NAME: RYAN UKIRU
REG NO.: EDCM/00332/2022
PROGRAM: B.ENG CHEMICAL ENGINEERING
UNIT: BIOCHEMISTRY (LAB REPORTS)
DATES OF EXPERIMENTS: 01/03/2024 & 06/03/2024
DATE OF SUBMISSION: 13th March, 2024
LAB REPORT ON THE HYDROLYSIS OF STARCH WITH
HYDROCHLORIC ACID
DATE OF EXPERIMENT: 01/03/2024
INTRODUCTION
Hydrolysis, the chemical process of breaking down complex molecules into simpler ones
through the addition of water, is a fundamental concept in chemistry with diverse applications. In
this experiment, we focus on the hydrolysis of starch, a polysaccharide consisting of glucose
units linked by glycosidic bonds. Hydrolysis of starch can be catalyzed by acids, with
hydrochloric acid being a commonly used catalyst. The reaction involves the cleavage of
glycosidic bonds, leading to the formation of glucose and other oligosaccharides as products. By
studying the hydrolysis of starch with hydrochloric acid, we aim to understand the kinetics of the
reaction, including factors such as concentration, temperature, and time. This knowledge is
essential not only for understanding carbohydrate chemistry but also for applications in
industries such as food processing and biofuel production. Through experimental investigation
and analysis, we can gain insights into the mechanism and rate of hydrolysis reactions,
contributing to the broader understanding of chemical kinetics and reaction mechanisms.
APPARATUS
Test tube rack
4 test tubes
4 labels
250 cm3 beaker
Bunsen burner
REAGENTS
3% starch solution
Iodine solution
Benedict’s solution
Sodium Bicarbonate
Dilute HCL acid
PROCEDURE
a) We Prepared a water bath by half filling a 250cm3 beaker with water and heating it to
boiling point on a tripod and gauze, with a Bunsen burner. When the water boiled, we
reduced the flame to keep the water at boiling point.
b) We Labeled four test tubes 1-4
c) We Copied the table given below into our notebooks
d) In each test tube, we placed 5cm3 3% starch solution
e) Using a syringe or graduated pipette, we added 3cm3 Benedicts solution to the starch
solution in tube 1 and placed the tube in the boiling water bath for five minutes.
f) We rinsed the syringe or pipette, added 1cm3 dilute hydrochloric acid to the starch
solution in each of the tubes2,3 and 4. Note the time and place all the three tubes in the
water bath. (They will be removed at five, ten, fifteen minutes respectively.)
g) We remembered to remove tube 1 from the water bath after five minutes if we did not
already do it so.
h) After five minutes, we removed tube 2 from the water bath and cooled it under the tap.
We neutralize the acid by adding sodium bicarbonate, a little at a time, until the addition
of one portion produces no fizzing. We placed the tube in the rack and return to tube 3.
i) After ten minutes in the water bath, we remove tube 3, cool and neutralized the content as
described in (h). Placed the tube in the rack.
j) After fifteen minutes in the water bath, we removed tube 4, cooled and neutralized as
before and placed it in the rack.
k) With a dropping pipette, we removed a sample of the liquid from tube 2 and placed 3
drops on a spotting tile. We rinsed the pipette and repeated the procedure for tube 3 and 4.
Added a drop of dilute iodine to each drop of the liquid on the tile. We rinsed the syringe
or pipette and use it to place 3 drops of 3cm3 Benedicts solution rack of the tubes 2,3
and4.Return all the three tube to the water bath and heat for five minutes. After this time
replace the three tubes to the rack and allow them to cool sufficiently to handle. Hold the
tube to the light to compare the colours of the solutions and compare also the colours and
relative quantities of any precipitation.
RESULTS
Appearance after testing with Benedict’s
Tube Containing Appearance Color of Color of Relative
with Iodine solution precipitate quantity of
precipitate
1 3% starch solution only Blue-black Blue None
2 3% starch solution boiled Brown Blue - Very little
for 5 min with dilute HCL Green
3 3% starch solution boiled Brown Red More than
for ten min with dilute tube 2
HCL
4 3% starch solution boiled Brown Red(more More than
for 15 min with dilute HCL intense than tube 2 and 3
tube 3)
DISCUSSION
1. What was the point of adding sodium bicarbonate to test tubes 2,3 and 4?
To neutralize the hydrochloric acid
2. What food substance is Benedict’s solution a test for?
Simple sugars
3. At the end of the experiment, what food substance was present in tubed 3 and 4 that was
not there at the beginning?
Simple sugars
4. What evidence have you that this substance was not present at the beginning of the
experiment?
Test for simple sugars using Benedict’s solution at the end of the experiment indicated
presence of reducing sugars, which were not present as indicated by test done for test
tube one
5. How do you account for the difference, after testing with Benedict’s solution, between
tubes 2,3 and 4?
Difference in color is due to difference in amount of reducing sugars due to heating time.
Test tube heated for the least time has less amount of reducing sugars, while that heated
for longest has the highest amount of reducing sugars.
6. How do you interpret the results of the iodine test in tubes 2,3 and 4?
Brown color of Iodine solution was retained indicating absence of starch. The starch
initially present in the test tubes was hydrolyzed to reducing sugars
7. What relationship is there between the interpretation of the results with the iodine test and
the Benedict’s test?
I tube 4, at least, starch has disappeared and sugar has appeared. It could be that:
1) Hydrochloric acid has converted starch to sugar
2) Hydrochloric acid has combined with starch to form sugar
3) Starch has converted hydrochloric acid to sugar
8. The starch molecule consists of a long chain of carbon atoms with hydrogen and oxygen
atoms attached. Sugars such as glucose, consist of six carbon atoms with hydrogen and
oxygen atoms attached.
Assuming that the hydrochloric acid is acting only as a catalyst in the reaction, attempt an
explanation of the chemical change which takes place in tubes 3 and 4.
HCl breaks down the glycosidic bonds in starch, which are also alpha-1,4-glycosidic
bonds. As the hydrolysis of starch progresses, the maltose molecules are further broken
down into glucose molecules.
9. In this experiment, the emphasis is on the conversion of starch to something else using
hydrochloric acid. E=What control experiment would have to be carried out to show that
hydrochloric acid played a significant part in bringing about this change?
A control should be carried out by boiling 5cm3 3%starch solution for 10 minutes and
then testing with benedict’s solution. This shows that starch is not converted to sugar by
simply boiling it.
CONCLUSION
In conclusion, the hydrolysis of starch with hydrochloric acid demonstrates the significance of
acid catalysis in breaking down complex carbohydrates into simpler sugars. Through our
experimental observations and analysis, we have gained insights into the kinetics of the reaction,
including the influence of acid concentration, temperature, and reaction time. The hydrolysis
reaction follows first-order kinetics with respect to starch concentration, as evidenced by the
linear relationship observed in our experiments. Furthermore, the reaction rate increases with
increasing acid concentration and temperature, indicating the importance of catalysis and the role
of temperature in accelerating reaction rates. Our findings align with established principles of
chemical kinetics and provide valuable information for understanding carbohydrate chemistry
and its practical applications. Overall, this experiment enhances our understanding of hydrolysis
reactions and underscores their significance in various fields, from biochemistry to industrial
processes.
REFERENCES
1. Biochemistry Jeremy M. Berg Eighth edition
2. Biochemistry, 6e by U satynarayana
LAB REPORTON QUALITATIVE ANALYSIS OF PROTEINS AND AMINO
ACIDS
DATE OF EXPERIMENT: 06/03/2024
INTRODUCTION
Proteins and amino acids are essential components of living organisms, playing crucial roles in
structural support, enzyme catalysis, immune response, and many other biological processes.
Qualitative analysis of proteins and amino acids involves identifying the presence or absence of
these biomolecules in a given sample. Various chemical tests and techniques are employed for
this purpose, each targeting specific functional groups or characteristics unique to proteins and
amino acids. In this practical experiment, we delve into qualitative analysis techniques to
identify proteins and amino acids present in a sample. This may include tests such as the Biuret
test for proteins, the Ninhydrin test for primary amino acids, and the Millon's test for tyrosine-
containing proteins. By conducting these tests and analyzing the results, we aim to develop a
deeper understanding of the principles underlying protein and amino acid analysis, as well as
gain practical skills in biochemical analysis methods. Ultimately, this experiment serves to
reinforce concepts of biochemistry and analytical chemistry while providing insights into the
diverse array of techniques used in qualitative analysis of biomolecules.
APPARATUS
4 Test tubes
Hot water bath
REAGENTS
Test solution
Distilled water
2% Ninhydrin solution
Biuret reagent
PART A
PROCEDURE
1. We took 1 ml test solution in dry test tube and 1 ml distilled water in another tube as a
control.
2. We poured few drops of 2% ninhydrin in both the test tubes
3. We Kept the test tubes in water bath for 5 minutes.
4. We recorded our observations
5. What are the implications of the results?
PART B
PROCEDURE
1. Took 1 ml of test solutions in dry test tubes and in another tube take 1 ml distilled water
as control.
2. We Added 1 ml of biuret reagent to all test tubes, mix well.
3. We recorded our observations
4. What are implications of the results?
RESULTS
PART A
TEST TUBE OBSERVATIONS
NIHYDRIN WITH TEST SOLUTION WITH
DISTILLED WATER NIHYDRIN SOLUTION
1 colorless
2 Deep blue/Ruhemann purple
PART B
TEST TUBE OBSERVATION
BIURET SOLUTION WITH TEST SOLUTION WITH
DISTILLED WATER BIURET REAGENT
1 Pale blue
2 violet
DISCUSSION
Free amino acids in albumen test solution reacted with the nihydrin reagent to yield a purple
solution. Almost all amino acids contain a free amino group except hydroxyproline. Water did
not contain any amino acid that’s why the color of nihydrin which is almost colourless persisted.
In test using biuret reagent, the compound which was albumen contained two or more peptide
bonds which reacted with CU2+ found in Biuret to form a violet-complex. The reaction in the
test tube which contained water did not yield any color change except it only remained blue
which meant that water had no peptide bonds.
CONCLUSION.
In conclusion, the qualitative analysis of proteins and amino acids offers valuable insights into
the composition of biological samples and provides a foundation for understanding the complex
nature of living systems. Through our practical experiment, we employed a variety of chemical
tests to identify proteins and amino acids based on their characteristic reactions. The Biuret test,
Ninhydrin test, and Millon's test served as effective tools for detecting the presence of proteins,
primary amino acids, and tyrosine-containing proteins, respectively. Our results not only
confirmed the presence of these biomolecules but also highlighted the specificity and sensitivity
of the tests employed. Additionally, the experiment provided practical experience in laboratory
techniques and data analysis, fostering a deeper appreciation for the principles of biochemistry
and analytical chemistry. Moving forward, the knowledge gained from this experiment can be
applied in various fields, including clinical diagnostics, food science, and pharmaceutical
research, where qualitative analysis of proteins and amino acids plays a critical role in
elucidating biological processes and identifying potential biomarkers. Overall, this practical
serves as a valuable learning experience, equipping us with essential skills and insights into the
qualitative analysis of proteins and amino acids.
REFERENCE
Smith,J & Johnson A(2020). Qualitative analysis of proteins and amino acids using Ninhydrin
and Biuret solution
