c



 



1. To test the presence of starch, reducing sugars, non -reducing sugars,

proteins and lipids in food samples.

Introduction: Glucose is an aldose which has aldehyde group as functional

group. Fructose is a ketose which has ketone group as functional group. All
monosaccharides are reducing sugars due to the presence of free aldehyde group or
ketone group.
The Benedict¶s reagent is a solution that contains copper (II) sulphate and
alkaline solution, and the copper (II) ions ar e reduced by the aldehyde or ketone
group to form copper (I) oxide precipitate. Besides, the yellow, green, and orange
color of precipitate also indicates the presence of reducing sugar.
Sucrose is the disaccharide formed from the enzymatic dehydration rea ction
between glucose and fructose. Lactose is the disaccharide formed from the
dehydration reaction between the glucose and galactose.
Lactose is reducing sugar due to the presence of aldehyde or ketone group in
one of the monomers that can gives the pos itive result on the Benedict¶s test.
Sucrose is non-reducing sugar, due to the absence of the aldehyde or ketone group
after the formation of the glycosidic linkage. Thus, sucrose gives negative result on
the Benedict¶s test.
Starch is a polysaccharide of glucose and thus cannot give positive result on the
Benedict¶s test, since all polysaccharides do not have any reducing ability. However,
starch gives positive result on the Iodine test. Starch forms the dark blue colo ur of
solution with iodine solution wh ich is the result from the complex ion formed between
the amylose and the iodine molecules.

Both disaccharides and polysaccharides can be hydrolyzed into their

constituents with addition of water and requiring specific enzyme that catalyze the
hydrolysis reaction. In the laboratory, these compound sugars can be hydrolyzed in
the test tube by addition of concentrated acid and heating process.

Proteins consist of one or more polypeptides coiled or folded into specific three
dimensional conformations. The fun ctional group of amino acids are the carboxyl
group (-COOH) and the amino group (-NH2). The carboxyl group and amino group of
the adjacent amino acids can be linked together to form a peptide linkage through
dehydration reaction.
The Biuret reagent is consisting of mixture of the copper (II) sulphate and an
alkaline solution, used to detect the presence of peptide linkages. A solution that
contains proteins will give violet coloration to the Biuret reagent.

The lipid is a group of macromolecules that are insoluble in water. The major
kinds of lipids in biological world include fats, phospholipids, and steroids.
 The fats or triacylglycerols are made up of glycerol (an alcohol) and fatty acids
(a monocarboxylic acid) which are bonded together by ester linkag es.

The lipids are greasy and would turn the area of dry lipids on a brown paper into
semi-transparent.
ã


Food sample A contains starch, food sample B contains non -reducing sugar
(sucrose), food sample C contains protein, food sample D contains reducing sugar
(glucose), food sample E contains lipid



a) Manipulated: Types of food samples

b) Responding: Changes colour of food samples
c) Fixed: Volume of the reagent used



400 B.C. -- Hippocrates, the "Father of Medicine", said to his students, "Let thy food
be thy medicine and thy medicine be thy food". He also said ³A wise man should
consider that health is the greatest of human blessings.´



 

Iodine solution, Benedict solution, sodium hydrogen carbonate solution, 20% sodium
hydroxide solution, hydrochloric acid, 1% copper (II) sulphate solution and food
samples (unknown to students)

(A-raisin/dates solution, B-pounded groundnut, C -milk, D-honey syrup, and E-

mayonnaise) the food samples can varies.

Test tubes, test-tube holders, beakers, Bunsen burner, dropper, wire gauze, tripod
stand, white tile and filter paper.

c

1. Five samples of food labelled A,B.C,D and E are prepared.

2. Food tests are carried out to determine presence of starch, reducing sugars,
non-reducing sugars, proteins and lipids in these food samples.



Food Test for Procedure Observation Inference

sample
A Starch 1. Pour 2ml of sample A blue black This is because
A into a test tube colour formed starch is present in
2. Add three drops of on the solution the solution.
iodine solution to
 the food sample
3. Observe what
happens
4. Repeat steps 1 to 3
with other food
samples.
B Non- 1. Pour 2 ml of sample Solution This is because
reducing B into a boiling tube. remains clear non-reducing sugar
sugar Then add a few (sucrose) is present
drops of dilute in the solution.
hydrochloric acid
2. Heat the mixture in
a water bath for
about five minutes
3. Remove the boiling
tube from water
bath and cool the
mixture under a
running tap.
4. Neutralise the acid
by adding sodium
hydrogen carbonate
solution until the
effervescence
stops.
5. Then, conduct
Benedict¶s test on
the mixture.
6. Observe any colour
change in the
mixture.
7. Repeat steps 1 to 6
using other food
samples.
C Protein, 1. Pour 2ml of sample A blue or lilac This is because
C into a test tube. purple form in protein is present in
2. Add 20% sodium the solution. the solution.
hydroxide solution
in excess to the
food sample and
shake well.
3. Slowly, add a few
drops of 1% copper
(II) sulphate solution
to the mixture.
4. Shake well and
allow the mixture to
stand.
5. Observe any colour
change in the
 mixture.
6. Repeat steps 1 to 5
using other food
samples.
D Reducing 1. Pour about 2ml of A brown or red This is because
sugar sample D into a test precipitate is reducing sugar
tube. formed in the (glucose) is present
2. Add about 1ml of solution in the solution
Benedict¶s solution
to the food sample
3. Shake the mixture.
Then, heat the test
tube by placing it in
water bath until the
mixture is brought to
a boil.
4. Observe any colour
change that takes
place.
5. Repeat steps 1 to 4
using other food
samples.
E Lipids 1. Rub a small amount A cloudy white This is because
of each food sample emulsion is lipids are present in
on a piece of filter formed on the the food sample.
paper. filter paper.
2. Dry the filter paper.
You may use a
hairdryer.
3. Hold the filter paper
against the light.
Record your
observations.




1. When a reducing sugar is heated with Benedict¶s solution, the reducing sugar
reduces the blue copper (II) sulphate in Benedict¶s solution to form a red
precipitate of copper (I) oxide.

2. A brick-red precipitate indicates that a large amount of reducing sugar is

present while an orange or green precipitate indicates the presence of a
lesser amount of reducing sugar. If the original pale blue colour of the solution
remains, this indicates that reducing sugar is not present.
 3. The different food samples are first heated with dilute hydrochloric acid to
hydrolyse the non-reducing sugar (sucrose) to its constituent
monosaccharides (glucose and fructose) which are reducing sugars.
4. The mixture of food sample and hydrochloric acid is first neutralised by adding
sodium hydrogen carbonate powder before adding Benedict¶s solution.

5. If the food sample provided is solid, the food needs to be cut into very small
piece or ground before adding a little water to provide a more concentrated
solution for testing.



The hypothesis is accepted. Food sample A contains starch, food sample B contains
non-reducing sugar, food sample C contains protein, food sample D contains
reducing sugar and food sample E contains lipids.

 2. To determine the vitamin C content in various fruit juices

c  Do imported fruits contain more vitamin C content in various fruit juices.

 : Vitamin C is found in green vegetables, fruits, and potatoes. It is

essential for a healthy diet. The chemical name for vitamin C is ascorbic acid.
Ascorbic acid is a good reducing agent and therefore it is easily oxidised. Methods
for the detection of vitamin C involve titrating it against a solution of an oxidising
agent.

ã
 Lime juice contains a higher concentration of vitamin C compared to

guava juice, papaya juice, and orange juice.

 :

a) Manipulated: Types of fruit juices

b) Responding: Volume of fruit juice needed to decolourise DCPIP solution
c) Constant: Volume of DCPIP solution



In 1912, the Polish -American biochemist Casimir Funk, while researching

deficiency diseases, developed the concept of vitamins to refer to the non -mineral
micro-nutrients which are essential to health. The name is a blend of "vital", due to
the vital role they play biochemically, and "amines" because Funk thought that all
these materials were chemical amines. One of the "vitamines" was thought to be the
anti-scorbutic factor, long thought to be a component of most fresh plant material.
 From 1928 to 1933, the Hungarian research team of Joseph L Svirbel y and
Albert Szent-Györgyi and the American worker Charles Glen King, first identified the
anti-scorbutic factor, calling it "ascorbic acid" for its vitamin activity. Szent -Györgyi
had isolated the chemical hexuronic acid from animal adrenal glands at the Mayo
clinic, and suspected it to be the antiscorbutic factor, but could not prove it without a
biological assay. At the same time, for five years King's laboratory at the University
of Pittsburgh had been trying to isolate the antiscorbutic factor in foods , using a
model of scorbutic guinea pigs, which proved to develop scurvy when not fed fresh
foods. In late 1931 Szent -Györgyi gave Joseph Svirbely, a former worker in King's
lab who had recently joined Szent -Györgyi's, the last of this hexuronic acid, with the
suggestion that it might be the anti -scorbutic factor. By the spring of 1932, King's
laboratory had proven this, but published the result without giving Szent -Györgyi
credit for it, leading to a bitter dispute over priority claims (in reality it had t aken a
teamwork effort by both groups, since Szent -Györgyi was unwilling to do the difficult
and messy animal studies). By 1932, Szent -Györgyi's group had discovered that
paprika peppers, a stable spice in the Hungarian diet, was a rich source of hexuronic
acid, the antiscorbutic factor, by then named ascorbic acid, in honor of its activity
against scurvy. Ascorbic acid turned out O
to be an amine, nor even to contain any
nitrogen.

For his accomplishment, Szent-Györgyi was alone awarded the 1937 Nobel
Prize in Medicine "for his discoveries in connection with the biological combustion
processes, with special reference to vitamin C and the catalysis of fumaric acid".



 

1.0% dichlorophenolindophenol solution (DCPIP), 0.1% ascorbic acid solution,

freshly prepared guava juice, papaya juice, orange juice and lime juice, specimen
tubes, syringes with needles (1ml and 5ml), beakers, gauze cloth and knife.

c

1. 1ml of 1.0 % DCPIP solution is placed in a specimen tube using a 1ml

syringe.
2. The 5ml syringe is filled with 0.1% ascorbic acid solution.
3. The needle of the syringe is placed into a DCPIP solution.
4. The ascorbic acid solution is added drop by drop to the DCPIP solution. The
mixture is gently stirred with the needle of the syringe. The ascorbic acid
solution is continuously added until the DCPIP solution is decolourised. The
volume of ascorbic acid solution used is recorded.
5. Steps 1 to 4 are repeated using freshly squeezed guava juice, papaya juice,
orange juice, and lime juice. The volume of fruit juice required to decolourise
the DCPIP solution in each case is recorded .
6. The results are tabulated. The percentage and the concentration of Vitamin C
in each of the fruit juices are calculated using the formulae given.
 

Solution/fruit juice Volume of solution/fruit juice Percentage Vitamin C

needed to decolourise 1 ml of of Vitamin concentration
DCPIP solution C in fruit in fruit juice
juice
1 2 3 Average
0.1% ascorbic acid 0.20 0.40 0.60 0.40 100% 1.00
guava juice 3.00 3.80 3.00 3.27 12% 0.12
papaya juice 3.00 3.00 3.00 3.00 13% 0.13
orange juice 3.90 4.10 4.30 4.10 10% 0.10
Lime juice 1.00 1.00 1.00 1.00 40% 0.40

 

Volume of 0.1% ascorbic acid used to decolourise DCPIP =x cm 3

Volume of fruit juice used to decolourise DCPIP = y cm 3

X cm3 0.1% ascorbic acid (concentration of 1 mg/ cm3) can decolourise 1 cm 3 of

DCPIP and y cm3 of fruit juice concentration of k mg/cm 3 can decolourise 1 cm 3 of
DCPIP

so,

The quantity of ascorbic acid in x cm3 of ascorbic acid 0.1%

= quantity of ascorbic acid in y cm3 of fruit juice

Thus,

x cm3 X 1mg/cm3 = y cm3 X k mg/cm3



k mg/cm3 = X mg/cm3



=  mg/cm3

Therefore the concentration of ascorbic acid in fruit juice = mg/cm3




šuestions:

1. Explain the need for our diet to contain: carbohydrates, sodium chloride, fresh
fruits and milk.
 We need to consume various types of food to stay healthy. Carbohydrate is
need for supplying energy to our body. Sodium chloride is used to maintain the
acid-base balance and water balance and also important component in blood
plasma. Fresh fruits are to make sure that we are consumes enough fibres to
provide bulk to the intestinal contents and aids in peristalsis. It also avoids us
from having constipation. Milk, which is rich with calcium, is used for formation of
bone and tooth and is needed in muscle an d nerve coordination. The most
important thing is by consuming milk, we can prevent from getting osteoporosis
when we become older.

2. How do vegetarians ensure that they receive a balanced diet?

A vegan is a strict vegetarian who does not eat any dairy products, eggs or
honey. Obtaining adequate protein on a vegan diet is not a problem. Nuts &
seeds, pulses, wholegrain and grain products and soya products all supply
protein. Previously, it has been thought that plant proteins are of a lower quality
than animal proteins in terms of their essential amino acid content. However, this
is no longer regarded as a problem and eating a balanced diet of plant foods will
provide all the essential amino acids in adequate amounts. p

There are two essential fatty acids which must be supplied by the diet. These
are linoleic acid and a -linolenic acid. Essential fatty acids are important for cell
membrane function, cholesterol metabolism and the synthesis of various
metabolites. Good sources of essential fatty acids are veg etable oils. It is
important to have the correct balance between linoleic acid and a -linolenic acid. It
has been suggested that vegans should use soyabean or rapeseed oils rather
than sunflower or corn oils as these help give a better dietary balance. p

Certain studies have found vegans to have a low intake of the vitamin,
riboflavin. Riboflavin is important in converting protein, fats and carbohydrates
into energy, and the synthesis and repair of body tissues. Good sources of
riboflavin include whole grains, mushrooms, almonds, leafy green vegetables and
yeast extracts.p

Vitamin B12 is found primarily in meat, dairy products and eggs and is absent
from plant foods. Considerable research has been carried out into possible plant
sources of B12. Fermented soya products, seaweeds and algae such as
Spirulina have all been proposed as containing significant amounts of B12.
However, the present consensus is that any B12 present in plant foods is likely to
be in a form unavailable to humans and so these foods should no t be relied upon
as safe sources. p

Vitamin B12 is important in the formation of red blood cells and the
maintenence of a healthy nervous system. When deficiency does occur it is more
likely to be due to a failure to absorb B12 from the intestine than a die tary
deficiency. Vegans can obtain B12 from a wide range of foods which have been
fortified with the vitamin. These include certain yeast extracts, veggieburger
mixes, breakfast cereals, vegetable margarines and soya milks. p
 Vitamin D is present in oily fish, eggs and dairy products in variable amounts.
It is not found in plant foods. However, vegans can obtain vitamin D from
vegetable margarines, some soya milks and certain other foods which are
fortified with the vitamin. Vitamin D is also synthesised by t he skin when exposed
to sunlight. Synthesis of vitamin D in this way is usually adequate to supply all the
body's requirements. Most vegans will obtain sufficient vitamin D providing they
spend time outdoors on bright days. Fortified foods further ensure a dequate
amounts. p

The major source of calcium in diets is generally milk and dairy products.
Vegans can obtain adequate calcium from plant foods. Good sources include
tofu, leafy green vegetables, watercress, dried fruit, seeds and nuts. Also, white
bread is fortified with calcium, as are some soya milks. Hard water can also
provide significant amounts of calcium. p
For iodine intake, vegan can consume seaweeds. Seaweeds are a good
source of iodine, and vegetables and grains can contain iodine depending on t he
amounts in the soil.

3. The body cannot digest fibre. Why is it still important in our diet?

Fibre consists mainly of cellulose. Although the body cannot digest fibre but
the dieticians recommended that about 25 -50g of fibre should be eaten each day
to ensure good health. Fibre provides bulk to the intestinal contents because of
its high water holding capacity. It also aids peristalsis and in a bowel movement.
This prevents the build-up of toxic substances in the rectum which can lead to
bowel cancer.

4. A slightly overweight friend decides to go on a crash diet. She/he tells you that
she/he is only going to eat grapefruit and drink black coffee for two weeks.
What advice would you give to your friend?

Black coffee contains a lot of caffeine while grapefruit rich in vitamin C. The
coffee diet suggests that people drink one cup of black coffee in the morni ng to
jump start their metabolism by up to ten percent and curb their appetite, resulting
in weight loss over time by burning calories faster than normal. The coffee diet is
not for everyone, and those who decide to follow the newest fad diet should
check with their health care provider before beginning, just as with any other diet
or fitness regime. Despite its seemingly fabulous effects, the coffee diet can
cause health problems and negative side effects that for some are simply not
worth the slight increase in metabolism.

The caffeine from a daily intake of black coffee can cause unstable blood
sugar, resulting in fatigue and drowsiness and in some instances, cause the body
to store other food as fat which would result in weight gain, the opposite desired
effect. The acidity in coffee can cause ulcers and airway corrosion along with
stained tooth enamel after prolonged use. Another side effect of caffeine is the
effect it has on the circulatory system, namely the heart.

Caffeine can cause the heart to beat quickly and abnormally, resulting in a
feeling of shakiness, fidgeting, hypertension, and ultimately a loss of
concentration. Keeping in contact with your physician while on the coffee diet is a
good idea, as problems can arise quickly and can be detrimental to one's
physical state.

Grapefruit is an excellent source of many nutrients and phytochemicals t hat

contribute to a healthy diet. Grapefruit is a good source of vitamin C, pectin fiber,
and the pink and red hues contain the beneficial antioxidant lycopene. Studies
have shown grapefruit helps lower cholesterol . Grapefruit forms a core part of
the "grapefruit diet", the theory being that the fruit's low glycemic index is able to
help the body's metabolism burn fat.

It was hypothesized that the grapefruit reduced insulin levels, encouraging fat
loss. However, the study involved a healthy diet with the a ddition of grapefruit, in
contrast to a normal "grapefruit diet".

Hence, I will tell my friends to stop the crash diet and if she wants to go on a
diet, she should meet dieticians to help her in consuming balanced intake of
nutrition.

5. Eskimos who eat mainly fish, seal oil and whale meat rather than beef and
butter have a low incidence of heart disease. How do you explain this?

p
Fish, seal oil and whale meat has lower cholesterol than the beef and butter. p
So, thepEskimos have low in fats especially in triglycerides and lipoproteins and in
cholesterol. The researchers concluded that the large amounts of fatty marine
food that the Eskimos were eating somehow protected them from heart disease.

The proof that I¶ve got from the internet goes as follow:

What excited the Danish researchers who visited Greenland in 1970 was the
Eskimos' low rate of heart disease in spite of their high -cholesterol, high-fat diet.
When the researchers studied health records from 1963 to 196 7, they found that
only two cases of atherosclerotic heart disease had occurred among the 1,300
inhabitants in an Eskimo settlement --one in a 78-year-old man, the other in
someone suffering from rheumatic heart disease. Death from heart disease
constituted only 3.5 percent of all deaths among Greenland Eskimos, whose
average lifespan was over 60 years. (In 1982, the American Heart Association
estimated that nearly I million Americans died of some form of cardiovascular
disease, or over half of all deaths fo r the year.) The researchers also observed
that arthritis and other chronic inflammatory diseases were almost unknown.
 They noted one other curious thing --the Eskimos tended to bruise easily and
bleed for a longer time when injured. When samples of their b lood were
analyzed, it was found to be low in fats, especially triglycerides (the main type of
fatty substance found in the fat tissue of the body and the main dietary fat) and
lipoproteins (proteins that are combined with cholesterol and triglycerides), a nd in
cholesterol. The researchers also found that the Eskimos bled more freely
because something was affecting the normal functioning of their platelets --tiny
disk-shaped cells that float in the blood and help in clotting. Their platelets
appeared to be less sticky and didn't clump together as readily as those of other
groups of people.

In May 1985, the New England Journal of Medicine created a stir in the
medical world by publishing the results of three studies which said that eating fish
was possibly beneficial to human health. Especially significant was a 20 -year
prospective Dutch study, which began in 1960 and involved 852 middle -aged
Dutch men initially free from heart disease. Their cooperative wives kept track of
what their husband ate for all those years. About 19 percent of the men didn't eat
fish at all--hard to believe of a people whose favo urite foods include raw herring
just off the fishing boat. In the others, fish intake ranged between one gram and
307 grams (about 11 ounces) a day. It was found that death from heart disease
was 50 percent lower among those who consumed as little as 30 grams (about
one ounce) a day than among those who did not eat fish. And it made no
difference whether the fish was lean or fatty. Thus, not eating fish at al l turned out
to be a risk factor for heart disease in this group.
The second New England Journal study, conducted at Harvard Medical School,
revealed that diets enriched with oils extracted commercially from the fatty
tissues of fish appeared to have anti -inflammatory effects on a group of seven
male volunteers. This is a possible explanation of the low rate of arthritis and
other inflammatory diseases among the Eskimos.



1. If the blue colour of the DCPIP disappears then vitamin C is present.

2. The water contained no vitamin C and so did not change the colour of the
DCPIP.


 !

http://www.waitrose.com/food/healthandnutrition/h ealthyeating/vegetarianandvegan i

http://vegetarian.about.com/od/healthnutrition/f/Vegnutrition.htm

http://www.addicted2coffee.org/coffee/diet.html

http://en.wikipedia.org/wiki/Grapefruit

http://www.whfoods.com/genpage.php?tname=foodspice&dbid=25

http://yourtotalhealth.ivillage.com/diet -fitness/grapefruit.html