Ana Quimpo

Biology 61
Dr. Seubert
2/2/17
Lab Report 1: Qualitative Analysis of Carbohydrates
INTRODUCTION
Cells, replication, evolution, information, and energy, these are the five
characteristics shared by all organisms. In order to sustain life, organisms must
continuously obtain and use energy. Plants have the ability to obtain energy
through photosynthesis. Animals, however, must obtain energy by ingesting food.
Carbohydrates, a food source to animals, and the fundamental component in this
experiment, are abundant in energy. Carbohydrates are the principal energy source
in the diets of most people and have a special role to play in energy metabolism
and homoeostasis (Mann, et. Al, 2007). People convert carbohydrates into sugars,
which provide chemical energy in cells and furnish some of the molecular building
blocks required for synthesis of larger, more complex compounds (Freeman, et. al,
108). Carbohydrates can be characterized as either monosaccharides, meaning one
sugar, or polysaccharides, meaning many sugars.
Monosaccharides, or simple sugars, are the monomers of carbohydrates (Freeman,
et. al, 108). The carbonyl groups in monosaccharides are trioses, ribose, and
glucose. Triose has a three-carbon structure, ribose has a five carbon structure
called pentose, and glucose has a six carbon structure called hexose. This structure
plays a key role in the functionality of cells, and its ability to convert carbohydrates
into energy.
When the structure allows these simple sugars to form long chains, a complex
carbohydrate is formed. Very long chains of complex carbohydrates are called
polysaccharides. Starch, glycogen, cellulose, and chitin are all examples of
polysaccharides.
Many studies indicate the difficulties in measuring carbohydrates. This is because
of the use of different energy systems such as combustible, digestible and
metabolizable. Furthermore, ingested macronutrients may not be fully available to
tissues and the tissues themselves may not be fully able to oxidize substrates made
available to them. Therefore, for certain carbohydrates the discrepancies between
combustible energy, digestible energy, metabolizable energy (ME) and net
metabolizable energy (NME) may be considerable (Mann, et. Al, 2007).
Due to the difficulties in measuring, this experiment called for a qualitative analysis
of carbohydrates. This experiment was designed to identify the unknown
carbohydrates in four different food items, and classify them as monosaccharides or
polysaccharides. The selected food items included a tomato, an apple, blueberries,
and piece of bread. Two tests were conducted on the food items. The first test used
the Benedicts Solution to test for monosaccharides, while the second test used
Luogols Iodine to test for the presence of polysaccharides.
Because glucose is present in monosaccharides, it was hypothesized that all four
carbohydrates: tomato, apple, blueberry, and bread, would test positive for small
amounts of monosaccharides. Additionally, because bread is made up of complex
carbohydrates, it was hypothesized that only this sample would yield a small
amount of polysaccharides (Table 1).

MATERIALS AND METHODS

The materials required to successfully conduct the experiment included test tubes,
a test tube holder, a 600 mL beaker, 24 mL of Benedicts Solution, 30 drops of
Lugols Iodine, water, a hot plate, two mortars and pestles, pipettes, 20% glucose,
and 2.5% starch.
500 mL of water was added to the 600 mL beaker, put on the hot plate, and brought
to a boil. The mortars and pestles were utilized to grind the food items to a
liquefied state. At this consistency, the food items were transferred, and used to fill
four individual test tubes. The filled test tubes were placed on the test tube holder,
and later used to pipette smaller samples into new, individual test tubes.
Test 1
Six additional test tubes were obtained and labeled as + for the positive control,
- for the negative control, 1 for the bread, 2 for the tomato, 3 for the
blueberries, and 4 for the apple. It was decided that the positive control for this
test would be the 20% glucose, while the negative control would be the water. The
colors for each test tube were recorded.
Four mL of Benedicts Solution was pipetted into the six labeled test tubes. An
additional four mL of the control groups was pipetted into their appropriately
labeled test tubes. To avoid contamination, separate pipettes were used for each of
the individual food samples to transfer 0.5 g into their respective tubes. Water was
added to test tubes containing the food samples, until the final volume reached
eight mL, the identical amounts for the control groups. The colors were again
recorded for each of the tubes after the Benedicts Solution was added.
Once the test tubes were identical in volume, each tube was placed in the boiling
water for two minutes. The colors for the individual test tubes were recorded after
the boiling process.
Test 2
Six additional test tubes were obtained and labeled as + for the positive control,
- for the negative control, 1 for the bread, 2 for the tomato, 3 for the
blueberries, and 4 for the apple. It was decided that the positive control for this
test would be the 2.5% starch, while the negative control would be the 20%
glucose. The colors for each test tube were recorded.
Four mL of the control groups was pipetted into their appropriately labeled test
tubes. To avoid contamination, separate pipettes were used for each of the
individual food samples to transfer 0.5 g into their respective tubes. Water was
added to test tubes containing the food samples, until the final volume reached four
mL. Five drops of Lugols Iodine was added to each individual test tube, then gently
swirled. After the solution in the tubes were swirled, the color was, again, recorded.

RESULTS
Test 1
In Test 1, Benedicts Solution was used to test for the presence of monosaccharides
in the food samples. If the sample turned orange in color after boiling, it was
positive for monosaccharides. If the sample turned blue after boiling, it was
negative for monosaccharides.
The water was used as the negative control group, while the glucose was used as
the positive control group. Unknown 1, 2, 3, and 4 were the bread, tomato,
blueberry, and apple, respectively.
Prior to adding the Benedicts Solution, the colors of the liquefied foods, and
controls were recorded. Both the water and the glucose had a starting color of
clear. The bread was a milky color, the tomato was pink, the blueberry peach, while
the apple yielded a yellow color (Table 2).
After adding the Benedicts Solution to the individual test tubes, the color was again
recorded. The color for the water, glucose, bread, and blueberry turned blue, while
the tomato and apple turned green (Table 1).
The test tubes were then placed in the boiling water for two minutes. After boiling,
the water turned blue. The glucose, bread, and blueberry, however, yielded a deep
orange color, while the tomato and apple turned a yellow/orange color (Table 2).
Based off of the richness of the color, it was found that the bread and the
blueberries had the highest concentration of monosaccharides. Because the tomato
and apple yielded a slightly yellow/orange color, it was determined that they have a
lower concentration of monosaccharides.
Test 2
In Test 2, five drops of Lugols Iodine was added to the food samples, and used to
test for the presence of polysaccharides. If the sample turned black after adding
the Lugols Iodine, the sample was positive for polysaccharides. If the sample
turned orange after adding the Lugols Iodine, the sample was negative for
polysaccharides.
In this test, glucose was used as the negative control group, while the starch was
used as the positive control group. Unknown 1, 2, 3, and 4 were the bread, tomato,
blueberry, and apple, respectively.
Prior to adding the Lugols Iodine, the colors of the liquefied foods, and the controls,
were recorded. The starting color for the glucose was clear. The starting color for
the starch was a cloudy/white. The bread, tomato, blueberry, and apple were milky,
pink, peach, and yellow, respectively, in color (Table 3).
Once the Lugols Iodine was added to the individual test tubes, they were gently
swirled, until a change in color occurred. The glucose, tomato, blueberry, and apple
turned orange. The starch and the bread, however, turned black (Table 3).
Because the change in color was used to indicate a high concentration of
polysaccharides, it was found that only bread and starch have a positive sample of
polysaccharides.

DISCUSSION
It was hypothesized that the tomato, apple, blueberry, and bread would all test
positive for concentrations of monosaccharides, while only the bread would test
positive for polysaccharides. After conducting the experiment, the results agreed
with the hypothesis.
The Benedicts Solution, which was blue in color was used to test for the presence of
monosaccharides in the four samples of carbohydrates. If the solution would turn
orange after boiling, monosaccharides are present. However, if the solution
remained blue after boiling, the sample is negative for monosaccharides. In the first
test, water was chosen as the negative control, because water does not have any
sugars to test positive for. Glucose, which is a monosaccharide, would test positive
for this, making it the best choice for a positive control.
As past studies indicated, measuring the exact amount of carbohydrates proved to
be difficult. This was evident in the color changes recorded. The original colors for
the water, glucose, bread, tomato, blueberries, and apple were clear, clear, milky,
pink, peach, and yellow, respectively. After adding the Benedicts solution, the
colors turned blue, blue, blue, green, blue, and green, respectively. The two
samples that turned green were the tomato and the apple, which also yielded a
yellow/orange color at the end of boiling. The bread and the blueberries, however,
yielded a deep orange color. This was interpreted as having a higher concentration
of monosaccharides.
The second test utilized Lugols Iodine to test for polysaccharides. If the solution
turned black after swirling, the sample was positive for polysaccharides. However,
if the solution turned orange after swirling, the sample was negative for
polysaccharides.
In the second test, starch was chosen as the ideal positive control because it is a
polysaccharide, and would test positive, as such. Glucose, however, was the
negative control because it also contains monosaccharides.
The starting color of the glucose, starch, bread, tomato, blueberry, and apple were
clear, cloudy/white, milky, pink, peach, and yellow, respectively. After adding the
five drops of Lugols Iodine, and swirling the solution together, the samples turned
orange, black, black, orange, orange, orange, respectively. Again, the results
agreed with the hypothesis, as bread was the only sample to test positive for
polysaccharides.
The classification of carbohydrates as monosaccharides and polysaccharides is
crucial in understanding how to obtain energy for cells. Although difficult to
determine, foods rich in polysaccharides yield more energy for the cells to store, or
convert. Energy is vital to sustaining life, as it allows organisms to transmit their
genetic information and do other cellular work.

Table 1. Qualitative analysis of concentrations of monosaccharides and

polysaccharides in carbohydrates.

Monosacchar Estimate Polysaccharid Estimate

Food
ides Present? Monosaccharid es Present Polysaccharid
Item
(Yes or No) e Amount (Yes or No) e Amount
Tomato Yes A little No None
Apple Yes A bit No None
Blueberr
y Yes A bit No None
Bread Yes A little Yes A little

Table 2. Comparison of controls groups, unknown carbohydrates, and yields of

monosaccharide concentration. An orange color after boiling, indicates a positive
sample for monosaccharides. A blue color after boiling, indicates the sample is
negative for monosaccharides.

Starting Color After Color After

Group Contents Color Benedict's Boiling
Negative Control
Group Water Clear Blue Blue
Positive Control
Group Glucose Clear Blue Orange
Unknown 1 Bread Milky Blue Orange
Unknown 2 Tomato Pink Green Yellow/Orange
Unknown 3 Blueberry Peach Blue Orange
Unknown 4 Apple Yellow Green Yellow/Orange

Table 3. Comparison of control groups, unknown carbohydrates, and yields of

polysaccharide concentration. A black color after adding the Iodine, indicates a
positive sample for polysaccharides. An orange color after adding the Iodine,
indicates the sample is negative for polysaccharides.

Starting Color After

Group Contents Color Iodine
Negative Control
Group Glucose Clear Orange
Positive Control Cloudy/Whi
Group Starch te Black
Unknown 1 Bread Milky Black
Unknown 2 Tomato Pink Orange
Unknown 3 Blueberry Peach Orange
Unknown 4 Apple Yellow Orange

Works Cited
Freeman, S. 2017. Biological Science, 6th Edition. 108.

Mann, J, Cummings, H, Englyst, N, Key, T, Liu, S, Riccardi, G, Summerbell, C, Uauy,

R , Van Dam, M, Venn, B, Vorster, Wiseman, M. 2007. FAO/WHO Scientific
Update on carbohydrates in human nutrition: conclusions. European Journal
of Clinical Nutrition. 61. 132137.