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Investigation on two types of orange juice, freshly squeezed and processed, which one has higher vitamin c concentration
Researcher question:
Research Question 1
From the two types of juice, Processed and freshly squeezed orange juice, which one has the higher vitamin C concentration?

Research Question 2
Will storing the Orange juice in a fridge or room temperature affect its overall vitamin C concentration? What’s the best option?

Introduction
Figure 1

Vitamin C structural formula

The aim of the experiment was to determine which type of orange juice processed or freshly squeezed had the higher vitamin c concentration. The second aim was being able to find out whether or not temperature effected the vitamin c concentration between the two types of juice. This experiment was important in determining how much vitamin c is in each juice, the effect temperature has on vitamin c and from the temperature determine which type of juice, freshly squeezed or processed had the higher vitamin concentration. Vitamin C, also known as, ascorbic acid is a water-soluble nutrient found in citrus fruits, example orange (figure 1). “Vitamin C is important for the body’s health because it helps maintain healthy cartilage and bones as well as healthy connective tissue,” (Zelman).

Temperature is a dependent variable and it is the measure of intensity of heat present in the particles of a substance. Temperature is important for this experiment because it has an effect on the changing concentration of vitamin C. “Since the vitamin is an acid, it is reasonably stable in acidic solutions, but in neutral or basic solutions it is easily and rapidly oxidized by dissolved oxygen,” (Oregon State University, n.d.). Therefore, factors like temperature and oxygen can oxidize the vitamin c compound and degrade its overall vitamin C concentration. “Vitamin C is water-soluble as such easily leached into the water and then degraded by heat.” (Imran, 2013) Heat can make vitamin C less stable by breaking down its compound through thermal expansion. “Thermal expansion is the tendency of matter to change in shape, area, and volume in response to a change in temperature.”(Elert, 2018) When temperature changes the overall volume of vitamin c concentration starts to change as well. For that reason, as temperature goes up vitamin c, concentration will go down (fig 2). An experiment done by the department of chemistry in Nigeria provides justification to the point as their results showed a 10.3 % decrease in vitamin c concentration in orange juice from a 50% increase in temperature. Ideally, vitamin c should be kept in a fridge because of its low temperature, the concentration will not be degraded by heat and therefore the vitamin c concentration will stay the same. It is better to keep the juice in a fridge with an average temperature of “1.6 degrees Celsius,” (Miller, 2013) which is a relatively low temperature, as oppose to leaving it at room temperature, which has varying temperature depending on the room itself. Vitamin C oxidizes when it comes into contact with air, Explain in lots of detail according to Elizabeth Kunkel, a faculty member of the Food Science and Human Nutrition Department at Clemson University. “Oxidation is a chemical change that results in the breakdown of the vitamin.”(Hunter, n.d.) Therefore when leaving Vitamin C at room temperature ensure that it should be covered so that, “the amount of oxygen the juice comes into contact with is reduced.”(Ipatenco, n.d.)

Between the two types of orange juice freshly squeezed and processed, freshly squeezed will have higher vitamin c concentration. Almost all store-bought juices are pasteurized. “Pasteurized juices have been heated to high temperatures for a short time in order to kill any bacteria or other microorganisms that may be present.” (Nichols, 2014) Freshly squeezed orange may contain harmful bacteria because it has not been pasteurized. Even though some manufacturers may add vitamin c after pasteurization, according to a science fair project done by Lindsay M. Harrison in 2008, the benefits of vitamin c are best found in a diet including fresh-squeezed orange juice. An experiment done by the University of Lahore showed that vitamin c in oranges had a much higher vitamin C concentration at 25 degrees, 50 degrees and at 55 degrees. Since heating destroys vitamin c by breaking its compound through thermal expansion and processed orange juice is being pasteurized, freshly squeezed orange juice will have higher concentration because in the process of being heated, processed orange juice will lose its overall vitamin c concentration due to thermal expansion. Since freshly squeezed orange juice is not being heated, its vitamin c concentration will remain the same and not degrade. However, concentration may be effected by other factors including the exposure to air because it results in the breakdown of the vitamin.
This reaction oxidizes ascorbic acid C6H8O6 to form dehydroascorbic acid while iodine is reduced to iodide ions.” Ascorbic acid+I2 ??2I?^-+dehydroascorbic acid The substances used in this redox titration were iodine and starch. The purpose for using Iodine and starch was because iodine reacts easily with starch to change color. “Amylose an enzyme in starch is responsible for the formation of a deep blue color in the presence of iodine when the iodine molecule slips inside of the amylose coil.” (Odette, 2016) (Libre Texts, 2017) Iodine does not dissolve in water easy because an “intermolecular bond in water is not very strong.” (Elmhurst College) To fix this problem iodine I2 reacts with I? iodide. “(University of Canterbury, 2018) Due to the iodine vitamin c reaction iodine will keep getting reduced to iodine ions while the ascorbic acid is oxidized to dehydroascorbic acid however when vitamin c runs out, the solution will react with starch and therefore the endpoint will be a black blue complex. The two types of orange juices reacting with the solution are processed and freshly squeezed. Both must react with the iodine starch solution in order for the groups completing the experiments to figure out which one has the higher vitamin c concentration.

1st Hypothesis: If the temperature increases, the vitamin c concentration decreases because
vitamin c is water soluble therefore its easily leached into the air and then degrade by heat. However since orange juice is in a fridge due to its low temperature the vitamin c concentration will not degrade because of the low temperature as appose to being kept on a table at room temperature there is a chance that temperature can go up therefore decreasing the overall vitamin c concentration.

2nd Hypothesis: Freshly squeezed orange juice will have a higher vitamin c concentration then processed. Once we have checked the concentration of both types of juice vitamin c and freshly squeezed, freshly squeezed will have higher concentration. “Processed orange juice is not natural and requires to be pasteurized which involves heating the juice to high temperature.” (Dubois) According to Nutrition Data Tables, one freshly squeezed orange juice has vitamin C over 200% of the Recommended Dietary Intake.

Materials
250 mL conical flask
250 mL beaker
100 mL beaker
Burette
Burette clamp
Small cooking knife
Oranges
Glass funnel
Glass pipette
Orange juice
Orange juice squeezer
Pipette filler
Retort stand
Glass measuring cylinder
Distilled water
Strainer
0.01 M iodine phosphate
5 % starch indicator

Method – Processed Orange juice
Used a pipette to take up 20 ml of aliquot solution and transferred it into a 250 ml conical flask.
Transferred 10 drops of 5 % starch indicator in the conical flask.
Used a measuring cylinder to measure 150 ml of distilled water and transferred it into the conical flask with the 10 drops of starch indicator.
Putted the 250 ml flask under the burette and poured in 0.01 M of iodine solution until a blue-black color appeared.
Repeated these steps for each of the processed orange juice experiments

Method – Fresh Orange juice
Slashed 10 oranges and then used an orange juice squeezer to squeeze oranges into 100 mL beaker
Poured the orange juice through a strainer and into another 100 mL or 250 mL beaker, ensuring no pulps are in the beaker.
Used a pipette to take up 20 ml of aliquot solution and dropped it into a 250 ml conical flask
Released in 10 drops of 5% starch indicator.
Used a measuring cylinder to measure 150ml of distilled water and transferred it into the conical flask with the 10 drops of starch indicator.
Putted the 250 ml flask under the burette and start pouring in 0.01 M of iodine solution until a blue-black color appeared.
Repeated these steps for each of the freshly squeezed orange juice experiments

Risk Management table
POTENTIAL RISK MANAGEMENT OF THE RISK
Potassium iodine, irritates eyes and skin Handle carefully and seal the lid tight when not in use. Immediately clean with tissue if spilt.
pipette, beaker, funnel, can break and cause cuts Place on the centre of table and while using it don’t fumble it. Don’t have too many things on the table. If glass shatters sweep it up with a broom
Retort stand quite heavy when picking it up from the shelve top. It can slip out of your hands and hurt anyone underneath it. Ensure if you can reach it, you have someone tall grab it for you. Carefully grab it and don’t fumble it in your hands. Use both hands when possible.
Slipping on the floor, wet floor A newspaper should be kept on the table and below the table to ensure water doesn’t go on the floor
Carrying too much equipment, dropping it and creating a mess Take turns in carrying equipment and ensure not to carry more than you can sustain. More than one person from each group should go to get equipment.

Calculation
Experiment 1 calculations did not count because we changed our variable from time to temperature
Freshly Squeezed Average:21.135
n=c ×v
n=0.005 ×0.021 L=10.5 ×?10?^(-5) moles
Ascorbic acid+ I_2 ?2I^-+dehydroascorbic acid
1 C6H806 1 I2
1:1 ratio between C6H8O6 and I2 therefore no. of moles in 20 ml is 10.5 x 10-5
Concentration in 100 ml of ascorbic acid= c= n/v
(10.5 ×?10?^(-5))/(0.1) = 0.00105

Processed Orange Average?15.95
n=c ×v=0.005 ×0.015
=75 × ?10?^(-6)
1 C6H8O8+1 I_(2 )
1:1 ratio therefore no. of moles in 20 ml is 75 x 10 -6 moles
Concentration in 100 ml of ascorbic acid =c= n/v
= (75 × ?10?^(-6))/(0.1) = 0.00075

Experiment 2 Calculations
Fridge Processed Orange Juice
Average: 14.13
n = c x v
n=0.005 ×0.01413 L =7.065 × ?10?^(-5) moles
1C6H8O6 + 1I2
1:1 ratio therefore no of moles in 20ml is 7.065 x 10-5 moles
Concentration in 100ml of ascorbic acid = c = n/v
= (7.065 ×?10?^(-5))/(0.1) =7.065 × ?10?^(-4)
Room temperature Processed Orange Juice
Average: 12.8
n = c x v – 0.005 x 0.0128
=6.4 × ?10?^(-5)
1C6H8O6 + 1 I2
1:1 ratio therefore no. of moles in 20ml in Vitamin c is 6.4 x 10-5
Concentration in 100ml of ascorbic acid = c = n/v
= (6.4 ×?10?^(-6))/(0.1)=6.4 × ?10?^(-4)

Experiment 3 Calculations
Fridge POJ
Average : 13
n = c x v
= 0.005 x 0.013
= 6.5 x 10-5 moles
1C6H806 + 1 I2
1:1 ratio therefore no. of moles in Vitamin C in 20ml is 6.5 x 10-5 moles
Concentration in 100ml of ascorbic acid = c = n/v
(6.5 ×?10?^(-5))/(0.1)= ?6.5×10?^(-4)

Room temperature POJ
Average: 43.2
n = c x v
= 0.005 x 0.0432
= 2.16 x 10-4 moles
1:1 ratio therefore no. of moles in Vitamin C in 20ml is 2.16 x 10-4 moles
Concentration in 100ml of ascorbic acid = c = n/v
= = (2.16 ×?10?^(-4))/(0.1)=?2.16 ×10?^(-3)
Experiment 4 Calculations
Fridge FOJ
Average: 27
n = c x v
= 0.005 x 0.027
= 1.35 x 10-4
1C6H8O6 + 1 I2
1:1 ratio therefore no. of moles in Vitamin C in 20 ml is 1.35 x 10-4 moles
Concentration in 100ml of ascorbic acid = c = n/v
(1.35 × ?10?^(-4))/(0.1 )= ?1.35 ×10?^(-3)

Room temperature FOJ
Average: 24
n = c x v
= 0.005 x 0.024
= 1.2 x 10-4
1C6H8O6 + 1 I2
1:1 ratio therefore no. of moles in Vitamin C in 20 ml is1.2 x 10-4 moles
Concentration in 100ml of ascorbic acid = c = n/v
(1.2 ×?10?^(-4))/(0.1) = ?1.2 ×10?^(-3)

Experiment 5
Fridge POJ
Average: 15.125
n = c x v
= 0.005 x 0.015125
= 7.5625 x 10-5
1C6H8O6 + 1 I2
1:1 ratio therefore no. of moles in Vitamin C in 20 ml is 7.5625 x 10-5
Concentration in 100ml of ascorbic acid = c = n/v
(7.5625×?10?^(-5))/(0.1)=7.5625 × ?10?^(-4)

Room temperature POJ
Average: 13.475
n = c x v
= 0.005 x 0.013475
= 6.7375 x 10-5
1C6H8O6 + 1 I2
1:1 ratio therefore no. of moles in Vitamin C in 20 ml is 6.7375 x 10-5
Concentration in 100ml of ascorbic acid = c = n/v
(6.7375 × ?10?^(-5))/(0.1)= ?6.7375 ×10?^(-4)
Results

Discussion
A negative to positive trend can be seen by the initial and final results in experiments 2, 3 and 5. The initial and final values always start from low and gradually move up, except for experiment 4 where we had very odd results because the experiment was not done carefully thus expanding the iodine starch reaction and taking it longer than necessary to even see a blink of a colour change. In Trials 1, 2 and 3 the initial values for freshly squeezed orange juice would be at 27.8, 1.2 and 26, whereas the final values would go up to about 51.6, 25.3 and 49.4 in trials 1, 2 and 3. A potential reason for this could be the lack of drops of starch indicator released into the conical flask. For the final volumes and opposite trend to the initial and final results can be seen with this time the trend being positive to negative in experiments 2,3,4and 5. The final volume values would start high up and gradually move down. However, in experiment 4 freshly squeezed orange juice at fridge temperature at a certain point in trial 2 the value of final volume sparks up near its original value. A reason for this occurrence could be that at trial 2 the group working on the experiment had to do it two times, because the iodine phosphate in the burette had ran out. The limitation for the research include better equipment such as the pipette filler because it took too long to accurately fill the pipette with orange juice without any bubbles appearing and some pipette fillers didn’t work at all. The orange juice squeezer was not very sharp, it took quite long to squeeze the oranges, a much sharper, resistant squeezer, and a stronger pipette filler would have made the experiment a lot speedier. In addition, a main setback for the group was that the 200 ml cylinders were unclear and this meant that the group had to use the 100ml glass cylinders and fill it with distilled water two times as oppose to one. A big limitation for this experiment was that the group hadn’t chosen their variables thoughtfully thus resulting in a wasted experiment, experiment 1 and also wasted efforts put into constructing an introduction with the wrong variables. Further research on the vitamin c concentration and the effect temperature has on the concentration of processed and freshly squeezed juice would have been great to get more detail about the topics. Since errors were made because of insufficient research like if temperature goes up, vitamin c concentration will also go up; in previous drafts, this mistake took up a lot of time and had to be fixed. Another crucial research mistake was on vitamin c itself and the structure of its bond. In previous draft, explaining what vitamin c is and how it works wasted a lot of time and effort because it was not chemistry related and did not link back to the original topic/research questions. This created an assignment on vitamin c and excluding other important aspects like talking about temperature and or why processed orange juice is better than freshly and vice versa.
Issue Modification
Oxidisation occurred in beakers The group had to expose the amount of oxygen in beakers therefore the beakers were sealed off with saran wraps
Lack of time The group started doing two titrations with 2 members from the group in each titration

Conclusion
The aim of this redox titration was to figure out which juice, processed or freshly squeezed had the higher vitamin c concentration. The second aim was to find out whether or not temperature affected vitamin c concentration. While doing research, an accurate hypothesis was created which proved that freshly squeezed orange juice has a higher concentration and temperature in fact does affect vitamin c concentration. The research well and truly supported the hypothesis and together the group were able to successfully complete the experiment efficiently and appropriately with only a few setbacks like materials, variable change and insufficient research.

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