CHEM 132.

2 – BIOCHEMISTRY (LABORATORY)
LABORATORY REPORT

Name: Monsanto, Rhey Christian O.__ Date Submitted: 06 – 25 – 21___

Lab Schedule: F 10:00 AM – 01:00 PM Date Performed: _____________

Exercise No. 9
Chemistry of Urine
I. OBJECTIVES:
1. To test urine for pH, specific gravity, and the presence of electrolytes and organic compounds.
2. To test urine for the presence of abnormally occurring compounds of proteins, glucose, and ketone
bodies

II. RESULTS AND DISCUSSION:

A. Color, pH and Specific gravity

Observation/Data
Color Amber
pH 5.9
Specific gravity 1.018

B. Urea

Color of solution Pink

C. Uric acid
Color of Solution Burnt Orange
Presence of Crystals (Yes or No) No
 D. Electrolytes

Electrolytes Presence of Electrolytes (Yes or No)

Chlorides (Cl-) Yes

Sulfate (SO42-) No

Phosphate (PO43-) Yes

Sodium (Na+) Yes

Potassium (K+) Yes

Calcium (Ca2+) No

E. Glucose

Color of solution Brick Red

Estimate of g% >2%

Estimate of mg/dL >2000mg/dL

F. Ketone Bodies
Color of Ring Purple

G. Protein
Observation/s The solution is coagulated

III. DISCUSSION
Urine contains hundreds of various body wastes, including waste from the kidneys. Urine tests can be
performed to determine the physical and chemical qualities of urine, as well as provide information about a
person's health and health issues. The color, pH, specific gravity, and presence of urea, uric acid, and
electrolytes in the urine are all tested. An unusually occurring substance, such as protein, glucose, or ketone
bodies, can also be detected through a urine test.
 Colour, pH and Specific gravity

Color, pH, and specific gravity of urine were examined in this part of the experiment. The color of one's
urine might vary based on what they eat, what medications they take, and how much water they drink. Urine's
standard hue is referred to as "urochrome," which refers to the presence of a yellow pigment in urine. The urine
color will be pale yellow or nearly clear if you stay hydrated. When you're dehydrated, your urine turns a deep
amber or even light brown color. Discolored urine, such as pink, red, blue, or green, is frequently produced by
medications, specific foods, or food colors taken by a person, and it does not indicate a significant illness. The
urine sample had an amber color in the experiment. This indicates that the person is dehydrated and needs to
drink more water.

The acidity or alkalinity of a person's urine can be determined by their diet. If a person consumes too
many acidic foods, their urine becomes acidic, and if they consume too many alkaline foods, their urine
becomes basic. The pH of urine should be between 7.0 and 7.5. Kidney issues might be indicated by a low pH
in the urine. Acidosis, dehydration, diabetic ketoacidosis, and diarrhea are some of the other health issues. A
urine pH that is higher than usual could indicate kidney failure, tubular acidosis in the kidneys, pyloric
obstruction, respiratory alkalosis, urinary tract infection, and vomiting. The urine pH level test, on the other
hand, has no major adverse effects. The pH of the urine sample was 5.9, as stated in the table above. This means
that the urine sample's pH is somewhat acidic. To have a neutral pH, changes in diet is recommended.

A urinometer was used to determine the urine's specific gravity. The principle of buoyancy is used in the
development of urinometers. The urinometer floats higher in urine than in water in this experiment. This
demonstrates that urine has a higher density than water.

The concentration of solutes in the urine is measured by urine specific gravity. It gives information on the
kidney's ability to concentrate urine by measuring the ratio of urine density to water density. The reference range
for urine specific gravity is 1.005-1.030, indicating that the kidneys are in good working condition. Results of
specific gravity greater than 1.010 may suggest moderate dehydration. A high specific gravity indicates that the
urine is particularly concentrated, which can be caused by not drinking enough water, excessive fluid loss
(excessive vomiting, sweating, or diarrhea), or compounds in the urine such as glucose and protein. A low specific
gravity indicates dilute urine, which can be caused by excessive fluid consumption or severe kidney disease. The
specific gravity of the urine sample was 1.018 based on the information gathered. This implies the person is mildly
dehydrated.
Urea
Ammonia is produced by the body when protein in foods is broken down. Nitrogen in ammonia reacts
with other elements in the body to generate urea, which is made up of carbon, hydrogen, and oxygen. When you
urinate, you're excreting urea, which is a waste product. A urease test was performed to identify urea in the urine
sample. Urease is a decomposition enzyme that breaks down urea into carbon dioxide and ammonia. When an
alkaline ingredient, such as sodium carbonate, is added to slightly acidic urine, it transforms into an alkaline
solution. The solution changed colour from yellow to pink as sodium carbonate (Na2CO3) was added. The
presence of ammonia was indicated by the pink colour in the solution. The chemical reaction is expressed as:
Na2CO3 + 3NH2CONH2 → 2NaOCN + 2CO2 + 4NH3
The pink color solution disappeared as acetic acid was added to the solution. This shows that the ammonia
produced in the chemical reaction was neutralized. The chemical reaction is summarized as:
NH3 + CH3COOH → CH3COO- + NH4+
After the ammonia was neutralized, soybean powder (urease) was added to the solution and heated to
speed up the reaction. The hue of the solution changed to pink, as shown in the table. The presence of ammonia
was responsible for the pink color noticed in this experiment. This indicates that urea was detected in the urine
sample. For the reaction, the urea was first hydrolyzed, then urease was used an enzyme to decomposed the urea
into carbon dioxide and ammonia. The chemical reaction is expressed as:

Uric acid
The breakdown of purines produces uric acid, which is a common waste product. Purines are compounds
found in the cells of the body as well as in some meals. Purines are found in foods such as liver, sardines, dried
beans, and beer. The majority of uric acid dissolves in the blood and travels to the kidneys, where it is excreted
in the urine. The urine sample was treated with HCl solution and left for 24 hours while the solution was covered
with plastic wrapping in this experiment. To decrease the degradation caused by changing handling and storage
conditions, hydrochloric acid was utilized as a preservative. Following that, the color changed to brown, and
crystals began to form. The urine sample turned light brown after some time and no crystals developed, indicating
that there is little to no uric acid in the urine sample. Low uric acid levels in the urine do not normally cause health
issues. High levels of uric acid, on the other hand, can cause crystals to accumulate in joints and lead to health
problems like gout, chronic renal disease, and kidney stones.

Electrolytes

Electrolytes are elements and compounds that occur naturally in the body. Electrolytes control important
physiologic functions such muscle functions, hydration, blood pH levels, blood pressure, etc. These substances
are present in the blood, bodily fluids, and urine. Electrolytes are also ingested with food, drinks, and supplements.
Excess electrolytes are filtered by the kidneys are carried out through the urine to be excreted. In this experiment,
presence of electrolytes in urine was conducted through precipitation reaction and flame test. These electrolytes
include, chlorides, sulfates, phosphates, sodium, potassium, and calcium was tested.
Chloride is necessary for maintaining the proper balance of bodily fluids. The presence of chloride in urine
was observed in this experiment as formation of silver chloride (AgCl) precipitate was yielded after the addition
of silver nitrate to the urine sample. The urine was acidified with nitric acid to prevent false positive results.

Urinary sulfate is a reflection of dietary protein intake, particularly meat, fish, and poultry, which are rich
in sulfur-containing amino acids methionine and cysteine. Urinary sulfate can be used to assess dietary protein
intake for nutritional purposes. The presence of sulphate in urine can be detected through precipitation reaction
with barium chloride to form a precipitate of BaSO4 (barium sulfate). As shown in the table, no precipitation
occurred in the reaction of urine sample and barium chloride. This implies that sulfates were not present in the
urine sample.

The kidneys, bones, and intestines work to balance phosphate levels in the body. Phosphate is necessary
for a wide variety of functions and interacts closely with calcium. The presence of phosphate in urine was detected
as yellow precipitate (phosphomolybdate) was formed after the addition of ammonium molybdate to the urine
sample.
Calcium is a vital mineral that your body uses to stabilize blood pressure and control skeletal muscle
contraction. The presence of calcium in urine can be detected through precipitation reaction with the urine and
ammonium oxalate solution. A positive result for the presence of calcium is indicated the formation of a white
precipitate of calcium oxalate. In this experiment, the urine sample used was found to have no calcium present in
the sample urine since there was no precipitation occurred in the reaction.
Sodium is necessary for the body to maintain fluid balance and is critical for normal body function. It also
helps to regulate nerve function and muscle contraction. Potassium is particularly important for
regulating heart function. It also helps maintain healthy nerves and muscles. For the detection of sodium and
potassium electrolytes, flame test was performed. After the nichrome wire was dipped to the urine solution and
was subjected to the flame, appearance of yellow flame and lilac flame (in cobalt glass) were observed. The
appearance of a yellow flame in the burner indicated the presence of sodium in the urine sample while the
appearance of lilac flame in the cobalt indicated the presence of potassium in the urine sample.

Glucose
Glucose is a type of sugar that the body requires and uses for energy. The body converts the carbohydrates into
glucose. Having too much glucose in body can be a sign of a health problem. The most common cause of
elevated glucose levels is diabetes, a condition that affects body’s ability to manage glucose levels. In this part
of the experiment, Benedict’s and Fehling’s test were conducted to detect reducing sugar (glucose) in the urine
sample. As shown in the table, the color of the urine solution after the addition of both the benedict’s and
Fehling’s reagent was brick-red. This shows that the Cu2+ in both reagents is reduced by the glucose to Cu2O
(brick-red precipitate). The color change of the solution indicates the amount of glucose present in the urine.
This indicates that there is a high concentration of glucose in the urine sample which is more than two percent
(>2%) and an estimate of >2000mg/dL.
Ketone Bodies
Ketone bodies are substances produced by the liver during gluconeogenesis, a process that creates glucose
in times of fasting and starvation. If the cells don't get enough glucose, the body burns fat for energy instead. This
produces a substance called ketones, which can show up in the blood and urine. Rothera’s test was performed in
this experiment to detect the presence of ketone bodies in the urine sample. As shown in the table, purple ring
was formed in the solution. This implies that ketone bodies were present in the urine sample. The ring purple
observed in the solution was the product of complex between the ketone and sodium nitropusside in the presence
of ammonia (alkaline solution). The presence of ketone bodies in the urine might indicate that the person has a
medical condition namely ketonuria. Ketonuria is a medical condition in which the body produces excess ketones
as an indication that it is using an alternative source of energy and can be seen during starvation or more
commonly in type 1 diabetes mellitus.

Protein
A protein in urine test measures how much protein is in the urine. Proteins are substances that are essential
for the body to function properly. Protein is normally found in the blood. If there is a problem with the kidneys,
protein can leak into the urine. A small amount is normal, but a large amount of protein in urine may
indicate kidney disease. For the presence of protein albumin in the urine sample, upper portion of the test tube
with the urine sample was heated. Acetic acid was then added to the sample to adjust the pH of the sample to the
isoelectric point of the albumin. During heating, coagulation was formed in the upper portion of the sample. This
indicates the positive results of protein albumin in the urine sample. However, medical condition cannot be
determine since the test is only qualitative and does not include the concentration of protein.
 IV. CONCLUSIONS:

This experiment was successful in determining the pH, specific gravity, and presence of urea, uric acid,
and electrolytes in the urine, as well as detecting abnormally formed chemicals using several tests. The urine
sample in this experiment was amber in color and had a specific gravity of 1.018, indicating that the subject was
somewhat dehydrated. The pH of the urine sample was 5.9, indicating that it was slightly acidic and that it needed
to be balanced. In addition, urea was identified in the urine sample using a urease test, which revealed the
production of a pink complex. This experiment only collected a small amount of uric acid in the urine sample.
Low uric acid levels do not normally cause health issues. Precipitation reaction and flame tests were also used to
detect electrolytes in the urine sample. Chloride, sodium, potassium, and phosphate were among the electrolytes
found in the urine sample. Calcium and sulfates were the only electrolytes absent from the urine sample. In
addition, abnormally compounds (ketone bodies, glucose, and proteins) were found in the urine. A high
concentration of glucose was identified using Benedict's and Fehling's tests. The development of a purple ring
under alkaline circumstances was used to detect ketone bodies. Finally, when the urine sample coagulated in the
upper area of the test tube, the protein albumin was detected. The protein albumin found in the urine denatured
after heating the urine sample.