BIOCHEMISTRY LABORATORY WORKSHEET
Name: Rating:
Course, Year and Section: Instructor:
Group Number: Date:
Urine
EXPERIMENT 7
Objectives:
At the end of the experiment, students should be able to:
1. Describe the characteristics and composition of normal urine
2. Determine the organic and inorganic constituents of urine.
3. Analyze and differentiate normal and pathological urine samples
Reagents/Materials
5% glucose 10% NaOH sulfur
5% (NH4)2SO4 10% HCl Benedict’s reagent
5% Sodium nitroprusside Concentrated HCl Acetone
5% albumin Concentrated NH4OH Litmus paper
5% BaCl2 Saturated picric acid
5% Acetic acid Saturated NaCl solution
Procedures:
First Batch
I. Physical Characteristics
1. Collect mid-stream urine sample in a container.
2. Describe the urine specimen as to color, odor, transparency and pH.
II. Test for Organic and Inorganic Constituents
A. Test for Creatinine
1. In a test tube, add 1 mL saturated picric acid and 1 mL of 10% NaOH to 5 mL urine.
2. Observe the color formed.
B. Test for Carbonates
1. In a test tube, add 10 drops of concentrated HCl to 5 mL urine.
2. Observe.
C. Test for Sulfates
3. In a test tube, add 1 mL of 10% HCl to 5 mL urine. Add a few drops of 5% BaCl2
4. Observe.
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BIOCHEMISTRY LABORATORY WORKSHEET
III. Test for Pathological Constituents
A. Test for Glucose – Benedict’s Test
1. To 1.0 mL of urine sample, add 5 mL of Benedict’s reagent.
2. Boil in a water bath for two minutes.
3. In another test tube, add 2 drops of 5% glucose to 1.0 mL of urine sample.
4. Add 5 mL of Benedict’s reagent and then boil in a water bath for two minutes.
Second Batch
B. Test for Ketone Bodies – Rothera’s Test
1. Place 5 mL of urine in a test tube. Add 3 drops of 5% (NH4)2SO4 and 3 drops of 5%
sodium nitroprusside.
2. Carefully add 1 mL of concentrated NH4OH and set aside for 2 to 3 minutes before
observing.
3. Repeat steps 1 and 2 but do not forget to add 5 drops of acetone to the urine sample
in step 1.
C. Test for Proteins – Osgood Ruskin Test
1. Place 5 mL of urine in a large test tube. Add 1 mL of 5% acetic acid and 3 mL of
saturated NaCl solution.
2. Using a test tube holder, heat the upper portion of the test tube over the flame.
3. Record your observation.
4. Repeat steps 1 and 2 but do not forget to add 5 mL of albumin to the urine sample in
step 1.
D. Test for Bile Acids
1. Place the urine sample in an ice bath for 30 minutes.
2. Sprinkle a small amount of powdered sulfur on the surface.
3. Observe what happens to the sulfur.
Note: Students in the second batch also need to collect their own samples and take note the
physical characteristics of their urine
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BIOCHEMISTRY LABORATORY WORKSHEET
Name: Rating:
Course, Year and Section: Instructor:
Group Number: Date:
Urine
EXPERIMENT 7
I. Physical Characteristics
Sample Color Odor Transparency pH
1st Sample of Urine Amber Mild Smell Clear 6
II. Organic and Inorganic Constituents of Urine
Result Explanation
5 mL of urine change its color to orange
Test for Creatinine when 1 mL of saturated picric acid and 1
mL of 10% NaOH are added. This indicates
POSITIVE (+) that the creatinine test result for the sample
ORANGE is positive. This occurs as a result of
creatinine being removed through urine
after being removed from the blood by the
kidneys.
A 5 mL urine sample was mixed with 10
Test for Carbonates drops of concentrated HCl, and the
resultant solution was dark yellow or amber
POSITIVE (+)
in color. The formation of calcium
AMBER
carbonate as a result of this indicates that
the urine sample tested positive for
carbonates.
The color of the solution did not change
Test for Sulfates when 1 mL of 10% HCl and a few drops of
NEGATIVE (-)
5% BaCl2 were added to 5 mL of urine. It
CLEAR
means that the sulfate test came out
negative.
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BIOCHEMISTRY LABORATORY WORKSHEET
III. Pathological Constituents
A. Test for Glucose
Sample Result Explanation
Nothing happens in the urine sample, and
Urine sample NEGATIVE (-)
the color doesn’t change.
Nothing happens in the urine when we put
Urine + 5% glucose
NEGATIVE (-) the 5% glucose and the color doesn’t
change just like the first one.
B. Test for Ketone Bodies
Sample Result Explanation
No purple ring formed in the urine sample
Urine sample
after adding 3 drops of 5% (NH4)2SO4, 3
NEGATIVE (-) drops of 5% sodium nitroprusside, and 1
mL of concentrated NH4OH, and letting
the mixture sit for 2 to 3 minutes.
1 mL of concentrated NH4OH should be
Urine + acetone
added to the 5 mL sample of urine after
adding 3 3 drops of 5% (NH4)2SO4 and 3
drops of 5% sodium nitroprusside and
POSITIVE (+)
letting it sit for 2 to 3 minutes. At the
junction, a purple ring developed,
indicating the presence of ketone in the
urine.
C. Test for Protein
Sample Result Explanation
Nothing happens and the color doesn’t
Urine sample NEGATIVE (-)
change.
The test was positive because coagulation
Urine + 5% albumin
POSITIVE (+) happens when we put the 5% albumin in
the urine.
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D. Test for Bile Acids
Sample Result Explanation
The test was positive because when the
Urine sample + sulfur
sulfur put in the sample it doesn’t fall in the
POSITIVE (+) bottom of the test tube, it stays in the top.
And even when you shake it the sulfur
doesn’t get wet by the urine.
Physical Characteristics of Urine
Urine + 5% Albumin
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Test for Glucose – Benedict’s Test
Test for Bile Acids: Urine Sample + Sulfur
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