Experiment: #1

Date: Title: Excretion & Osmoregulation- Urine Test

Rationale: FIVE urine samples were provided to a health clinic but were mixed up in the
laboratory. The samples came from 5 patients who have a variety of health issues and even some
who may be healthy. Diabetics with unmanaged blood glucose levels may have protein in their
urine. A person with kidney damage could have protein in their urine. A person who has too
much acid in their body may also have ketones in their urine.

Aim: To compare the urine samples of the five patients after testing for glucose and ketones in
the urine samples.

Materials & Apparatus: ‘Urine’ samples A, B, C, D and E, Benedict’s solution, distilled water,
potassium hydroxide, dilute copper sulphate, test tests, test tube holder, test tube rack, measuring
cylinders.

Method:
1. Five test tubes were labelled A, B, C, D, and E and 2cm3 of the corresponding ‘urine’
samples were added to each test tube using syringe.

2. 2cm3 of potassium hydroxide and 2cm3 of dilute copper dilute were mixed to create the
Biuret solution and added to the test tubes. Thus, the biuret test was carried out on each
2cm3 sample of urine that was syringed into the test tubes.

3. The observations were then recorded in a table.

4. Five fresh test tubes were labelled A, B, C, D and E and 2cm3 of the corresponding
‘urine’ samples were added to the test tubes by using a syringe.

5. Benedict’s test was carried on each sample. This was done by adding 2cm 3 of the ’urine’
samples into the test tube then adding 2cm3 of Benedict’s solution to each sample using a
syringe. The test tubes were then placed in a beaker of boiling water using clamp. It was
placed there for 5 minutes.

6. After the 5 minutes were up, the test tubes were taken out of the boiling bath and
observed. The observations were recorded then each sample was tested with glucose test
strip and the color and concentration of the sample were recorded.
 7. Five fresh tubes were labelled A, B, C, D, and E. 2cm3 of corresponding ‘urine’ samples
were then added to each test tube.

8. The ketone strips were dipped into the ‘urine’ samples to test for ketones.

9. The color of the strip and its corresponding concentration was observed and recorded in a
table along with the inferences.

Results:

Table Showing Results for Tests on Urine Samples

Sample Biuret Test Benedicts Test and Glucose test Ketones Test strip
Strips
Results Inferences Results Inferences Results Inferences
A Blue solution No protein Negative Not present Negative beige
with blue present
precipitate

B Dark blue No protein 500 Glucose present. Negative beige

solution with present mmol/l (+ Strip color turned
blue precipitate +) 30 from aqua to
green
C Purple solution Protein Negative Glucose not Negative beige
with dark blue present present
precipitate

D Light blue Protein not Negative Glucose not 0.5 peach

solution with present present mmol/l
light blue
precipitate

E Dark purple Protein 250 Glucose present. negative beige

solution with present mmol/l (+) Strip color turn
dark blue 15 from aqua to
precipitate green
 Discussion:

Excretion is the process by which removal of metabolic waste and harmful substances produced by
chemical reactions occurring inside body cells (metabolic processes) are removed from the body.
The kidney has numerous coiled in them called nephrons. They are the functional unit of the
kidney. The nephron has various parts. They are the glomerulus, the Bowman’s capsule
(glomerulus capsule), the Proxima convoluted tubule, the loop of Henle, distal convoluted tubule,
and the collecting duct. In the glomeruli, ultrafiltration occurs. Due to the increase of high
hydrostatic pressure small molecules in the blood are forced into the bowman’s capsule or
glomerulus capsule. It also performs selective absorption which occurs in the Proxima convoluted
tubule. This is process, the reabsorption of a significant portion of vital substances that were first
filtered out of the blood in the glomerulus. These essential substances are glucose, amino acids,
salts, vitamins, hormones, and water. The Loop Henle maintains sodium ion region, the distal
convoluted tubule reabsorbs sodium and water as needed to preserve electrolyte balance and blood
pressure regulation. Lastly, the collecting duct. It is under influence of ADH as its walls become
more permeable to water when it’s present and when absent, the walls are less permeable to water
thus resulting in dilute urine. ADH Anti diuretic hormone, is created by the hypothalamus and
secreted by the pituitary gland of the brain. It plays a cardinal role in the body’s ability to conserve
water.

The results of the test indicate whether the patients have ketones, protein, and glucose in their
urine and thus will indicate or determine underlying health conditions related to the kidney. In
patient A, there was no glucose nor protein in his urine as the Biuret and Benedict’s solution
remained aqua and so did the glucose test strip. The ketone test was also negative. It can be
inferred that this patient is healthy and has healthy kidney function. In the next patient, patient B,
their sample was also negative for proteins as the solution remained blue. However, the urine
sample tested positive for glucose which was indicated in the olive green color change of the test
strip and the benedict’s solution turning brown. The corresponding amount of glucose for this color
change was 500 mg/dl or 30 mmoL/L is but negative for ketones. This result indicates that Patient
B is a diabetic patient as patients with diabetes have glucose present in their urine Glucose of
course, isn’t supposed to be present in the urine because glucose is 100% absorbed back into the
blood in the Proxima convoluted tubule. The amount of glucose detected was 500 mmol/ which is
reflected in the change of strip from aqua to olive green. Patient C had protein present in his urine
as the urine sample turned purple which indicates a positive result. However, the glucose test and
ketones test were negative as there was no color change in the Benedict’s solution, Glucose test
strip nor ketone test strip. . In patient D’s urine sample, the biuret test was negative therefore, no
protein in the urine sample. The Benedict’s test and glucose strip test was also negative as they
remained their respective colors indicating there was no glucose present in the urine sample either.
However, the urine sample did test positive for ketones indicating the presence of ketones.
The amount of ketones detected was 0.5 mmol/l which changed the color of strip to beige to peach.
Ketones are too large to fit through the tiny slits in the walls of the capillaries of the glomerulus,
however oddly enough, a trace amount of ketones in urine isn’t alarming, however it can indicate
several health conditions and lifestyle conditions including diabetes however the amount of the
ketones in the urine detected doesn’t reflect that health condition. Another factor contributing to
ketone presence in urine is the consumption of low carbohydrate foods for long periods of time the
body being too acidic. Lastly, patient E’s urine sample test positive for protein which is indicated
by the purple color change. The benedict’s test was test positive as the color changed to orange
well as glucose strip test was positive. The positive test was indicated in the color of glucose test
change from aqua to green which indicated the amount of glucose was 250 mg/dl or 15mmoL/L
The ketone test was negative as the ketone strip remained beige. With both glucose and protein
found in the urine, this indicates that patient E has both diabetes and kidney failure. Ketones are
chemicals that the liver produces when it breaks down fats. Ketosis is a metabolic state that occurs
when your body burns fat for energy instead of glucose. This is seen as individuals who are on a
ketogenic diet. However, gluconeogenesis can also occur as glucose molecules are created from
the non-carbohydrate sources. For the patients that have protein in their urine as a result of kidney
damage, it would be best to do a kidney transplant or have the patient go under dialysis. Moreover,
the patients with glucose in their urine, it would be best for them to maintain a healthy lifestyle and
diet. Lastly, the patient with ketones in their urine would just need to control the amount of ketones
produced by the body as it can be problematic when there’s a high concentration of ketones in the
body. This can be done by consuming more alkaline foods such as vegetables.

Precautions:

It was made sure gloves were used as the experiment involved the handling of urine.

It was ensured that the instructions from the manufacturer were followed to avoid error.

Sources of errors:

Since the color changes of the of the ketone strips were so close to one another, the color along it’s
corresponding concentration of ketones, may have been recorded incorrectly due to the individuals
perspective of the color. The same goes for the glucose test with the glucose strips.

The volume of the benedict’s solution and biuret test may have been measured incorrectly.

Limitations:

The experiment was limited to that particular environment which was cold.

The assumption that each "urine" sample is homogeneous, and representative of the entire sample
may not hold true in real-world scenarios. Variability within the samples can impact the results.
Conclusion:

In conclusion, each patient’s urine was tested for the presence of proteins, glucose and ketones.
Patient A’s ‘urine’ sample test negative for proteins, glucose and ketones respectively. The color of
‘urine’ sample remained blue in the Biuret test and same as the Benedict’s test. The glucose strip
remained aqua and the same was observed by the beige color of the ketone strip did not change. In
Patient B’s ‘urine’ sample tested negative protein as the biuret solution mixed with the urine
remained blue. However, the urine sample did test positive for glucose as the strip turned olive
green indicating there was 500 mg/dl or 30 mmoL/L. The ketone test was negative as indicating
the test strip remaining beige. Patient C’s urine sample tested positive for protein as the Biuret
solution turned purple but the glucose test was negative and so was ketones test with the strips
remaining aqua and beige respectively. Moreover, in patient D’s urine sample, protein wasn’t
present and so was glucose as the solution remained blue. However, urine sample did test positive
for ketones. This amount detected was 0.5 mmol/l which was reflected in the peach color change
of the test strip. Lastly, patient E, their urine tested positive for protein which was indicated in the
purple color change of the biuret solution, and glucose was also detected as the test strip did turn
green which indicates a 250 mg/dl or 15 mmoL/L. The ketone test was negative.