BIOCHEMICAL TESTS OF DIFFERENT FOOD PRODUCTS

USED FREQUENTLY BY THE HUMAN POPULATION

A DISSERTATION

Submitted to

MJRP Academy of Science

MAHATMA JYOTI RAO PHOOLE UNIVERSITY

Jaipur, Rajasthan

Submitted

by

Santosh Choudhary

For the partial fulfillment of the Degree of

MASTER OF SCIENCE (BOTANY)

(2016)
 CERTIFICATE-1

This is to certify that the project entitled as “BIOCHEMICAL

TESTS OF DIFFERENT FOOD PRODUCTS USED
FREQUENTLY BY THE HUMAN POPULATION”
submitted for the degree of M.Sc. in Botany of the Mahatma Jyoti
Rao Phoole University, Jaipur is a bonafide research work carried
out by Santosh Choudhary under my supervision.

The assistance & help received during the course of

investigation have been fully acknowledged.

Date: Advisor

Designation: Name
 DECLARATION

I hereby declare that the project entitled “BIOCHEMICAL

TESTS OF DIFFERENT FOOD PRODUCTS USED
FREQUENTLY BY THE HUMAN POPULATION”
submitted by myself to the Mahatma Jyoti Rao Phoole University,
Jaipur in the partial fulfillment of the degree of Master of Science in
Botany is a marginal work done to the best of my knowledge during
the period February to April 2016.
This work is not similar to any other research work of any
candidate of any other University.

Place : Jaipur Santosh Choudhary

Date:
 ACKNOWLEDGEMENT

I take opportunity to thank to all those without whose efforts

the completion of the project would not have been possible.

I express my thanks and deep sense of gratitude to Shri Nirmal

Panwar (The Chancellor). I would like to thank Dr. Yamini Yadav
for her cooperation. My sincere regards to Dr. Ruby Grover for
permitting me to utilize all the facilities available in the University.

I am grateful to Dr. Neha Sharma whose valuable guidance

helped me in undertaking this project. I am thankful to Ms Ritika
Singh and Dr. Richa Joshi for providing me laboratory facilities as
the incharge.

I would be failing in my duty if I don’t acknowledge the

assistance received from technical staff of Botany Department.
 CONTENTS

 ABSTRACT

 INTRODUCTION

 REVIEW OF LITERATURE

 MATERIALS & METHODS

 OBSERVATIONS

 RESULTS & DISCUSSION

 CONCLUSION & FUTURE SCOPE

 BIBLIOGRAPHY
ABSTRACT
 ABSTRACT

The present investigation implements the testing of various common food products used

by general human population to assess the nutritional quality. Thus, assessment of nutrients in

food gives a scope for their quality to be used by humans. Proper balanced diet plans and healthy

meals can be maintained by better knowledge of food and nutrients we take regularly. This saves

us from different malnutritions and diseases.

The analytical tests were made taking 3 major macronutrients under consideration, viz.

carbohydrates, proteins and fats. Analytical tests were done in vitro (laboratory) to know whether

the food product is a carbohydrate, protein or fat. The main food samples used by general human

population taken for testing were- rice, wheat flour, gram flour, sugar, milk and groundnut oil.

It was analysed that rice, sugar and wheat were main sources of carbohydrate, gram flour

had protein alongwith milk sample, and groundnut oil had maximum fat content.

Thus we can carry out food testing to analyse the nutrients in food for healthy balanced

diet maintenance in human adult.

INTRODUCTION
 INTRODUCTION

The Definition of Food

Food is any substance consumed to provide nutritional support for the body. It is usually
of plant or animal origin, and contains essential nutrients, such as fats, proteins, vitamins, or
minerals. The substance is ingested by an organism and assimilated by the organism's cells to
provide energy, maintain life, or stimulate growth. Historically, people secured food through two
methods: hunting and gathering and agriculture. Today, the majority of the food energy required
by the ever increasing population of the world is supplied by the food industry (Audrey and
Ensminger, 1994).

Food safety and food security are monitored by agencies like the International
Association for Food Protection, World Resources Institute, World Food Programme, Food and
Agriculture Organization, and International Food Information Council. They address issues such
as sustainability, biological diversity, climate change, nutritional economics, population growth,
water supply, and access to food. The right to food is a human right derived from the
International Covenant on Economic, Social and Cultural Rights (ICESCR), recognizing the
"right to an adequate standard of living, including adequate food", as well as the "fundamental
right to be free from hunger"

Nutrition in food

Nutrition is the science that interprets the interaction of nutrients and other substances in
food (e.g. phytonutrients, anthocyanins, tannins, etc.) in relation to maintenance, growth,
reproduction, health and disease of an organism. It includes food intake, absorption, assimilation,
biosynthesis, catabolism and excretion. The diet of an organism is what it eats, which is largely
determined by the availability, the processing and palatability of foods. A healthy diet includes
preparation of food and storage methods that preserve nutrients from oxidation, heat or leaching,
and that reduce risk of food-born illnesses (Donatelle and Rebecca, 2005).

Macronutrients

The macronutrients are carbohydrates, fats, protein, and water. The macronutrients
(excluding fiber and water) provide structural material (amino acids from which proteins are
built, and lipids from which cell membranes and some signaling molecules are built) and energy.
Some of the structural material can be used to generate energy internally, and in either case it is
measured in Joules or kilocalories (often called "Calories" and written with a capital C to
distinguish them from little 'c' calories). Carbohydrates and proteins provide 17 kJ approximately
(4 kcal) of energy per gram, while fats provide 37 kJ (9 kcal) per gram, though the net energy
from either depends on such factors as absorption and digestive effort, which vary substantially
from instance to instance. Vitamins, minerals, fiber, and water do not provide energy, but are
required for other reasons (Matthews et al., 1999).

Micronutrients
Dietary minerals are inorganic chemical elements required by living organisms, other
than the four elements carbon, hydrogen, nitrogen, and oxygen that are present in nearly all
organic molecules. The term "mineral" is archaic, since the intent is to describe simply the less
common elements in the diet. Some are heavier than the four just mentioned, including several
metals, which often occur as ions in the body. Some dietitians recommend that these be supplied
from foods in which they occur naturally, or at least as complex compounds, or sometimes even
from natural inorganic sources (such as calcium carbonate from ground oyster shells). Some
minerals are absorbed much more readily in the ionic forms found in such sources. On the other
hand, minerals are often artificially added to the diet as supplements; the most famous is likely
iodine in iodized salt which prevents goiter (Nelson and Cox, 2005).

Malnutrition
Malnutrition refers to insufficient, excessive, or imbalanced consumption of nutrients by
an organism. In developed countries, the diseases of malnutrition are most often associated with
nutritional imbalances or excessive consumption. In developing countries, malnutrition is more
likely to be caused by poor access to a range of nutritious foods or inadequate knowledge. In
Mali the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) and the
Aga Khan Foundation, trained women's groups to make equinut, a healthy and nutritional
version of the traditional recipe di-dèguè (comprising peanut paste, honey and millet or rice
flour). The aim was to boost nutrition and livelihoods by producing a product that women could
make and sell, and which would be accepted by the local community because of its local
heritage.

Although there are more organisms in the world who are malnourished due to insufficient
consumption, increasingly more organisms suffer from excessive over-nutrition; a problem
caused by an over abundance of sustenance coupled with the instinctual desire (by animals in
particular) to consume all that it can.

Nutritionism is the view that excessive reliance on food science and the study of nutrition
can lead to poor nutrition and to ill health. It was originally credited to Gyorgy Scrinis, and was
popularized by Michael Pollan. Since nutrients are invisible, policy makers rely on nutrition
experts to advise on food choices. Because science has an incomplete understanding of how food
affects the human body, Pollan argues, nutritionism can be blamed for many of the health
problems relating to diet in the Western World today.

Nutrients and their functions

Foods provide us with energy in the form of calories (Kcal). Calories effectively act as
the fuel that powers our bodies and enables us to function, in the same way that petrol fuels a car.
Some foods provide us with more energy than others, but by eating a wide variety of foods in the
correct balance we can meet our requirements. Foods that provide many nutrients relative to the
amount of energy they contain are known as “nutrient rich” foods e.g. milk and dairy foods.
Energy requirements vary depending on age, sex, size, metabolic rate and activity level. If we
consume more energy/calories than we need, we deposit the excess energy in the form of fat or
adipose tissue. Conversely if we use more energy than we consume we use up fat to provide us
with more energy.

The Present Investigation

This study is based on analysis of basic macronutrients viz. carbohydrates, proteins and
fats in food products used frequently by human population. It provides an incentive and
knowledge about the daily intake of nutrients by common people and regulation of diet. The
basic knowledge about carbohydrates, proteins and fats source is also essential for healthy diet
program.
Aims and Objectives:

1. Samples of food products are collected by knowledge of frequent usage by humans.

2. Qualitative biochemical test of each food product is done.

3. Identification of food product as main source of carbohydrate, protein or fat.

4. Significance of carbohydrates to be outlined

5. Significance of proteins to be outlined

6. Significance of fats to be outlined

REVIEW OF LITERATURE
 REVIEW OF LITERATURE

Primary macronutrients and their importance

The three macronutrients of protein, fat, and carbohydrates all perform essential roles in
the human body. Macronutrients are the main components of our diet. Our bodies require others
nutrients as well, such as vitamins and minerals. However, these are needed in much smaller
quantities, and thus are referred to as micronutrients. All three macronutrients are needed in the
diet, as each perform vital functions in the body (David et al., 2009).

Carbohydrates and their significance

Carbohydrates are the main energy source of the body. They are chains of small, simple
sugars that are broken down and enter the body as glucose. Glucose is essential for the body, as it
is the preferred source of energy in our brain, heart and central nervous system. For this reason,
your diet should consist of at least 45 to 65 percent carbohydrates.

Carbohydrates are found in many foods. However, you should stay away from sweet
foods with excess sugar. Instead, opt for healthier options like fruits, vegetables, beans and nuts,
as well as whole grains. These foods also contain fiber. Fiber cannot be digested by you body.
However, fiber aids our intestine in expelling waste and can help lower cholesterol (Jenkins et
al., 1986).

Significance of Protein

Protein should consist of about 10 to 35 percent of your diet. Amino acids are the
building blocks of protein. They are linked together in complex formations. There are 20
different amino acids, 9 of which are considered essential because our bodies cannot produce
them naturally, and therefore they must be obtained through the diet. Proteins that contain all 20
amino acids are called complete proteins. Complete proteins are found in animal products such
as meat, eggs and milk (Pehrsson et al., 2000).

Proteins crucial role in the body includes building, maintaining and repairing body tissue.
It is especially important to physically active individuals whose muscle tissue is constantly in
need of repair. Protein has other roles in the body. All enzymes and hormones, which perform
vital functions, are proteins. In addition, proteins are used to aid in the immune process.
 Significance of fats

The misconception about fat is that it is always bad for you. In fact, fat is essential for
maintaining a healthy body. Your diet should consist of about 20 percent fat. The trick is to eat
more of the good fats and less of the bad fats. Saturated and trans fats should be avoided while
increases levels of unsaturated and the essential fatty acids, such as omega 3 and omega 6, can be
good for you. Replacing sweets and high fat meats with foods such as nuts, avocados, and olive
oil will help shift the balance away from unhealthy towards those fats that are useful to the body.

Fat has many roles in the human body. One of fats main functions is protection. This
includes insulation to keep body temperature and cushioning to protect body organs. It also
promotes growth and development, as well as maintaining cell membranes. Fat, in addition,
plays a vital role in the digestion of vitamins. Vitamins A, D, E, and K are fat soluble vitamins,
meaning they need fat in order to be absorbed into the body (Pichon et al., 2006).

Carbohydrates and their Deficiency

Carbohydrates are fuel for metabolism.

Chemical reactions in cells require fuel. Muscles, from heart to digestive system to biceps
and quads all use carbohydrate, in the form of glucose. Brain cells need twice the energy as other
cells.

The brain runs almost exclusively on glucose, using up about 20 percent of your daily
calorie needs. Because glucose is so critical to brain function, your body maintains blood glucose
above a certain level, with a complex set of hormone signaling systems.

Carbohydrate in food comes in two forms: sugar and starch. The basic building blocks of
both are simple sugars like glucose and fructose. More complex carbohydrates are made by
linking simple sugar molecules together. Fructose and glucose join to make sucrose. Long chains
of glucose are called starch. Digestion breaks these carbohydrates into single sugar molecules,
which are absorbed into the blood.
 There is no set RDA for carbohydrate, as there is for vitamins or protein. Carbohydrate is
used for energy needs, so daily intake will depend on how many calories you need. Since basic
fuel needs, for brain function and metabolism must be met, a minimum amount of carbohydrate
should be eaten everyday. That level, in the chart below, is 130 grams/day for children and
adults.

Carbohydrate has 4 calories per gram, so 130 grams equals 520 calories. The basal
energy needs of most adults are at least twice that. Additional calories can come from
carbohydrate, fat or protein in food.

The best way to think of carbohydrate intake is as a percent of your total daily calorie
intake. The recommended range is 45 – 65% of calories as carbohydrate. For example, if you eat
1600 calories per day, and 50% of those calories are carbohydrate, you would eat 200 grams per
day. Athletes need to eat at the high end of the carbohydrate range, to fuel training and
competition.

Carbohydrate deficiency symptoms due to poor intake are rare. A minimum blood
glucose level must be maintained to support brain function and basic metabolism. If you eat few
carbohydrates, glycogen, the storage form of glucose, is broken down to keep blood glucose in
the normal range.

In extreme situations, such as starvation, muscle protein is metabolized, and the amino
acids from that protein are modified to make glucose (Robert, 2008).

Blood glucose can fall below normal levels for reasons unrelated to dietary intake of
carbohydrate. Insulin controls blood glucose, and excess insulin, due to metabolic abnormalities
or excess dosing for diabetics, can cause blood glucose to drop. Symptoms include shakiness,
confusion, anxiety and vision disturbance. Again, such symptoms are more likely to be caused
by metabolic abnormalities than by eating too little carbohydrate.
 Another sign of eating few carbohydrates might be weight loss. If you cut calorie intake
by cutting out carbohydrate foods, your body will have to use fat stores to meet energy needs.
Strictly speaking, this isn’t so much a carbohydrate deficiency as a fuel deficit (Westman, 2002).

Proteins and their Deficiency

Even with a wide variety of protein sources available, some people experience protein
deficiency symptoms due to a lack of protein intake. Severely restrictive diets, lack of knowledge
about nutrients, and even poverty can contribute to protein deficiency. Protein deficiency is
nothing to fool around with either; deficiency can lead to gallstones, arthritis, and muscle
deterioration, as well as heart problems. Here are some of the most common symptoms. The
symptoms of protein deficiency are:-

Edema: Edema is a collection of fluid under the skin, which most commonly affects the legs,
feet, and ankles but can occur anywhere on the body. Protein is essential for maintaining a
balance of water in your body; without it, you may store water improperly.

Weight Loss: Severe weight loss is one symptom of a protein deficiency. It may be attributed to
muscle wastage, as your body breaks down your muscles in an attempt to get protein from them.

Thinning or Brittle Hair: Hair loss and thinning or brittle hair can also be caused by a protein
deficiency. Hair is made up of protein, so a deficiency may lead your hair to lack the amount of
protein it needs to stay healthy. If you become deficient enough, your hair will even begin to fall
out.

Ridges in Nails: Ridges or white lines in both finger and toe nails can be caused by a lack of
protein the diet. Ridges that run from top to bottom on the nail can indicate an ongoing protein
deficiency, while a ridge that runs transverse may indicate a deficiency that has now passed.

Pale Skin: Skin that loses pigment and burns more easily in the sun can be caused by a lack of
iron, as well as protein. Frequently, foods that are rich in iron also contain protein, and protein is
necessary for the body to utilize iron properly. Anemia or lack of iron may result in pale skin.
Skin Rashes: Skin rashes, which may be accompanied by dry or flaking skin are a symptom of a
protein deficiency. These rashes are caused by extreme protein deficiency and may resemble
eczema or other dry skin rashes.

General Weakness: Weakness and lethargy may be caused by a lack of protein in the diet. Over
time, as your stores of protein are depleted, your body will begin to break down muscle tissue,
which can lead to a feeling of general weakness or lethargy.

Slow Healing: Amino acids, the building blocks of protein, are crucial in wound healing. When
your body heals itself, it needs to repair or produce tissues. These tissues use amino acids in the
repairs, and a lack of protein can lead to a slower building process.

Difficulty Sleeping: Difficulty in sleeping could be caused by a serotonin deficiency, which is

caused by a lack of certain amino acids. These amino acids are produced when protein is broken
down, and a diet with insufficient protein could lead to difficulty in sleeping.

Headache: Headaches can be caused by many things, including a protein deficiency. A lack of
protein may lead to headaches in one of a couple ways:

By causing anemia, By causing low blood sugar, Fainting

Fainting and general weakness may be caused by a protein deficiency as well. This may be
attributed to low blood sugar or to your body's need to break down muscle tissue to get the
nutrients it needs.

Other Symptoms: Not all of the symptoms of protein deficiency are physical. Some are
emotional or mental, and include the following:

Crankiness, moodiness, Problems with conflict resolution, Severe depression, Anxiety

Lack of energy, no desire to do things (Whitney et al., 2005)

Fats and their deficiency

You need at least a small amount of fat in your diet to provide the essential omega-3 and
omega-6 fatty acids your body can't make and to help with the absorption of fat-soluble vitamins.
If you don't get at least 15 percent of your calories from fat, you could develop an essential fatty
acid deficiency or a deficiency of one or more of the fat-soluble vitamins A, D, E and K. An
essential fatty acid deficiency can be corrected by getting more omega-3 and omega-6 fats in
your diet, and the combination of fat and fat-soluble vitamins can resolve fat-soluble vitamin
deficiencies.

You can develop deficiencies of vitamins A, D, E and K if you don't get enough fat in
your diet because your body can only absorb these nutrients in the presence of fat. Vitamin A
deficiency symptoms can include dry skin, night blindness, increased susceptibility to infections
and problems with bone and tooth development. Not absorbing enough vitamin D can cause soft
and weak bones, and too little vitamin K can lead to increased bleeding.

You need the essential fatty acids for keeping your skin healthy, so not getting enough fat
in your diet can cause skin problems. An essential fatty acid deficiency increases the loss of
water from your skin, which results in a dry, scaly rash. This type of deficiency can also make it
harder for your wounds to heal.

Two types of omega-3 fatty acids, eicosapentaenoic acid and docosahexaenoic acid, are
essential for brain development. Not getting enough DHA may result in learning deficits and
increase the risk for certain types of dementia, including Alzheimer's disease. The omega-6 fat
arachidonic acid is also an important component of brain tissue (World bank, 1994).

EPA and DHA are also important components of the retina, so an essential fatty acid
deficiency could affect your vision. DHA helps form the pigment rhodopsin, which you need for
your brain to turn the light hitting your retina into the images you see. The retina can recycle and
conserve DHA, however, so even low intakes of essential fatty acids may be enough to protect
your vision (WHO, 2006).
MATERIALS & METHODS
 MATERIALS AND METHODS

The various food samples were taken which were tested for essential macronutrients (primary).
These were:

Gram flour, sugar, groundnut oil, milk, wheat flour, rice

Test for carbohydrates :

Fehling’s test

All the samples were tested for presence of carbohydrate. Those with it were notified.
This forms the reduction test of carbohydrates. Fehling’s solution contains blue alkaline cupric
hydroxide solution, heated with reducing sugars gets reduced to yellow or red cuprous oxide and
is precipitated. Hence, formation of the yellow or brownish-red colored precipitate helps in the
detection of reducing sugars in the test solution.

Benedict’s Test

In the test tube with 2 ml of Benedict's reagent, add 5-6 drops of the test carbohydrate
solution and mix well. Place the test tube in a boiling water bath for 5 minutes and observe any
change in color or precipitate formation. Cool the solution. Observe the colour change from blue
to green, yellow, orange or red depending upon the amount of reducing sugar present in the test
sample.

Molisch’s Test

In a test tube, add 2 ml of the test carbohydrate solution and 2 drops of α-naphthol
solution. Carefully incline the tube and pour dropwise conc. H2SO4, using a dropper,along the
sides of the tube. Observe the violet colour at the junction of the two liquids.
Test for Proteins :

Biuret test

Add an equal amount of NaOH to a solution of the food, mix carefully. Add a few drops of
1% CuSO4, do not shake the mixture. A purple/mauve color is a positive result: protein is
present.

Xanthoproteic test

To 1ml of protein solution add 1ml of concentrated HNO3. Boil for 30 seconds. Cool under tap
water and add 2-3 ml of 40% NaOH.

Test for lipids :

Sudan dye test

To 1 ml of sample add a pinch of sudan iv dye powder. If it dissolves completely, it shows red
solution, positively presence of fat.

The food samples were discriminated according to the macronutrient test and hence
analysed.
OBSERVATIONS
 OBSERVATIONS

Food samples chosen for qualitative testing

1. Gram flour

Gram flour, also known as garbanzo bean flour, or besan, is a pulse flour made
from ground chickpeas known in a number of Asian countries as gram. It is a staple
ingredient in Indian, Pakistani, Nepali and Bangladeshi cuisines. Gram flour can be made
from either raw or roasted gram beans. The roasted variety is more flavorful, while the
raw variety has a slightly bitter taste.

Fig.1: Gram flour

2. Sugar

Sugars are found in the tissues of most plants, but are present in sufficient
concentrations for efficient extraction only in sugarcane and sugar beet. Sugarcane refers
to any of several species of giant grass in the genus Saccharum that have been cultivated
in tropical climates in South Asia and Southeast Asia since ancient times. A great
expansion in its production took place in the 18th century with the establishment of sugar
plantations in the West Indies and Americas. This was the first time that sugar became
available to the common people, who had previously had to rely on honey to sweeten
foods.
 Fig.2: Sugar crystals

3. Rice

Rice is the seed of the grass species Oryza sativa (Asian rice) or Oryza glaberrima
(African rice). As a cereal grain, it is the most widely consumed staple food for a large part of
the world's human population, especially in Asia. It is the agricultural commodity with the third-
highest worldwide production, after sugarcane and maize (Juliano and Bienvenido, 1993).

Fig.3: Rice
 4. Wheat flour

Wheat flour is a powder made from the grinding of wheat used for human
consumption. More wheat flour is produced than any other flour. It is a blended wheat
with a protein content lower than bread flour, ranging between 9% and 12%. Depending
on brand or the region where it is purchased, it may be composed of all hard or soft
wheats, but is usually a blend of the two, and can range from low protein content to
moderately high. It is marketed as an inexpensive alternative to bakers' flour which is
acceptable for most household baking needs (Belderok et al., 2000).

Fig.4: Wheat flour

5. Milk

Milk is a pale liquid produced by the mammary glands of mammals. It is the primary source
of nutrition for infant mammals before they are able to digest other types of food. The females of
all mammal species can by definition produce milk, but cow's milk dominates commercial
production. In 2011, FAO estimates 85% of all milk worldwide was produced from cows (Goff,
2010).

Human milk is not produced or distributed industrially or commercially; however, human

milk banks collect donated human breastmilk and redistribute it to infants who may benefit from
human milk for various reasons (premature neonates, babies with allergies, metabolic diseases,
etc.) but who cannot breastfeed (Hemme and Otte , 2010).
Fig.5: Milk

6. Groundnut Oil

Peanut oil, also known as groundnut oil or arachis oil, is a mild-tasting vegetable
oil derived from peanuts. The oil is available with a strong peanut flavor and aroma,
analogous to sesame oil. It is often used in Chinese, South Asian and Southeast Asian
cuisine, both for general cooking, and in the case of roasted oil, for added flavor. Peanut
oil has a high smoke point relative to many other cooking oils, so is commonly used for
frying foods (Anyasor et al., 2009).

Fig.6: Groundnut oil

Table 1: Presence of primary macronutrients in different food products

S.No. Food product Carbohydrate Protein Fat

1. Sugar + - -

2. Gram flour - + -

3. Wheat flour + - -

4. Milk - + +

5. Groundnut oil - - +
Table 2: Biochemical tests performed to detect different food products

S.No. Food product Biochemical test Observation Type of

primary
macronutrient

1. Sugar Molish test Brown ring Carbohydrate

present
Fehling’s test Reddish brown ppt.

Benedict’s test Red ppt.

2. Gram flour Biuret test Purple solution Protein present

Xanthoproteic test Orange ppt.

3. Wheat flour Molish test Brown ring Carbohydrate

present
Fehling’s test Reddish brown ppt.

Benedict’s test Red ppt.

4. Milk Biuret test Purple solution Protein present

Xanthoproteic test Orange ppt.

5. Groundnut oil Sudan dye test Red solution Fat present

Fig.7: Molish test of sugar solution showing brown ring

Fig.8: Fehling’s test of sugar solution showing dark red ppt.

Fig.9: Positive biuret test for proteins showing purple color

Fig.10: Xanthoproteic reaction showing yellow to orange ppt.

Fig. 11: Sudan IV dye powder dissolves in lipids

RESULTS & DISCUSSION
 RESULTS AND DISCUSSION

Analytical tests for Carbohydrates with detailed Biochemistry

Molisch’s test

Molisch’s reagent is 10% alcoholic solution of α-naphthol. This is a common chemical

test to detect the presence of carbohydrates. Carbohydrates undergo dehydration by sulphuric
acid to form furfural (furfuraldehyde) that reacts with α-naphthol to form a violet coloured
product.

Fehling's test

This is an important test to detect the presence of reducing sugars. Fehling’s solution A is
copper sulphate solution and Fehling’s solution B is potassium sodium tartrate. On heating,
carbohydrate reduces deep blue solution of copper (II) ions to red precipitate of insoluble copper
oxide.

Benedict's test

Benedict’s test distinguishes reducing sugar from non-reducing sugar. Benedict’s

reagent contains blue copper (II) ions (Cu2+, cupric ions) that are reduced to copper (I) ions (Cu
+
, cuprous ions) by carbohydrates. These ions form precipitate as red coloured cuprous (copper
(I) oxide (Kenneth, 2012).
Analytical tests for Proteins with detailed Biochemistry

Biuret test

The biuret test is a chemical test used for detecting the presence of peptide bonds. In the
presence of peptides, a copper(II) ion forms violet-colored coordination complexes in an alkaline
solution. Several variants on the test have been developed, such as the BCA test and the
Modified Lowry test.

The biuret reaction can be used to assess the concentration of proteins because peptide
bonds occur with the same frequency per amino acid in the peptide. The intensity of the color,
and hence the absorption at 540 nm, is directly proportional to the protein concentration,
according to the Beer-Lambert law.

Despite its name, the reagent does not in fact contain biuret ((H2N-CO-)2NH). The test is
so named because it also gives a positive reaction to the peptide-like bonds in the biuret molecule
(Bruckdorfer et al., 2004).

The Biuret reagent is made of sodium hydroxide (NaOH) and hydrated copper(II) sulfate,
together with potassium sodium tartrate.[4] Potassium sodium tartrate[5] is added to complex to
stabilize the cupric ions. The reaction of the cupric ions with the nitrogen atoms involved in
peptide bonds leads to the displacement of the peptide hydrogen atoms under the alkaline
conditions. A tri or tetra dentate chelation with the peptide nitrogen produces the "buret" color.
This is found with dipeptides.
Xanthoproteic test

The xanthoproteic reaction is a method that can be used to determine the amount of
protein soluble in a solution, using concentrated nitric acid. The test gives a positive result in
those proteins with amino acids carrying aromatic groups, especially in the presence of tyrosine.
If the test is positive the proof is neutralized with an alkali, turning dark yellow. The yellow
colour is due to xanthoproteic acid which is formed due to nitration of certain amino acids, most
common examples being tyrosine and tryptophan.[1] This chemical reaction is a qualitative test,
determining the presence or absence of proteins.

Add 1 ml of conc HNO3 to 1 ml of the test sample. Heat the mixture and cooled it.
Slowly add sodium hydroxide (40% w/v in water) solution till the mixture become alkaline and
color changes is noted. If color changes from yellow to orange. It indicates the presence of
aromatic amino acid (Dobson, 2000).
Analytical tests for Lipids with detailed Biochemistry

Sudan dye test

Sudan stains and Sudan dyes are synthetic organic compounds that are used as dyes for
various plastics and are also used to stain sudanophilic biological samples, usually lipids. Sudan
II, Sudan III, Sudan IV, Oil Red O, and Sudan Black B are important members of this class of
compounds.

Sudan dyes have high affinity to fats, therefore they are used to demonstrate triglycerides,
lipids, and lipoproteins. Alcoholic solutions of Sudan dyes are usually used, however pyridine
solutions can be used in some situations as well.

Since they are characteristically "oil-and fat-soluble," Sudan dyes are also useful for
dying plastics and fabrics.[1] Sudan dyes I-IV and Sudan Red G consist of arylazo-substituted
naphthols. Such compounds are known to exist as a pair of tautomers.
Analytical tests of food products has great significance

Food analysis is the discipline dealing with the development, application and study of
analytical procedures for characterizing the properties of foods and their constituents. These
analytical procedures are used to provide information about a wide variety of different
characteristics of foods, including their composition, structure, physicochemical properties and
sensory attributes. This information is critical to our rational understanding of the factors that
determine the properties of foods, as well as to our ability to economically produce foods that are
consistently safe, nutritious and desirable and for consumers to make informed choices about
their diet. The objective of this course is to review the basic principles of the analytical
procedures commonly used to analyze foods and to discuss their application to specific food
components, e.g. lipids, proteins, water, carbohydrates and minerals (Hayes, 2005).

Foods are analyzed by scientists working in all of the major sectors of the food industry
including food manufacturers, ingredient suppliers, analytical service laboratories, government
laboratories, and University research laboratories (Grover et al., 2009).

Government regulations and recommendations

Government regulations and recommendations are designed to maintain the general

quality of the food supply, to ensure the food industry provides consumers with foods that are
wholesome and safe, to inform consumers about the nutritional composition of foods so that they
can make knowledgeable choices about their diet, to enable fair competition amongst food
companies, and to eliminate economic fraud. There are a number of Government Departments
Responsible for regulating the composition and quality of foods, including the Food and Drug
Administration (FDA), the United States Department of Agriculture (USDA), the National
Marine Fisheries Service (NMFS) and the Environmental Protection Agency (EPA). Each of
these government agencies is responsible for regulating particular sectors of the food industry
and publishes documents that contain detailed information about the regulations and
recommendations pertaining to the foods produced within those sectors. These documents can be
purchased from the government or obtained on-line from the appropriate website (Webm.com,
2009).

Nutrient Labeling

In 1990, the US government passed the Nutritional Labeling and Education Act (NLEA),
which revised the regulations pertaining to the nutritional labeling of foods, and made it
mandatory for almost all food products to have standardized nutritional labels. One of the major
reasons for introducing these regulations was so that consumers could make informed choices
about their diet. Nutritional labels state the total calorific value of the food, as well as total fat,
saturated fat, cholesterol, sodium, carbohydrate, dietary fiber, sugars, protein, vitamins, calcium
and iron. The label may also contain information about nutrient content claims (such as low fat,
low sodium high fiber fat free etc), although government regulations stipulate the minimum or
maximum amounts of specific food components that a food must contain if it is to be given one
of these nutrient content descriptors. The label may also contain certain FDA approved health
claims based on links between specific food components and certain diseases (e.g., calcium and
osteoporosis, sodium and high blood pressure, soluble fiber and heart disease, and cholesterol
and heart disease). The information provided on the label can be used by consumers to plan a
nutritious and balanced diet, to avoid over consumption of food components linked with health
problems, and to encourage greater consumption of foods that are beneficial to health.
CONCLUSIONS
&
FUTURE SCOPE
 CONCLUSIONS AND FUTURE SCOPE

The big 3 macronutrients are- fats, proteins and carbohydrates

We often see or hear claims about a food product being “low in carbs”, “high in protein”
or “fat free”. We thought it would be a good idea to get back to the basics and do a post on these
3 macronutrients. They are the source of practically all calories in food. They are called macro-
nutrients because we need these in large quantities to function properly.

Foods generally contain different proportions of fats, protein, carbs and water. However,
some foods are made up of only one macronutrient such as oil (pure fat) or sugar (pure
carbohydrate).

Carbohydrates (Carbs)

Contrary to many fad diets out there – Carbs are the main source of energy for our
bodies. The most simple form of carbohydrate is glucose, the only substance our brains use for
energy. Our kidneys, heart and nervous system also use glucose to function properly. Out of our
total energy intake for the day, it is recommended to get around 45-65% of energy from carbs.
So in a 2000 calorie diet this is 900 to 1,300 calories a day, or 225 – 325 grams (One gram of
carbohydrate provides 4 calories). Our body can use the energy from carbs immediately, or store
it for later use. Common sources of carbohydrates include breads, cereals, pasta, rice, potatoes,
fruits and vegetables (Welsh et al., 2011).

When choosing a carbohydrate it is important to select something that is high in fiber,

low glycemic index and as least processed as possible. Examples include wholegrain bread
(Slaffer and Satorre, 1999), apples, sweet potatoes, brown rice and oats (Tighe et al., 2010).

In the present study it was tested that wheat flour, sugar and rice are sources of
carbohydrates.
Proteins

We only need about 10-35% of our diets to come from protein – most Americans get
more than ample protein every day (Reynolds and Tanford, 2003). Proteins are important for
repair and growth of tissues, building muscle, enzymes, hormones, amino acids and our immune
system. Protein isn’t used immediately for energy – it’s more of a backup for our body. One
gram of protein, like carbs, also provides 4 calories (Sleator, 2012).

Choose foods that have high quality protein: lean meats, fish, poultry, low fat cheese,
milk, yoghurt, eggs, legumes, soy, nuts and seeds (Schwarzer and Cole, 2005).

In this study, gram flour and milk are main source of proteins.

Fats

Fats should make up about 20-35% of our diet. Unfortunately, they’ve been getting a bad
reputation for decades. Nonetheless, fats are essential in our diet. Fats are important for
protecting our organs, maintaining cell membranes, promoting growth and development and
absorbing essential vitamins. One gram of fat contains about 9 calories (more than double a gram
of carb or protein, hence the vilification) (Liu et al., 2011).

Choose foods that contain unsaturated fats over saturated fats, such as oily fish, nuts and
seeds, olive oil, avocado and canola oil instead of full cream milk, deep fried foods, cream,
butter and animal fat (Prentice et al., 2005).

In the present study groundnut oil is main source of fat.

Balanced diet can be planned by knowledge of nutritional intake

A balanced diet is one that gives your body the nutrients it needs to function correctly. In
order to get the proper nutrition from your diet, you should obtain the majority of your daily
calories from fresh fruits, fresh vegetables, whole grains, legumes, nuts, lean
proteins. A balanced diet is important because your organs and tissues need proper nutrition to
work effectively (Molina et al., 2011). Without good nutrition, your body is more prone to
disease, infection, fatigue, and poor performance. Children with a poor diet run the risk of
growth and developmental problems and poor academic performance. Bad eating habits can
persist for the rest of their lives.

Daily calorie intake

The number of calories in a food is a measurement of the amount of energy stored in that
food (UNICEF committee, 2006). Your body uses calories from food for walking, thinking,
breathing, and other important functions. The average person needs to eat about 2,000 calories
every day to maintain their weight. However, a person’s specific daily calorie intake can vary
depending on their age, gender, and physical activity level. Men generally need more calories
than women, and people who exercise need more calories than people who don’t (Ashworth,
2003).

The following examples of daily calorie intake are based on United States Department of
Agriculture (USDA) guidelines:

1. Children ages 2 to 8: 1,000 to 1,400 calories

2. Active women ages 14 to 30: 2,400 calories

3. Sedentary women ages 14 to 30: 1,800 to 2,000 calories

4. Active men ages 14 to 30: 2,800 to 3,000 calories

5. Sedentary men ages 14 to 30: 2,000 to 2,600 calories

6. Active men and women over 30: 2,200 to 3,000 calories

7. Sedentary men and women over 30: 1,800 to 2,200 calories

Thus, by knowing detailed calorific value, nutritional supplements in food we can maintain a
healthy balanced diet.
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