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BIOCHEMISTRY MANUAL

Prepared by Admer Rey C. Dablio, v2012

Experiment 8
Score:

Test for Nutrients in Foods


Name: Joann H. Justiniane
Groupmates:
Marla C. Basa
Ana
Margarita
Baytion

Date Performed: September 6 ,


2016
Instructors Signature:
L.

Objectives:

II

To apply all the analytical tests conducted on the different macrobiomolecules


like carbohydrates, lipids, proteins and nucleic acids.
To determine the presence of peptide bonds in the food sample.
To determine the presence to carbohydrates, fats and minerals in the food
sample.
Chemicals:

III

Ammonium Oxalate (NH4)2C2O4


Nitric acid (HNO3)
Silver Nitrate (0.1N AgNO3)
Acetic acid (CH3COOH)
Molisch reagent

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

Schematic Diagram of the Procedure:


Please write on a short bond paper.

Summary of Theory

The nutrients known to be essential for human beings are proteins,


carbohydrates, fats and oils, minerals, vitamins, and water. Most essential nutrients
are needed only in small quantities, and are stored and reused by the body. The
absence of essential nutrients usually leads to the development of deficiency
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

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

Table VI.a Description of Results of Tests


Description of Results of Tests
Food
Mineral
Fat
Carbohydrat
Sample
Matter
es
(Molisch)
Apple Fruit
Slightly
+
juice
translucent
Margarine
Translucent
Raw
egg
white
Raw egg yolk

Cooked rice

Slightly
translucent
Slightly
translucent
Burned

Proteins
(Biuret)

-, yellow
green
-, bluish
green
+, violet

+, purple

-, blue

Table VI.b Extraction of fat from food sample


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

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

absorption spectroscopy, which identify the electromagnetic frequencies a sample


will absorb, allow testers to quantify the concentration of protein in a sample.

The reaction of the peptide bond in Biurets reagent is shown below.

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

foods, and is a major source of energy in our diets. Carbohydrates are


the major constituents of plants, and are also found in some animal
tissues. Its presence in food products varies in quantities. The most common
and abundant form of carbohydrates are sugars, fibers, and starches . Proteins are
essential nutrient for the body. They are the building blocks of body tissue,
and can also serve as a fuel source. Proteins usually come from fruits,
vegetables, grains, nuts and animals. Food products contain complex organic
compounds. Fats & fatty acids are also important in the body. They act as
cushion in cells, stores energy in triacylglycerols, provide protection from the
mechanical shock and prevents excessive loss of heat energy.
For further experimentation of the test for nutrients in food, the
experimenter recommends to determine the quality, sensory attributes and
their behavior during production, storage and consumption.

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

D. L. Morris, Colorimetric determination of Glycogen, Disadvantages of Iodine


Method Received for publication July 15, 1998.
Stoker S. H., Biochemistry, 1st Edition, pp. 110-137.
Labastida S.C., Testing the Presence of Food Nutrients. Retrieved from
http://www.slideshare.net/seanclarkluinor/testing-the-presence-of-foodnutrients
Analyzing Fruit and Vegetable Juices, Retrieved from 123HelpMe.com. 11
Sep 2016. http://www.123helpme.com/view.asp?id=120780
Gunnars K., Fruit Juice is Just as Unhealthy as a Sugary Drink. Retrieved from
https://authoritynutrition.com/fruit-juice-is-just-as-bad-as-soda/
Ware M., Butter vs Margarine: Which is Healthier, December 21, 2015.
Retrieved from http://www.medicalnewstoday.com/articles/304283.php
Nutritional Facts of Apple Juice. Retrieved from
http://nutritiondata.self.com/facts/fruits-and-fruit-juices/2075/2
Arnarson A., Rice 101: Nutrition Facts and Health Effects, 2004. Retrieved from
https://authoritynutrition.com/foods/rice/
Ulmer, G., Carbohydrates in an Egg White, October 29, 2015. Retrieved from
http://www.livestrong.com/article/460143-carbohydrates-in-an-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

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 9. Egg White


Proteins & Amino
Acids

Figure 8. Egg
White
D. Egg Yolk

Figure 11. Egg


Yolk
Fats & Fatty
Figure 10. Egg
Yolk
Minerals
Figure 13. Egg
Yolk
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.

BIOCHEMISTRY MANUAL
Prepared by Admer Rey C. Dablio, v2012

Figure 12. Egg


Yolk
Carbohydrates

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.

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