Name: Date Performed:

Course and Year: Group #: 1 Date Submitted:

Exercise No. 8

Vitamins

OBJECTIVES:

At the end of the experiment, student should be able to:

1. Identify a vitamin as water or fat-soluble.

2. Determine the vitamin C content in a variety of citrus juices and other solutions.

3. Determine the effect of heat upon vitamin C.

RESULTS

Table 1. Vitamin Solubilities

SAMPLE WATER METHYLENE CHLORIDE

Vit B1 + Vit B6 + Vit B 12 Slightly soluble Insoluble
500 mg Vit C Soluble Insoluble
Vit D Insoluble Soluble
Vit E Insoluble Soluble

Table 2. standardization of Vitamin C

Mass of Erlenmeyer flask (g) 135.30

Mass of Erlenmeyer flask + Vit. C (g) 145.74
Mass of Vitamin C (mg) 440
Total I2 solution dispensed (mL) 50.53
Mg Vit. C/ 1mL I2 solution 8.71
Table 3. Determination of Vitamin C in Fruit Juices

TYPE OF MASS OF JUICE (g) TOTAL I2 SOLUTION MASS OF VITAMIN C AVERAGE

JUICE DISPENSED (mL) (mg) MASS
Trial 1 Trial 2 Trial 1 Trial 2 Trial 1 Trial 2 VITAMIN C
(mg)
Tang 1.03 1.00 4.5 2.77 39.195 24.127 31.66
Sundays 1.02 1.04 9.85 12.03 85.794 104.781 95.29

Table 4. Heat Destruction of Vitamin C

TIME OF HEATING TOTAL I2 SOLUTION MASS OF VITAMINS C MASS OF VITAMIN C

(mins.) DISPENSED (mL) (mg) LOST (mg)
0 1.18 10.28 0.00
10 0.87 7.58 2.70
20 0.53 4.62 5.66
30 0.38 3.31 6.97
Juice used was Tang (mango flavor)

DISCUSSION

Vitamins play a vital role in many biochemical functions in the human body and are essential

components for maintaining optimal health. There are two main groups of vitamins – fat-soluble

(easily stored in fat upon absorption) and water-soluble (washed out and not easily stored) (Lykstad

& Sandeep, 2023). In the experiment, it was observed that vitamin C was water soluble and vitamin

B was slightly soluble; on the other hand, vitamins D and E were fat soluble. It was noticed that

Vitamin B (Vit B1 + B6 + B12) was slightly soluble in water. A possible reason for vitamin B's slight

solubility was that vitamins B6 and B12 are moderately soluble. These vitamins are still water-

soluble; however, the structure, charges, and presence of other molecules might be the possible

cause of their moderate solubility affecting the whole vitamin B tested for solubility. For vitamin C, it

was water soluble because it readily dissociated when in contact with water. Meanwhile, vitamins D

and E readily dissociate when in contact with an organic solvent, methylene chloride, or DCM.
 Ascorbic acid determination was carried out by iodine titration. When iodine is added to a starch

solution, it reacts to produce a purple color. However, if there is any vitamin C in the solution, it

“neutralizes” the iodine, preventing the formation of the purple color. Thus, the amount of vitamin C

present in a solution may be measured by first adding a small amount of acidified starch (called

“reaction mix”), and then adding iodine drop wise until the solution turns purple. The end point is

the appearance of the blue starch-iodine color (IKEWUCHI & IKEWUCHI, 2010). The reaction of

iodine and ascorbic acid is shown below:

Figure 1. Reaction of Iodine with Vitamin C

The standardization of vitamin C was performed by measuring the ratio of iodine volume (in mL)

required to oxidize a predetermined quantity of standard vitamin C (in mg) during titration. The

standard value was determined to be 8.71 mg of vitamin C per 1 mL of iodine. The formula that was

used in the calculation of mg of vitamins per 1 mL of iodine is shown below:

𝑚𝑔 𝑉𝑖𝑡𝑎𝑚𝑖𝑛 𝐶 𝑚𝑔 𝑉𝑖𝑡𝑎𝑚𝑖𝑛 𝐶
=
𝑚𝐿 𝐼2 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 1 𝑚𝐿 𝐼2 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛

In the determination of vitamin C in fruit juices, there were two juices, Tang and Sundays

juice (melon flavor) that was used. It was determined in table 3 that Sundays juice have more

vitamin C content which has 95.29 mg average mass of vitamin C than of Tang juice which has 31.66

mg average mass of vitamin C. The determination of the mass of fruit juices can be calculated during

the formula below:

𝑚𝑔 𝑉𝑖𝑡𝑎𝑚𝑖𝑛 𝐶
𝑚𝐿 𝑜𝑓 𝐼2 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛 𝑥 = 𝑚𝑔 𝑉𝑖𝑡 𝐶 𝑠𝑎𝑚𝑝𝑙𝑒
1 𝑚𝐿 𝐼2 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
 Vitamin C is a water-soluble and temperature-sensitive vitamin, so is easily

degraded during cooking, and elevated temperatures and long cooking times have been

found to cause particularly severe losses of vitamin C (Lee, Suknark, Kluvitse, Phillips, &

Eitenmiller, 1999). The Vitamin C flasks were subjected to heat destruction, with

temperatures ranging from 100-150°C, leading to gentle boiling. Three flasks were

subjected to varying durations of heat exposure (10 minutes, 20 minutes, 30 minutes),

whereas one flask containing vitamin C was kept at room temperature without heating.

It was shown that increasing the duration of heating resulted in a more significant loss of

vitamin C. This occurred because higher temperatures resulted in increased destruction

of vitamin C.

CONCLUSION

During the experiment, four different vitamins were tested to determine their solubility.

These are vitamins B, C, D, and E. It was observed that when these vitamins were added with water

and methylene chloride, vitamins B and C indicated water soluble, whereas vitamins D and E showed

fat solubility. During the analysis of vitamin C levels in fruit juices, it was found that Sundays juice

contained more vitamin C (95.2874 mg) than Tang juice (31.66 mg). During heat destruction, the

duration of heating correlates directly with the amount of vitamin C loss.

REFERENCES

IKEWUCHI, C., & IKEWUCHI, C. (2010). IODOMETRIC DETERMINATION OF THE ASCORBIC ACID
(VITAMIN C) CONTENT OF SOME FRUITS CONSUMED IN A UNIVERSITY COMMUNITY IN
NIGERIA. GLOBAL JOURNAL OF PURE AND APPLIED SCIENCES VOL. 17, 47-49.

Lee, J., Suknark, K., Kluvitse, Y., Phillips, R., & Eitenmiller, R. (1999). Rapid liquid chromatographic
assay of vitamin E and retinyl palmitate in extruded weaning foods. Journal of Food Science,
64:968–972. doi:10.1111/j.1365-2621.1999.tb12261.x
Lykstad, J., & Sandeep, S. (2023, March 6). Biochemistry, Water Soluble Vitamins. Retrieved from
National Library of Medicine:
https://www.ncbi.nlm.nih.gov/books/NBK538510/#:~:text=There%20are%20two%20main%
20groups,nature%20of%20water%2Dsoluble%20vitamins.