Indian Institute of Technology Jammu

Department of Chemical Engineering

SBP001U3M
BIOLOGY FOR ENGINEERS
III YEAR, FIFTH SEMESTER, 2021

EXPERIMENT #3

Experiment title
Detection of sugar, protein and bile salt in urine

INSTRUCTOR:
Instructor Name

MEMBERS: PIYUSH VERMA

Experiment carried on: September 20, 2021

Report submitted on : September 27, 2021

EXECUTIVE SUMMARY :-

(1) Protein :- The urinary protein test is a screening test to look for protein in the urine. One of
these proteins is called albumin.

(2) Sugar:- The Benedict test is used to detect the amount of sugar in the urine. The Benedict
test uses Benedict's solution as a reagent. Benedict's reagent is a combination of sodium
carbonate, sodium citrate, and copper (II) sulfate pentahydrate (CuSO4.5H2O).

(3) Bile Salts :- The presence of bile salts in a given urine sample is indicated by the formation
of a green ring at the intersection of both layers. This experiment uses the Smith test.

INTRODUCTION :-
The urine protein test measures the amount of protein present in the urine. 4,444 healthy people do
not have significant amounts of protein in their urine. However, if the kidneys are not functioning
properly, or if certain proteins are present at high levels in the bloodstream, the proteins can be
excreted in the urine. Urine is a liquid waste product produced by a pair of kidneys that needs to be
removed from our body. When you urinate, urine is drained from the urethra. In humans, urine is a
pale yellow liquid that contains water and a variety of other chemicals such as uric acid, urea, trace
enzymes, hormones, and carbohydrates.
OBJECTIVES :-

The only way to tell if your urine contains protein is to do a urine test. This test measures the amount
of protein in the urine. Urine is a liquid waste product produced by a pair of kidneys that needs to be
removed from our body. When you urinate, urine is drained from the urethra. In humans, urine is a
pale yellow liquid that contains water and a variety of other chemicals such as uric acid, urea, trace
enzymes, hormones, and carbohydrates. Bile is usually a yellow-green liquid. It is composed of
organic molecules such as bile acids, bilirubin and cholesterol, and water.

SCOPE :-

Laboratory measurements of urinary albumin and total urinary protein in the screening of proteinuria
for chronic kidney disease. Under abnormal conditions, trace amounts of sugar may be present in the
urine, in addition to proteins, ketone bodies, blood and bile. Profiling of circulating bile acids and uric
acid in patients with biliary atresia before and after biliary stent placement.

PROCEDURE :-
(A) Protein :- Prepare a test tube that has been washed and dried. Place your
favorite food sample in a test tube. Add 2 ml of sodium hydroxide and 5-
6 drops of copper sulphate solution. Gently shake the test tube to mix the
ingredients thoroughly and leave the mixture for 4-5 minutes. If a bluish
purple color is displayed, it indicates the presence of protein.

(B) Sugar :- Take a clean and dry test tube. Measure exactly 2 ml of
the specified urine sample in a graduated cylinder. Take a clean
and dry test tube. Measure exactly 2 ml of the specified urine
sample in a graduated cylinder. Pour the measured urine sample
into. Place exactly 2 ml of Fehling's solution A in a tube containing
a urine sample and shake well. Add exactly 2 ml of Fehling's
solution B to the same tube and mix all the solutions slowly. Next,
attach the test tube holder, bring the test tube near the Bunsen
burner, and heat it for 2 minutes.Continue to stir the tube while it
heats. Please be aware of the changes. Pour the measured urine sample
into a test tube. Add exactly 2 ml of Fehling’s solution A to the tube
containing the urine sample and shake well. Add exactly 2 ml of
Fehling’s solution B to the same tube and mix all the solutions slowly.
Next, fix the test tube holder, bring the test tube close to the Bunsen
burner, and heat it for 2 minutes. Continue to stir the tube while heating.
Please be aware of the changes.

(C) Bile salts :- Take a clean and dry test tube. Using a graduated cylinder, add 1 ml
of Smith's reagent to the test tube. Use a pipette to take a few drops of the
collected urine sample from the bottle. Then hold the test tube at an angle and
slowly add the urine sample to the side or wall of the test tube with a pipette.
Observe and note the changes.

RESULT AND DISCUSSIONS :-

(A) test :- Benedict's Test is used to test for simple carbohydrates. The Benedict's test
identifies reducing sugars (monosaccharide's and some disaccharides), which have
free ketone or aldehyde functional groups. Benedict's solution can be used to test for
the presence of glucose in urine.
(B) Sulphocylic acid test :- A method in which
sulfosalicylic acid (that is, the SSA reagent) is
added to a small and equal volume of clear urine.
The acidification causes precipitation of protein in
the sample (seen as turbidity), which can be
subjective quantitated visually or more precisely
quantitated using photometry.
a. NITRIC ACID RING TEST :- Concentrated
nitric acid is added to a protein solution from
the side of the test tube to form two layers. A
white ring appears between the two layers if
the test is positive. Heller's test is commonly
used to test for the presence of proteins in
urine.
b. Heat coagulation test :- Take about 6 to 8 mL of
urine in a test tube. Incline the test tube at an angle

and heat the upper one-third of the test tube by a

low flame. Turbidity develops in the heated portion
of the urine. Add 1% acetic acid drop by drop and
boil or simply add a drop of 33% acetic acid.

c. Smith’s test :-Smith’s Reagent is a reddish-

brown solution containing methylene iodine
and zinc reagent. It Is used to detect the
presence of bile salt in the urine sample by
forming a green colored ring At the surface.
All three samples showed a formation of a
green colored ring, confirming the Presence
of bile salt.
 d. Pettenkofer test :- The Pettenkofer’s test is
used to prove the presence of bile acids. We
use the interaction of bile acids with
phurphural and the formation of colored
products. … In the Salkowski’s test,

concentrated sulphuric acid should be

carefully added to a chloroformic solution of
cholesterol.

CONCLUSION :-
 (A) Protein :- The given sample of urine appears as a cloudy turbid solution or
whitish which indicates that albumin is present in the sample.
(B) Sugar :- Upon heating the sample, gradually, a green precipitate is formed in
the test tube, which indicates the presence of sugar in the given urine sample. Different
precipitates are formed depending upon the sugar concentration in urine which can be yellow,
green, brick red.
(C) Bile salts :- The presence of bile salt in the given urine sample is indicated by
the formation of a red ring is between both layers.

REFERENCES :-
WEBSITES :-
i. https://www.google.com/search?
q=detection+of+protein+in+urine+by+byjus
ii. https://www.google.com/url?
q=https://byjus.com/biology/test-for-presence-
of-bile-salts-in-
urine/&usg=AOvVaw1rv5uw3h1BwXNiDlZU4
mTO
iii. https://www.google.com/url?
q=https://byjus.com/biology/test-for-presence-
of-sugar-in-
urine/&usg=AOvVaw0BE6WIQPyXaeURvd9c
cDXa

APPENDIX :-
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