are detected
with routine testing before symptoms appear. If total protein is abnormal, further testing
must be performed to identify which specific protein is abnormally low or high (DW
Landry, 2016).
Thus, the researchers conclude that Heller’s ring test is a great test for proteins.
55 | P a g e
For the detection of glucose, the researchers used the Benedict’s reagent test. The
Benedict’s reagent was added to 5ml of urine, after adding the reagent, the solution was
boiled.
Visualization
The adding of Benedict’s reagent to the simulated urine
The heating of the solution
56 | P a g e
The result
The researchers observed that when the Benedicts reagent is added with the
simulated urine, it turned a little bit green, but when it was heated it turned to a yellow-
orange color. The color indicates the presence of glucose. In the Benedict’s reagent
spectrum it is seen that yellow-orange solution contains about 2.6-3.5% of glucose.
The most common cause of elevated glucose levels is diabetes, a condition that
affects your body’s ability to manage glucose levels. It’s important to monitor your glucose
levels if you have already been diagnosed with diabetes (B. Holloway, n.d.). The normal
amount of glucose in urine is 0 to 0.8 mmol/L (millimoles per liter) which is less than 1%
mmol/L. A higher measurement could be a sign of a health problem.
In conclusion, the researchers conclude that the Benedict’s reagent can be used in
detection of glucose in urine.
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Further Analysis of Data
Ammoniacal Zinc Chloride Test
This test resulted a positive giving away a light-yellow solution, indicating the
presence of little urobilin. Urobilin is formed from the reduction of bilirubin. Bilirubin is a
yellowish substance found in your liver that helps break down red blood cells. Normal
urine contains very minimal urobilin, if the contrary were to happen, that would be one
indicator that the patient has liver disease such as cirrhosis (J. Hinkle, 2014).
Based on the National Institute of Diabetes and Digestive and Kidney Diseases, low levels
of urobilin (clear – greenish fluorescent - light yellow) may indicate:
• A blockage in the structures that carry bile from your liver
• A blockage in the blood flow of the liver
• A problem with liver function
While, a higher-than-normal level of urobilin (dark yellow – red) may indicate:
• Hepatitis
• Cirrhosis
• Liver damage due to drugs
• Hemolytic anemia, a condition in which red blood cells are destroyed before they
can be replaced. This leaves the body without enough healthy red blood cells
Normal urobilin levels (light yellow- medium dark yellow) may indicate:
• normal metabolic functions
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Urinary sulphate is a reflection of dietary protein intake, particularly meat, fish, and
poultry, which are rich in sulfur-containing amino acids methionine and cysteine. Urinary
sulphate can be used to assess dietary protein intake for nutritional purposes. A protein-
rich diet has been associated with an increased risk for stone formation, possibly due, in
part, to an increase in urinary calcium excretion caused by acid production from
metabolism of sulfur-containing amino acids (W. Tschope, 1985) . The normal excretion
of sulphates is 1 gram/L
High sulphate levels in urine indicate:
• high protein diets
• kidney inflammation.
Low sulphate levels in urine indicate:
• renal impairment
• homocystinuria
Sulfate is a major anion in the urine that has significant affinity for cations and
modulates the availability of cations for reacting with other anions in the urine. It thus is
an important factor of urinary supersaturation for various crystals or stones such as calcium
oxalate, hydroxyapatite, and brushite. Urinary sulfate also has a major impact on buffering
or providing hydrogen ions and as such modulates the supersaturation of uric acid.
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The urine chloride test measures the amount of chloride in a certain volume of
urine. Chloride is an electrolyte. It is a negatively charged ion that works with other
electrolytes, such as potassium, sodium, and bicarbonate, to help regulate the amount of
fluid in the body and maintain the acid-base balance (D. Batlle, 2013) . The normal range
is 110 to 250 mEq/L per day. This range depends on the amount of salt and fluid you take
in.
A higher than normal urine chloride level may be due to:
• Low function of the adrenal glands
• Inflammation of the kidney that results in salt loss (salt-losing nephropathy)
• Production of an unusually large amount of urine (polyuria)
• Too much salt in the diet
Decreased urine chloride level may be due to:
• Body holding in too much salt (sodium retention)
• Cushing syndrome
• Decreased salt intake
• Fluid loss that occurs with diarrhea, vomiting, sweating, and gastric suction
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A phosphate in urine test measures the amount of phosphate in your urine.
Phosphate is an electrically charged particle that contains the mineral phosphorous.
Phosphorous works together with the mineral calcium to build strong bones and teeth. It
also plays an essential role in nerve function and how the body uses energy.
Your kidneys control the amount of phosphate in your body. If you have a problem
with your kidneys, it can affect your phosphate levels. Phosphate levels that are too low or
too high can be a sign of a serious health problem (J. Hopkins, 2018).
If test shows high phosphate/phosphorous levels, it may indicate:
• Kidney disease
• Too much vitamin D in your body
• Hyperparathyroidism
If test shows low phosphate/phosphorous levels, it may indicate:
• Liver disease
• Malnutrition
• Alcoholism
• Diabetic ketoacidosis
• Osteomalacia (also known as rickets), a condition that causes bones to become soft
and deformed. It's caused by a vitamin D deficiency.
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Measurement of urine albumin is an important tool for diagnosing kidney disease,
and monitoring the progression of kidney disease. Health care providers regularly test
people for albuminuria as part of a routine medical exam and will closely monitor urine
albumin in people with kidney disease (M. Afkarian, 2016).
A urine albumin level that stays the same or goes down may mean that treatments
are working. Treatment that lowers the urine albumin level may lower the chances that
kidney disease will progress to kidney failure.
People who have diabetes, high blood pressure, heart disease, or a family history of
kidney failure are at risk for kidney disease such as proteinuria. Talk with your health care
provider about how often you should get a urine test for albumin.
Low albumin levels may be a sign of:
• Trauma, such as a burn injury
• Chronic illness
• Liver disease
• Certain infections
• Inflammation
High prealbumin levels may be a sign of Hodgkin disease, kidney problems, or other
disorders, but this test is not used to diagnose or monitor conditions related to high
prealbumin. Other types of lab tests will be used to diagnose these disorders.
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Detection of Proteins
A protein in urine test measures how much protein is in your urine. Proteins are
substances that are essential for your body to function properly. Protein is normally found
in the blood. If there is a problem with your kidneys, protein can leak into your urine. While
a small amount is normal, a large amount of protein in urine may indicate kidney disease.
If a large amount of protein is found in your urine sample, it doesn't necessarily
mean that it has a medical problem needing treatment. Strenuous exercise,
diet, stress, pregnancy, and other factors can cause a temporary rise in urine protein levels.
Your health care provider may recommend additional urinalysis tests if a high level of
protein is found This testing may include a 24-hour urine sample test (MJ Manary, 2016).
If urine protein levels are consistently high, it may indicate kidney damage or other
medical condition. These include:
• Urinary tract infection
• Lupus
• High blood pressure
• Preeclampsia, a serious complication of pregnancy, marked by high blood pressure.
If it is not treated, preeclampsia can be life-threatening to the mother and baby.
• Diabetes
• Certain types of cancer
63 | P a g e
A glucose in urine test measures the amount of glucose in your urine. Glucose is a
type of sugar. It is your body's main source of energy. A hormone called insulin helps move
glucose from your bloodstream into your cells. If too much glucose gets into the blood, the
extra glucose will be eliminated through your urine. A urine glucose test can be used to
help determine if blood glucose levels are too high, which may be a sign of diabetes (FF
Ferri, 2018).
Glucose is not normally found in urine. If results show glucose, it may be a sign of:
• Diabetes
• Pregnancy. As many half of all pregnant women have some glucose in their urine
during pregnancy. Too much glucose may indicate gestational diabetes.
• A kidney disorder
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Summary
To sum it all up, the researchers conclude that all 7 tests are utilizable for detection
of the different constituents of urine.
TEST PRODUCT
Ammoniacal Zinc Chloride Test Urobilin
Detection of Sulphates Barium Sulphate
Detection of Chlorides Free radical Chloride
Detection of Phosphorus Ammonium Phosphate
Detection of Albumin Albumin
Heller’s ring test Proteins
Benedict’s reagent test Oxidized Copper
Here is a tabulates summary of the tests. For the ammoniacal zinc chloride test, it
resulted a positive, giving away light yellow urobilin, which indicates the pigments in
urine. High urobilin levels may indicate cirrhosis, liver damage, etcetera. For the detection
of sulphates, barium sulphate was the product of the reaction. High levels of sulphates may
indicate kidney inflammation. For the test for chlorides, the reactant side formed a free
radical chloride atom. High levels of chloride may indicate inflammation of the kidney.
For the detection of phosphorus, ammonium phosphate was the bi-product. High levels of
phosphorus in urine may indicate kidney diseases such as UTI. For the Detection of
65 | P a g e
albumin, albumin was really formed. High levels of albumin in urine may indicate kidney
diseases such as proteinuria. For the Heller’s ring test, protein was found to be present.
High levels of protein in urine may indicate UTI and/or diabetes. For the Benedict’s reagent
test, glucose was found to be present. High levels of glucose may indicate diabetes.
All the tests that were utilized by the researchers resulted a positive. They are all
usable for the detection of the different constituents of urine. This now brings a new light
to the field of chemical examination of urine. For the different types of chemicals used by
the researchers are all just available and usable by any laboratory.
For the future good of the world, this study now concludes that all the tests are
utilizable and doable. A simple, specific, sensitive, and speedy tests have been conducted
by the researchers, thus creating the new light for chemical examination of urine.
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CHAPTER V:
SUMMARY, DISCUSSION, RECOMMENDATIONS
Preamble
For the greater good of the future, the researchers conducted this research to create
a simple laboratory test for urine. This test would be able to indicate diseases such as
urinary tract infection, cirrhosis, diabetes, protein in urine (proteinuria), and kidney
inflammation (glomerulonephritis). The Philippines being a 3rd world country suffers from
bad health care systems, which is one of the reason why the researchers opted to create this
study. The researchers hypothesized that by using different kinds of simple reagents they
can determine different constituents of urine. The sample was the simulated urine made by
the researchers, and the data collection technique that was used was the tallying observation
technique.
This chapter aims to give a recap on the things that were found on the tests. This
chapter also aims to help readers understand more the outcome of the research. In addition,
this chapter aims to highlight major findings that have been stated in the previous chapter.
This chapter further aims to refresh the minds of the readers, to reorganize their thoughts
and remind them that it is doable to do a simple laboratory test for urine.
The main sections of this chapter include the summary of the findings, the
discussion of the researchers, and the researchers recommendation and suggestions.
67 | P a g e
Summary
In totality, all seven experiments prove to accomplish a positive outcome. Below
are the tests that were used and the results they formed:
• By using the ammoniacal zinc chloride test, the researchers concluded that
it determines the presence of urobilin
• By adding acetic acid solution, the researchers concluded that it determines
the presence of sulphates.
• By the adding of nitric acid, silver nitrate, and ammonium hydroxide
solution, the researchers concluded that it determines the presence of
chlorides.
• By the adding of ammonium hydroxide solution then followed by heating,
the researchers concluded that it determines the presence of phosphates.
• By the adding of acetic acid solution then followed by boiling, the
researchers concluded that it determines the presence of albumin.
• By the adding of nitric acid, the researchers concluded that determines the
presence of proteins.
• By the adding of Benedict’s reagent, the researchers concluded that it
determines the presence of glucose.
68 | P a g e
In addition, here is the tabulated form of the constituents and their indicated diseases:
CONSTITUENTS MAY INDICATE
Urobilin Cirrhosis and/or Liver Damage
Sulphates Kidney Inflammation
Chlorides Kidney Inflammation
Phosphates UTI
Albumin Proteinuria
Proteins UTI and/or Diabetes
Oxidized Copper (colored ppt. from test) Diabetes
The researchers, based on previous medicinal studies, concluded that the presence
of urobilin in urine may indicate cirrhosis and/or liver damage. The researchers also
concluded that high presence of sulphates and chlorides may indicate that the sample has
kidney inflammation. In connection, high levels of phosphates in urine may indicate
urinary tract infection (UTI). Furthermore, high presence of albumin may indicate that the
sample has proteinuria. Also, high levels of proteins in urine, may indicate that the sample
has UTI or diabetes. In addition, high level of glucose in urine may indicate that he/she has
diabetes.
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Discussion
In the world today, there are many competing health care products. But the
researchers of this study has proudly claimed that this study is an investment to look into.
This study not only makes chemical laboratory much easy, it shines a new light in the field
of medicine, specifically in the field of urinalysis.
The researchers of this study, guarantee that this research is a simple, specific,
sensitive and speedy test for urine.
This research his at edge compared to other researches. This research is relevant
nowadays because it aims to also provide cheaper health care services, not only for the
Filipino but also for the world, because not only does it use cheaper chemicals. The
experiments utilized were also very easy to use.
For example the making of the zinc chloride test, usually what normal laboratories
do, they use amino benzaldehyde, searching this up in the internet, one can buy this for
150.43 pesos per liter bases on the Indian market. Comparing it to buying ammonium
hydrazide and zinc chloride, which is just 150 for 5 liters and 200 for 3 liters, respectively,
based on Lazada’s medical supplies. If I were to buy 3 liters of each compounds, the
70 | P a g e
ammoniacal zinc chloride solution would be much cheaper than buying 3 liters of amino
benzaldehyde.
For the detection of sulphates and albumin, this can be done in our own very homes,
acetic acid can just be found I your home simply as vinegar.
For the detection of chlorides and proteins, there were no much differences because
these two tests are still the best simple tests we have for the detection of glucose and
proteins, but the tests utilized by the researchers are still very fruitful in terms of data and
analysis.
For the detection of glucose, the researcher used the Benedict’s reagents. Usually,
the reagent used is the glucose oxidase peroxidase test. But since the glucose oxidase test
is very expensive, the researcher opted to use the Benedict’s solution as a replacement.
Though there are differences, the produce is still to detect glucose. The data from
Benedict’s reagent is relatively similar to that of glucose oxidase test.
We are not here on Earth to just spectate. This study could save a life. This study
could bring life to those who need it. Prevention is better than cure. This research aims to
make a simple laboratory test for the detection of different kinds of constituents, and help,
not only the Filipinos, but the whole world.
71 | P a g e
Recommendations
The researchers would like to recommend this study to the following:
To the aspiring-to-be medical students who will be enrolling in the Science,
Technology, Engineering and Mathematics (STEM) strand. This study aims to show the
readers, or the world rather, that one can utilize simple chemicals to perform a simple
laboratory test for urine. We recommend the future students to incorporate paper
chromatography into the study, for the researchers could not do it, because the researchers
were lacking of time.
We would also like to recommend that this study, it’s data, and it’s analyzations,
be further looked into by scientist, and medical technicians. Thought the research proved
to obtain a positive result, this study was still conducted by amateur students. May they
also expound further the concepts of using a simple laboratory test for urine in the field of
medicine.
Lastly, to the government, specifically Department of Health. We the researchers
would like to recommend this study to the DOH for them to further use this as a stepping
stone to creating cheap and affordable urinalysis for our fellow country-men who are
poverty situations, who cannot afford good quality health care.
72 | P a g e
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Simple Laboratory Test for Urine

THE PROBLEM AND ITS SCOPE

Background of the Study

protein in urine (proteinuria), and kidney inflammation (glomerulonephritis) have risen,

people still lose their lives to it.

of UTI increases with increasing duration of catheterization. In non-institutionalized

nearly 25% of all infections (B. Foxman, 2002).

liver is injured — whether by disease, excessive alcohol consumption or another cause —

cirrhosis). Advanced cirrhosis is life-threatening. The liver damage done by cirrhosis

steps being taken to stem the tide (Castilo, R. 2018).

Protein in urine — known as proteinuria — is excess protein found in a urine sample

often a sign of kidney disease (Khatri MD, 2018).

In addition, other rampant diseases include kidney inflammation

(glomerulonephritis). Glomerulonephritis is the inflammation of the tiny filters in kidneys

diabetes. Severe or prolonged inflammation associated with glomerulonephritis can

(Murrel MD, 2018).

diseases costs less than actually treating it.

STATEMENT OF THE PROBLEM

leads us to thinking that it is better to have health than wealth.

the world (D.O.H. 2015).

more problems if left undiscovered and untreated.

on understanding the indicators of urinary tract infection, cirrhosis, diabetes, protein in

urine, and kidney inflammation.

specifically, it aims to find the answers of the following questions:

1. What are the tests that will be utilized by the researchers?

3. What is the significance of the said laboratory tests?

on the field of medicine.

by visual examinations, by chemical examinations, and by microscopic examination

(Urinalysis, 2017). Under chemical examinations, the researchers hypothesize that a

oldest tests known in clinical chemistry (Alfred H. et al 1957).

out what they indicate to the health factor of our body.

Laboratory Tests for Urine

Different types of Laboratory

Coagulation Test for albumin

Heller’s Ring Test for proteins

Benedict’s Reagent Test for glucose.

- used as a simple test for urine.

In alignment with the world’s different kinds of urinalysis,

the researchers decided to use different reagents as listed

The outcome of the research

outcome of the tests itself. On the other hand, the

independent variable found are: the different reagents, the

way of testing their urine.

incorporate modern technology, thus creating more advanced medicinal approaches to

reagents, to test urine.

LIMITATIONS OF THE STUDY

underage. In addition, urine, being a waste product of humans, is strictly prohibited to be

resources were not sufficient to back up the study.

In lieu of this paper, the researchers have found 23 highfalutin words of

which definitions are given below.

Benedict’s Reagent - is used as a simple test for reducing sugars.

Conceptual framework- is an analytical tool with several variations and contexts. It is

used to make conceptual distinctions and organize ideas.

Devastation- great destruction or damage.

Diminish- make or become less.

Epidemic- The occurrence of more cases of a disease than would be expected in a

community or region during a given time period.

individuals at the same time.

Factors - matters affecting other medical conditions diagnosed when attitudes or

Healthcare- the organized provision of medical care to individuals or a community.

Highfalutin- expressed in or marked by the use of language that is elaborated or heightened by

artificial or empty means.