Biomolecules

Urine Biomarkers

Urine Biomarkers: Qualitative Determination of the Inorganic, Organic,

and Pathological Organic Constituents of Urine via Urinalysis
2019. Valdez, L; Catama, D; Miguel, M; Balangay, R; Yabut, C; Esporo, E.
Department of Biology, School of Natural Sciences,
Saint Louis University, Baguio City, Philippines

Abstract
An estimated 5 to 10 million people died from kidney disease in 2018 alone and about 150 million people are infected
annually with urinary tract infection. Statistically, kidney and urinary tract diseases ranked as the 12 th among the causes
of death and 17th among the causes of disability annually worldwide. In the Philippines, end-stage renal disease is the
7th leading cause of death. In Baguio City, 9.87% of 700 pupils was tested positive for urinary tract infection using the
dipstick method. Detection of these diseases can be easily done via urinalysis, a diagnostic tool that determines the
presence of certain urine constituents. Hence, this study aimed to determine the presence of certain inorganic, organic,
and pathological organic constituents in the urine samples as biomarkers to suggest indications of it to human health.
The scope of this study, however, was delimited to the qualitative tests in urinalysis. As such, data interpretation for the
inorganic and organic constituents that are normally found in the body was adjusted; pathological organic constituents,
however, was not adjusted. A total of randomly sampled 12 individuals (6 males and 6 females) aged 20 to 21 years
old participated in the study and about 200ml urine samples per individual were collected 24 hours before testing, using
toluene (C7H8) as preservative and amber bottle as the container. Based on the results of the tests for physical
properties, both of the randomly selected urine samples exhibited normal condition. For the results of the qualitative
tests for urine biomarkers, generally, most of the participants are also in normal condition. However, there is still a
probability of exhibiting diseases of the kidney and urinary tract, hepatobiliary diseases, diabetes, and heart diseases
for some of the participants. Overall, the results of the tests for physical properties and the qualitative tests for urine
biomarkers are influenced by certain lifestyle factors such as the use of diuretics, dehydration, starvation, increased or
decreased intake of foods having certain constituents found in urine, and others. The researchers recommend to
reproduce the qualitative tests results using the methodology used in the study and via the dipstick method; to conduct
quantitative tests in urinalysis using spectroscopy methods such as the near-infrared spectroscopy, surface-enhanced
Raman spectroscopy, or the Fourier transform infrared vibrational spectroscopy; and to have a follow-up diagnostic
screening tests such as the magnetic resonance imaging, computerized-tomography scan, plasma chemistry profile,
and complete blood count analysis among others.
Keywords: Biomarkers, Urinalysis, Urine

Introduction deaths, 19 million disability-adjusted life-years

and 18 million years of life lost from
Approximately 830,000 deaths and
cardiovascular diseases were directly attributable
18,467,000 disability-adjusted life years annually
to reduced glomerular filtration rates
are accounted for the diseases of the kidney and
(Kassebaum, et al., 2016). Moreover,
urinary tract (Jamison, Breman, & Measham,
approximately 146 million people (30%) of
2006). A significant increase in mortality was
diabetic patients in 2014 alone (Bethesda, 2014)
observed throughout the 21st century that in
are found to have diabetic nephropathy (Coresh,
kidney disease alone, an estimated 5 to 10 million
Astor, Greene, Eknoyan, & Levey, 2003). Renal
people died in 2018 (Luyckx, Tonelli, & Stanifer,
diseases affect both men and women (King,
2018); urinary tract infection is also considered as
Aubert, & Herman, 1998), and they can progress
the second most common infection, accounting to
to end-stage renal disease (ESRD) that requires
150 million people infected annually (Sewify, et
renal replacement therapy to maintain
al., 2016). Statistically, it ranked as the 12th
physiological function affecting 1.5 million people
among the causes of death and 17th among the
worldwide (Weening, 2004) and has a mortality of
causes of disability annually worldwide (John,
2.3 to 7.1 million people in 2010 worldwide
Giuseppe, Susan, Arrigo, & Adibul, 2006). The
(Luyckx, Tonelli, & Stanifer, 2018). In the
total prevalence of chronic kidney diseases,
Philippines, ESRD is the 7th leading cause of
however, should also account patients suffering
death; every hour, one Filipino develops chronic
from cardiovascular or cerebrovascular disease
renal failure; and more than 70,000 Filipino
(Hostetter, 2014), because according to the
patients are currently under dialysis (National
Global Burden Study 2015 study, 1.2 million

1
 Biomolecules
Urine Biomarkers
Kidney and Transplant Institute, 2019). According
to Dr. Rowena Galpo, the health officer of Baguio
City, 9.87% of 700 pupils was tested positive for
urinary tract infection using the dipstick method
(Refuerzo, 2018). Last year, the Baguio General
Hospital Medical Center was able to document
some 105 individuals who acquired kidney
ailment due to hypertensive nephrosclerosis
while there were some 83 diabetic nephropathy
patients who were also treated to be suffering
from kidney ailment. This year, Dr. Jennifer Pira,
the medical officer of the Cordillera Office of the
Department of Health, said that for the first
quarter of this year, there were already 96
individuals who were treated with kidney ailment
triggered by hypertensive nephrosclerosis while
there were some 87 cases of diabetic
nephropathy which showed that more
hypertension and diabetes patients are now
prone to acquiring kidney ailment. To address the
aforementioned statistics, the significance of
having an early diagnosis using an easy, efficient,
but reliable diagnostic tool such as
urinalysis is very important to lessen the
progressiveness of these diseases.
The blood flows into the kidney via the
renal artery and the wastes from the blood
plasma are filtered in the glomerulus of the
kidney’s nephron in order to rid the body of toxic
substances and other wastes (National Institute
of Diabetes and Digestive and Kidney Diseases,
2019), the filtered blood will then flow out of the
kidney via the renal vein and the wastes filtered
from the blood will be excreted in the body as
urine (U.S. National Library of Medicine, 2019).
Urine is an aqueous waste byproduct excreted
from the body via urination that is usually
composed of 95% water and different inorganic
and organic constituents such as urea (9.3 g/L),
chloride (1.87 g/L), sodium (1.17 g/L), potassium
(0.750 g/L), creatinine (0.670 g/L), and other
dissolved ions and compounds such as proteins,
hormones, and metabolites (Boundless, 2019).
Urine was started to be used as a diagnostic tool
6000 years ago through the use of a technique
called uroscopy (Kamaledeen & Vivekanantham,
2015). Physicians before even described urine as
a ‘divine fluid’ or a ‘window to the body’ because
of its efficacy to diagnose a wide diversity of
diseases (Armstrong, 2007). The different
constituents of the urine can be used as
biomarkers to indicate different health conditions.
Biomarkers are any substances, properties, or
constituents in a physiological system that can
indicate abnormalities or health conditions
including diseases.
Nowadays, analysis of urine or urinalysis
is still widely used to diagnose different types of
diseases such as diabetes, kidney diseases, liver
diseases, and urinary tract infection (Krans &
Jewell, 2018). Urinalysis is even considered as
the third major diagnostic screening test in the
clinical laboratory, only preceded by plasma
chemistry profiles and complete blood count
analysis because of its efficiency in diagnosing
health conditions including possible diseases an
individual may have (Coppens, Speeckaert, &
Delanghe, 2010). Detection of these diseases
can be efficiently done in urinalysis by
determining the presence of certain urine
biomarkers that serve as biomarkers of health
conditions. According to the study of Tzimenatos,
L. et al. (2018) entitled “Accuracy of the Urinalysis
for Urinary Tract Infections in Febrile Infants 60
Days and Younger” urinalysis is highly sensitive
and specific for diagnosing urinary tract infection
(Tzimenatos, et al., 2018). For other health
conditions and diseases, urinalysis may indicate
the possibility of having a certain disease by
analyzing abnormalities on the urine constituents.
The presence of certain inorganic,
organic, and pathological organic constituents
are determined to identify abnormalities in the
urine sample, which can be used as biomarkers
of health conditions. Inorganic urine biomarkers
such as calcium (Ca), magnesium (Mg), chlorides
(Cl), sulfates (SO4), and phosphates of Ca, Mg,
Na, K, and NH4; organic urine biomarkers such
as urea (CH₄N₂O), uric acid (C5H4N4O3), and
creatinine (C4H7N3O); and pathologic organic
urine biomarkers such as protein, glucose
(C6H12O6), acetone bodies (C3H6O), bile salts and
bile acids (C24H40O5), and blood were all tested.
This study aimed to determine the
physical properties of the urine samples and the
2
 Biomolecules
Urine Biomarkers
presence of certain inorganic, organic, and
pathological organic constituents as biomarkers
to suggest human health conditions.
However, the scope of the study is
delimited to qualitative urinalysis tests. As such,
quantitative data that can be used to analyze the
urine biomarkers as diagnostic tools more
effectively in terms of sensitivity and specificity is
not within the scope of the study.
Methodology
A total of randomly sampled 12
individuals (6 males and 6 females) aged 20 to 21
years old participated in the study. A total of
approximately 200ml urine samples per individual
were collected 24 hours before testing to prevent
contamination which may result in false-positive
or false-negative results. Toluene (C7H8) was
used as a preservative to prevent metabolic
changes of urine analytes and bacterial
overgrowth. The urine samples were stored in an
amber bottle to protect light-sensitive metabolites
such as porphyrins and urobilinogen (Coppens,
Speeckaert, & Delanghe, 2010).
Physical properties such as the color,
odor, transparency, and specific gravity was
determined using 2 randomly selected urine
samples, 1 from males and another from females.
The color, odor, and transparency of the urine
samples was tested empirically. The specific
gravity of the urine samples was quantified using
a urinometer.
The presence or absence of various
inorganic, organic, and pathological organic
constituents of the urine samples was then
tested.
Inorganic constituents such as Ca, Mg,
Cl, SO4, earthly phosphates (Ca and Mg), and
alkaline phosphates of (Na, K, and NH4) were
tested.
The test for Ca was done by mixing 2ml
of the urine sample with 3 drops of 2% potassium
oxalate (C2K2O4) solution in a test tube. The
solution was filtered using a standard filter paper
with an 11um pore size. The filtrate was then
tested for further precipitation by adding the
C2K2O4 solution dropwise.
C2K2O4 is a reducing reagent that inhibits
the formation of turbidity; the presence of Ca in
the urine, however, results in the formation of
turbidity (D'Sa, 2015).
The test for Mg was done by filtrating
again the filtrate from the test for Ca. The solution
was then made alkaline by adding 10% ammonia
(NH3) water dropwise and few drops of
ammonium oxalate (C2H8N2O4) was added. It
was then set aside and was observed for
precipitate formation.
By making the pH of the urine alkaline
before the addition of C2H8N2O4 after the addition
of C2K2O4, the coprecipitation of Mg occurs if it is
present in the urine sample (D'Sa, 2015).
The test for Cl was done by acidifying a
2ml urine sample with nitric acid (HNO3) solution
and then adding 3 drops of silver nitrate (AgNO3)
solution.
Chloride is precipitated as silver chloride
(AgCl) with AgNO3 in presence of HNO3 (D'Sa,
2015).
The test for SO4 was done by acidifying
a 2ml urine sample with hydrochloric acid (HCl)
solution and then adding 10% barium chloride
(BaCl2) drop by drop in excess until a precipitate
was formed.
When the BaCl2 is added to the urine, it
combines with the radicals of SO4 forming a
precipitate of barium sulfate (BaSO4) (D'Sa,
2015).
The test for the phosphates of Ca and Mg
was done by alkalifying a 5ml urine sample with
dilute ammonium hydroxide (NH4OH). It was set
aside and the formation of precipitates was
observed. The solution was then warmed gently
in a hot water bath and filtered using the standard
filter paper.
If the urine is acidic, it will readily accept
hydrogen ions (H+) of NH4OH to balance the free
hydroxide ions (OH-). When urine contains a
3
 Biomolecules
Urine Biomarkers

large amount of Ca in proportion to the PO4
present, it results in the formation of turbidity and
more magnesium citrate (C6H6MgO7) must be
added by the addition of NH4OH (D'Sa, 2015).
The test for the phosphates of Na, K, and
NH4 was done by adding 3 drops of magnesia
mixture to the filtrate in the test for the
phosphates of Ca and Mg. The mixture was then
warmed gently in a hot water bath, the nature and
amount of the precipitate were observed.
Magnesium hydroxide (Mg(OH)2), when
dissolved in water, dissociates into ions. When
magnesia reacts with the ions upon warming,
inorganic phosphates results in the formation of
turbidity (D'Sa, 2015).
Organic constituents such as urea
(CH₄N₂O), uric acid (C5H4N4O3), and creatinine
(C4H7N3O) were also tested.
The presence of CH₄N₂O was tested by
evaporating a 10ml urine sample until its volume
was reduced to one-third (3.33ml). The solution
was cooled and about 2ml of 20% sodium
hydroxide (NaOH) was added. The mixture was
shaken gently and 3 drops of 0.05% copper
sulfate (CuSO4) were added.
Formation of brisk effervescence when
copper sulfate (CuSO4) decomposes CH₄N₂O to
give nitrogen (N2) gas (D'Sa, 2015).
The presence of C5H4N4O3 was tested by
microscopy and the murexide test.
For microscopy, C5H4N4O3 crystals were
first isolated by mixing 50ml filtered urine sample
with 1ml of 25% HCl and setting it aside for
approximately 48 hours. C5H4N4O3 crystals that
settle out were collected and were examined
under a compound light microscope.
HCl is a strong acid that increases the
concentration of H+, making the solution more
acidic. If HCl is added to the urine, it results in the
formation of C5H4N4O3 crystals that precipitate at
a pH less than 5.4 (D'Sa, 2015).
For the murexide test, a small amount of
C5H4N4O3 was placed in an evaporating dish with
3 drops of concentrated HNO3. It was then
evaporated in a water bath to dryness until all
HNO3 has been removed and a reddish or
yellowish residue remains. The residue was
cooled and a drop of dilute NH3 water was added.
C5H4N4O3 is a reducing agent in a strong
alkaline condition that reduces colorless HNO3
forming a colored complex (D'Sa, 2015).
The presence of C4H7N3O was tested by
Jaffe’s test and nitroprusside test.
For Jaffe’s test, 1ml of the saturated
picric acid (C6H3N3O7) solution was added to a
3ml urine sample in a test tube and was made
alkaline by adding 1% NaOH.
NaOH dissociates in water to produce
OH-. The C4H7N3O in the urine reacts with
C6H3N3O7 solution in the alkaline medium to form
a reddish colored complex of creatinine picrate. It
turns yellow when acidified since it is the original
color of urine when it is acidic (D'Sa, 2015).
For the nitroprusside test, 3 drops of
freshly prepared nitroprusside (C5FeN6Na2O)
solution were added to 2ml urine and were made
alkaline by adding 10% NaOH.
The addition of NaOH makes the solution
alkaline. C5FeN6Na2O in an alkaline medium
reacts with urine resulting in the formation of
orange coloration due to C5H4N4O3in the
presence of purines (D'Sa, 2015).
Pathologic organic constituents such as
protein, glucose (C6H12O6), acetone bodies
(C3H6O), bile salts and bile acids (C24H40O5), and
blood were also tested.
The presence of protein was tested by
the coagulation test, Heller’s ring test, and
Robert’s test.
For the coagulation test, a 2ml urine
sample was heated at an angle of 45o directly on
a Bunsen burner, boiling the upper half by
passing it over the flame. The development of
turbidity was observed and was confirmed by
acidifying with 2 drops of acetic acid (C2H4O2).

4
 Biomolecules
Urine Biomarkers
In an alkaline medium, proteins may not
be precipitated owing to the formation of alkaline
meta-proteins, thus, acidification is necessary by
adding C2H4O2. The heated part shows turbidity
due to the denaturation of proteins (D'Sa, 2015).
For the Heller’s ring test, 1.5 ml
concentrated HNO3 was placed in a tested and
2ml of urine was added carefully along the inner
walls of the tube without mixing. A white ring
appears in the junction between the two layers in
the addition of HNO3.
The concentrated HNO3 causes protein
denaturation, hence the precipitation of proteins
due to the decomposition into oxides of nitrogen
and water (D'Sa, 2015).
For Robert’s test, 2ml of urine was mixed
with 2ml of Robert’s reagent, and setting it aside
for observation.
HNO3 in Robert’s reagent (concentrated
HNO3 and saturated magnesium sulfate (MgSO4)
solution) causes the precipitation of albumin
resulting in the formation of a white ring (D'Sa,
2015).
The presence of C6H12O6 was tested by
Benedict’s test. About 2ml of Benedict’s reagent
was mixed thoroughly with 4 drops of the urine
sample. The solution was boiled in a water bath
for 3 minutes and was set aside to cool for
observation.
Cuprous ions (Cu+) combine with OH- to
form yellow cuprous hydroxide (CuOH) which
upon heating is converted to red cuprous
oxide(Cu2O) (D'Sa, 2015).
The presence of C3H6O was tested by
the legal test. A freshly prepared 5%
C5FeN6Na2O solution was used to treat a 2ml
urine sample. The solution was mixed thoroughly
and was made alkaline by adding NaOH and NH3
water.
Acetone (C3H6O) and acetonic acid
(C4H8O3) react with 5% sodium nitroprusside
solution in the presence of alkali to produce a
purple ring at the junction (D'Sa, 2015).
The presence of bile salts (C24H40O5)
was tested by Gmelin’s test. About 1ml of
concentrated HNO3 was placed in a test tube and
3ml of urine was added by pipetting it carefully
along the inner sides of the tube, without mixing,
and was set aside for observation.
HNO3 oxidizes bilirubin (C33H36N4O6) to
biliverdin (C33H34N4O6) giving different colors
from green to violet (D'Sa, 2015).
The presence of bile acids (C24H40O5)
was tested by Petten Kofer’s test. About 3ml urine
sample was mixed with 3 drops of 5% sucrose
(C12H22O11) solution in a test tube. About 3 ml of
concentrated sulfuric acid (H2SO4) was poured
down along the inner sides of the test tube
carefully. The formation of a red ring was
observed and was then stirred and set aside for
additional observation.
Bile acids will react with
hydroxymethylfurfural (C6H6O3) to form a red
solution. C6H6O3 is formed from sugar that was
dehydrated by H2SO4 (Selengkapniya, 2013).
The presence of blood was tested by
benzidine reaction by bringing a 2ml urine sample
into a boil. It was then cooled and about 2ml of
saturated benzidine (C12H12N2) solution glacial
C2H4O2 was added and another 1 ml of 3%
hydrogen peroxide (H202) was added.
H2O2 liberates oxygen from hemoglobin.
Liberated oxygen from this reaction is allowed to
react with C12H12N2 which undergoes oxidation to
produce a blue or a green colored product
(Agarwal, Anand, & Anand, 2016).
5
 Biomolecules
Urine Biomarkers
Results and Discussion
The inorganic, organic, and pathological
organic constituents of the urine were determined
to be used as biomarkers of health conditions.
This study, however, focused only on the
qualitative tests in urinalysis, as such, the
inorganic and the organic constituents that are
normally present in the urine but are indicative of
health conditions only when it exhibited abnormal
concentrations can’t be used effectively because
of the unavailability of the quantitative data for the
concentrations of constituents. Qualitative tests
in urinalysis indicate only the presence or
absence of a certain constituent without providing
any data of its concentration.
Since the all of the included inorganic
and organic constituents are normally present in
the urine, to make use of the data collected as
biomarkers of health conditions, however, the
Table 1: Results of the Tests for the Physical Properties of 2 Randomly Selected Urine Samples
Results of the Tests for the Physical Properties of 2 Randomly Selected Urine Samples
Physical Properties Male
Color Slight yellow
Odor Slightly aromatic
Transparency Clear
Specific Gravity 1.002
As per the data presented in table 1, the
2 randomly selected urine samples exhibited
normal color, odor, and transparency, and is
within the normal range of specific gravity.
The color of the urine sample is
dependent on the pigments present in the urine.
The observed yellow color for both of the male
and urine samples is due to the presence of
urochrome and urobilin, which are the pigments
responsible for the normal yellow color of urine;
intensity of the color is directly proportional to the
concentration of these pigments, mainly due to
differences in diet and water consumption.
For the odor, the more aromatic the urine
is, the more concentrated are the volatile acids
researchers would preliminarily interpret the
‘presence’ of either inorganic or organic
constituent as ‘present at a high level’, while
‘absence’ as ‘present at a low level’. This
adjustment, however, will be included in the
recommendation section to conduct follow-up
diagnostic screening tests to validate the results
of the study. For the pathological organic
constituents, no adjustment was made because
its presence is already an indication of disease.
The following sections of this paper will
present the results of the tests for physical
properties and qualitative tests in urinalysis for
determining the inorganic, organic, and
pathological organic biomarkers of urine, along
with the interpretation and analysis of the results
and its implications to human health.
Female Interpretation
Yellow to orange Normal
Aromatic Normal
Clear Normal
1.003 Slightly below normal
present in the urine sample such as ammonia
from the decomposition of urea.
For the transparency, turbidity of the
urine may be indicative of conditions such as the
formation of pus, bacterial infection, cellular
damage, acidosis, alkalosis etc. The more turbid
a urine is, the more it is indicative of the
aforementioned different health conditions. It is
also directly correlated to the amount of total
dissolved solids in the urine, which is also directly
related to specific gravity.
The specific gravity of urine, the normal
range of which is from 1.015 to 1.025. The results
for both the urine samples from male (1.002) and
female (1.003) were slightly below normal. This
6
 Biomolecules
Urine Biomarkers

can be due to decreased salt intake, decreased more concentrated the urine is because it gives a
protein intake, or increased water intake because rough estimate of the total amount of dissolved
the value of specific gravity is directly proportional solids in the urine and the state of hydration of a
to the amount of salt, urea, and protein in the patient.
urine and is inversely proportional to the amount
of water. Thus, the higher the specific gravity, the
Table 2.1: Inorganic Urine Biomarkers for Specific Health Conditions

Inorganic Urine Biomarkers for Specific Health Conditions

Urine Biomarker Health Conditions
Calcium (Ca) High level:
High level or low level Dehydration, too much vitamin D intake, too much
calcium intake, acidic urine, use of loop diuretics,
hyperparathyroidism, hypercalciuric, or hypocalcemia
Low level:
Vomiting, diarrhea, vitamin D deficiency, use of thiazide
diuretics, hypoparathyroidism, hypocalciuric, or
hypercalcemia
Magnesium (Mg) High level:
High level or low level Dehydration, use of thiazide diuretics, use of laxatives,
hypothyroidism, hypercalciuric, hypocalcemia, or
hypomagnesia
Low level:
Vomiting, diarrhea, use of diuretics, use of antibiotics,
hyperthyroidism, hypocalciuric, hypercalcemia, or
hypermagnesia
Chlorides (Cl) High level:
High level or low level Starvation, dehydration, increased salt intake, polyuria,
salt wasting nephropathy, adrenocortical insufficiency, or
Addison disease
Low level:
Decreased salt intake, diarrhea, vomiting, excessive
sweating, increased salt retention, Cushing syndrome,
Conn syndrome, or congestive heart failure
Sulfates (SO4) High level:
Low level High protein intake
Low level:
Low protein intake or diabetic kidney disease
Phosphates of Ca High level:
High level Dehydration, increased salt intake, increased oxalate
intake (e.g. in spinach), intake of calcium supplements,
intake of vitamin D, or kidney stones
Low level:
Decreased salt intake, decreased oxalate intake
Phosphates of Mg High level:
High level or low level Dehydration, acidic urine, or hypomagnesemia
Low level:
Citrate consumption (e.g. citrus), use of thiazide
diuretics, alkaline urine, or hypermagnesemia
Phosphates of Na High level:
High level or low level High salt intake, vomiting, use of water pills,
inflammation of kidneys, hypothyroidism, or
hypoaldosteronism
Low level:
Low salt intake, dehydration, diarrhea, use of thiazide
diuretics, kidney problems, hyperthyroidism,
hyperaldosteronism, heart failure, or cirrhosis

7
 Biomolecules
Urine Biomarkers

Phosphates of K High level:

High level or low level Dehydration, lack of magnesium (green leafy vegetables
and fruits), eating disorders (anorexia or bulimia),
infection, or hyperkalemia
Low level:
Vomiting, diarrhea, lack of potassium (dried fruits,
banana, oranges etc.) or hypokalemia caused by acute
kidney failure or chronic kidney disease
Phosphates of NH4 High level:
High level or low level High protein intake, dehydration, vomiting, diarrhea,
infection, or chronic metabolic acidosis such as
hyperchloremic acidosis and liver infections and
diseases
Low level:
Low protein intake

Table 2.2: Results of the Qualitative Tests for Inorganic Constituents

Results of the Qualitative Tests for Inorganic Constituents

Results
Male Female Both
Test % of % of %of
1st 2nd 3rd 4th 5th 6th Positive 1st 2nd 3rd 4th 5th 6th Positive Positive
Results Results Results
Inorganic Constituents
Calcium - + + + + + 83% - + + + + + 83% 83%
Magnesium - + + + + + 83% - + + + + + 83% 83%
Chlorides + + + + + + 100% + + + + + + 100% 100%
Sulfates + + + + + + 100% + + + + + + 100% 100%
Phosphates
+ + + - + + 83% + + + - - + 67% 75%
of Ca
Phosphates
+ + + - + + 83% + + + + + + 100% 92%
of Mg
Phosphates
- - + + + + 67% - - + - - + 33% 50%
of Na
Phosphates
- - + + + + 67% - - + - + + 50% 58%
of K
Phosphates
- - + - + + 50% - - + - + + 50% 50%
of NH4

The data presented in table 2.1 shows
the different inorganic biomarkers determined in
the qualitative tests in urinalysis and the health
conditions or diseases they are indicative of. As
per the result of the tests for inorganic
constituents in the urine samples of 12 individuals
(6 males and 6 females) presented in table 2.2,
the following percentages of positive results to
each test and their implications to human health
are presented in this section.
For the presence of Ca, 83% of the urine
samples of males and 83% of the urine samples
of females exhibited positive results, a total of
83% of the urine samples from both males and
females exhibited positive results.
It is normal to find Ca in the urine
because it is widely used in the body. However, if
urine Ca levels are too high or too low, it is
indicative of health conditions and diseases. High
urine Ca level is indicative of dehydration, too
much vitamin D intake, too much calcium intake,
acidic urine, use of loop diuretics,
hyperparathyroidism, hypercalciuric, or
hypocalcemia (Murrell, 2018) (Blocka, 2018).
Low urine Ca level is indicative of vomiting,
diarrhea, vitamin D deficiency, use of thiazide

8
 Biomolecules
Urine Biomarkers
diuretics, hypoparathyroidism, hypocalciuric, or
hypercalcemia (Murrell, 2018) (Brown, 2019).
In using urine Ca as a biomarker as
presented in table 2.1 and the result of the
qualitative test for the presence of Ca presented
in table 2.2, about 83% of the male participants
are in normal condition. Using the adjustment
made by the researchers to use qualitative data
as biomarkers of health conditions, however,
about 83% of the male participants may exhibit
health conditions such as dehydration, too much
vitamin D intake, too much calcium intake, acidic
urine, use of loop diuretics, hyperparathyroidism,
hypercalciuric, or hypocalcemia; the remaining
17% may exhibit health conditions such as
vomiting, diarrhea, vitamin D deficiency, use of
thiazide diuretics, hypoparathyroidism,
hypocalciuric, or hypercalcemia. For female
participants, about 83% are in normal condition.
In adjustment, however, about 83% of the female
participants may exhibit health conditions such as
dehydration, too much vitamin D intake, too much
calcium intake, acidic urine, use of loop diuretics,
hyperparathyroidism, hypercalciuric, or
hypocalcemia; the remaining 17% may exhibit
health conditions such as vomiting, diarrhea,
vitamin D deficiency, use of thiazide diuretics,
hypoparathyroidism, hypocalciuric, or
hypercalcemia. In total, about 83% are in normal
condition. In adjustment, however, about 83% of
all the participants may exhibit health conditions
such as dehydration, too much vitamin D intake,
too much calcium intake, acidic urine, use of loop
diuretics, hyperparathyroidism, hypercalciuric, or
hypocalcemia; the remaining 17% may exhibit
health conditions such as vomiting, diarrhea,
vitamin D deficiency, use of thiazide diuretics,
hypoparathyroidism, hypocalciuric, or
hypercalcemia.
For the presence of Mg, 83% of the urine
samples from males and 83% of the urine
samples from females exhibited positive results,
a total of 83% of the urine samples from both
males and females exhibited positive results.
The normal concentration of intracellular
Mg cation should be maintained for normal
neuromuscular activity. Furthermore, Mg is an
important cofactor for various enzymes,
transporters, and nucleic acids that are essential
for cellular function, replication, and metabolism.
(Bringhurst, Krane, & Kronenberg, 2012)
(Devkota, 2019). About 1.3 to 2.1 mEq/L is the
normal urine Mg level. High urine Mg level can be
due to dehydration, use of thiazide diuretics, use
of laxatives, hypothyroidism, hypercalciuric,
hypocalcemia, or hypomagnesia. While too low
urine Mg level can be due to vomiting, diarrhea,
use of diuretics, use of antibiotics,
hyperthyroidism, hypocalciuric, hypercalcemia,
or hypermagnesia (Brown, 2019) (Devkota,
2019).
In using urine Mg as a biomarker as
presented in table 2.1 and the result of the
qualitative test for the presence of Mg presented
in table 2.2, about 83% of the male participants
are in normal condition. Using the adjustment
made by the researchers to use qualitative data
as biomarkers of health conditions, however,
about 83% of the male participants may exhibit
health conditions such as dehydration, use of
thiazide diuretics, use of laxatives,
hypothyroidism, hypercalciuric, hypocalcemia, or
hypomagnesia; the remaining 17% may exhibit
health conditions such as vomiting, diarrhea, use
of diuretics, use of antibiotics, hyperthyroidism,
hypocalciuric, hypercalcemia, or hypermagnesia.
For female participants, about 83% are in normal
condition. In adjustment, however, about 83% of
the female participants may exhibit health
conditions such as dehydration, use of thiazide
diuretics, use of laxatives, hypothyroidism,
hypercalciuric, hypocalcemia, or hypomagnesia;
the remaining 17% may exhibit health conditions
such as vomiting, diarrhea, use of diuretics, use
of antibiotics, hyperthyroidism, hypocalciuric,
hypercalcemia, or hypermagnesia. In total, about
83% are in normal condition. In adjustment,
however, about 83% of all the participants may
exhibit health conditions such as dehydration,
use of thiazide diuretics, use of laxatives,
hypothyroidism, hypercalciuric, hypocalcemia, or
hypomagnesia; the remaining 17% may exhibit
health conditions such as vomiting, diarrhea, use
of diuretics, use of antibiotics, hyperthyroidism,
hypocalciuric, hypercalcemia, or hypermagnesia.
9
 Biomolecules
Urine Biomarkers
For the presence of Cl, 100% of the urine
samples from males and 100% of the urine
samples from females exhibited positive results,
a total of 100% of urine samples for both males
and females exhibited positive results.
Cl is an anion that works with other
electrolytes to help regulate the amount of fluid in
the body and maintain acid-base balance.
Normal urine Cl level range from 25 to 40 mEq/L.
An increased level of urine Cl can indicate
starvation, dehydration, increased salt intake,
polyuria, salt wasting nephropathy,
adrenocortical insufficiency, or Addison disease
(Goldman & Schafer, 2011) (American
Association for Clinical Chemistry, 2016). A
decreased level of urine Cl, however, can indicate
decreased salt intake, diarrhea, vomiting,
excessive sweating, increased salt retention,
Cushing syndrome, Conn syndrome, or
congestive heart failure (Goldman & Schafer,
2011) (University of Rochester Medical Center
Rochester, 2019).
In using urine Cl as a biomarker as
presented in table 2.1 and the result of the
qualitative test for the presence of Cl presented
in table 2.2, 100% of the male participants are in
normal condition. Using the adjustment made by
the researchers to use qualitative data as
biomarkers of health conditions, however, 100%
of the male participants may exhibit health
conditions such as starvation, dehydration,
increased salt intake, polyuria, salt wasting
nephropathy, adrenocortical insufficiency, or
Addison disease. For female participants, 100%
are in normal condition. In adjustment, however,
100% of the female participant may exhibit health
conditions such as starvation, dehydration,
increased salt intake, polyuria, salt wasting
nephropathy, adrenocortical insufficiency, or
Addison disease. In total, 100% are in normal
condition. In adjustment, however, all the
participants may exhibit health conditions such as
starvation, dehydration, increased salt intake,
polyuria, salt wasting nephropathy,
adrenocortical insufficiency, or Addison disease.
For the presence of SO4, 100% of the
urine samples from males and 100% of the urine
samples from females exhibited positive results,
a total of 100% of the urine samples from both
males and females exhibited positive results.
SO4 should be present in the urine
because higher urinary SO4 concentration is
associated with a more beneficial profile of renal
risk markers and is independently associated with
a reduced risk for diabetic kidney disease in type
2 diabetes individuals. Each doubling of urinary
SO4 was associated with 19% lower risk of
diabetic kidney disease (van den, et al., 2016).
High SO4 indicate high protein intake, while low
SO4 indicate low protein intake or diabetic kidney
disease. (Laboratory Corporation of America®
Holdings, 2019)
In using urine SO4 as a biomarker as
presented in table 2.1 and the result of the
qualitative test for the presence of SO4 presented
in table 2.2, 100% of the male participants are in
normal condition. Using the adjustment made by
the researchers to use qualitative data as
biomarkers of health conditions, however, about
100% have high protein intake. For female
participants, 100% are in normal condition. In
adjustment, however, 100% have high protein
intake. In total, about 100% are in normal
condition. In adjustment, however, 100% have
high protein intake.
For the presence of phosphates of Ca,
83% of the urine samples from males and 67% of
the urine samples from females exhibited positive
results, a total of 75% of the urine samples from
both males and females exhibited positive
results.
Phosphates of Ca are usually present in
urine with a pH above 6.5. The formation of
calcium phosphate (Ca3(PO4)2) crystals can be
caused by a combination of factors including
decreased urine volume, urine alkalization, or a
diet rich in Ca. In individuals with kidney stones,
a repeated presence of (Ca3(PO4)2) crystals
could indicate the probable nature of stones (U.S.
National Library of Medicine, 2019). High urine
phosphates of Ca level indicates dehydration,
increased salt intake, increased oxalate intake
(e.g. in spinach), intake of calcium supplements,
10
 Biomolecules
Urine Biomarkers
intake of vitamin D, or kidney stones. While low
urine phosphates of Ca level indicates decreased
salt intake or decreased oxalate intake (Biggers,
2019).
In using urine phosphates of Ca as a
biomarker as presented in table 2.1 and the result
of the qualitative test for the presence of
phosphates of Ca presented in table 2.2, about
83% of the male participants are in normal
condition. Using the adjustment made by the
researchers to use qualitative data as biomarkers
of health conditions, however, about 83% of the
male participants may exhibit health conditions
such as dehydration, increased salt intake,
increased oxalate intake, intake of calcium
supplements, intake of vitamin D, or kidney
stones; the remaining 17% may exhibit health
conditions such as decreased salt intake or
decreased oxalate intake. For female
participants, about 67% are in normal condition.
In adjustment, however, about 67% of the female
participants may exhibit health conditions such as
dehydration, increased salt intake, increased
oxalate intake, intake of calcium supplements,
intake of vitamin D, or kidney stones; the
remaining 33% may exhibit health conditions
such as decreased salt intake or decreased
oxalate intake. In total, about 75% are in normal
condition. In adjustment, however, about 75% of
all the participants may exhibit health conditions
such as dehydration, increased salt intake,
increased oxalate intake, intake of calcium
supplements, intake of vitamin D, or kidney
stones; the remaining 25% may exhibit health
conditions such as decreased salt intake or
decreased oxalate intake.
For the presence of phosphates of Mg,
83% of urine samples of males and 100% of urine
samples of females exhibited positive results, a
total of 92% of the urine samples from both males
and females exhibited positive results.
The normal urine phosphates of Mg level
is 3.0-4.5 mg/dL or 0.97-1.45 mmol/L (Pagana,
Pagana, & Pagana, 2019). Too high urine
phosphates of Mg level indicates dehydration,
acidic urine, or hypomagnesemia. Too low urine
phosphates of Mg level, on the other hand,
indicates citrate consumption (e.g. citrus), use of
thiazide diuretics, alkaline urine, or
hypermagnesemia (Suller, et al., 2005) (WebMD
LLC, 2019).
In using urine phosphates of Mg as a
biomarker as presented in table 2.1 and the result
of the qualitative test for the presence of
phosphates of Mg presented in table 2.2, about
83% of the male participants are in normal
condition. Using the adjustment made by the
researchers to use qualitative data as biomarkers
of health conditions, however, about 83% of the
male participants may exhibit health conditions
such as dehydration, acidic urine, or
hypomagnesemia; the remaining 17% may have
had recent citrate consumption, use of thiazide
diuretics, alkaline urine, or hypermagnesemia.
For female participants, about 100% are in
normal condition. In adjustment, however, about
100% of the female participants may exhibit
health conditions such as dehydration, acidic
urine, or hypomagnesemia. In total, about 92%
are in normal condition. In adjustment, however,
about 92% of all the participants may exhibit
health conditions such as dehydration, acidic
urine, or hypomagnesemia; the remaining 8%
may have had recent citrate consumption, use of
thiazide diuretics, alkaline urine, or
hypermagnesemia.
For the presence of phosphates of Na,
67% of the urine samples from males and 33% of
the urine samples from females exhibited positive
results, a total of 50% of the urine samples from
both males and females exhibited positive
results.
The normal phosphates of Na value
ranges from 40 to 220 mEq/L per day. High
phosphates of Na level in the urine indicates high
salt intake, vomiting, use of water pills,
inflammation of kidneys, hypothyroidism, or
hypoaldosteronism. Low phosphates of Na levels
in the urine indicate low salt intake, dehydration,
diarrhea, use of thiazide diuretics, kidney
problems, hyperthyroidism, hyperaldosteronism,
heart failure, or cirrhosis (Felson, 2019).
11
 Biomolecules
Urine Biomarkers
In using urine phosphates of Na as a
biomarker as presented in table 2.1 and the result
of the qualitative test for the presence of
phosphates of Na presented in table 2.2, about
67% of the male participant are in normal
condition. Using the adjustment made by the
researchers to use qualitative data as biomarkers
of health conditions, however, about 67% of the
male participants may exhibit health conditions
such as having a high salt intake, vomiting, use
of water pills, inflammation of kidneys,
hypothyroidism, or hypoaldosteronism; the
remaining 33% may exhibit health conditions
such as having a low salt intake, dehydration,
diarrhea, use of thiazide diuretics, kidney
problems, hyperthyroidism, hyperaldosteronism,
heart failure, or cirrhosis. For female participants,
about 33% are in normal condition. In adjustment,
however, about 33% of the female participants
may exhibit health conditions such as having a
high salt intake, vomiting, use of water pills,
inflammation of kidneys, hypothyroidism, or
hypoaldosteronism; the remaining 67% may
exhibit health conditions such as having a low salt
intake, dehydration, diarrhea, use of thiazide
diuretics, kidney problems, hyperthyroidism,
hyperaldosteronism, heart failure, or cirrhosis. In
total, about 50% are in normal condition. In
adjustment, however, about 50% of all the
participants may exhibit health conditions such as
having a high salt intake, vomiting, use of water
pills, inflammation of kidneys, hypothyroidism, or
hypoaldosteronism; the other 50% may exhibit
health conditions such as having a low salt intake,
dehydration, diarrhea, use of thiazide diuretics,
kidney problems, hyperthyroidism,
hyperaldosteronism, heart failure, or cirrhosis.
For the presence of phosphates of K,
67% of the urine samples from males and 50% of
the urine samples from females exhibited positive
results, a total of 58% of the urine samples from
both males and females exhibited positive
results.
The normal K range for an adult is 35-125
mEq/L per day. Having too much or too little K in
the body is detrimental, exceeding the normal
range (indicated by having high urine phosphates
of K level) results in hyperkalemia, which if left
undetected or untreated can be fatal. However,
too low K level in the body (indicated by low urine
phosphates of K level) results in hypokalemia
(Holland, 2016). High urine phosphates of K level
indicates dehydration, lack of magnesium (green
leafy vegetables and fruits), eating disorders
(anorexia or bulimia), infection, or hyperkalemia
(Crop, Hoorn, Lindemans, & Zietse, 2007). While
low urine phosphates of K level indicates
vomiting, diarrhea, lack of potassium (dried fruits,
banana, oranges etc.) or hypokalemia caused by
acute kidney failure or chronic kidney disease
(Huang & Kuo, 2007).
In using urine phosphates of K as a
biomarker as presented in table 2.1 and the result
of the qualitative test for the presence of
phosphates of K presented in table 2.2, about
67% of the male participants are in normal
condition. Using the adjustment made by the
researchers to use qualitative data as biomarkers
of health conditions, however, about 67% of the
male participants may exhibit health conditions
such as dehydration, lack of magnesium, eating
disorders, infection, or hyperkalemia; the
remaining 33% may exhibit health conditions
such as vomiting, diarrhea, lack of potassium or
hypokalemia caused by acute kidney failure or
chronic kidney disease. For female participants,
about 50% are in normal condition. In adjustment,
however, about 50% of the female participants
may exhibit health conditions such as
dehydration, lack of magnesium, eating
disorders, infection, or hyperkalemia; the
remaining 50% may exhibit health conditions
such as vomiting, diarrhea, lack of potassium or
hypokalemia caused by acute kidney failure or
chronic kidney disease. In total, about 58% are in
normal condition. In adjustment, however, about
58% of all the participants may exhibit health
conditions such as dehydration, lack of
magnesium, eating disorders, infection, or
hyperkalemia; the remaining 42% may exhibit
health conditions such as vomiting, diarrhea, lack
of potassium or hypokalemia caused by acute
kidney failure or chronic kidney disease.
12
 Biomolecules
Urine Biomarkers
For the presence of phosphates of NH4,
50% of the urine samples from males and 50% of
the urine samples from females exhibited positive
results, a total of 50% of the urine samples from
both males and females exhibited positive
results.
To maintain acid-base balance, the
kidney must generate new bicarbonate by
metabolizing glutamine and excreting NH4. High
level of phosphates of NH4 in the urine is
indicative of high protein intake, dehydration,
vomiting, diarrhea, infection, or chronic metabolic
acidosis such as hyperchloremic acidosis and
liver infections and diseases. While low level of
phosphates of NH4 indicates low protein intake
(Carlisle, Donnelly, & Halperin, 1991) (Lee, et al.,
2015) (Barhum, 2018).
In using urine phosphates of NH4 as a
biomarker as presented in table 2.1 and the result
of the qualitative test for the presence of
phosphates of NH4 presented in table 2.2, about
50% of the male participants are in normal
Table 3.1: Organic Urine Biomarkers for Specific Health Conditions
Organic Urine Biomarkers for Specific Health Conditions
Urine Biomarker
Urea (CH₄N₂O)
High level or low level
Uric acid (C5H4N4O3)
High level or low level
Creatinine (C4H7N3O)
High level or low level
condition. Using the adjustment made by the
researchers to use qualitative data as biomarkers
of health conditions, however, about 50% of the
male participants may exhibit health conditions
such as high protein intake, dehydration,
vomiting, diarrhea, infection, or chronic metabolic
acidosis such as hyperchloremic acidosis and
liver infections and diseases; the remaining 50%
have low protein intake. For female participants,
about 50% are in normal condition. In adjustment,
however, about 50% of the female participants
may exhibit health conditions such high protein
intake, dehydration, vomiting, diarrhea, infection,
or chronic metabolic acidosis such as
hyperchloremic acidosis and liver infections and
diseases; the remaining 50% have low protein
intake. In total, about 50% are in normal
condition. In adjustment, however, about 50% of
all the participants may exhibit health conditions
such as high protein intake, dehydration,
vomiting, diarrhea, infection, or chronic metabolic
acidosis such as hyperchloremic acidosis and
liver infections and diseases; the remaining 50%
have low protein intake.
Health Conditions
High level:
High protein intake, high protein metabolism (low
carbohydrate intake), fasting, kidney disease, urinary
tract obstruction, congestive heart failure, or
gastrointestinal bleeding
Low level:
Low protein intake, low protein metabolism (high
carbohydrate intake), Kwashiorkor disease, or kidney
disease
High level:
Overweight, intake of foods high in purines (shellfish, red
meat, organ meat etc.), kidney disease, hyperthyroidism,
psoriasis, renal insufficiency, or diabetes
Low level:
Recent alcohol intake, acute kidney injury, or
hypothyroidism
High level:
High protein intake
Low level:
Low protein intake or kidney disease
13
 Biomolecules
Urine Biomarkers

Table 3.2: Results of the Qualitative Tests for Organic Constituents

Results of the Qualitative Tests for Organic Constituents

Results
Male Female Both
Test % of % of % of
1st 2nd 3rd 4th 5th 6th Positive 1st 2nd 3rd 4th 5th 6th Positive Positive
Results Results Results
Organic Constituents
Urea + - - - + + 50% + - - - - + 33% 42%
Uric Acid + - - + + + 67% + - - + + + 67% 67%
Jaffe’s test
+ + + + + + 100% + + + + + + 100% 100%
(Creatinine)
Nitroprusside
test + + - + + + 83% + + - + + + 83% 83%
(Creatinine)

The data presented in table 3.1 shows
the different organic biomarkers determined in
the qualitative tests in urinalysis and the health
conditions they are indicative of. As per the result
of the tests for organic constituents in the urine
samples of 12 individuals (6 males and 6
females) presented in table 3.2, the following
percentages of positive results to each test and
their implications to human health are presented
in this section.
carbohydrate intake), fasting, kidney disease,
urinary tract obstruction, congestive heart failure,
or gastrointestinal bleeding (Stephens, 2018)
(Mayo Foundation for Medical Education and
Research, 2019). Its presence at low level,
however, is indicative of low protein intake, low
protein metabolism (high carbohydrate intake)
(Stephens, 2018), Kwashiorkor disease
(Fogoros, 2018) or kidney disease (Gabbey,
2018).

For the presence of urea (CH₄N₂O), 50% In using CH₄N₂O as a biomarker as

of the urine samples from males and 33% of the
urine samples from females exhibited positive
results, a total of 42% of the urine samples from
both males and females exhibited positive
results.
The main nitrogenous waste byproduct of
metabolism is CH₄N₂O and it is also generated
from protein catabolism. Its production occurs
mainly in the hepatocytes and the only other
source is the renal cells. Less than 10 % of the
total CH₄N₂O in the body is eliminated via sweat
and the gut, but most of the urea produced in the
liver is transported in the blood to the kidneys
where it is eliminated from the body as urine
(Higgins, 2016). As such, it is normal to find
CH₄N₂O in the urine and around 7 to 20 mg/dL
(2.5 to 7.1 mmol/L) is considered normal (Mayo
Foundation for Medical Education and Research,
2019). At high levels, it indicates high protein
intake, high protein metabolism (low
presented in table 3.1 and the result of the
qualitative test for the presence of CH₄N₂O
presented in table 3.2, about 50% of the male
participants are in normal condition. Using the
adjustment made by the researchers to use
qualitative data as biomarkers of health
conditions, however, about 50% of the male
participants may have high protein intake, high
protein metabolism (low carbohydrate intake),
fasting, kidney disease, urinary tract obstruction,
congestive heart failure, or gastrointestinal
bleeding; the remaining 50% may have low
protein intake, low protein metabolism,
Kwashiorkor disease, or kidney disease. For
female participants, about 33% are in normal
condition. In adjustment, however, about 33% of
the female participants have high protein intake,
high protein metabolism (low carbohydrate
intake), fasting, kidney disease, urinary tract
obstruction, congestive heart failure, or
gastrointestinal bleeding; the remaining 67% may

14
 Biomolecules
Urine Biomarkers
have low protein intake, low protein metabolism,
Kwashiorkor disease, or kidney disease. In total,
about 42% are in normal condition. In adjustment,
however, about 42% of all the participants may
have high protein intake, high protein metabolism
(low carbohydrate intake), fasting, kidney
disease, urinary tract obstruction, congestive
heart failure, or gastrointestinal bleeding; the
remaining 58% may have low protein intake, low
protein metabolism, Kwashiorkor disease, or
kidney disease.
For the presence of uric acid (C5H4N4O3),
67% of the urine samples from males and 67% of
the urine samples from females exhibited positive
results, a total of 67% of the urine samples from
both males and females exhibited positive
results.
Catabolism of purines (Adenine [C5H5N5]
and Guanine [C5H5N5O]) leads to the formation of
C5H4N4O3. Purines are compounds that enter the
bloodstream during the natural breakdown of
cells in the body. Some foods such as sardines,
mushrooms, and peas also contribute to the
formation of purines (Gabbey, Uric Acid Test
(Urine Analysis), 2018). Once purines release
C5H4N4O3, it is carried by the blood to the kidneys
to remove it from the body through urination. As
such, the presence of C5H4N4O3 in the urine is
normal. However, too much C5H4N4O3 or
hyperuricemia (above 2.4-6.0 mg/dL for female
and 3.4-7.0 mg/dL for male) or too little C5H4N4O3
or hypouricemia (below 2.4-6.0 mg/dL for female
and 3.4-7.0 mg/dL for male) (Chemocare, 2019).
High level C5H4N4O3 is indicative of being
overweight, intake of foods high in purines
(shellfish, red meat, organ meat etc.), kidney
disease, hyperthyroidism, psoriasis, renal
insufficiency, or diabetes (Chemocare, 2019)
(Mayo Foundation for Medical Education and
Research, 2019). Low level C5H4N4O3, on the
other hand, can be due to recent alcohol intake,
acute kidney injury, or hypothyroidism (Mount,
2019).
In using C5H4N4O3 as a biomarker as
presented in table 3.1 and the result of the
qualitative test for the presence of C5H4N4O3
presented in table 3.2, about 67% of the male
participants are in normal condition. Using the
adjustment made by the researchers to use
qualitative data as biomarkers of health
conditions, however, about 67% of the male
participants may exhibit health conditions such as
being overweight, intake of foods high in purines,
kidney disease, hyperthyroidism, psoriasis, renal
insufficiency, or diabetes; the remaining 33% may
exhibit health conditions such as being drunk,
acute kidney injury, or hypothyroidism. For
female participants, about 67% are in normal
condition. In adjustment, however, about 67% of
the female participants may exhibit health
conditions such as being overweight, intake of
foods high in purines, kidney disease,
hyperthyroidism, psoriasis, renal insufficiency, or
diabetes; the remaining 33% may exhibit health
conditions such as being drunk, acute kidney
injury, or hypothyroidism. In total, about 67% are
in normal condition. In adjustment, however,
about 67% of all the participants may exhibit
health conditions such as being overweight,
intake of foods high in purines, kidney disease,
hyperthyroidism, psoriasis, renal insufficiency, or
diabetes; the remaining 33% may exhibit health
conditions such as being drunk, acute kidney
injury, or hypothyroidism.
For the presence of creatinine (C4H7N3O)
using Jaffe’s test, 100% of the urine samples from
males and 100% of the urine samples from
females exhibited positive results, a total of 100%
of the urine samples from both males and
females exhibited positive results; using
nitroprusside test, 83% of the urines samples
from males and 83% of the urine samples from
females exhibited positive results, a total of 83%
of the urine samples from both males and
females exhibited positive results.
C4H7N3O is a waste byproduct of muscle
metabolism that is filtered in the blood by the
kidney to remove it from the body through
urination (Pietrangelo, 2017). Normal urine
C4H7N3O values generally range from 955 to
2,936 milligrams (mg) per 24 hours for males, and
601 to 1,689 mg per 24 hours for females (Mayo
Foundation for Medical Education and Research,
2019). High C4H7N3O level indicate high protein
15
 Biomolecules
Urine Biomarkers

intake. Low C4H7N3O level indicate low protein
intake or a kidney disease because as the
kidneys become impaired for any reason, the
ability of the kidney to filter C4H7N3O from the
blood decreases (Stephens, Creatinine Urine
Test (Urine 24-Hour Volume Test), 2017)
(Michigan Medicine, University of Michigan,
2018).
In using C4H7N3O as a biomarker as
presented in table 3.1 and the result of the
qualitative tests for the presence of C4H7N3O
presented in table 3.2, in using Jaffe’s test 100%
of the male participants are in normal condition,
in using the nitroprusside test about 83% of the
male participants are in normal condition. Using
the adjustment made by the researchers to use
qualitative data as biomarkers of health
conditions, however, in using Jaffe’s test 100% of
the male participants have high protein intake, in
using the nitroprusside test about 17% of the
male participants have high protein intake; the
remaining 83% have low protein intake or kidney
disease. For female participants, in using Jaffe’s
test 100% of the female participants are in normal
condition, in using the nitroprusside test about
83% of the female participants are in normal
condition. In adjustment, however, in using
Jaffe’s test 100% of the female participant have
high protein intake, in using the nitroprusside test
about 17% of the female participants have high
protein intake; the remaining 83% have low
protein intake or a kidney disease. In total, using
Jaffe’s test 100% of all the participants are in
normal condition, in using the nitroprusside test
about 83% are in normal condition. In adjustment,
however, using Jaffe’s test 100% have high
protein intake, in using the nitroprusside test
about 17% have high protein intake; the
remaining 83% have low protein intake.

Table 4.1: Urine Pathological Organic Biomarkers for Specific Diseases

Urine Pathological Organic Biomarkers for Specific Diseases

Urine Biomarker Diseases
Protein Chronic kidney disease, diabetes, heart disease,
Presence pyelonephritis, nephrotic syndrome, and
glomerulonephritis
Glucose (C6H12O6) Diabetes
Presence
Acetone (C3H6O) Bodies Diabetes
Presence
Bile Salts (C24H40O5) Hepatobiliary diseases such as cholestatic liver diseases
Presence and cholangiopathies
Bile Acids (C24H40O5) Cholestatic diseases
Presence
Blood Urinary tract infection, acute cystitis, pyelonephritis,
Presence kidney stones, glomerulonephritis, polycystic kidney
disease, and sickle cell disease

Table 4.2: Results of the Qualitative Tests for Pathological Organic Constituents

Results of the Qualitative Tests for Pathological Organic Constituents

Observed Result
Male Female Both
Test % of % of % of
1st 2nd 3rd 4th 5th 6th Positive 1st 2nd 3rd 4th 5th 6th Positive Positive
Results Results Results
Pathological Organic Constituents
Coagulation
test + - - - + + 50% + + + - + + 83% 67%
(Protein)

16
 Biomolecules
Urine Biomarkers

Heller’s ring
test - - - - - - 0% - - - - - - 0% 0%
(Protein)
Robert’s test
- - - - - - 0% - - - - - - 0% 0%
(Protein)
Benedict’s
test - - + - - - 17% - - - - - - 0% 8%
(Glucose)
Legal test
(Acetone + + - - - + 50% + - - - - + 33% 42%
bodies)
Gmelin’s
test + - + + - + 67% + + - + - + 67% 67%
(Bile salts)
Petten
Kofer’s test + + + - + + 83% + + - - + + 67% 75%
(Bile acids)
Benzidine
test - - - - - - 0% - - - - - - 0% 0%
(Blood)

The data presented in table 4.1 shows
the different pathological organic biomarkers
determined in the qualitative tests in urinalysis
and the diseases they are indicative of. As per the
result of the tests for pathologic organic
constituents in the urine samples of 12 individuals
(6 males and 6 females) presented in table 4.2,
the following percentages of positive results to
each test and their implications to human health
are presented in this section.
For the presence of protein using the
coagulation test, 50% of the urine samples from
males and 83% of the urine samples from
females exhibited positive results, a total of 67%
of the urine samples from both males and
females exhibited positive results; in using
Heller’s ring test, 0% of the urine samples of
males and 0% of the urine samples of females
exhibited positive results, a total of 0% of the
urine samples from both males and females
exhibited positive results; and in using Robert’s
test, 0% of the urine samples from males and 0%
of the urine samples from females exhibited
positive results, a total of 0% of the urine samples
from both males and females exhibited positive
results.
Glomerular filtration in the kidney filters
waste products in the blood while retaining those
that are needed by the body, including proteins
(American Kidney Fund, 2019). However, some
conditions or diseases allow proteins to pass
through the filtration system of the kidney causing
the presence of protein in the urine. A temporary
rise in urine’s protein level can be observed but
doesn’t necessarily indicate a disease, after
having a strenuous exercise, fever, exposure to
extreme cold, emotional stress, and dehydration.
Persistently elevated protein levels in the urine,
however, is indicative of diseases such as, but
not limited to, chronic kidney disease, diabetes,
heart disease, pyelonephritis, nephrotic
syndrome, and glomerulonephritis (Mayo
Foundation for Medical Education and Research,
2018).
In using the presence of protein as a
biomarker as presented in table 4.1, together with
the result of the qualitative tests for the presence
of protein using the coagulation test presented in
table 4.2, about 50% of the male participants are
in normal condition and the remaining 50% may
have chronic kidney disease, diabetes, heart
disease, pyelonephritis, nephrotic syndrome, or
glomerulonephritis. For female participants,
about 17% are in normal condition and the
remaining 83% may have chronic kidney disease,
diabetes, heart disease, pyelonephritis, nephrotic
syndrome, or glomerulonephritis. In total, about
33% of all the participants are in normal condition
and the remaining 67% may have chronic kidney

17
 Biomolecules
Urine Biomarkers
disease, diabetes, heart disease, pyelonephritis,
nephrotic syndrome, or glomerulonephritis.
In using the presence of protein as a
biomarker, determined by Heller’s ring test, 100%
of the male participants are in normal condition.
For female participants, about 100% are in
normal condition. In total, 100% of all the
participants are in normal condition.
In using the presence of protein as a
biomarker, determined by Robert’s test, 100% of
the male participants are in normal condition. For
female participants, about 100% are in normal
condition. In total, 100% of all the participants are
in normal condition.
For the presence of glucose (C6H12O6)
using Benedict’s test, 17% of the urine samples
from males and 0% of the urine samples from
females exhibited positive results, a total of 8% of
the urine samples from both males and females
exhibited positive results.
When the blood C6H12O6 concentration is
10mmol/L (180mg/dL), which is the blood
C6H12O6 level of diabetic patients (Freeth, 2019),
the kidney starts to remove extra C6H12O6 in the
bloodstream resulting in the presence of C6H12O6
in the urine. Thus likewise, the presence of
C6H12O6 in the urine is indicative of having a high
blood C6H12O6 level due to diabetes ( Institute for
Quality and Efficiency in Health Care, 2018).
In using the presence of C6H12O6 as a
biomarker as presented in table 4.1, together with
the result of the qualitative tests for the presence
of C6H12O6 using Benedict’s test presented in
table 4.2, about 83% of the male participants are
in normal condition and the remaining 17% may
have diabetes. For female participants, 100% are
in normal condition. In total, about 92% of all the
participants are in normal condition and the
remaining 8% may have diabetes.
For the presence of acetone (C3H6O)
bodies using the legal test, 50% of the urine
samples from males and 33% of the urine
samples from females exhibited positive results,
a total of 42% of the urine samples from both
males and females exhibited positive results.
Normally, the human body uses C6H12O6
as its primary source of energy. When C6H12O6
levels, however, are low or because the body
lacks enough insulin to help tissues absorb
C6H12O6, the physiology turns into breaking down
fats for energy, byproducts of which are ketone
(U.S. National Library of Medicine, 2019). C3H6O
is the simplest ketone, it can be found in the urine
when C3H6O levels in the blood are way above
the normal range of 0.6 to 1.5 mmol/L; its
presence indicates diabetes which can develop to
serious complications such as diabetic
ketoacidosis (DKA), which may lead to cerebral
edema, loss of consciousness, diabetic coma,
and even death (Nall, 2017).
In using the presence of C3H6O bodies as
a biomarker as presented in table 4.1, together
with the result of the qualitative tests for the
presence of C3H6O bodies using the legal test
presented in table 4.2, about 50% of the male
participants are in normal condition and the
remaining 50% may have diabetes. For female
participants, about 67% are in normal condition
and the remaining 33% may have diabetes. In
total, about 58% of all the participants are in
normal condition and the remaining 42% may
have diabetes.
For the presence of bile salts (C24H40O5)
using Gmelin’s test, 67% of the urine samples
from males and 67% of the urine samples from
females exhibited positive results, a total of 67%
of the urine samples from both males and
females exhibited positive results.
There should be no presence of
C24H40O5 in the urine because it only appears in
the urine when there is an obstruction to the
biliary tract causing cholestasis, symptoms of
which include jaundice skin itching, dark urine,
and foul-smelling, light-colored stools (Herrine,
2018). Moreover, this obstruction will lead to an
increase in the bile acids in the blood resulting in
hepatobiliary diseases such as cholestatic liver
diseases and cholangiopathies (Fickert &
Wagner, 2017).
In using the presence of C24H40O5 as a
biomarker as presented in table 4.1, together with
18
 Biomolecules
Urine Biomarkers
the result of the qualitative tests for the presence
of C24H40O5 using Gmelin’s test presented in table
3.2, about 33% of the male participants are in
normal condition and the remaining 67% may
have a hepatobiliary disease such as cholestatic
liver diseases or cholangiopathies. For female
participants, about 33% are in normal condition
and the remaining 67% may have a hepatobiliary
disease such as cholestatic liver diseases or
cholangiopathies. In total, about 33% of all the
participants are in normal condition and the
remaining 67% may have hepatobiliary disease
such as cholestatic liver diseases or
cholangiopathies.
For the presence of bile acids (C24H40O5)
using Petten Kofer’s test, 83% of the urine
samples from males and 67% of the urine
samples from females exhibited positive results,
a total of 75% of the urine samples from both
males and females exhibited positive results.
C24H40O5 have diverse physiological
functions that include the elimination of
cholesterol, fat absorption, energy expenditure
regulation, and sugar and lipid metabolism
(Monte, Marin, Antelo, & Vazquez-Tato, 2009). In
high concentrations, however, it exhibits
pathological effects due to their direct detergent
effects on biological membranes, apoptotic and
necrotic effects, and cancer-promoting effects
(Beuers, 2010). As such, the presence of bile
acids in the urine serves as a biomarker for
cholestatic diseases (Sinakos & Lindor, 2010).
In using the presence of C24H40O5 as a
biomarker as presented in table 4.1, together with
the result of the qualitative tests for the presence
of C24H40O5 using Petten Kofer’s test presented in
table 4.2, about 17% of the male participants are
in normal condition and the remaining 83% may
have a cholestatic disease. For female
participants, about 33% are in normal condition
and the remaining 67% may have a cholestatic
disease. In total, about 25% of all the participants
are in normal condition and the remaining 75%
may have a cholestatic disease.
For the presence of blood using
benzidine test, 0% of the urine samples from
males and 0% of the urine samples from females
exhibited positive results, a total of 0% of the
urine samples from both males and females
exhibited positive results.
The presence of blood in the urine or
hematuria is a biomarker for serious conditions
that include urinary tract infection, acute cystitis,
pyelonephritis, kidney stones,
glomerulonephritis, polycystic kidney disease,
and sickle cell disease (National Institute of
Diabetes and Digestive and Kidney Diseases,
2018).
In using the presence of blood as a
biomarker as presented in table 4.1, together with
the result of the qualitative tests for the presence
of blood using the benzidine test presented in
table 4.2, 100% of the male participants are in
normal condition and 0% may exhibit urinary tract
infection, acute cystitis, pyelonephritis, kidney
stones, glomerulonephritis, polycystic kidney
disease, or sickle cell disease. For female
participants, 100% are in normal condition and
0% may exhibit urinary tract infection, acute
cystitis, pyelonephritis, kidney stones,
glomerulonephritis, polycystic kidney disease, or
sickle cell disease. In total, 100% of all the
participants are in normal condition and 0% may
exhibit urinary tract infection, acute cystitis,
pyelonephritis, kidney stones,
glomerulonephritis, polycystic kidney disease, or
sickle cell disease.
In conclusion, based on the results of the
tests for physical properties, both of the randomly
selected urine samples exhibited normal
condition. For the results of the qualitative tests
for urine biomarkers, generally, most of the
participants are also in normal condition.
However, there is still a probability of exhibiting
diseases of the kidney and urinary tract,
hepatobiliary diseases, diabetes, and heart
diseases for some of the participants. Overall, the
results of the tests for physical properties and the
qualitative tests for urine biomarkers are
influenced by certain lifestyle factors such as the
use of diuretics, dehydration, starvation,
increased or decreased intake of foods having
certain constituents found in urine, and others.
19
 Biomolecules
Urine Biomarkers

The researchers recommend to infrared vibrational spectroscopy (Oliver, et al.,

reproduce the qualitative tests results using the
methodology used in the study and via the
dipstick method; to conduct quantitative tests in
urinalysis using spectroscopy methods such as
the near-infrared spectroscopy (Shaw, Kotowich,
Mantsch, & Leroux, 1996), surface-enhanced
Raman spectroscopy (Premasiri, Clarke, &
Womble, 2001), and the Fourier transform
2016); and to have a follow-up diagnostic
screening tests such as magnetic resonance
imaging, computerized-tomography scan,
plasma chemistry profile, and complete blood
count analysis among others (National Health
Service, 2017) (Lights & Boskey, 2017) (Mayo
Foundation for Medical Education and Research,
2019) (Matheny, Long, & Roth, 2019).

Bibliography

Journals

Armstrong, J. (2007). Urinalysis in Western culture: A brief history. Kidney International, 384-387.

Bethesda, M. (2014). 2014 USRDS annual data report: Epidemiology of kidney disease in the United
States. United States Renal Data System. National Institutes of Health, National Institute of
Diabetes and Digestive and Kidney Diseases, 188-210.

Beuers, M. (2010). Bile salts and cholestasis. Digestive and Liver Disease, 42,409–418.

Carlisle, E., Donnelly, S., & Halperin, M. (1991). Renal tubular acidosis (RTA): recognize the ammonium
defect and pHorget the urine pH. Pediatric Nephrology, 5(2):242-8.

Coppens, A., Speeckaert, M., & Delanghe, J. (2010). The pre-analytical challenges of routine urinalysis.
Acta Clinica Belgica, 65(3):182-9.

Coresh, J., Astor, C., Greene, T., Eknoyan, G., & Levey, S. (2003). Prevalence of Chronic Kidney
Disease and Decreased Kidney Function in the Adult U.S. Population: Third National Health and
Nutrition Examination Survey. American Journal of Kidney Diseases, 41:1–12.

Crop, M., Hoorn, E., Lindemans, J., & Zietse, R. (2007). Hypokalaemia and subsequent hyperkalaemia in
hospitalized patients. Nephrology Dialysis Transplantation, 22(12):3471.

Fickert, P., & Wagner, M. (2017). Biliary bile acids in hepatobiliary injury – What is the link? Journal of
Heaptology, 619-631.

Hostetter, T. (2014). Chronic Kidney Disease Predicts Cardiovascular Disease. New England Journal of
Medicine, 351(13):1344–46.

Huang, C., & Kuo, E. (2007). Mechanism of Hypokalemia in Magnesium Deficiency. Journal of the
American Society of Nephrology, 18(10):2649.

Kamaledeen, A., & Vivekanantham, S. (2015). The rise and fall of uroscopy as a parable for. Journal of
the Royal College of Physicians of Edinburgh, 45: 63–6.

Kassebaum, N., Arora, M., Barber, R., Bhutta, Z., J, B., & A., C. (2016). GBD 2015 DALYs and HALE
Collaborators. Global, regional, and national disability-adjusted life-years (DALYs) for 315
diseases and injuries and healthy life expectancy (HALE), 1990-2015: a systematic analysis for
the Global Burden of Disease Study 2015. Lancet, 10 8;388(10053):1603–58.

King, H., Aubert, H., & Herman, W. (1998). Global Burden of Diabetes, 1995–2025. Prevalence,
Numerical Estimates, and Projection. Diabetes Care, 21:1414–31.

20
 Biomolecules
Urine Biomarkers

Lee, H., Osis, G., Handlogten, M., Guo, H., Verlander, J., & Weiner, D. (2015). Effect of dietary protein
restriction on renal ammonia metabolism. American Journal of Physiology-Renal Physiology,
308(12): F1463–F1473.

Luyckx, V., Tonelli, M., & Stanifer, J. (2018). The global burden of kidney disease and the sustainable
development goals. Bulletin of the World Health Organization, 96:414-422D. doi:
http://dx.doi.org/10.2471/BLT.17.206441.

Monte, M. J., Marin, J. J., Antelo, A., & Vazquez-Tato, J. (2009). Bile acids: chemistry, physiology, and
pathophysiology. World Journal of Gastroenterology, 15,804–816.

Oliver, K., Vilasi, A., Maréchal, A., Moochhala, S., Unwin, R., & Rich, P. (2016). Infrared vibrational
spectroscopy: a rapid and novel diagnostic and monitoring tool for cystinuria. Scientific Reports,
34737.

Premasiri, W., Clarke, R., & Womble, M. (2001). Urine analysis by laser Raman spectroscopy. Lasers in
Surgery and Medicine, 330-4.

Sewify, M., Nair, S., Warsame, S., Murad, M., Alhubail, A., Behbehani, K., . . . Tiss, A. (2016). Prevalence
of Urinary Tract Infection and Antimicrobial Susceptibility among Diabetic Patients with Controlled
and Uncontrolled Glycemia in Kuwait. Journal of Diabetes Research , Article ID 6573215, 7
pages.

Shaw, R., Kotowich, S., Mantsch, H., & Leroux, M. (1996). Quantitation of protein, creatinine, and urea in
urine by near-infrared spectroscopy. Clinical Biochemistry, 11-19.

Sinakos, E., & Lindor, K. D. (2010). Bile acid profiles in intrahepatic cholestasis of pregnancy: is this the
solution to the enigma of intrahepatic cholestasis of pregnancy? American Journal of
Gastroenterology, 105,596–598.

Suller, M., Anthony, V., Mathur, S., Feneley, R., Greenman, J., & Stickler, D. (2005). Factors modulating
the pH at which calcium and magnesium phosphates precipitate from human urine. Urological
Research, 33(4):254-60.

Tzimenatos, L., Mahajan, P., Dayan, P., Vitale, M., Linakis, J., Blumberg, S., . . . Kuppermann, N. (2018).
Accuracy of the Urinalysis for Urinary Tract Infections in Febrile Infants 60 Days and Younger.
Pediatrics, 141(2).

van den, B., Frenay, A., Bakker, S., Pasch, A., Hillebrands, J., Lambers, . . . van Goor, H. (2016). High
urinary sulfate concentration is associated with reduced risk of renal disease progression in type
2 diabetes. Nitric oxide: Biology and Chemistry, 1;55-56:18-24.

Weening, J. (2004). Advancing Nephrology around the Globe: An Invitation to Contribute. Journal of the
American Society of Nephrology, 15:2761–62.

Books

Boundless. (2019). Map: Fundamentals of General Organic & Biological Chemistry (McMurry et. al.). In J.
McMurry, V. Peterson, & C. Hoegar, Fundamentals of General Organic & Biological Chemistry.

Bringhurst, F., Krane, S., & Kronenberg, H. (2012). Harrison's Principles of Internal Medicine. New York:
McGraw-Hill.

Goldman, L., & Schafer, A. (2011). Cecil Medicine. 24th ed. Philadelphia, Pa: Saunders Elsevier.

21
 Biomolecules
Urine Biomarkers

Institute for Quality and Efficiency in Health Care. (2018). Type 2 diabetes: Measuring sugar levels in
blood and urine yourself. Retrieved from National Center for Biotechnology Information:
https://www.ncbi.nlm.nih.gov/books/NBK279508/

Jamison, D., Breman, J., & Measham, A. (2006). Disease Control Priorities in Developing Countries. 2nd
edition. Washington (DC); New York: The International Bank for Reconstruction and Development
/ The World Bank; Oxford University Press.

John, D., Giuseppe, R., Susan, H., Arrigo, S., & Adibul, H. (2006). Diseases of the Kidney and the Urinary
System. National Center for Biotechnology Information, Chapter 36.

Matheny, S., Long, K., & Roth, J. (2019). Chapter 33: Hepatobiliary Disorders. Retrieved from Access
Medicine:
https://accessmedicine.mhmedical.com/content.aspx?bookid=1415&sectionid=77058047

Pagana, K., Pagana, T., & Pagana, T. (2019). Mosby's Diagnostic & Laboratory Test Reference. 14th
Edition. St. Louis, Mo: Elsevier.

Web Sites

Agarwal, R., Anand, U., & Anand, C. (2016). Test for Occult Blood. The Benzidine Reaction: a Text Book
Error. Retrieved from Wiley: https://iubmb.onlinelibrary.wiley.com/doi/pdf/10.1016/0307-
4412(94)90014-0

American Association for Clinical Chemistry. (2016). Chloride. Retrieved from Labtestsonline:
https://labtestsonline.org/tests/chloride

American Kidney Fund. (2019). Protein in Urine. Retrieved from Kidney Fund:
https://www.kidneyfund.org/kidney-disease/kidney-problems/protein-in-urine.html

Barhum, L. (2018). Why does my urine smell like ammonia? Retrieved from Medical News Today:
https://www.medicalnewstoday.com/articles/320903.php

Biggers, A. (2019). Crystals in the Urine: What You Need to Know. Retrieved from Healthline:
https://www.healthline.com/health/urine-crystals

Blocka, K. (2018). Urine Calcium Level Tests. Retrieved from healthline:

https://www.healthline.com/health/calcium-urine

Brown, E. (2019). Hypercalcemia and autosomal dominant hypocalcemia. Retrieved from UpToDate:
https://www.uptodate.com/contents/disorders-of-the-calcium-sensing-receptor-familial-
hypocalciuric-hypercalcemia-and-autosomal-dominant-
hypocalcemia?search=Urine%20magnesium%20excretion&source=search_result&selectedTitle=
7~25&usage_type=default&display_ra

Chemocare. (2019). Hyperuricemia (High Uric Acid). Retrieved from Chemocare:

http://chemocare.com/chemotherapy/side-effects/hyperuricemia-high-uric-acid.aspx

Devkota, B. (2019). Magnesium . Retrieved from Medscape:

https://emedicine.medscape.com/article/2088140-overview#a2

D'Sa, J. (2015). Normal constituents of urine. Retrieved from Slideshare:

https://www.slideshare.net/janicedesa/normal-constituents-of-urine)

Felson, S. (2019). What is a Phosphate Blood Test? Retrieved from WebMD: https://www.webmd.com/a-
to-z-guides/phosphate-blood-test#1

22
 Biomolecules
Urine Biomarkers

Fogoros, R. (2018). The Importance of Amino Acids With Protein Deficiency. Retrieved from verywellfit:
https://www.verywellfit.com/what-are-the-effects-of-protein-deficiency-4160404

Freeth, A. (2019). Diabetes Causes, Myths, Treatment, and Home Care. Retrieved from eMediHealth:
https://www.emedihealth.com/manage-diabetes.html

Gabbey, A. E. (2018). Uric Acid Test (Urine Analysis). Retrieved from healthline:
https://www.healthline.com/health/uric-acid-urine

Gabbey, A. E. (2018). Urine Urea Nitrogen Test. Retrieved from healthline:

https://www.healthline.com/health/urea-nitrogen-urine

Herrine, S. (2018). Cholestasis. Retrieved from MSD Manual Consumer Version:

https://www.msdmanuals.com/home/liver-and-gallbladder-disorders/manifestations-of-liver-
disease/cholestasis

Higgins, C. (2016). Urea and the clinical value of measuring blood urea concentration. Retrieved from
acutecaretesting.org: https://acutecaretesting.org/en/articles/urea-and-the-clinical-value-of-
measuring-blood-urea-concentration

Holland, K. (2016). Potassium Urine Test. Retrieved from healthline:

https://www.healthline.com/health/potassium-urine

Krans, B., & Jewell, T. (2018). Urinalysis. Retrieved from Healthline:

https://www.healthline.com/health/urinalysis

Laboratory Corporation of America® Holdings. (2019). Sulfate, Quantitative, 24-Hour Urine. Retrieved
from LabCorp: https://www.labcorp.com/test-menu/35251/sulfate-quantitative-24-hour-urine#

Lights, V., & Boskey, E. (2017). Everything You Need to Know About Urinary Tract Infection. Retrieved
from healthline: https://www.healthline.com/health/urinary-tract-infection-adults

Mayo Foundation for Medical Education and Research. (2018). Protein in Urine. Retrieved from Mayo
Clinic: https://www.mayoclinic.org/symptoms/protein-in-urine/basics/causes/sym-20050656

Mayo Foundation for Medical Education and Research. (2019). Blood urea nitrogen (BUN) test. Retrieved
from Mayo Clinic: https://www.mayoclinic.org/tests-procedures/blood-urea-nitrogen/about/pac-
20384821

Mayo Foundation for Medical Education and Research. (2019). Chronic kidney disease. Retrieved from
Mayo Clinic: https://www.mayoclinic.org/diseases-conditions/chronic-kidney-disease/diagnosis-
treatment/drc-20354527

Mayo Foundation for Medical Education and Research. (2019). High uric acid level. Retrieved from Mayo
Clinic: https://www.mayoclinic.org/symptoms/high-uric-acid-level/basics/causes/sym-20050607

Mayo Foundation for Medical Education and Research. (2019). Test ID: CTU . Retrieved from Mayo Clinic
Laboratories: https://www.mayocliniclabs.com/test-catalog/Clinical+and+Interpretive/8513

Michigan Medicine, University of Michigan. (2018). Creatinine and Creatinine Clearance. Retrieved from
uofmhealth.org: https://www.uofmhealth.org/health-library/hw4322

Mount, D. (2019). Hypouricemia: Causes and clinical significance. Retrieved from UpToDate:
https://www.uptodate.com/contents/hypouricemia-causes-and-clinical-significance

Murrell, D. (2018). Urine Calcium Level Tests. Retrieved from Healthline:

https://www.healthline.com/health/calcium-urine

23
 Biomolecules
Urine Biomarkers

Nall, R. (2017). Checking Ketone Levels. Retrieved from healthline:

https://www.healthline.com/health/type-2-diabetes/facts-ketones#complications

National Health Service. (2017). Diagnosis. Retrieved from NHS: https://www.nhs.uk/conditions/coronary-

heart-disease/diagnosis/

National Institute of Diabetes and Digestive and Kidney Diseases. (2018). "Hematuria (Blood in the
Urine).". Retrieved from WebMD: https://www.webmd.com/digestive-disorders/blood-in-urine-
causes#1

National Institute of Diabetes and Digestive and Kidney Diseases. (2019). Your Kidneys & How They
Work. Retrieved from NIH: https://www.niddk.nih.gov/health-information/kidney-disease/kidneys-
how-they-work

National Kidney and Transplant Institute. (2019). KIDNEY HEALTH PLUS. Retrieved from nkti.gov.ph:
http://www.nkti.gov.ph/index.php/patients-and-visitors/kidney-health-plus

Pietrangelo, A. (2017). Creatinine Urine Test (Urine 24-Hour Volume Test). Retrieved from healthline:
https://www.healthline.com/health/creatinine-clearance

Refuerzo. (2018). Health Office Refutes UTI cases. Retrieved from pressreader:
https://www.pressreader.com/philippines/sunstar-baguio/20180714/281479277188406

Selengkapniya, B. (2013). Pettenkofer Test. Retrieved from Professor Biology:

http://birdingpark.blogspot.com/2013/11/pettenkofer-test.html

Stephens, C. (2017). Creatinine Urine Test (Urine 24-Hour Volume Test). Retrieved from Healthline:
https://www.healthline.com/health/creatinine-clearance

Stephens, C. (2018). Urine Urea Nitrogen Test. Retrieved from Healthline:

https://www.healthline.com/health/urea-nitrogen-urine#process

U.S. National Library of Medicine. (2019). Ketones in Urine. Retrieved from Medline Plus:
https://medlineplus.gov/lab-tests/ketones-in-urine/

U.S. National Library of Medicine. (2019). Phosphate in Urine. Retrieved from MedlinePlus:
https://medlineplus.gov/lab-tests/phosphate-in-urine/

U.S. National Library of Medicine. (2019). ToxTutor. Retrieved from Urinary Excretion:
https://toxtutor.nlm.nih.gov/13-002.html

University of Rochester Medical Center Rochester. (2019). Chloride (Urine). Retrieved from University of
Rochester Medical Center:
https://www.urmc.rochester.edu/encyclopedia/content.aspx?contenttypeid=167&contentid=chlorid
e_urine

WebMD LLC. (2019). What Is a Urine Phosphate Test? Retrieved from WebMD:
https://www.webmd.com/kidney-stones/urine-phosphate-test#1

24