BIOLOGY PRACTICUM REPORT 97

Course : Experimental of Biochemistry

VITAMIN C TEST

BY :

NAME : DINDA ANGGITA MAYSARAH

ID NUMBER : 4183141061

DEPARTMENT : BIOLOGY
: BILINGUAL BIOLOGY EDUCATION
PROGRAM
2018
GROUP :2

DAY/DATE : WEDNESDAY/ MARCH 11th, 2020

FACULTY OF MATHEMATICS AND NATURAL SCIENCE

UNIVERSITAS NEGERI MEDAN

MEDAN
 I. EXPERIMENT TITLE : Vitamin C Test (5)

II. EXPERIMENT PURPOSE : (5)

2.1 Student can find out what the samples that contain of vitamin C.
2.2 Student can find out the effect of betadine in the practicum activity.
2.3 Student can compare fruit or juice that more healthy to consume.
2.4 Student can find out the vitamin C level that needed in human body.
2.5 Student can find out what is the differences of vitamin and minerals.

III. THEORITICAL REVIEW : (20)

Vitamins fall into two categories: fat soluble and water soluble. The fat-soluble
vitamins A, D, E, and K dissolve in fat and can be stored in your body. The water-
soluble vitamins C and the B-complex vitamins (such as vitamins B6, B12, niacin,
riboflavin, and folate) need to dissolve in water before your body can absorb them.
Because of this, your body can't store these vitamins. Any vitamin C or B that your body
doesn't use as it passes through your system is lost (mostly when you pee). So you need a
fresh supply of these vitamins every day. Whereas vitamins are organic substances (made
by plants or animals), minerals are inorganic elements that come from the soil and water
and are absorbed by plants or eaten by animals. Your body needs larger amounts of some
minerals, such as calcium, to grow and stay healthy. Other minerals like chromium,
copper, iodine, iron, selenium, and zinc are called trace minerals because you only need
very small amounts of them each day. Although they are all considered micronutrients,
vitamins and minerals differ in basic ways. Vitamins are organic and can be broken down
by heat, air, or acid. Minerals are inorganic and hold on to their chemical structure
(Combs,2017).
Vitamins and minerals are essential nutrients because they perform hundreds of
roles in the body. There is a fine line between getting enough of these nutrients (which is
healthy) and getting too much (which can end up harming you). Eating a healthy diet
remains the best way to get sufficient amounts of the vitamins and minerals you need.
Although they are all considered micronutrients, vitamins and minerals differ in basic
ways. Vitamins are organic and can be broken down by heat, air, or acid. Minerals are
inorganic and hold on to their chemical structure. Antioxidants are defined as compounds
that can delay, slow down and prevent lipid oxidation. In a special sense, antioxidants are
substances that can prevent the formation of free radical reactions (peroxide) in lipid
oxidation. Synthetic antioxidants such as BHA (butylated hydroxy aniline) and BHT
(butylated hydroxy toluene) have been known to have major side effects, including
causing liver damage. On the other hand, nature provides an effective and relatively safe
source of antioxidants such as flavonoids, vitamin C, beta carotene and Vitamin E. This
encourages more and more exploratory research to be done on natural ingredients to
become sources of antioxidants. Free radicals are compounds or molecules that contain
one or more unpaired electrons in their outer orbitals. The existence of unpaired electrons
causes the compound to be very reactive looking for a partner, by attacking and binding
to the electrons around it so that it can trigger disease (Lung and Dika,2017).
Vitamin C, also known as L-ascorbic acid, is a water-soluble vitamin C6H8O6
that is naturally present in some foods, added to others, and available as a dietary
supplement. Humans, unlike most animals, are unable to synthesize vitamin C
endogenously, so it is an essential dietary component.Vitamin C is an organic acid in the
form of white crystals and feels acidic and odorless. In the solution of Vitamin C is easily
damaged by oxidation by oxygen from the air. This vitamin becomes more stable when in
the form of dry crystals. The total body content of vitamin C ranges from 300 mg (at
near scurvy) to about 2 g. High levels of vitamin C (millimolar concentrations) are
maintained in cells and tissues, and are highest in leukocytes (white blood cells), eyes,
adrenal glands, pituitary gland, and brain. Relatively low levels of vitamin C (micromolar
concentrations) are found in extracellular fluids, such as plasma, red blood cells, and
saliva. Deficiency of vitamin C causes scurvy, widespread connective tissue weakness
and capillary fragility. Among chemists, it is used as a reagent for the preparation of fine
chemicals, enzymatic reagent and nanomaterials. Consequently, the detection and
quantification of ascorbic acid in food samples, products and nutraceuticals is receiving
overwhelming importance among researchers, medical practitioners and also in the
pharmaceutical and food industry (Devaki,2017).
Vitamin C is an important antioxidant food and significantly decreases side
effects of reactive species such as reactive oxygen which can cause damage by oxidative
reactions to macromolecules such as lipids, DNA, and proteins, which are involved in
chronic diseases including neurodegenerative diseases. Moreover stress has been shown
to cause a decrease in GSH and vitamin C levels, which protect tissues from oxidative
damage. Besides polyphenols, especially vitamin E, carotenoids and vitamin C have been
linked to antioxidant properties Fruits contain a variety of vitamins needed by the body,
one of which is vitamin C. Vitamin C acts as an antioxidant and is effective in dealing
with free radicals that damage cells or tissues. Vitamin C is easily dissolved in water,
therefore, when experiencing the process of slicing, washing and boiling food ingredients
that contain vitamin C will decrease in levels. The content of vitamin C in fruit and food
will be damaged by the oxidation process by the outside air, especially if it is heated.
Therefore, storage is carried out at low temperatures (in the refrigerator) and cooking
does not cause color changes in foods containing vitamin C (Putri and Yunita,2017).
Vitamin C can additionally restore vascular responsiveness to
vasoconstrictors,preserve endothelial barrier by maintaining cyclic guanylate phosphatase
and occludin phosphorylation andpreventing apoptosis. Vitamin c needs of each person is
different. For adults, the recommended daily amount for vitamin C is 65 to 90 milligrams
(mg) a day, and the upper limit is 2,000 mg a day. Although too much dietary vitamin C
 is unlikely to be harmful, megadoses of vitamin C supplements might cause: Diarrhea,
Nausea, Vomiting, Heartburn, Abdominal cramps, Headache, Insomnia. Finally, high-
dose vitamin C can augment antibacterial defense. These protective effectsagainst
overwhelming oxidative stress due to ischemia/reperfusion, sepsis or burn seems to
mitigate organ injuryand dysfunction, and promote recovery after cardiac
revascularization and in critically ill patients, in the latterpartially in combination with
other antioxidants. Of note, several questions remain to be solved, including optimaldose,
timing and combination of vitamin C with other antioxidants. Vitamin C also has other
effects. Vitamin C improvesimmune function, and facilitates enteral uptake of non-heme
iron, reduction of folic acid intermediates and syn-thesis of collagen (wound healing),
cortisol, catecholaminesand carnitine. These effects are beyond the scopeof this review
(Oudemans-van,2014).

IV. APPARATUS AND MATERIALS : (5)

4.1 Apparatus
No. Apparatus Quantity
1. Bowl 1 pcs
2. Grinder 1 pcs
3. Knife 1 pcs
4. Spoon 1 pcs

4.2 Materials
No Materials Chemical Color Phase Quantity
. formula
1. Apple - Cream Solid Sufficient
2. Buavita guava - Light pink Liquid Sufficient
3. Guava - Light pink Solid Sufficient
4. Avocado - Light Green Solid Sufficient
5. Tomato - Red Solid Sufficient
6. Banana - Cream Solid Sufficient
7. Orange - Orange Solid Sufficient
8. Markisa - Orange Solid Sufficient
9. Betadine C6H9I2NO Brown Liquid Sufficient

V. WORK PROCEDURE : (5)

No. Work Procedure Result
Apple
1. Cut the fruit and take some of the flesh The color of apple changes to be light
brown

2. Mash the fruit until smooth

3. Drop the fruit with betadine sufficiently,

stir until color change occur

Buavita Guava
1. Pour the Buavita into a bowl The color of Buavita guava changes to
light brown

2. Drop the Buavita with betadine

sufficiently, stir until color change occur
 Water apple
1. Cut the fruit and take some of the flesh The color of water apple still pink but
2. Mash the fruit until smooth a more thick

3. Drop the fruit with betadine sufficiently,

stir until color change occur

Avocado
1. Cut the fruit and take some of the flesh The color of avocado changes to deep
2. Mash the fruit until smooth green with brown

3. Drop the fruit with betadine sufficiently,

 stir until color change occur

Tomato
1. Cut the fruit and take some of the flesh The color of tomato changes to
reddish brown and clear

2. Mash the fruit until smooth

3. Drop the fruit with betadine sufficiently,

stir until color change occur

Banana
1. Cut the fruit and take some of the flesh The color of banana changes to
 blackish brown

2. Mash the fruit until smooth

3. Drop the fruit with betadine sufficiently,

stir until color change occur

Orange
1. Press the orange until the water comes The color of orange changes to
out reddish orange

2. Drop the fruit with betadine sufficiently,

stir until color change occur
 Markisa
1. Cut the fruit and take some of the flesh The color of markisa changes to
reddish orange

2. Drop the fruit with betadine sufficiently,

stir until color change occur

VI. EXPERIMENT RESULT : (27)

VI.1 Table of experiment
Amount of Color changes
No. Materials
betadine drops Before After
1. Apple 21 drops Cream Light brown
2. Buavita Guava 23 drops Light pink Light brown
3. Guava 10 drops Light pink Pink
4. Avocado 20 drops Light green Deep green with brown
5. Tomato 13 drops Red Reddish brown and clear
6. Banana 11 drops Cream Blackish brown
7. Orange 20 drops Orange Reddish orange
8. Markisa 21 drops Orange Reddish orange
VI.2 Discussion
Vitamin C is ascorbic acid, a chemical compound that is soluble in water. This
type is used by the body, although it is acidic but the strength of the acid is much
lower than stomach acid. This type is cheaper than other types. Ascorbyl palmitate is
an ascorbic acid that is related to fatty acids to make a fat-soluble delivery system for
vitamin C. In this test, using betadine as an indicator of the presence of vitamin C.
The betadine packaging stated that betadine contains 10% povidone iodine which is
equivalent to iodine 1 %. Iodine is actually the indicator, because the reaction
between ascorbic acid in vitamin C and iodine will remove the color of iodine, so the
brighter the color of the fruit when it has been dropped by betadine, the fruit has a lot
of vitamin C. (Suprayogi and Mia,2011).
In this practicum there are 8 samples that use to test the vitamin C levels with
using betadine, namely :
VI.2.1 Apple
One example of a high source of vitamin C from fruits is apples, in
addition to having good nutritional content, the fruit also has properties to cure
various diseases. Storage of apples is the most crucial period in maintaining
the quality of vitamin C in the fruit. in pure ascorbic acid and apple samples to
be analyzed showed the same results, namely the appearance of a brownish
white color. This shows that qualitatively apples that have been stored for 0
days, 1 day, 2 days, 3 days contain vitamin C. Brownish white color indicates
the formation of dehydro acid L-ascorbic from oxidation of ascorbic acid by
permanganate ions (Asmara dan Hanik,2019).
Based on the practicum that has been done, after the sample is drop by
betadine, the color of apple change to be light brown, almost the same as the
theory conveyed, it’s mean the practicum was successful and the apple that
contain of vitamin C is correct.

VI.2.2 Buavita Guava

The advantage in guava juice is the high content of vitamin C which is a
natural antioxidant that is found in the flesh of red guava fruit that is ripe.
Antioxidants are compounds that can protect the body from various diseases
related to the presence of free radicals when routinely consumed, but vitamin
C from fruit can last longer in the body than vitamin C found in juice or
supplements. Besides staying longer, taking vitamin C from fruit is also more
beneficial because there is fiber consumed. This is a minimum course or even
absent in juice products or supplements. By consuming fiber, one can feel full
longer and maintain digestive health (Aufa,dkk.,2020).
For the color changed, I didn’t found the article that specific discuss about
it, but as we know if there are color changed that occur in the experiment, it’s
mean the samples contains of the vitamin C. Based on the practicum we can
 show that the buavita guava changed the color to be light brown, it’s mean the
practicum was successful and the buavita guava that contain of vitamin C is
correct.

VI.2.3 Guava
Another local fruit that is very high in vitamin C is guava. In every 100g
guava contains 116mg vitamin C. Other nutrients are 61 kcal energy, 0.9g
protein, 15.4g carbohydrates, 0.3g fat, 31mg calcium, 41mg phosphorus and
0.2mg iron. The benefits of this guava include to help control diabetes, ulcers,
diarrhea (colic), colds, reduce cholesterol levels, as an antioxidant to prevent
cancer, ambient and thrush. High potassium, iron and zinc make guava very
well consumed by people with dengue fever. In tests that use betadine as an
indicator of the presence of vitamin C. Iodine is actually the indicator, because
the reaction between ascorbic acid in vitamin C and iodine removes the color
from iodine. (Suprayogi and Mia,2011).
Based on the practicum that has been done, after the sample is drop by
betadine, the color of guava change to be more pink, it’s mean the practicum
was successful and the guava that contain of vitamin C is correct.

VI.2.4 Avocado
Avocados contain high fat content, distinctive flavor and soft flavor,
causing avocados have a high taste. Avocados also have minerals such as
calcium 10 mg, phosphorus 20 mg, 0.9 gram protein, caloric value 85, vitamin
A 180 IU, vitamin C 13 mg and vitamin D 20 IU (Anova dan kamsina,2013).
For the color changed, I didn’t found the article that specific discuss about
it, but as we know if there are color changed that occur in the experiment, it’s
mean the samples contains of the vitamin C. Based on the practicum the
avocado changes to be deep green with brown after drops by betadine, it’s
mean this sample contain of vitamin C, but maybe not as much as tomatoes or
oranges, it is shows at the color not clear or bright.

VI.2.5 Tomato
Tomatoes are among the foods that are rich in vitamin C, Vitamin C
deficiency can cause bleeding gums, wounds difficult to heal, canker sores
and others.. In addition to vitamins, other tomato content is solanin (0.007%),
saponins, folic acid, malic acid, citric acid, bioflavonoids, proteins, fats,
minerals and histamine. The results of a qualitative analysis where tomatoes
are dropped into a test tube containing a betadine solution will produce a clear
color (Anggreani,2019).
 Based on the practikum that has been done, after the sample is drop by
betadine, the color of markisa change to be brown, although not too clear, it
has shown that tomatoes contain vitamin C. There may be errors when
undergoing work procedures so that the fruit does not turn to be as clear and
bright as possible.

VI.2.6 Banana
Bananas have iron and vitamin C levels which can help increase and
absorb iron in the body. The higher the content of vitamin C in food the higher
the absorption and use of iron in the body. Bananas contain iron which will
stimulate the production of hemoglobin in the blood and also helps prevent
anemia. Vitamin c contained in bananas is also good for health to help rebuild
the immune system. In the analysis of vitamin C content in bananas, bananas
will experience changes in color becoming brighter and clearer
(Mahardika,2016).
Based on the practicum that has been done, after the sample is drop by
betadine, the color of banana change to be blackish brown and clear, it has
shown that banana contain vitamin C. But, maybe the drops of betadine is
over, it’s make the banana changes to blackish.

VI.2.7 Orange
The vitamin C content of citrus fruits ranges from 20-60 mg / 100 ml of
fruit juice. Citrus fruit as a source of vitamin C, the benefits are very large on
health. Vitamin C acts as an antioxidant that can neutralize free radicals
resulting from fat oxidation, so it can prevent several diseases such as cancer,
heart disease and premature aging. Vitamin Amun is very easy to experience
oxidation, so it can be lost or reduced during processing and storage, storage
of orange juice in the refrigerator can maintain vitamin C levels and
acceptability of orange juice. To see the vitamin C content, iodine titration
test, the reaction between ascorbic acid in vitamin C and iodine removes the
color from iodine (Wariyah,2010).
Based on the practicum that has been done, after the sample is drop by
betadine, the color of orange just change a little to be reddish orange, it has
shown that orange contain vitamin C. But, maybe in the procedure work, the
drops of iodine is over, it make the orange have a little reddish color.

VI.2.8 Markisa
Markisa have sweet taste and fresh when ripe becomes the choice for
consumption. In addition to the taste of fruit that is an allure, passion fruit also
contains compounds that are important for the health of the human body.
Active compounds found in fruits are generally able to act as antioxidants.
 Vitamin C contained in fruits and vegetables is a good source of antioxidants
for the body. Titration using iodine aims to reduce strong compounds such as
vitamin C. The results of the titration test with iodine solution are given until
the solution changes color (Ngginak,dkk.,2019)
Based on the practikum that has been done, after the sample is drop by
betadine, the color of markisa change to be reddish orange from orage color,
it’s mean the practicum was successful and the markisa that contain of vitamin
C is correct.

VII. CONCLUSION : (10)

VII.1 This practicum is to test the vitamin C levels in the samples, based on the
practicum, all of samples (orange, apple, guava, avocado, banana, markisa, tomato,
buavita guava) contains vitamin C, but the levels maybe have different, this condition
can show in the color changed that occur in every samples.
VII.2 Vitamin C can be oxidized by iodine (which is found in betadine), when the
betadine drops, fusion occur. This oxidation causes iodine to change into iodine. So
when Vitamin C is added to betadine which contains iodine (clear) color changes will
occur.
VII.3 Vitamin C from fruit can last longer in the body than vitamin C found in juice or
supplements. Besides staying longer, taking vitamin C from fruit is also more
beneficial because there is fiber consumed. This is of course minimal or even absent
in juice products or supplements. By consuming fiber, one can feel full longer and
maintain digestive health.
VII.4 For adults, the recommended daily amount for vitamin C is 65 to 90 milligrams
(mg) a day, and the upper limit is 2,000 mg a day. Although too much dietary vitamin
C is unlikely to be harmful, megadoses of vitamin C supplements might cause:
Diarrhea, Nausea, Vomiting, Heartburn, Abdominal cramps, Headache, Insomnia.
VII.5 Minerals and vitamins are important catalysts in different kind of chemical
reactions that are going into your body and so they support your health. Vitamins and
minerals differ in basic ways. Vitamins are organic substances, meaning they are
made by plants or animals. Minerals, however, are inorganic elements, which mean
they come from the earth, soil and water and are absorbed by plants.

VIII. QUESTION AND ANSWER : (10)

1. Based on the practicum done, what is the difference between vitamins and
minerals?
Answer :
Vitamins and minerals differ in basic ways. Vitamins are organic and can be broken
down by heat, air, or acid, processes like cooking or improper storage can cause
vitamins to break down in foods, so either plants or animals make vitamins. So, for
 instance, vitamin D or vitamin E or vitamin C, you've heard of vitamin C coming
from oranges, well oranges make vitamin C out of chemicals they get out of the soil.
While minerals are inorganic and hold on to their chemical structure, trace minerals
found in soil or water will stay intact in the plants, fish, meat, or other foods you
consume. So, calcium, potassium, zinc, selenium, those are mineral.

2. Give examples of fruits that are rich in vitamin C based on the practicum?
Answer :
All our sample contain of vitamin C, example : orange, apple, guava, avocado,
banana, markisa, tomato, buavita guava. But the vitamin C level is different.

3. Why does the color change in the fruit or juice when given a reaction or dropped
with betadine?
Answer :
Vitamin C is also called ascorbic acid. This acid can be oxidized by iodine (which is
found in betadine), when the betadine drops, fusion occur. This oxidation causes
iodine to change into iodine. So when Vitamin C is added to betadine which contains
iodine (clear) color changes will occur. The function of iodine standard solution is
also as a reagent to show the amount of vitamin C contained in the sample into
dehydro ascorbate compounds so that it will change color.

4. Write the chemical formula from povidone iodine (betadine)!

Answer :
C6H9I2NO (Povidone Iodine)

IX. BIBLIOGRAPHY : (10)

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VARIETAS BUAH TOMAT YANG DIKONSUMSI MASYARAKAT
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Anova, Inda Three, and Kamsina Kamsina.K. (2013). Efek Perbedaan Jenis Alpukat dan
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Asmara, Anjar Purba dan Hanik Khuriana Amungkasi. (2019). Kajian Kinetika Pengaruh
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Aufa, Muhamad Raihan, Wendry Setiyadi Putranto, and Roostita Lobo Balia. (2020).
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Combs, Gerald F. and James P. Mc Clung. (2017). The Vitamin 5th Edition. USA :
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Ngginak, James, Anggreini Rupidara, and Yanti Daud. (2019). Analisis Kandungan
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Vol, 1(1) : 50 - 55

MEDAN, MAY 14th 2020

LABORATORY ASSISTAN T PRACTICAN

NIDA ANDZAR SHOFIYYAH DINDA ANGGITA MAYSARAH

ID NUMBER : 4163141032 ID NUMBER : 4183141061