CHM 4203 Lab #3.

1 Name: RAMNARINE, Awadnarine

Group #1 Reg. # 13/0705/2171

Title: Determination of Ascorbic Acid Content in Fruit Juice and Investigation of the Effect of

Pasteurization on the Retention of Ascorbic Acid (Vitamin C) in Fruit Juice.

Aim: To determine the ascorbic acid (vitamin C) content in fresh and pasteurized fruit juices and

to standardize indophenol dye solution-to evaluate the concentration of dye solution: also to

evaluate the ascorbic acid content in fresh fruit juices (lime, orange, tangerine, grapefruit) and to

evaluate the ascorbic acid content in pasteurized fruit juices and to assess the effect of

pasteurization on the retention of ascorbic acid in these fruit juices.

Principle: Vitamin C (L-Ascorbic Acid) occurs in all living tissues, where it influences

oxidation-reduction reactions. Ascorbic acid is a strong reducing agent. It is oxidized to

dehydroascorbic acid (DHAA) by 2, 6-dichlorophenol indophenol dye. The reaction is given in

the figure below:

The major source of L-ascorbic acid in foods is vegetables and fruits. The human requirement of

vitamin C is not well defined. According to De Man, figures ranging from 45 to 75 mg per day

have been listed.

Vitamin C is the least stable of all vitamins and is easily destroyed during processing and

storage. The rate of destruction is increased by the action of metals, especially copper and iron,

and by the action of enzymes. Exposure to oxygen, prolonged heating in the presence of oxygen,

and exposure to light are all harmful to the vitamin C content of foods. Enzymes containing

copper or iron in their prosthetic groups are efficient catalysts of ascorbic acid decomposition. In

fruit juices, the enzymes can be inhibited by pasteurization, deaeration, or holding at low

temperature for a short period.

Materials & Apparatus: As per lab handout.

Procedure: Same as per lab handout with the following modifications:

 Four citrus fruits were used. They were lemon, lime, orange and grapefruit.

 Oxalic Acid was used instead of metaphosphoric acid

Results:

Group # Type of Conc. Of AA Content AA Content Loss in AA %

Citrus Fruit Dye Soln. in Fresh Juices in Pasteurized Content after Loss
(mg (mg/100mL) Juices Pasteurizing of AA
AA/mL (mg/100mL) by heat
ID)

1 Lime 1.08 55.08 28.10 26.98 48.98

2 Lemon 1.43 38.61 34.32 4.29 11.11
3 Grapefruit 1.05 29.40 14.70 14.70 50.00
4 Orange 1.60 62.40 31.00 31.40 50.32
Table 1 showing the various concentrations of vitamin C in citrus juices after various treatments.
Discussions: It was necessary to carry out a standardization of the indophenol dye because the

concentration which was determined based on the standardization, was needed to calculate the

ascorbic acid content for the fresh and pasteurized fruit juices. As was stated in the theory,

vitamin C is the least stable of all the vitamins. It undergoes oxidation and this is accelerated in

the presence of heat. Because of this, it is expected that pasteurization would lead to a

loss/decrease of ascorbic acid content. This is exactly what was observed for each of the four

fruit juices studied in this experiment. From the table above, it can be seen that after

pasteurization, orange had the greatest loss of ascorbic acid followed by lime and then grapefruit.

The lemon had the least amount losses of ascorbic acid.

Theoretically the highest vitamin C content should be found in oranges followed by grapefruit,

lemon and finally lime. This was particularly true for oranges which showed the highest

concentration for vitamin. However the other three did not corresponded to literature, instead

they exhibit the opposite trend from literature. The trend was in the order grapefruit< lemon<

lime, with lime having more. The reason this deviation may have arisen was because the

experiment done was completed by different analysts, whom may have interpreted the endpoint

differently during the titration. The endpoint was difficult to detect since the juices were opaque

and the first trace of a pink colour was needed to determine the end of the reaction and hence this

may have resulted in excess indophenol dye being added to the solution.

The fruit juice that loses the most vitamin C is least stable to heat and vice versa. Based on the

results obtained, it can be said that orange, which had the most loss of ascorbic acid, was the

least heat stable fruit juice, followed by grapefruit, then lime and lastly lemon. Therefore it can

be said based on the results that lemon was the most stable juice.
Conclusions: it can therefore be concluded that the greatest loss of ascorbic acid was in the

orange and the least loss of ascorbic acid was found in the lemon.

References:

• de Man, J. 1999. Principles of Food Chemistry. USA: Aspen Publishers.

• Fennema, OR. 1996. Food Chemistry 3rd Edition. USA: Marcel Dekker Inc.
Appendix

I. Standardisation of Indophenol Dye Solution

Using data from group 1: lime

10mL of Standard Ascorbic Acid solution was used.

Mass of AA Used (mg)

Mass of Ascorbic Acid ( AA )∈10 mL= × Volume of Analyte (mL)
TotalVolume (mL)

(0.2 g AA ×1000)
¿ × 10 mL Sample=2.0 mg AA
1000 mL Sample

Concentration ( mL
mg
)= Volume
Mass of AA (mg)
of Titrant (mL)

2.0 mg Ascorbic Acid

¿ =1.08 ( mg AA /mL ID )
1.85mL Indophenol Dye

II. Determination of Vitamin C Content in Fresh Juice

Using data from group 1

Mass of AA in 10mL Sample= Average Titre Value ⨯ Concentration

mg AA
Mass of AA ∈10 mL Sample=2.55 mL ID ×1.08 mg AA /mL ID=2.75
10 mL

Concentration ( 100 mL )
mg AA
=( mg AA ∈10 mL AA ) × Dilution Factor ×10

c=( 2.75 ) × ( ) ×10=55.0

mg AA 50 mg
10 mL Sample 25 100 mL

The dilution factor was taken as the total volume of solution divided by the volume of juice in
solution. The number 10 was included in the calculation since it was the volume of solution used
for the titration.
All other calculations for the vitamin C of fresh juices were done the same way for the other
groups.

III. Determination of Vitamin C in Pasteurised Juices

Using data from group 1

Mass of AA in 10mL Sample= Average Titre Value ⨯ Concentration

mg AA
Mass of AA ∈10 mL Sample=1.30 mL ID ×1.08 mg AA /mL ID=1.404
10 mL

Concentration ( 100 mL )
mg AA
=( mg AA ∈10 mL AA ) × Dilution Factor ×10

c=(1.404 ) ×( ) ×10=28.1
mg AA 50 mg
10 mL Sample 25 100 mL

The dilution factor was taken as the total volume of solution divided by the volume of juice in
solution. The number 10 was included in the calculation since it was the volume of solution used
for the titration.

All other calculations for the vitamin C content of pasteurised juice were done the same way for
the other groups.