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Experiment 8
Score:
Objectives:
II
III
Biuret reagent
Antimony trichloride (SbCl3)
Sulfuric acid (conc. H2SO4)
Chloroform (CHCl3)
Water bath
Test tube rack
2 Test tube holder
Hot plate
Wash bottle
Fume Hood
Thermometer
20mL test tubes
Raw egg white
Cooked rice
Brown paper
Apparatus/Materials/Equipment:
50 mL beaker
100 mL beaker
Stirring rod
10 Test tubes
Wire Gauze
10 mL Graduated cylinder
Fruit juice
Margarine
Raw egg yolk
Fourier Transform Infrared
Spectroscopy (FTIR)
Crucible
IV
Summary of Theory
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
disease. The three major function of nutrients is that it provides energy, promotes
growth and development and regulates body functions.
Carbohydrates are the most abundant class of bioorganic molecules on planet
earth. It provides energy, stores energy in the form of glycogen, supplies carbon
atoms for the synthesis of other biochemical substances and links lipids and proteins.
Carbohydrates are compounds composed of carbon, hydrogen and oxygen containing
an aldehyde or ketone functional group and many alcohol groups, and can also be
called polyhydroxy aldehydes or polyhydroxy ketones. Molecules that when
hydrolyzed yield polyhydroxy aldehydes or ketones are also considered
carbohydrates. (Stoker, 2011)
Proteins from food is broken down into amino acids by the digestive system.
These amino acids are then used for building and repairing muscles, red blood cells,
hair, and other tissues. Main sources of proteins are animal products like meat, fish,
milk, cheese and eggs and vegetable sources like legumes and seeds. (Stoker, 2011)
The fat in food includes a mixture of saturated and unsaturated fat. Animalbased foods such as meats and milk products are higher in saturated fat whereas
most vegetable oils are higher in unsaturated fat. Dietary fat plays an important role
in a healthy diet. Fat maintains skin and hair, cushions vital organs, provides
insulation, and is necessary for the production and absorption of certain vitamins and
hormones. (Labastida S.C)
Vitamins helps regulate chemical reactions in the body. There are 13 vitamins,
including vitamins A, B complex, C, D, E, and K. Because most vitamins cannot be
made in the body, we must obtain them through diet. (Labastida S.C)
Minerals are components of foods that are involved in many body functions.
For example, calcium and magnesium are important for bone structure, and iron is
needed for our red blood cells to transport oxygen. Minerals are not a source of
energy and are best obtained through a varied diet rather than supplements. There
are two main categories of the essential minerals needed for survival. Macro minerals
include calcium, magnesium, chloride, phosphorus, sulfur, potassium, and sodium.
The other group is called trace minerals. This includes copper, cobalt, iodine, iron,
fluoride, manganese, selenium, zinc, and molybdenum. (Labastida S.C)
This experiment is an application of all the analytical test conducted on the
different macrobiomolecules like carbohydrates, lipids, proteins and nucleic acids
which can be detected in food.
VI
Observations
Cooked rice
Slightly
translucent
Slightly
translucent
Burned
Proteins
(Biuret)
-, yellow
green
-, bluish
green
+, violet
+, purple
-, blue
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
Egg yolk
Egg white
Apple fruit extract
Margarine
Mashed cooked rice
Test for
VII
Observations
Yolk is at the lower layer (aqueous)
Crystal-like lower layer
The extract is at the lower layer
At the lower layer
Clumped at the lower layer
Test for
Extraction of
Analysis
The principle of Molischs Test is that the solution is first treated with a strong
acid. This hydrolyzes the carbohydrates to monosaccharide. When the water is
remove from a monosaccharide, a compound called furfural is formed. This furfural is
condensed with alpha naphthol to form a violet ring and other colored products. All
monosaccharides, disaccharides, and polysaccharides will give a positive reaction,
and nucleic acids and glycoproteins will also give a positive reaction. (Morris, 1998)
Example of Molischs Test is the reaction of glucose to Molischs reagent
shown in below.
The principle of Biuret test is that the copper (II) ion forms a violet-colored
complex in an alkaline solution. The biuret test uses an alkaline mixture, or reagent,
composed of potassium hydroxide and copper sulfate. The normal color of biuret
reagent is blue. The reagent turns violet in the presence of peptide bonds which the
chemical bonds that hold amino acids together. The proteins detected must have at
least three amino acids, which means that the protein must have at least two peptide
bonds. The reagents copper ions, with a charge of +2, are reduced to a charge of +1
in the presence of peptide bonds, causing the color change. The techniques of
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
The Biuret Test works for any compound containing two or more of the
following groups.
Based on the results, apple juice, raw egg white, raw egg yolk and cooked rice
gave a positive result in molisch test while margarine gave a negative result. This
means that all food samples except margarine contains carbohydrates. In the test for
fat, margarine, apple juice, raw egg white and raw egg yolk gave a positive result. In
the test for proteins, raw egg white and raw egg yolk gave a positive result.
Carbohydrates are classified on the basis of molecular size as
monosaccharides, oligosaccharides and polysaccharides. A large amount of sugar
present on fruits is fructose, a monosaccharide that is a polyhydroxy ketone. (Stoker,
2011) Margarine was developed as a substitute for butter and is made from plantbased oils such as canola oil, palm fruit oil and soybean oil. (Ware, 2015) Egg whites
are a rich source of protein, vitamins and minerals. Although egg whites are
composed entirely of water and protein, they also contain trace carbohydrates and
fat. (Ulmer, 2015) The yolk or the yellow protion of an egg makes up about 34% of
the liquid weight of the egg. It contains all of the fat in the egg. (Ulmer, 2015) The
cooked rice used in the experiment was the white one. Rice is a staple food for more
than half of the worlds population, particularly those living southern and eastern
asia. White rice is the most commonly consumed type, but brown (whole grain) rice is
becoming increasingly popular in some Western countries due to its health benefits.
(Arnarson, 2004)
The experiment was not accurate due to personal and experimental errors. In
Table VI.a, the rice in test for fat was burned due to mishandling of the sample. The
cooked rice in test for proteins and minerals should give a positive result since rice
has minerals and proteins.
Theoretically, the fat, protein and carbohydrate content of apple juice is
shown on Figures 1, 2 and 3. The fat content of margarine is shown on Figure 4.
The carbohydrate, mineral, fat and protein content of egg white is shown in Figures
5, 6, 7 and 8. The mineral, fat, carbohydrate and protein content of egg yolk is
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
shown in Figures 9, 10, 11 and 12. The mineral, fat, carbohydrate and protein
content is shown in Figures 13, 14, 15 and 16.
Note: All Figures can be seen in the Appendix A.
VIII
Conclusion
Nutrients obtain from food are important to the body. The basic
essential nutrients, vitamins and minerals plays a significant role in metabolic
processes. Carbohydrates are one of the three principal classes of
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
IX
References
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
Appendix A
A. Apple Juice
Figure 3. Apple
Juice
Proteins & Amino
Figure 4. Apple
Juice
Carbohydrates
Figure 2. Apple
Juice
B. Margarine
Figure 5.
Margarine
Fats & Fatty
C. Egg White
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
Figure 6. Egg
White
Carbohydrates
Figure 7. Egg
White
Minerals
Figure 8. Egg
White
D. Egg Yolk
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.
BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012
E. Cooked Rice
Figure 14.
Cooked Rice
Minerals
Figure 16.
Cooked Rice
Carbohydrates
Figure 15.
Cooked Rice
Fats & Fatty Acids
Figure 17.
Cooked Rice
Proteins & Amino
No part of this manual may be reproduced without written permission from the Chemistry Department of
the College of Arts and Sciences, Mindanao University of Science and Technology, Cagayan de Oro City.