EXPERIMENT : 1

OBJECT:
To study the effect of Temperature and PH on hydrolysis of sugar.

THEORY:
Hydrolysis is the process of cleavage of molecules with the help of water molecules. It can
be carried out in an acidic or basic medium. Hydrolysis of sugars breaks them into smaller
molecules by cleaving the glycosidic bonds that hold the monomer units together. A
glycosidic bond is formed between two monosaccharides when they are near enough to
each other with the leaving of a water molecule. It is a condensation reaction. So when
water is added to them in the reaction mixture they separate to their former states.
Hydrolysis of sugar results in forming of smaller units which may be polysaccharides,
disaccharides or the smallest of all- monosaccharides.

EFFECT OF TEMPERATURE:
The Brix of sugar solution is hardly depended upon temperature. Brix measurement of a
liquid fluctuates according to temperature. The Brix of a cold sample will measure higher
than the same sample at room temperature. When measured hot, the Brix reading will be
lower.

PROCEDURE:
 Made 25% sugar solution in 100 ml water. ( Ambient temperature & neutral PH )
 Add 1 drop conc. HCl in 10 ml sugar solution ( Acidic PH)
 Add 1 palette NaOH in another 10 ml sugar solution ( basic PH)
 Then Place 10 ml sugar solution on hot plate at 80C.
 Note the Brix value and PH of the solution.

OBSERVATION:

ph

NEUTRAL ACIDIC BASIC

22.5 23.6 24.3
Temperature

AMBIENT TEMPERATURE HIGH TEMPERATURE

22.5 28.6

1
RESULT:
Brix is a measuring scale which determines the pure sucrose content in aqueous solution
with the help of Refractometer. In this experiment, observed that the value of Brix is high
on basic side of Ph and higher temperature. While, Brix value is lower on neutral side and
ambient temperature.

DICUSSION:
Degree Brix is a measure to determine the total soluble solids in a sample using
refractometer. At ambient temperature we found 22.5 ̊B of 25% sugar solution. 1% of pure
sugar solution gives 1̊B at room temperature but not for all sugars because it depends upon
the purity level of sugar used in solution, some sugar may carry non sugar particles thus
not give exactly level of brix.

REFERENCE:
Adams, J. P., Simunovic, J., & Smith, K. L. (1984). Temperature histories in a UHT indirect heat
exchanger. Journal of Food Science, 49, 273.

2
 EXPERIMENT : 02
OBJECT:
To study the effect of degree of saturation and seeding on the recrystallization of sucrose.

THEORY:

CRYSTALLIZATION:
Crystallization is a Physical separation technique in which mass transfer of a solute from
the liquid solution converted in to a solid crystalline state.

SEEDING:
Crystal seeding is the process of adding homogenous or heterogeneous crystal to a
crystallizing solution to nucleate or grow more crystals. Seeding has emerged as one of the
most critical steps in optimizing the crystallization process. Seeds could be added into the
super saturated solution or saturated solution. Seed crystal used to promote the
crystallization process, avoids the otherwise slow randomness of natural crystal growth
and decrease the necessary amount of time needed for nucleation to occur.

PROCEDURE:
 Prepare different types of sugar solution of different concentration that are
saturated, unsaturated and supersaturated.
 First of all prepare 50% of unsaturated solution by dissolving 15ml of sugar in 30ml
of H20.
 Then prepare the 200% of saturated sugar solution in two batches by dissolving
120gm of sugar in 60ml of H20.
 Now prepare the 233% of supersaturated solution in two batches by dissolving the
140gm of sugar in 60ml of H2O with heating
 Again prepare the 266% of supersaturated solution in two batches by dissolving
160gm of sugar in 60ml of H20 with heating.
 200%, 233% and 266% samples of one batch are required for seeding and other
used for non-seeding.
 Seeding can be carried out by adding 1 pinch of sugar crystals in seeding samples
when the solution is hot and dispersed them with the help of glass rod.
 Formation of crystals is observed in 3-4 days or upto 1 week.

OBSERVATION:
3
READING OF SEEDING CRYSTAL:

SAMPLE SEEDING CRYSTAL

200% Least crystal formation
233% more crystal occur
266% Uniformity or more crystal formation

READING WITHOUT SEEDING:

SAMPLE WITHOUT SEEDING

50% Irregular pattern of crystal
200% Less crystal formation occur
233% Random or large crystal formation
266% non uniform or more crystal formation

RESULT:
From the experiment it is observed that the maximum no of sugar crystals are formed in
266% and 233% super saturated solution.

DISCUSSION:
FOR SUPER SATURATED SOLUTION:

From experiment it is observed that the highest no of crystals were formed in

supersaturated solution. A supersaturated solution is a solution that contains more than
the maximum amount of solute that is capable of being dissolved at a given temperature.
The recrystallization of the excess dissolved solute in a super saturated solution can be
initiated by the addition of tiny crystal of solute, called seed crystals..

FOR SATURATED SOLUTION:

In contrast to super saturated solutions, saturated solution is slightly or little tendency to

form or grow crystal because saturated solution is a solution that contain the maximum
amount of solute that is capable of being dissolved.

FOR UNSATURATED SOLUTION:

Unsaturated solution is a solution in which the amount of dissolved solute is less than the
saturation point at fixed temperature. When you add a crystal of a solute to an unsaturated
solution, the crystals dissolves, becoming part of the solution. An unsaturated has the
capacity to dissolve more solute, so any solute added below the saturation point, dissolves.

4
 EXPERIMENT: 03

OBJECT:
Determination of invert % in refined sugar.

THEORY:

INVERT SUGAR:
Invert sugar is a mixture of glucose and fructose (reducing sugar) and it is obtained by
splitting sucrose (non-reducing sugar) or by hydrolyzing sucrose with high temperature or
acid. It is thought to be sweeter than table sugar and foods that contain it retain moisture
and crystallize less easily. Sucrose is a non-reducing sugar and dextrorotatory. Mixture of
glucose and fructose as it rotates the plane of polarized light and becomes levorotatory
because fructose has a greater molar rotation than dextrorotatory glucose.

PROCEDURE:
● Prepare sugar solution by dissolving 5g sugar in 5ml water
● Add 2ml of Alkaline copper reagent
● After thoroughly mixing, Place the solution in water bath at 100C for 5 min
● Then cold test tube in running water
● Transfer the test tube in conical flask with water
● Add 0.1 gm of indicator Titrate the solution with EDTA solution
● The color change is form green through grey to purple, the grey stage appearing just
before the end point which should be approached fairly rapidly since the purple
color tends to be fading due to oxidation.
● Calculate the % of inversion by formula.

OBSERVATION:
Observe colour from green to grey during titration via addition of EDTA solution along with
Munecsite Indicator.
Samples Burette reading (ml)

Murexide indicator 3.4

EDTA solution 2.5

5
 Alkaline copper reagent 1.9

CALCULATIONS:
Mean = (3.4 + 2.5 + 19)/3 = 2.6ml
RESULT/ DISCUSSION:
As per the value of titration we found 0.016% of invert sugar present in our sucrose
samples. This method of knight and Allergen method is suitable for the determination of
low invert sugar (0.01-0.02%). If the percentage of reducing sugar/ invert become equal or
more than 0.02% than it would reflect inversion.
REFERENCES:
1. Ellborg, A., & Tragardh, C. (1990). An experimental method for evaluation of the
thermal time distribution in continuous ¯ow sterilization. In W. E. L. Spiess, & H.
Schubert, Engineering and food, vol. 2 (pp. 136±144). London: Elsevier. Ellborg, A.,
& Tragardh, C. (1994).
2. A method for measuring thermal time distributions in continuous ¯ow heat
exchanger. Journal of Food Process Engineering, 17, 279±298.

6
 EXPERIMENT : 04
OBJECT:
To determine the concentration of an unknown sugar solution through spectrophotometer.

THEORY:

SPECTOPHOTOMETER:

Spectrophotometry is a method to measure how much a chemical substance

absorbs light by measuring the intensity of light as a beam of light passes
through sample solution. The basic principle is that each compound absorbs
or transmits light over a certain range of wavelength.

ABSORBANCE: Absorbance is a measure of the quantity of light absorbed by a sample. It

is also known as optical density, extinction, or decadic absorbance.

PROCEDURE:
● Prepare six standard glucose solutions of 0.5%, 1%, 1.5%, 2%, 2.5%, 3 % and one is
unknown solution.
● Observe the absorbance of the standard solution and unknown solution at 420nm
● Plot the standard curve between concentration and absorbance of the solutions

OBSERVATION:
Plot a graph in between concentration of sugar solution and their absorbance and also find
the concentration of unknown sugar solutions by using absorbance.

CONCENTRATION(g/ml) ABSORBANCE(A)

0.5% 0.053

1% 0.021

1.5% 0.006

2% 0.094

2.5% 0.094

7
 3% 0.098

Unknown sample 0.068

GRAPH:

RESULT:
The sugar concentration of an unknown solution has appeared to be 5.80% .

DISCUSSION:
From the experiment it is observed that the lighter the concentration of the solution, the
higher will be the absorbance value which means absorbance is directly proportional to
concentration of the substance. This is because the proportion of light that get absorbed is
affected by no of molecule that it interact with. Solution that are more concentrated has
large no of molecule that interact with the light that enters, thus increasing its absorbance.
That is why concentration and absorbance are directly proportional to each other.
REFERENCES:
1. Gore, Michael. Spectrophotometry & Spectrofluorimetry. New York: Oxford
University Press, 2000.
2. Price, Nicholas and Dwek, Raymond and Wormald, Mark. Principles and Problems in
Physical Chemistry for Biochemists. R. G. Ratcliffe. New York: Oxford University
Press, 1997.

8
 EXPERIMENT: 05
OBJECT:
Decolorization of cane juice by using active carbon column of various lengths and compare
the brix, density, and color intensity.

THEORY:

Activate carbon
Activated carbon, also called activated charcoal, is a form of carbon processed to have
small, low-volume pores that increase the surface area available for adsorption or chemical
reactions.

Activated carbon filters

Activated carbon filters are generally employed in the process of removing organic
compounds and/or extracting free chlorine from water, thereby making the water suitable
for discharge or use in manufacturing processes. Eliminating organics in potable water,
such as humic and fulvic acid, prevents chlorine in the water from chemically reacting with
the acids and forming trihalomethanes, a class of known carcinogens.

PROCEDURE:
1. Take three burette stands in which three different columns are filled with different
quantities of activated carbons.
2. Fill first one up to 10ml, the second one up to 20ml, and the third one up to 30ml.
3. Now take sugar cane juice and filter them to remove fiber components.
4. On the other side take three similar sized conical flasks and take their pre weight.
5. Then fill each one with 60ml sugar cane juice and weigh them again.

9
 6. Also check pre brix of sugar cane juice before experiment to be performed and
calculate density.
7. Now label the conical flasks as 10ml, 20ml, and 30ml.
8. Now transfer the juice to another empty beaker and place the labeled and pre weigh
conical flasks to collecting place of each respective burette and pass the sugar cane
juice from each one.
9. The collected juice will be free from colored compounds, now take after weight and
check brix of each one.
10. Compare the results before and after taken results of experiment

OBSERVATION:

Activated Initial and Final Observations Color

Carbons in ml reduction
Initial Final Density Initial Brix Final Brix
Density
D=m/v
D=m/v
10ml 1.06 0.996 100 3.60 +

20ml 0.97 0.989 100 4.60 ++

30ml 0.97 0.916 100 40 +++

RESULT:
Higher color reduction in cane juice found in 30ml active carbonated column.

DISCUSSION:
Activated carbon is used to filter liquids and gases in a range of applications, such as
municipal drinking water, food and beverage processing, odour elimination, and industrial
pollution management, as we all know. We employed activated carbon to decolorize and
purify the cane juice in this experiment. The density of the cane juice is reduced when it is
treated with activated carbon, and when the amount of activated carbon is raised, more
mass is lost in the form of impurities, resulting in a further reduction in density. Although
there is a significant drop in brix, the trend with the amount of activated carbon is not
consistent. This brix reading must contain a mistake. Because all three columns were not
given adequate time, the brix values were not uniform.

10
 EXPERIMENT : 06
OBJECT:
To determine the purity of honey.

THEORY:
Honey is a sweet, viscous food substance made by bees and some related insects. Honey
gets its sweetness from the monosaccharide fructose and glucose. Due to its attractive
chemical properties it is used in baking and as a natural sweetener. Honey has a relatively
low water activity and thus can be used even after thousands of years as most of
microorganisms can’t grow in it.

Honey provides 46 calories in a serving of one tablespoon (15ml).viscosity of honey is

greatly affected by temperature and water content. At 25C, it usually possess 14% water
content and 400 poise viscosity. Natural honey has Brix of about 83 degree B and PH about
3.9. Honey has high acidicity and higher sugar content about 82%. Normally natural
moisture content poses by honey is about 17.8% but the moisture content should be below
20% for pure honey.

PROCEDURE:
METHOD 1 (Absorption on Paper):

● To check whether honey is diluted with water, drop honey on a paper towel
● Observe its absorbance whether it leaves mark or absorbs on the surface of paper.

METHOD 2 (Paper Burning):

● To check honey is pure or artificial, wick of paper dip in honey then flame it
● Observe the speed of burning

METHOD 3 (Cotton Burning):

11
 ● Take small piece of cotton and soak in honey then flame it with candle
● Observe how much ash is remaining after burning

METHOD 4 (Dispersion Test):

● Take half beaker of water then add ½ tp honey in it

● Observe honey is settle down in the bottom or disperse in water

METHOD 5 (Thumbnail Test):

● One drop of honey place on thumbnail

● Observe the honey dripping

METHOD 6 (Foaming Test):

● Take 1 ml honey add 2 ml acetic acid solution

● Observe the foam layer on the surface of sample

OBSERVATION:
Sample Method 1 Method Method 3 Method 4 Method 5 Method 6
2
(Absorptio ( Cotton ( Dispersio ( thumbnai ( Foamin
n on paper) ( Paper Burning) n test) l Test ) g test)
burning)

Marhaba Positive Negative Negative Positive Negative Positive

Honey result result result result result result

RESULT/DISCUSSION:
Honey is a sweet, viscous food substance made by bees and some related insects. Honey
gets its sweetness from the monosaccharide fructose and glucose. Due to its attractive
chemical properties it is used in baking and as a natural sweetener. Honey has a relatively
low water activity and thus can be used even after thousands of years as most of
microorganisms can’t grow in it. As per performed experimental methods to check the
purity of honey. We found that the positive result came in only three methods such as
absorption on paper, dispersion test and foaming test and in other methods found to be
negative result. After observing the all results we conclude that the Marhaba honey is not
pure as some adulteration or impurities present in it that’s why it give the negative result
in several methods.

12
REFERENCES:
1. Irwin H. Segel, Biochemical Calculations (How to Solve Mathematical Problems in
General Biochemistry), 2nd edition, John Wiley & Sons, 1975
2. http://www.nist.gov/pml/div685/grp03/spectrophotometry.cfm

EXPERIMENT : 07
OBJECT:
To study the effect of different sweeteners on dough rising.
THEORY:
The most commonly used yeast such as baker yeast or brewer’s yeast utilizes hexose sugar,
mostly glucose and gives out by product such as CO2 and ethanol. Sugar effect the rate of
fermentation reaction. Over 3 % sugar, however fermentation rate no longer increases.
Above 6%, sugar actually decreases the rate this is because sugar begins to dehydrate the
yeast cells. Jiggery is a nutrient rich sweetener, but this react alternate also contains about
60 to 85% of sucrose. Honey is made up of around 75% sugar of which roughly half is
glucose and half is fructose. Brown sugar contains at least 88% of sucrose plus invert sugar.
Commercial brown sugar contain from 3.5% molasses( light brown sugar) to 6.5%
molasses ( dark brown sugar) based on total volume. White sugar is made from raw sugar
that has undergone a refining process to remove the molasses. Raw sugar is sucrose which
is extracted from sugar cane to sugar beet.

PROCEDURE:
1. Take different types of sweeteners (brown sugar, glucose, white sugar and jaggery).
2. Add each sweetener in separate bowl containing flour and knead to make dough by
using the recipe as under.
3. Flour = 50g, yeast = 2g, salt = 1g, water = 12ml and sugar = 6g.
4. Place the dough in cylinder and mark them A, B, C, and D.
5. Observe the effect of each type of sweetener in dough rising after each 10 min at
least for one hour and compare their results.

13
OBSERVATION:
Table: 01: Change in volume observed by the effect of type of sugar after every 10 minutes:

Volume Change (ml)

Time
No Brown
(min) Jiggery Sucrose Glucose
Sugar Sugar

0 65 75 78 83 80

10 67 77 82 86 83

20 70 78 87 88 87

30 73 79 90 93 92

40 76 79 98 105 98

50 80 80 104 110 108

60 85 80 109 122 112

Table: 02: Difference in volume by the effect of sugar:

Type of Sugar VF-VI

No sugar 20

Brown sugar 5

Jiggery 31

Sucrose 39

Glucose 32

14
RESULT / DISCUSSION:
In this experiment, take different sweeteners to check the effect of volume on dough. After
see all the observation we conclude that, sucrose gave the highest volume on dough as
compare to others like white sugar, brown sugar and glucose, because sucrose is a
complete sugar when it is break down it convert into two monosaccharide one is glucose
and other is fructose. When it is added in flour, yeast readily ferment the sugar and release
CO2 and alcohol. When CO2 release it rise the volume in the dough.

EXPERIMENT: 8
OBJECT: DNS colorimetric method of available carbohydrates in food.

THEORY:
A sound, reliable, time- and cost-efficient method for detection of reducing sugars, based on
the well-known dinitrosalicylic acid (DNS) colorimetric method, adapted for micro titer
plates, in a modified water bath with microwave treatment is proposed.

The Beer-Lambert Law

The Beer-Lambert Law (also called Beer’s Law) is a relationship between the attenuation of
light through a substance and the properties of that substance. Beer-Lambert Law
which states that the amount of light absorbed is directly proportional to the concentration
of the solute in the solution and thickness of the solution under analysis.

PROCEDURE:
DNS reagent preparation:

Dissolve 10g DNS in 200ml of 2M NaOH with warming and vigorous stirring. Dissolve 300g
sodium potassium tartrate tetra hydrate in 500ml distilled water (color stabilizer). Mix
these two solutions and make up to 1L with distilled water.
Food preparation:
Hydrolysis of available carbohydrate (solid food, breakfast, and cereals):
Grind the food to a fine powder. Weight above 0.1-0.2-point 2 grams of the powder food
into a boiling tube add 10:00 AM alarm 1.5 M sulfuric acid and heat in a boiling water bath
for about 20 minutes stirring occasionally to hydrolyze polysaccharide and other non-

15
 reducing sugar cool and carefully add 12 ml of 10% NaOH mix and filter into our 100ml
volumetric flask washing the two into the flask with distilled water make up the volume
with distilled water and mix well by inversion.
OBSERVATIONS:
Standard Solution
Tube No Glucose Absorbance Colour Intensity
1 0 0.025  

2 0.25 0.05 *

3 0.5 0.15 **

4 0.75 0.2 ***

5 1 0.25 ***

6 1.25 0.3 ****

7 1.5 0.35 *****

Unknown Solution 2.507 0.59 *******

DISCUSSION:
In this experiment we used absorbance to find out the sugar concentration of our unknown
samples. We first made a standard glucose solution whose reducing sugar concentration
was known to us and by using a spectrophotometer we found out its absorbance. Now
using the absorbance and reducing sugar concentration of this standard solution we
plotted a graph on excel. On the contrary, we used spectrophotometer to find out the
absorbance of our sample. Now using this absorbance and the graph we plotted, we can
find the sugar content of our unknown sample. From the absorbance of our unknown
sample of the glucose concentration and from the graph using excel sheet, the glucose
concentration is 2.507.
REFERENCES:

  Melander, C.; Andersson, E.; Axelsson, S.; Gorton, L.; Anal. Bioanal.

Chem. 2007, 387, 2585.

16
 2. European Community; Official J. Europ. Commun., Directive 2001/110/EC,
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?
uri=OJ:L:2002:010:0047:0052:EN:PDF accessed on July 2, 2012.

17