CHEM 137.

2 – BIOCHEMISTRY (LABORATORY)
LABORATORY REPORT

Name: Reynand E. Corpuz Date Submitted: 02-07-2021

Lab Schedule: TTh 8:00 – 10:00 Date Performed: ____________

Exercise No. 8
Vitamins
I. OBJECTIVES:
In the end of the experiment, the students should be able,
1. To identify vitamins as water or fat-soluble,
2. To determine the vitamin C content in a variety of citrus juices and other solutions, and
3. To determine the effect of heat to the vitamin C.

II. RESULTS AND DISCUSSION:

A. Solubility of Vitamins
Solubility (Water-soluble or
Vitamins Solubility in Water
Fat-soluble)

Vitamin A - Fat-soluble

Vitamin D - Fat-soluble

Vitamin B12 X Water-soluble

Discussion:
Vitamins are groups of organic molecules that are essential for normal growth and nutrition
which organism need in trace amounts to help catalyse many of the biochemical reactions in the body.
It came from the word “vita” and “amine” because the first vitamin being discovered contained amino
group in its molecule. Vitamins are essential micronutrient, that means vitamins cannot be synthesized
by humans (or synthesized but in very little quantity) and must be obtained through diet. The vitamins
that we know today are not chemically, functionally and structurally related to each other, but they are
group through their solubility in polar and nonpolar solvents. The first part of the experiment
investigates the solubility of vitamins such as vitamin A, vitamin D and vitamin B 12 in water. Based
on the result, it was observed that vitamin B 12 was the only one that was dissolved in water while
vitamin A and D were not soluble in water. Vitamin B12 or Cobalamin able to dissolved in water
because vitamin B12 is a very large polar molecule with many available O, N and H terminals for
hydrogen bonding with another polar molecule such as water.

Figure 1. Molecular structure of Cobalamin (Vit. B 12)

On the other hand, vitamin A and D are nonpolar compounds and since ‘like dissolves like’ is
still applicable in biological systems, then only nonpolar solvents can dissolve vitamins A and D and
not polar solvent like water. The long aliphatic chain of vitamin A and the prominent aliphatic and
aromatic groups of vitamin D is the cause of its solubility.

B. Standardization of Vitamin C
mg of Vitamin C
Volume of Iodine
Mass of Vitamin C mL of Iodine

28.1 mg 3.4 mL 8.26 mg/mL

NOTE: Express your answer up to two decimal points

Discussion:
The second part of the experiment is the standardization of Vitamin C. Since vitamin C is our
focus, one will expect that the concentration of the vitamin C is one that are determined here. However,
the experiment used a known concentration of ascorbic acid (vitamin C) and doing so, the iodine is
the one that was being standardized to get its molarity, not the ascorbic acid. Regardless of that, it can
give us an idea in standardizing Vitamin C through using an iodine solution with known
concentration— the reversed condition of what was performed in the experiment. In the experiment,
the ascorbic acid, with starch indicator, was titrated with iodine solution. As iodine was added during
the titration, the ascorbic acid is oxidized to dehydroascorbic acid, while the iodine is reduced to iodide
ions (Note: further explanation is on Part C).
𝑎𝑠𝑐𝑜𝑟𝑏𝑖𝑐 𝑎𝑐𝑖𝑑 + 𝐼2 → 2𝐼 − + 𝑑𝑒ℎ𝑦𝑑𝑟𝑜𝑎𝑠𝑐𝑜𝑟𝑏𝑖𝑐 𝑎𝑐𝑖𝑑
 The volume of the iodine consumed at the end point of the titration of ascorbic acid was
determined to be 3.4 mL. With that, the ratio of the mass of Vitamin C to the mL of iodine was
calculated to be 8.26 mg/mL. This ratio is advantageous as it can be used to get the mass of vitamin C
sample in Part C.

C. Determination of Vitamin C in Fruit Juices

Mass (mg) of Vitamin C in
Sample
Volume of Iodine (mL) sample

Minute Maid 13.98 115.47

Lemon (Soda) 3.64 30.07

NOTE: Express your answer up to two decimal points. Use your answer on B to calculate
the mass (mg) of Vitamin C present in your samples.
Discussion:
Fruits, as well as vegetables, are good source of vitamins C. Vitamin C is a very potent
antioxidant, as well as promotes healthy skin and immune function. The human body cannot produce
or store vitamin C, so it is necessary to consume it regularly in sufficient amounts to prevent vitamin
C deficiency or scurvy. As a substitute for fresh fruit, processed fruit juices such as Minute Maid and
Lemon (Soda) provides a decent amount of vitamin C that is required by our body. In the experiment,
the processed fruit juice of Minute Maid and Lemon (Soda) underwent iodine test to determine the
amount of ascorbic acid (vitamin C) in each 10 mL of these samples. In the titration, the ascorbic acid
is oxidized to dehydroascorbic acid, while the iodine is reduced to iodide ions. The presence of
ascorbic acid makes the iodine to change into its reduced form (see reaction in Part B). If all ascorbic
acid in the solution has been oxidized, the excess iodine will now free to react with the starch indicator,
forming a blue-black complex— this is the end point of the titration.
Based on the results, it was observed that the volume of iodine consumed in Minute maid and
Lemon (Soda) samples were 13.98 mL and 3.64 mL, respectively. Utilizing the ratio that we
previously calculated in part B and multiply it to volume used, the mass of the vitamins can be
determined. Doing the math, it was calculated the Minute Maid and Lemon (Soda) have a vitamin C
mass of 115.47 mg and 30.07 mg, respectively. This means that Minute Maid juice is a better source
as it has higher vitamin C content than with Lemon (Soda).
 D. Heat Destruction of Vitamin C
Test tube Temperature Condition Observations

A (Tomato Juice) Water Bath set at 75˚C The solution turned dark blue

The solution has portion with

B (Tomato Juice) Room Temperature
faint blue coloration

NOTE: Temperature condition: Room temperature or Water bath

Discussion:
The destruction of vitamin C by heat was investigated in this part of the experiment. Here, 2
sample of 10 mL tomato juice were prepared and was exposed to different temperature for five
minutes. With a starch indicator, the samples were then added with iodine solution, only a drop for
every 3 minutes as indicated in the video. Results shows that test tube A, which was set at elevated
temperature produced dark-blue color after the addition, while the test tube B, which was set at room
temperature produce faint blue coloration, not as intense and dominating as with test tube A. This is
due to the fact that heating destroyed most of the vitamin C in test tube A, and since there are now less
number of vitamin C to be oxidized by the added iodine to form iodide, the excess iodine are free to
react with the starch indicator, hence forming a blue-black complex which was observed in the
experiment. The test tube B, which was left at room temperature, has more vitamin C molecule that
are able to keep reducing the added iodine to iodide, which prolonged the time for the starch to react
with iodine thus delaying the formation of blue-black complex.

III. CONCLUSIONS:
The experiment focuses to identify vitamins as water or fat-soluble, to determine the vitamin C
content in a variety of citrus juices and other solutions, and to determine the effect of heat to the
vitamin C. It was observed in the first part of the experiment that only vitamin B12 is water soluble
while vitamins A and D were fat soluble vitamins. In part B, the ratio of the mass of Vitamin C to the
mL of iodine was calculated to be 8.26 mg/mL. Furthermore, it was determined in Part C of the
experiment that Minute Maid and Lemon (Soda) fruit juices have 115.47 mg and 30.07 mg of Vitamin
C, respectively. Lastly, it was confirmed that the vitamin C in heated test tube A was destroyed as the
solution turned dark blue while, the test tube B, which was left at room temperature remained the
number of Vitamin C as the solution has a portion with faint blue coloration.