MAICA KRISTINE M.

DICION CHEMBIO3
BSBIO-2B

PRE-LABORATORY NO.4
CARBOHYDRATES

INTRODUCTION
Carbohydrates is a biomolecule composed of carbon, hydrogen, and oxygen atoms. Has the empirical
formula C6H12O6. Technically, carbohydrates are carbon hydrates; structurally, more accurate to see
them as aldoses and ketones. Carbohydrates have a significant crucial role in the energy metabolism of
living organisms. They serve as an energy source, help control blood sugar and insulin metabolism, and
fermentation. One of the three macronutrients in human diet, along with protein and fat.
The term is most frequent in biochemistry, where it is a synonym of saccharide, a group that consists of
sugar and starch and cellulose. The word saccharide comes from the Greek word (sákkharon), meaning
"sugar” (Avenas, 2012). The saccharides are divided into four chemical groups: monosaccharides,
disaccharides, and polysaccharides. As explained monosaccharides and disaccharides are the smallest
carbohydrates usually known as sugar (Flitsch and Ulijin, 2003). While the scientific nomenclature of
carbohydrates is complex, the names of monosaccharides and disaccharides very often end in the suffix -
ose, as in the monosaccharide’s fructose (fruit sugar) and glucose (starch sugar) and the disaccharides
sucrose (cane or beet sugar) lactose (milk sugar).
One of the primary components of insoluble dietary fiber is cellulose, a polymer found in all plant cell
walls. Insoluble dietary fiber, which is not digested, helps to a healthy digestive tract by
reducing defecation. Other polysaccharides found in dietary fiber include resistant starch and insulin,
which feed some microorganisms in the microbiota of the large intestine, and these bacteria metabolize
and generate short-chain fatty acids (Bymes, Chambers, Morrison, 2015).

OBJECTIVES
1. The objective of this laboratory experiment was to identify and characterized carbohydrates in various
biochemical test and observe and understand the process of fermentation, specifically alcoholic
fermentation and its underlying metabolic reactions.
MATERIALS/REAGENTS

 Test tube brush  Glucose

 Beaker  Maltose
 Test tube holder  Lactose
 Pipette  Fructose
 Aspirator  Galactose
 Fehlings A&B  Sucrose
 Seliwanoff’s Reagent  Starch
 I2KI solution  Distilled
 Arabinose water

MATERIALS FOR FERMENTATION

 1 kg of turnip  Knife  Cling wrap

 1 kg refined sugar  Measuring cup  1 egg white
 1 packs of yeast  1 plastic bottle
(saccharomyces
cerevisiae)
 1 foot plastic tubing  Casserole
 Blender  cotton

METHODS
The student initiated the solution preparation of different biochemical test, the experimental methods
employed include test for monosaccharides, disaccharides, and polysaccharides. Along with necessary;
laboratory materials and reagents for test preparation. In this experiment the student used different test to
test the reducing sugar of carbohydrates. Fehling’s solution consists of two separate solutions known as
Fehling’s A and B are both blue aqueous solution of copper sulfate pentahydrate crystals, while Fehling’s
B is a transparent solution of aqueous potassium sodium tartrate. In each test tubes, the student put 5% of
different sugar solution then added 1 mL of Fehling’s reagent in A and B tests, then the student heated
continuously in water until the solution changed in color to observe and identify the reducing groups like
aldehyde function. The formation of a colored precipitate indicates the presence of reducing sugar. To test
the specific test for some carbohydrates, the student prepares test tubes and different sugar solution to test
the sugar solution contain ketoses particularly fructose, from hydroxymethyl by the action of HCL test
and condenses with resorcinol to form a red color. Using with Seliwanoff’s test the student will identify
the appearance of a cherry- red color indicates the presence of ketoses. The student prepared test tube
with 5% of different sugar solution then added 3 mL of Seliwanoff’s reagent. In iodine test, the student
prepares test tube with different 5% sugar solution then using pipette, add 1 mL of 0.1 M I2KI solution.
The student used this test to indicates and identify that Amylose interacts with starch to form a blue-black
colored complex with the iodine.
In fermentation, the student uses Saccharomyces cerevisiae as a catalyst in the alcoholic fermentation
process. These microorganisms are often used in the production of wines, beer, and cider. A process that
releases energy from a sugar or another chemical molecule, which does not required oxygen or an
electron transport mechanism and uses an organic molecule as the final electron acceptor occurs only in
anaerobic condition.

PROCEDURE
A. General Tests
Fehling’s Test
Individually, 2mL of each sample solutions were added to a clean test tube. Then add 2mL of Fehling’s
solution A and B were added. And placed the solution into a beaker with boiling water and boiled for 10
minutes to observe the solution change in red color precipitate that indicates the presence of
carbohydrates.
Seliwanoff’s Test
In a clean test tube, placed 1 mL of sample solution were added 3 mL of Seliwanoff’s reagent then mix
and heat the mixture into a beaker with boiling water for 10 minutes to observe the appearance of cherry
red precipitates without separation of a brown to red precipitates that indicates the presence of a ketoses.
Iodine Test
In a nine clean test tube, put 1 mL of sample solution. Using a pipette add 1 mL of 0.1 M l2Kl solution
add to sample solution. Then observe the changes in color from red to blue.

FERMENTATION
1. Wash the fruits, then peel and remove any seeds. Dice the flesh and then purée it with food.
processor.
2. Boil the water to blanch the huge fermentation bottle. Dispense within the fruit purée. For each 2 cups
of fruit puree, add 1 cup of distilled water and 1 cup of sugar.
3. Pasteurize the solution for 30 minutes in a water bath. Allow the solution to cool before adding.
the yeast. Shake the solution thoroughly.
4. To allow saccharification, place a cotton stopper on the bottle.
5. After two days, remove the cotton plug and replace it with a cork. Put the plastic tube in the middle.
of the cork; one end should be within the bottle without contacting the solution, and the other. Then end
should be soaked in water using a tiny plastic container.
6. Allow the solution to ferment undisturbed for 14 days. Check for bubbles to confirm that
Fermentation occurs.
7. After 14 days, pour the produces clear liquid into a clean, blanched bottle. To removed contaminants,
add 1 egg white to the wine and let it sit for 24 hours.
8. Pasteurize to destroy any leftover yeast and harden the egg white. Separate the clear wine.
Label the bottle, seal it, and store it.

RESULT
A. General Tests
Fehling’s Test

Test tube No. Sugar Solution Solution A Solution B

1 5% fructose Green Orange
2 5% galactose Blue Violet
3 5% glucose Blue Orange
4 5% maltose Blue Green and in time
turns to orange
5 5% sucrose Blue Orange
6 5% lactose Blue Orange
7 5% starch Blue Blue
8 water Blue Blue
B. Specific Test for some Carbohydrates
Seliwanoff’s Test

Test tube No. Sugar Solution Color

1. 5% fructose Cherry red
2. 5% galactose Orange (rusty)
3. 5% glucose Orange (rustier)
4. 5% maltose Red
5. 5% sucrose Cheery red
6. 5% lactose Orange (rusty)
7. 5% starch Red
8. water Yellowish

C. Iodine Test

Test tube No. Sugar Solution Color

1. 5% fructose Red
2. 5% galactose Red
3. 5% glucose Red
4. 5% maltose Red
5. 5% sucrose Red
6. 5% lactose Red
7. 5% starch Black
8. water Red
D. Fermentation

DISCUSSION
The chemical properties of the saccharides vary depend on the quantity if OH groups and the presence or
absence of CHO/=CO groups. These variances serve as the foundation for the creation of the color
reaction used to identify saccharides. The general carbohydrate test aid in identifying the properties of the
known samples in the experiment.
In Fehling’s test, Fehling’s solution A and B were added to the carbohydrates. After the solution added in
the solution, the carbohydrates turned blue except test tube no. 1 contains 5% of fructose turned green.
The test tubes were then heated for 10 minutes. After heating, test tube A and B developed an orange to
reddish precipitate, but test tube no. 7&8 that contains 5% of starch and water remained blue.
Carbohydrates can be classified based on the reactivity as reducing sugar and non-reducing sugars. All
monosaccharides and the most of monosaccharides are reducing sugars. Fehling’s test determines the
presence of reducing sugar. Recuing sugar is capable of acting as a reducing agent (Pratt&Cornley, 2012).
Reducing sugars turns the deep blue solution of copper ll in the test reagent into a red precipitate of
insoluble cuprous oxide. Thus, the orange to reddish precipitate in test tubes contains 5% of fructose,
glucose, maltose, and lactose reveals the presence of reducing sugar. Upon the addition of reagent, it was
observed that in 8 test tubes the test tube with 5% galactose turned violet colored. Thus, this indicates the
presence of carbohydrates while the absence of orange to reddish precipitates in starch and water
confirms that they are non-reducing sugars.
In Seliwanoff’s test is a chemical test for distinguishing between aldose and ketoses sugars. If sugar has a
ketone group, it is a ketose. If a sugar contains an aldehyde group, it is an aldose. This test relies on the
principles that when heated, ketoses dehydrate faster than aldoses. The test also yielded the same result as
the Fehling’s test wherein test-tube A and B formed a reddish precipite. Upon the addition of solution to
fructose, maltose, sucrose, and starch then boiled for 10 minutes, the mixture formed a red-cherry red
precipitate. This is because when added to a solution containing ketones, a red color formed rapidly
indicating a positive test indicates that Selowanoff’s reagent, resorcinol and HCI, whill dehydrate ketones.
Aldoses maty react with the reagent, but it will take a longer reaction time to do so.
In iodine test used to detect the presence of starch. In this test the student drops iodine solution was added
to the carbohydrates. Iodine reacts with starch to form a starch/iodine complex which gives a
characteristic black color indicates the confirmation of positivity. Amylose in starch is responsible for the
formation of a deep blue color in the presence of iodine. The iodine molecule slips inside the amylose
coil. Iodine - KI Reagent: Because iodine is not highly soluble in water, the iodine reagent is prepared by
dissolving it in water in the presence of potassium iodide. This produces a soluble linear triiodide ion
complex, which slides into the starch coil, resulting in an intense blue-black hue (Libretext, 2012). Other
carbohydrates do not give this reaction. Thus, the appearance of black coloration in the suspension of
starch confirms the presence of amylose.
The last conducted was the fermentation which is a natural process that humans have utilized for
thousand of years to produce alcoholic beverages. It is a central process in which an organism turns a
carbohydrate such as starch or sugar, into alcohol or acid. The wine fermentation was made successfully
accomplished by following standard processes and using the proper tools and ingredients with strict
adherence to the method, no molds formed in the container and no spoilage occurred. After 14 days, there
are no contaminations, tangy or sour smells, or a change in general appearance. Using saccharomyces
cerevisiae yeast responsible to produce wines, beers, and ciders. In this natural environment, yeasts can
carry out their metabolism and fermentation activity satisfactorily as they have the necessary nutrients
and substrates. Yeast is essential for the production of alcoholic beverages it plays an important role in the
production of alcoholic beverages and the selection of suitable yeast strains is crucial not only to
minimize alcohol yield, but also to maintain beverage sensory quality (Walker & Stewart, 2016). Ethanol
is formed from acetaldehyde in the absence of oxygen, whereas two moles of ATP are generated. This is
not a completely acceptable reaction for cells because they must consume large amounts of glucose to
supply enough ATP to the system. As a result, ethanol accumulates, and the fermentative action stops
(Puligundla, Smogrovicova & Obulam, 2011)

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https://en.wikipedia.org/wiki/Seliwanoff%27s_test