Escience labs test tube rack

Experiment 2 Urinalysis Experiment Inventory

Materials 25 mL Simulated Urine Sample A

25 mL Simulated Urine Sample B

25 mL Simulated Urine Sample C

25 mL Simulated Urine Sample D

(4) pH Test Strips

15 mL Benedict’s Solution

10 mL 3% Hydrogen Peroxide, H2O2

10 mL Biuret Solution

*Stopwatch/Timer

*Permanent Marker

Labware (4) Glass Test Tubes

Test Tube Rack

100 mL Graduated Cylinder

(16) Pipettes

*Hot Water Bath

*Hot Pad or Towel

Note: You must provide the materials listed in *red.

EXPERIMENT 2: URINALYSIS Urine is the waste product filtered by the kidney. It is composed of various bodily wastes as well as excess water. Urine is also a helpful tool for doctors when diagnosing various conditions in patients. In this experiment, you will perform a urinalysis on four different urine samples, testing a variety of components. When all components have been tested, you will determine which urine samples are “abnormal” using Table 3.

PROCEDURE Part 1: Testing pH

1. Use the permanent marker to label the test tubes A, B, C, and D.

2. Place the test tubes in the test tube rack.

3. Use a pipette to add 5 mL of Simulated Urine Sample A to the corresponding test tube.

4. Repeat Step 3 with samples B, C, and D. Use a new pipette for each sample.

5. Dip the reaction pad on one pH strip into Sample A for 5 – 10 seconds, and then remove it. Wait approximately 30 seconds, and then compare the resulting color on the pad to the color key (color key provided with the pH strips).

6. Record the pH of Sample A in Table 4.

7. Repeat Steps 5 and 6 with samples B, C, and D. Use a new pH test strip each time.

Part 2: Glucose Test 1. Wash and dry test tubes A – D. Relabel the tubes if the letters wash off.

2. Replace the test tubes in the test tube rack.

3. Use a pipette to add 5 mL of Simulated Urine Sample A to the corresponding test tube.

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4. Repeat Step 3 with samples B, C, and D. Use a new pipette each time.

5. Add 3 mL of Benedict’s solution to each test tube. Gently swirl each tube to mix the solutions.

6. Record the color of the urine samples in Table 5.

7. Prepare a hot water bath. To do this, pour enough water into a pot or deep microwaveable bowl to cover the solution in the test tubes. For example, if the solutions in the tubes are approximately 6 cm deep, fill the pot or bowl with at least 6.1 cm of water.

8. Heat the water on a stove or in a microwave until boiling.

9. Use a hot pad or towel to carefully remove the pot or bowl from the heat source.

10. Place all four tubes into the boiling water bath for 3 minutes. If you do not want to hold the test tubes in the water, you may place the test tubes in a container. However, monitor the set-up to ensure that the tubes do not tip over.

11. Use a hot pad or towel to carefully remove the test tubes from the hot water. Place them in the test tube rack, then record the color change in Table 5.

Note: A reducing sugar is present in the sample if a red, yellow, or green precipitate forms.

Part 3: Protein Test 1. Wash test tubes A – D. Relabel the tubes if the letters wash off.

2. Replace the test tubes in the test tube rack.

3. Use a pipette to add 5 mL of Simulated Urine Sample A to the corresponding test tube.

4. Repeat Step 3 with samples B, C, and D. Use a new pipette each time.

5. Record the color of each sample in Table 6.

6. Add 25 drops of Biuret solution to each test tube.

7. Gently remove sample A from the rack and swirl the solution. Watch for a color change as you swirl. Record any color changes in Table 6.

8. Repeat Step 7 with samples B, C, and D.

Part 4: Yeast Test 1. Wash test tubes A – D. Relabel the tubes if the letters wash off.

2. Replace the test tubes in the test tube rack.

3. Use a pipette to add 5 mL of Simulated Urine A to the corresponding test tube.

4. Repeat Step 3 with samples B, C, and D. Use a new pipette each time.

5. Record the presence or absence of bubbles in Table 7.

6. Add 2 mL of hydrogen peroxide to each test tube.

7. Gently swirl each tube to mix the solutions.

8. Observe the test tubes, and record the presence or absence of bubbles.

Part 5: Ketone Test 1. Wash test tubes A – D. Relabel the tubes if the letters wash off.

2. Replace the test tubes in the test tube rack.

3. Use a pipette to add 5 mL of Simulated Urine A to the corresponding test tube.

4. Repeat Step 3 with samples B, C, and D. Use a new pipette each time.

5. Using a wafting motion (pull your hand over the tube without bringing the tube directly to your nose), note the odor of each sample. Record your observations in Table 8.

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Part 6: Determination of Normal and Abnormal Urine Samples 1. Determine if each result was normal or abnormal for the urine specimens using Table 3. If abnormal, include supporting data

from your experiment.

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Data Sheet Experiment 2 Data Sheet Table 3: Urine Test

Test Normal Abnormal

pH 4.5 - 7.5

Acidic Urine (below 4.5) - Diabetes, starvation, dehydration, respiratory acidosis. Alkaline Urine (above 7.5) - Kidney disease, kidney failure, urinary tract infection, respiratory alkalosis.

Glucose None Glucose present (red or green color after test); diabetes mellitus.

Protein None Protein present (violet color after test); kidney disease.

Yeast None Yeast present (bubbles form after test); yeast infection in urinary tract.

Ketones Little or None Large amount of ketones present (sweet smell of urine); starvation, prolonged vomiting, diabetes, hyperthyroidism, and other metabolic disorders.

Table 4: Simulated Urine pH Test

Simulated Urine Sample pH A B C D

Table 5: Simulated Urine Glucose Test

Simulated Urine Sample Color Before Hot Water Bath Color After Hot

Water Bath A B C D

Table 6: Simulated Urine Protein Test

Simulated Urine Sample Color Before Biuret Solution Color After Biuret

Solution A B C D

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Table 7: Simulated Urine Yeast Test

Simulated Urine Sample Bubbles Before Hydrogen Peroxide? Bubbles After Hydrogen

Peroxide? A B C D

Table 8: Simulated Urine Ketone Test

Simulated Urine Sample Odor Observation

A

B

C

D

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