URINE INGREDIENT ANALYSIS

Name: Yunita Punarisma

Class: XI IA 3/ 15

BIOLOGY LABORATORY

STELLA DUCE SENIOR HIGH SCHOOL

YOGYAKARTA
I. Introduction
Urine is a liquid product of the body that is secreted by the kidneys by a process
called urination and excreted through the urethra. Cellular metabolism generates numerous
waste compounds, many rich in nitrogen, which require elimination from the bloodstream.
This waste is eventually expelled from the body in a process known as micturition, the
primary method for excreting water-soluble chemicals from the body. These chemicals can
be detected and analyzed by urinalysis. Amniotic fluid is closely related to urine, and can be
analyzed by amniocentesis.

Urine formation begins with the movement of plasma ultra filtrate into the kidneys.
This plasma ultra filtrate is an essentially protein-free fluid which passively passes from the
glomerular capillaries into the Bowman's space. This process is driven by Starling forces.
Glomerular filtration is followed by reabsorbtion of water and solutes from the different
parts of the renal tubules, then by the secretion of selected solutes into the renal tubules.

II. Problems:
a. What are the physical and chemist feature in urine?

b. What are the substances kinds that implied in urine?

III. Objectives:
a. To detect physical feature and chemist feature in urine.

b. To detect kinds of substances that implied in urine.

IV. Literature
Blood Cleaning by the Kidneys

This section is about processes performed by the kidneys in order to filter (clean) blood. 
These are:

1. Glomerular Filtration also called "Ultra-filtration",

2. Tubular Reabsorption also called "Selective Re-Absorption" and

3. Tubular Secretion.

(1) Glomerular Filtration

Blood enters the kidney via the renal artery. 

This separates many times (Renal Artery -> Segmental Arteries -> Interlobar Arteries -> Arcuate Arteries
-> Interlobular Arteries -> Afferent Arterioles), eventually forms manyafferent arterioles, each of which
delivers blood to an individual kidney nephron.

The diameter of the afferent (incoming) arteriole is greater than the diameter of the efferent arteriole
(by which blood leaves the glomerulus). The pressure of the blood inside the glomerulus is increased
due to the difference in diameter of the incoming and out-going arterioles. 
This increased blood pressure helps to force the following components of the blood out of the
glomerular capillaries:

 Most of the water;

 Most/all of the salts;

 Most/all of the glucose;

 Most/all of the urea.

The above are filtered in preference to other components of blood based on particle size. (Water and
solutes of relative molecular mass less than 68,000 form the filtrate.) Blood cells and plasma proteins
are not filtered through the glomerular capillaries because they are relatively larger in physical size.

The water and salts that have been forced out of the glomerular capillaries pass into the Bowman's
Capsule and are called the glomerular filtrate. This glomerular filtrate is formed at a rate of above 125
cm3 per minute in humans. This volume is approx. 20% of the plasma delivered during that time.
(Again: It contains all the materials present in the blood except blood cells and most proteins - which are
too large to cross the basement membrane of the glomerulus.)

(2) Tubular Reabsorption

Only about 1% of the glomerular filtrate actually leaves the body because the rest (the other 99%) is
reabsorbed into the blood while it passes through the renal tubules and ducts. This is called tubular
reabsorption and occurs via three mechanisms. They are:

 Osmosis

 Diffusion, and

 Active Transport.

Reabsorption varies according to the body's needs, enabling the body to retain most of its nutrients.
The processes of tubular reabsorption occur in the following order :

In the PCT
Most of the volume of the filtrate solution is reabsorbed in the proximal convoluted tubule (PCT). This
includes some water and most/all of the glucose (except in the case of diabetics).

Most of the energy consumed by the kidneys is used in the reabsorption of sodium ions (Na +), which
are solutes - that is, they are dissolved in the water component of the filtrate solution. 
As the concentration of Na+ in the filtrate solution are high (about the same as the concentration of
Na+ in blood plasma), Na+ moves from the tubular fluid into the cells of the PCT. In the cases of many
Na+ ions this occurs with the help of symporters. Symporters simultaneously facilitate passage through
the PCT membrane of both Na+ and other substances/solutes. Other such substances that are
reabsorbed with Na+ in this way include glucose (an important type of sugar), amino acids, lactic acid,
and bicarbonate ions (HCO3-). These then move on through cells via diffusion and/or other transport
processes.
A short way to summarize the above is to say that solutes are selectively moved from the glomular
filtrate to the plasma by active transport. (However, almost all glucose and amino acids, and high but
variable amounts of ions, are reabsorbed again later - see the next section, below).

Following the movement of solutes (including Na +), water is then also reabsorbed by osmosis. About
80% of the filtrate volume is reabsorbed in this way. As this part of the reabsorption process is not
controlled by the proximal tubule itself, it is sometimes called obligatory water reabsorption.

In the Loop of Henle: the remaining water (together with the dissolved salts and urea) passes from the
PCT into the descending limb of Henle. It then passes along the Loop of Henle, and up the ascending
limb of Henle.

The different permeability properties of the two limbs of the Loop of Henle, together with their counter
flow arrangement, allows a countercurrent multiplication to generate a high solute concentration in
the tissue fluid of the medulla (that is, outside of the tubules). The highest solute concentrations are
generated deep in the medulla. This is explained as follows:

1. Descending Limb of Loop of Henle

The epithelium lining of the descending limb of Henle is relatively permeable to water - but
 much  less  permeable to the salts Na+  and Cl-, and to urea. Therefore water gradually
moves from the descending limb and into the interstitium (surrounding the tubules) as fluid
flows through this part of the system of renal tubules.

2. Thin Ascending Limb of Loop of Henle

The thin ascending limb of Henle differs from the descending limb in that it is impermeable to
water (so the water that is inside the tubule at this stage generally remains inside it), but is
highly permeable to Na+  and Cl-, and somewhat permeable to urea. Therefore while the tubular
fluid flows back towards the renal cortex, Na + and Cl- (which are more concentrated in the
tubular fluid than in the interstital fluid) diffuse from the tubules into the interstitium. Some
urea also enters the tubules at this stage - but the loss of NaCl from the tubular fluid greatly
exceeds the gain in urea.

3. Thick Ascending Limb of Loop of Henle

the thick ascending limb of Henle (and its continuation into the first part of the DCT), reabsorbs
NaCl from the tubular fluid via a different transport process from that of the thin ascending
limb of Henle.

The overall effect of the processes outlined above is that the concentration of the fluid inside the renal
tubules that form the Loop of Henle is highest at the deepest part of the renal medulla, and is less
concentrated in the renal cortex. This is what is meant by the "concentration gradient" of the Loop of
Henle. The term "counter-current" is also used in descriptions of the Loop of Henle - and refers to the
tubular fluid flowing in opposite directions along the descending and ascending limbs (as indicated by
the thin red arrows in the diagram above.

In the DCT
The water, urea, and salts contained within the ascending limb of Henle eventually pass into the distal
convoluted tubule (DCT).

The DCT reacts to the amount of anti-diuretic hormone (ADH) in the blood:

 The more ADH is present in the blood, the more water is re-absorbed into it. This happens
because the presence of ADH in the blood causes the cells in the last section of the DCT (and
associated tubules and collecting ducts) to become more permeable to water, therefore they
allow more water to pass from the tubular fluid back into the blood. This results in more
concentrated urine.

 The opposite is also true, i.e. if the level of ADH in the blood is reduced then the cells in the
latter sections of the DCT (and associated tubules and collecting ducts) becomes less permeable
to water therefore less water is able to pass from the tubular fluid back into the blood - which
results in less concentrated urine.

The amount of ADH in the blood may be affected by conditions such as diabetes insipidus, or by
consumption of diuretics* in the diet (*substances that occur in some foods and drinks).
(3) Tubular Secretion
The third process by which the kidneys clean blood (regulating its composition and volume) is
called tubular secretion and involves substances being added to the tubular fluid. This removes
excessive quantities of certain dissolved substances from the body, and also maintains the blood at a
normal healthy pH (which is typically in the range pH 7.35 to pH 7.45).

The substances that are secreted into the tubular fluid (for removal from the body) include:

 Potassium ions (K+),

 Hydrogen ions (H+),

 Ammonium ions (NH4+),

 Creatinine,

 urea,

 some hormones, and

 Some drugs (e.g. penicillin).

Tubular secretion occurs from the epithelial cells that line the renal tubules and collecting ducts.

It is the tubular secretion of H+ and NH4+ from the blood into the tubular fluid (i.e. urine - which is then
excreted from the body via the ureter, bladder, and urethra) that helps to keep blood pH at its normal
level. The movement of these ions also helps to conserve sodium bicarbonate (NaHCO 3).
The typical pH of urine is about 6. Urine formed via the three processes outlined above trickles into the
kidney pelvis. At this final stage it is only approx. 1% of the originally filtered volume but includes high
concentrations of urea and creatinine, and variable concentrations of ions.

The typical volume of urine produced by an average adult is around 1.5 - 2.0 dm 3 per day.

Urine physical characteristics:

Volume is one of the physical characteristics of urine. Other physical characteristics that can apply to
urine include color, turbidity (transparency), smell (odor), pH (acidity - alkalinity), and density.

 Color: Typically yellow-amber but varies according to recent diet and the concentration of the
urine. Drinking more water generally tends to reduce the concentration of urine, and therefore
cause it to have a lighter color. (The converse is also true.)

 Smell: The smell (or "odour", which is the more clinical term, American spelling "odor") of urine
may provide health information. For example, urine of diabetics may have a sweet or fruity
 odour due to the presence of ketones (organic molecules of a particular structure). Generally
fresh urine has a mild smell but aged urine has a stronger odour, similar to that of ammonia.

 Acidity: pH is a measure of the acidity (or alkalinity) of a solution. The pH of a substance

(solution) is usually represented as a number in the range 0 (strong acid) to 14 (strong alkali,
also known as a "base"). Pure water is "neutal" in the sense that it is neither neither acid nor
alkali, it therefore has a pH of 7. The real significance of pH in terms of physical chemistry is
that pH is a measure of the activity of hydrogen ions (H+) in a solution.
The pH of normal urine is generally in the range 4.6 - 8, a typical average being around 6.0.
Much of the variation is due to diet. For example, high protein diets result in more acidic urine,
but vegetarian diets generally result in more alkaline urine (both within the typical range 4.6 -
8).

 Density: Density is also known as "specific gravity". This is the ratio of the weight of a volume of
a substance compared with the weight of the same volume of distilled water. 
Given that urine is mostly water, but also contains some other substances dissolved in the
"water", its density is expected to be close to, but slightly greater than, 1.0. This is true - the
density of normal urine is in the range 0.001 to 0.035.

V. Hypothesis:
a. Physically, urine`s color should be yellow (pH=4,8 – 7,5)

b. Substances, that may implied in urine:

 NaCl

 KCl

 C6H12O6

 Protein

 Ion Cl-

 Ion PO43+

 Ion SO42-

 Ion H2O

 2% Urea
  2% other metabolism product

VI. Tools and Materials:

 Tools:

 Reaction tube

 Pipette

 Chemistry glass

 Bunsen Lamp

 Test Tube Clamp

 Materials:

 Urine 250 ml every student

 Liquid AgNO3

 Liquid HCl

 Benedict

 Fehling

 Paint color indication

 Acetate acid

 Ammonium oxalate

 Sugar

 Water

 Label

VII. Procedures:

(1) Color and Smell Test:

 Take 5 ml urine and pack into reaction tube.

 Place in an available place.

  Check the color urine of available standard urine`s color.

 Note the result.

 Smell the urine and note the result.

(2) pH measurement:

 Take 5 ml of urine and pack it into reaction tube.

 Insert paper pH or pH stick into urine during 30 second.

 Lift pH stick and check the color with color indicator in box pH of pH stick.

 Note the result.

(3) Dilution measurement:

 Take 5 ml urine and pack into reaction tube.

 Keel over reaction tube up to flat

 Count the time urine that gets down to genuinely finished by animate
stopwatch moment urine begin to drip and kill stopwatch when urine genuinely
finished.

 Note the result.

(4) Urine ingredient analysis:

 Glucose Analysis

 Sugar solution test with solution Benedict as color change standard

that happened. Its way, make sugar solution from one sugar
tablespoon that melted into half water glass.

 Put 20 gram of sugar solution drops into reaction tube.

 Drop the sugar solution with 5ml solution of Benedict, shake it until
dissolved.

 Clamp the reaction tube with pincers. Heat the end of tube reaction
above Bunsen lamp`s flame. Every second u take the reaction tube up,
so it won`t burnt. Not until boiling, in 3-5 minutes.

 Attention the color changes. If it`s contains glucose, then it should

appeared a red brick color.

 The, test urine with the same procedure started from second point, we
 only change the glucose solution with urine. Better be students are
checking their own urine narrowly.

 Protein analysis:

 Put 5 ml of urine into the reaction tube.

 Add 5 ml of Biuret.

 Attention the color`s changing (if reddish or purple colors mean it is

positively contains protein)

 Note the result.

 Ion NH4 analysis:

 Put 2 ml of urine.

 Add 5 drops of urine into Na2CO3 and placed the litmus paper above
the tube.

 Heat carefully

 If the red litmus color changed into blue, means there was some
reaction

2 NH4Cl + Na2CO3 ↔ 2 NaCl + (NH4)2CO3

(NH4)2CO3 + H2O ↔ 2 NH4OH + H2CO3

 Ion Ca test:

 Add 1 ml of acetic acid and 1 ml of ammonium oxalate into 5 ml of

urine.

 Attention the appearing of colorless sediment (calcium oxalate)

 Note the result.

 Ion Mg test:

 Solution that contained a calcium oxalate from the result of the test
above was continued by heating it until boil.

 Filter it in hot state with filter paper.

 Take the filtrate and add 2 ml of ammonium until smell something

 typical.

 Attention the ammonium magnesium ammonium phosphate

(MgNH4PO4)

 Ion Cl test:

 Add 3 ml of AgNO3 into 5 ml of urine.

 Attention the sediment.

 Note the result.

 Ion PO4 test:

 Add ammonium molybdate then add 1 ml of concentrated HNO3.

 Heat until boiled.

 Attention the yellow sediment.

 Note the result.

 Ion SO4 test:

 Add 3 ml of BaCl2 and 2 ml of sloppy HCl into 5 ml of urine.

 Attention the sediment

 Note the result.

(5) Pregnancy test:

 Patch all urine in a clean and dry place.

 Ripped the sachet and take the test`s tool.

 Put the end of absorbent test tool in urine container in 5 seconds (don’t run
over the narrow sign that located in the test`s tool)

 The test result can be read in 3 minutes.

Procedure to read the test result: Positive result is when there appeared two
color lines or if the color line of the test area (T) was darker of equal than in
control area (C). Negative result is when there appeared one color line or if the
color line of the test area (T) was lighter than the red color line in control area
or (C)
VIII. Observation data:

A. Physical and chemist characteristic of urine.

Name Color pH Smell Dilution

Fani +++ 6 ++ 20 s
Feli + 6 + 10 s
Tami ++ 6 +++ 10 s

B. Substances that contained in urine.

No Substance Fan Feli Tami Explanation

tested i
1 Glucose - - - Unchanged
2 Protein - - - Unchanged
3 NH4+ - + - Litmus turned into blue
4 Ca+ - - - No sediment
5 Mg+ - - - No sediment
6 Cl- + + + Color changing and sediment
7 Po4- - - - No yellow sediment
8 SO4- + + + White sediment

C. Pregnancy test.

Fani -
Feli -
Tami -

IX. Discussion

 Physical and chemist characteristic of urine.

 Color of urine: Fani got +++, Feli +, and Tami ++. The more + they have
means darker urine`s colors they have. Colors of humans are all
different to each others. Each of them has their own information of
what substances that contained in their body. The three main causes
of abnormal urine color are: Foods you have eaten, Drugs you have
taken, Health issues. The most common colors are:

 Orange: There are many foods that may cause an orange urine
color. Blackberries can give your urine orange hue.
Beets may give your urine a red or orange urine color.
 Rhubarb can cause an orange urine color as well. Senna herbs
have also been known to cause an orange color in urine. Other
foods or medications with orange dye may cause an orange
urine color. There are some serious health issues to consider
that may cause orange urine. Dehydration causes a darker
urine color that may sometimes appear to be orange urine.
Jaundice can cause bile to show up in your urine. The bile can
give urine an orange color.

 Light yellow: A light yellow urine color typically signifies very

healthy urine. The light straw yellow urine will have a low odor,
few bubbles in the urine, little to no foam in urine, and have a
clear urine consistency.

So, we can conclude that the healthiest urine above three

people is Feli`s urine.

 pH of Urine: between three people, they have the same pH of urine in

6. Means they have acidity of solution.

 Urine pH is used to classify urine as either a dilute acid or base

solution. Seven is the point of neutrality on the pH scale. The
lower the pH, the greater the acidity of a solution; the higher
the pH, the greater the alkalinity. The glomerular filtrate of
blood is usually acidified by the kidneys from a pH of
approximately 7.4 to a pH of about 6 in the urine. Depending
on the person's acid-base status, the pH of urine may range
from 4.5 to 8.

 Smell of urine: Fani got ++, Feli got +, and Tami got +++. Actually, smell
doesn’t give you any information accurately from your body. It is
mostly the result of hat you have eaten. But if you have smelled some
sting odor frequently, you better check to doctor to see hat happened.

 Urine`s dilution: Fani had 20 s, Feli 10 s, and Tami 10 s. The faster urine
to flow, the dilute urine that they have. Te more dilutes urine means
you are not thirsty and have no dehydration. It is good, because you
also have a good kidney.

 Urine`s composition

 Glucose: we test them by using Benedict that had been heated with
urine fro 3-5 minutes. We have the result all negative. We didn’t find
any color changing in reaction tube. Means they have no glucose in
 their urine. This is good, cause glucose containing in urine means they
have some disease that caused by glucose contents inside. It prevents
them from being diabetes disease.

 Protein: in protein test, same like glucose, we didn’t find any color`s
changing in reaction tube. Proteins are large molecules which help
make up our muscles, important parts of our immune system, and
many other portions of our bodies. Most proteins are also too large to
pass through the filtering system of the kidney. And since they are not
supposed to pass into the kidney, there is no mechanism for proteins
to be reabsorbed if they make it in there. Therefore, if protein is
detected in the urine, it means there is something going on with the
filter (called the glomerulus) that is allowing the proteins to pass. So,
we can conclude that students have no problems with their internal
excretion organs.

 NH4+: we only have 1 student that positive contained ammonia in her

urine test, who was Feli. As the literature shown above, ammonia
should contain in urine. Because, the third step of kidney`s procedure
(tubular secretion) the ammonia is removed from the blood flow. It is
also caused the odor of the urine. The students that have no ammonia
in their urine may have some trouble with their tubular secretion.

 Ca+: we didn’t find any sediment inside the reaction tube. Means the
students have no calcium oxalate in their body. Calcium oxalate crystals
in the urine are the most common constituent of human kidney stones,
and calcium oxalate crystal formation is also one of the toxic effects
of ethylene glycol poisoning. The urine picture can be shown below as

my opinion supports.

 Mg+: in our experiment, we again had no found any sediment that

indicates any magnesium inside the urine. Though actually, magnesium
in urine means that the person is in health conditions. So magnesium in
our body is so important because Magnesium is needed for more than
300 biochemical reactions in the body. It helps maintain normal muscle
and nerve function, keeps heart rhythm steady, supports a healthy
immune system, and keeps bones strong. Magnesium also helps
 regulate blood sugar levels, promotes normal blood pressure, and is
known to be involved in energy metabolism and protein synthesis.
There is an increased interest in the role of magnesium in preventing
and managing disorders such as hypertension, cardiovascular disease,
and diabetes. Dietary magnesium is absorbed in the small intestines.
Magnesium is excreted through the kidneys.

 Cl-: we have all students positively have Chlorine in their test urine.
Because the color changing and sediment appearing. And it should be
appearing because, the reabsorption do remove chlorine from blood
because it has no use for body anymore. We can conclude that there
was no problem with their glomerulus.

 PO4-: we have all negative result for phosphate contents in the reaction.
Because we didn’t see any sediment inside the reaction tube. It is good,
because phosphate contents indicate a crystal in kidney that can cause
kidney stone.

 SO4-: we have all positive result, because there was white sediment in
reaction tube. Sulfate should be removed from the blood from
reabsorption.

 Pregnancy test: in this test, we have all negative result because appeared one
color line or if the color line of the test area (T) was lighter than the red color
line in control area or (C)

X. Conclusion:

1. Urine`s physical and chemist feature:

 Physical: urine`s color is yellow - dark yellow, it`s dilution, and

has a typical smell from ammonia.

 Chemist: urine has pH between 4.8 and 7.5. Otherwise, is the

health pH one.

2. Urine`s composition: glucose and calcium oxalate only exist when the
person got some infection or problems with their excretion organs.

 NH4+

 Cl-

 SO4-
  Mg+

 PO4 3-

XI. References:

 http://en.wikipedia.org/wiki/Sulfate

 http://en.wikipedia.org/wiki/Urine

 http://id.wikipedia.org/wiki/Fosfat

Guide, Yogyakarta, May 3rd, 2010

Student,

Mr. Gun Yunita Punarisma