Extended Experimental

Investigation: Stage 2
An Investigation of Vitamin C Content in Orange
Juice
Andrew Zhao

Equipment:
-

2 Conical Flasks
2 Pipettes (20ml)
1 Pipette (10ml)
2 Pipette Fillers
250ml Volumetric Flask
1 Funnel
3 Beakers
1 Measuring Cylinder
1 Burette
1 Retort Stand
1 Clamp
2% Starch Solution

Sodium Thiosulfate (

Potassium Iodate (

Potassium Iodide ( KI )

1M Sulphuric Acid
Distilled Water
Orange Juice
1 Sieve
2 Droppers
Heat Plate

Na 2 S2 O3 )

KI O3 )

Method
Preparing the 0.01M Iodine Solution (250ml) for Standardization
1. Weigh out 0.18grams of potassium iodate, 2.1 grams of potassium iodide
and 5ml of sulphuric acid
2. Put the potassium iodate into a 250ml volumetric flask and fill the flask up
with distilled water until it is 1/3 full
3. Add the potassium iodide and sulphuric acid, and proceed to fill the
volumetric flask with distilled water with a dropper until it reaches the line
(see Figure 0.1)
4. Shake the volumetric flask
Preparing of 0.25M Sodium Thiosulfate Solution (250ml) for
Standardization
5. Weigh out 18.625 grams of sodium thiosulfate
6. Add the sodium thiosulfate into a 250ml volumetric flask and fill it with
distilled water with a dropper until it reaches the line (see Figure 0.1)
7. Shake the volumetric flask
Standardization of Sodium Thiosulfate
8. Attach the burette to the retort stand with a clamp
9. Fill the burette up with the sodium thiosulfate solution
10.Use the pipette to draw 20ml of iodine solution into a conical flask

11.Begin titrating the sodium thiosulfate solution into the flask

12.Add 3 drops of starch solution when the iodine solution changes colour
into urine yellow
13.End titration when iodine solution turns colourless
14.Measure amount of sodium thiosulfate used, looking at where the
meniscus touches the line
15.Repeat steps 10 to 14, washing the conical flasks with distilled water when
necessary
Determining Vitamin C Concentration in Orange Juice
1. Weigh out 0.18grams of potassium iodate, 2.1 grams of potassium iodide
and 5ml of sulphuric acid
2. Add the potassium iodate into the 250ml volumetric flask and fill it up with
distilled water until the line, using a dropper if necessary (see Figure 0.1)
3. Shake the volumetric flask
4. Filter the orange juice into a beaker
5. Attach the burette to the retort stand with a clamp
6. Fill the burette up with the sodium thiosulfate solution
7. Use the 10ml and 20ml pipette to draw 30ml of iodate solution into the
conical flask
8. Use the 20ml pipette to draw the orange juice into the conical flask
9. Add the potassium iodide and sulphuric acid (see Figure 0.2)
10.Begin titrating the sodium thiosulfate solution into the flask
11.Add 3 drops of starch solution when the orange juice solution changes
colour into light green (see Figures 0.3, 0.4 and 0.5)
12.End titration when iodine solution turns milky blue (see Figure 0.6)
13.Measure amount of sodium thiosulfate used, looking at where the
meniscus touches the line
14.Repeat steps 1 to 13, washing the conical flasks with distilled water when
necessary
Note: See logbook for modifications to the method

Figure 0.1 Adding distilled water into

the iodine solution

Figure 0.3 Midpoint of Titration

Figure 0.2 General setup of

experiment

Figure 0.4 Adding of Starch Solution

Figure 0.5 Midpoint of Titration after

Starch added

Figure 0.6 Endpoint of Titration

Risk Assessment
-

Sulphuric acid is highly corrosive and any contact with skin must be
washed with water
Iodine solution is slightly toxic and an irritant, and any contact must be
washed with water
Glassware can be easily broken and are extremely sharp. Report all broken
equipment to the teacher
Hot surfaces are used for certain parts of the experiment, and can cause
burns if it contacts skin

Results
Experiment 1: Effects of exposure on Vitamin C content in orange juice stored in
fridge
Figure 1.1 Vitamin C content and amount of sodium thiosulfate required in
different days of exposure
Da
y

Volume of Sodium
Thiosulfate Required (litres)

Vitamin C Content
(mg/100ml)

0
4
5
7
11
12

0.00140
0.00157
0.00177
0.00223
0.00247
0.00203

162.0304
89.29
67.01
77.62
57.55
94.36

Absolute
Uncertainty
(mg/100ml)
39.50
57.24
62.43
59.96
64.63
56.07

Figure 1.2 Graph of average Vitamin C content by the number of days exposed

Vitamin C Content by Day

180
160
140
120
Vitamin C content (mg/100ml)

100

R = 0.37

80

Vitamin C Content
(mg/100ml)
Linear (Vitamin C Content
(mg/100ml))

60
40
20
0
0 4 5 7 11 12
Day

As seen in Figure 1.2, there is a general decrease of Vitamin C content in orange

juice as the number of days stored increases, aligning with Hypothesis 1 that if
the time of exposure increases, then the Vitamin C content will decrease because
the ascorbic acid will oxidize in presence of oxygen in the air, producing
dehydroascorbic acid. However, an anomaly is seen on day 12, where there is a
drastic increase of Vitamin C content. This is likely due to experimental error and
should not be used. Therefore, to truly show the relationship between Vitamin C
content and time of exposure, another graph without the results on Day 12 must
be plotted.

Figure 1.3 Modified graph of average Vitamin C content by the number of days
exposed

Vitamin C Content by Day

Vitamin C Content (mg/100ml)

180
160
140
120
100
80
60
40
20
0

R = 0.7

Vitamin C Content (mg/100ml)

Linear (Vitamin C Content
(mg/100ml))

7 11

Day

Once the anomaly has been cut, a better trend line can be drawn (R increases)
and a definite relationship is isolated.
Experiment 2: Effects of exposure on Vitamin C content in orange juice stored in
cupboard
Figure 2.1 Vitamin C content and amount of sodium thiosulfate required in
different days of exposure
Da
y

Volume of Sodium
Thiosulfate Required (litres)

Vitamin C Content
(mg/100ml)

0
4
5
7
11
12

0.00140
0.00158
0.00202
0.00273
0.00267
0.00250

162.0304
88.17
39.16
35.79
40.82
55.04

Absolute
Uncertainty
(mg/100ml)
39.50
57.50
68.9
69.69
68.52
65.21

Vitamin C Content by Day

Vitamin C Content (mg/100ml)

180
160
140
120
100
80
60
40
20
0

R = 0.55

Vitamin C Content
(mg/100ml)
Linear (Vitamin C Content
(mg/100ml))

Day

Figure 2.2 Graph of average Vitamin C content by the number of days exposed
As seen in Figure 2.2, Experiment 2 followed a similar trend to Experiment 1 (ie.
Figure 1.2), as there is a gradual decrease in Vitamin C content as the time of
exposure increases, affirming Hypothesis 1. There is an increase by the end of
the graph which is due to experimental error.
Experiment 3: Effects of temperature on Vitamin C content in orange juice
Figure 3.1 Vitamin C content and amount of sodium thiosulfate required in
different temperatures
Temperat
ure (C)
5
10
15
20
25
30
35
40
45
50

Volume of Sodium
Thiosulfate Required
(litres)
0.00130
0.00130
0.00140
0.00145
0.00135
0.00150
0.00170
0.00165
0.00175
0.00175

Vitamin C Content
(mg/100ml)
172.60
172.60
165.55
162.03
169.08
158.51
144.42
147.94
140.90
140.90

Absolute
Uncertainty
(mg/100ml)
37.87
37.87
39.50
40.32
38.69
41.14
44.42
43.60
45.34
45.34

Figure 3.2 Graph of average Vitamin C content by temperature

Vitamin C Content by Temperature

180
170

R = 0.88

160
VItamin C Content (mg/100ml)

150
140

Vitamin C Content (mg/100ml)

Linear (Vitamin C Content
(mg/100ml))

130
120

Temperature (C)

As seen in Figure 3.2, there is a definite relationship seen between the

temperature of the orange juice and Vitamin C content. As the temperature of
the juice increases, the Vitamin C content will decrease in a constant fashion.
This confirms Hypothesis 2: If the temperature, the Vitamin C content will
therefore decrease, since the heat will break the bonds, causing it to denature.
There is a large anomaly on 30C, which is due to external errors, but doesnt
affect the general trend of the graph.
Others

Vitamin C Content by Days

180
160
Fridge Vitamin C Content
(mg/100ml)

140
120

Linear (Fridge Vitamin C

Content (mg/100ml))

100
Vitamin C Content (mg/100ml)

80

Cupboard Vitamin C Content

(mg/100ml)

60

Linear (Cupboard Vitamin C

Content (mg/100ml))

40
20
0
0 4 5 7 11 12
Day

Figure 4.1 Graph of average Vitamin C content by the number of days exposed
(fridge and cupboard)