Practical Chemistry Investigation

Achievement Standard 3.1

1. Purpose To investigate the degradation of Vitamin C in: 1. Store bought orange juice 2. Store bought lemon juice 3. Oranges 4. Lemons over time in different storage conditions and to obtain whether or not Vitamin C is influenced by juice being previously squeezed and bottled. 2. Background Information Vitamin C (ascorbic acid) is a colourless crystalline water soluble vitamin found in particularly in citrus fruits and green vegetables. Though it is know Vitamin C plays essential roles within the body, the full extent of its properties are still unknown. It plays an essential role in metabolic processes, and the absence of Vitamin C in ones diet relates to various sicknesses, most notably scurvy. Vitamin C was first recognised as an essential vitamin in 1947 when James Lind discovered that oranges cured scurvy. This observation was proven in 1930 when two scientists proved that ascorbic acid was responsible in preventing the disease, and held importance in many bodily functions. Most species are able to convert glucose to vitamin C using the specific enzyme gulonolactone oxidase. Humans do not have this specific enzyme present to make Vitamin C, and so must obtain it via food sources containing Vitamin C on a daily basis. Vitamin C (ascorbic acid) is a water soluble compound so is easily absorbed, but cannot be stored within the body because of this. The recommended daily intake of Vitamin C is between 100- 120mg per day. The degradation of Vitamin C is therefore important to consumers as it can be significantly reduced by exposure to air or sunlight. Current research shows that Vitamin C may prevent the development of cancer; however this is not yet proven. It is known that Vitamin C is essential for; the absorption of iron, healing, the formation of bone, teeth, collagen and intercellular material. Neurons within the brain are regulated by Vitamin C and it is important in the formation and repair of the nervous system. It is also vital in aiding metabolic activities in the formation of amino acids, and in aiding cellular respiration. The institution of Research at Otago University has recently released new found information on the oxidation rates of Vitamin C. They are now able to test the amount of Vitamin C present before any exposure with air and have found that the concentration of the vitamin is much higher than previously thought. Simply cutting into an orange exposes it to air and their research shows that this is all it takes to significantly reduce Vitamin C present. Constraints of the school laboratory make it impossible to carry out this experiment, though the idea of immediate rapid oxidation is of considerable importance to the value of consuming Vitamin C containing products. Vitamin C is an antioxidant, protecting tissues and cells against the damage of oxidation. Antioxidants are able to aid the treatment of diseases, as well as retaining muscle development, explaining why Vitamin C deficiency leads to muscle weakness. Ascorbic acid is a labile molecule as it is lost from foods during processing or cooking and is readily destroyed by heat and light, despite its ability to preserve foods as it is a reducing agent.. Due to this fact I am interested to compare the effects that bottling/ processing has on the amount of Vitamin C in juice, as well as degradation rates in the presence of sunlight. Vitamin C (C6H8O6) is a reducing agent and so is easily oxidised in the air. In previously squeezed and bottled juices ascorbic acid is oxidised to dehydroascorbic acid (C6H804) by loosing 2 oxygen. Trapped air within juice containers leads to this gradual breakdown of Vitamin C over time. The degradation of vitamin

C occurs as air is constantly let into the bottles every time it is opened. Juice manufactures compensate for this loss by recording less Vitamin C amounts in the nutritional information than is actually present within the juice. Loss of Vitamin C is also greatly reduced by the addition of preservatives. Simply Squeezed orange and lemon Juice contain preservative 202, potassium sorbate which is the sodium salt of sorbic acid (preservative 200). It plays a similar role in the preservation of food to Sorbic acid, however is more soluble so is suitable in the preservation of Vitamin C in juice. Sulfur dioxide is present in many preservatives as it is a reluctant, oxidising to sulphate ions in aqueous solutions in replacement of Vitamin C, hence reducing degradation. Sulfate ions are antioxidants and help to prevent the oxidation of Vitamin C. Stabilisers can also be added to juice to inhibit a chemical reaction (negative catalyst), in this case the oxidation of Vitamin C. Buffers resist changes in the pH, and so can retain the amount of Vitamin C (ascorbic acid) in juice. Buffer solutions may be made up to maintain a specific pH value, so retaining ascorbic acid. They are made up of a weak acid and a salt that supplies the conjugate base of the weak acid. In my investigation I would like to compare the amount of Vitamin C in bottled Simply Squeezed Orange Juice and lemon juice to that of picked oranges and lemons. Simply Squeezed Orange juice contains the preservative 202, however no buffer solutions or stabilisers to prevent the degradation of Vitamin C. Simply Squeezed orange juice contains; squeezed orange juice, imported reconstituted orange juice, sucrose, preservative 202 and Vitamin C 300. It claims to have 350 mg/L. Simply Squeezed Lemon Juice contains; squeezed lemon juice, imported reconstituted lemon juice concentrate and preservative 202. It does not claim to contain any Vitamin C. According to http://en.wikipedia.org/wiki/Vitamin_C oranges contain around 500mg/L. According to http://en.wikipedia.org/wiki/Vitamin_C lemon contain around 400mg/L. In comparing the initial amount of Vitamin C between bottled and freshly squeezed juice results will show whether or not Vitamin C is affected by being previously squeezed and bottled. There has been some conflict over the labelling of Simply squeezed juice as it is not freshly squeezed juice, containing concentrates and preservatives also. Those criticising the company hold views that this is a health based issue. I would like to determine the amount of Vitamin C present in the juice to obtain the Vitamin C content in order to see if this product has health benefits. I shall also be looking at the degradation of these four samples over a time period of 4 days, comparing the rate of degradation between juice stored in bottles (remembering that bottled juice contains preservative, so Vitamin C loss should be slower) and within freshly squeezed fruit juice on a daily basis. The presence of sunlight greatly increases the rate of vitamin C degradation. Radicals or oxidants are produced in cells by sunlight and are responsible for rapid oxidation as they are very reactive due to the presence of an unpaired electron. I will leave the 4 samples on a bench in full sunlight to record the rate of Vitamin C loss, compared to the loss on Vitamin C when 4 other samples are stored within a fridge (4C ). To find the amount of Vitamin C (ascorbic acid) present within the samples, the method of back titration shall be used. This method is based on the fact that Vitamin C is a reducing agent. Iodine is used as it is reduced by the Vitamin C, in turn the ascorbic acid (C6H806) is oxidised to dehydroascorbic acid.

+ I2

2H+

2I-

The number of moles of Vitamin C can therefore be found by knowing the initial moles of iodine (by titration with sodium thiosulfate), and taking away the moles of iodine which has reacted with sodium thiosulfate in titration.

n(I2) total

n(I2)reacted with thiosulfate in titration = n(Vitamin C)

The technique of excess iodine is used is used- iodate is reacted with iodide to produce a solution of iodine. IO3+ 5I+ 6H+ 3I2 +S4O62-

Vitamin C reduces the equivalent amount of iodine, therefore it reacts in a 1:1 ratio and so the moles of iodine reacted with the Vitamin C equals the moles of Vitamin C found in the added 50mL of juice. The remaining iodine present in the solution is then found by titration with sodium thiosulfate. This process is aided by the addition of starch when the solution lightens. It turns the solution dark blue so a clear colour change/end point can be determined with the further titration of sodium thiosulfate. Colour change can be hard to detect. To ensue the end point has been reached add excess sodium thisulfate, the solution will stay the same colour if the end point of titration was obtained. 2S2O32- + I2 2I+ S4O62-

Method: Back titration Equipment Simply Squeezed Orange juice (used by date, 4th September 07)- brought on the 27th of August. Simply Squeezed Lemon juice (used by date, 23rd October 07)- brought on the 27th of August. 12 Oranges- picked on the 27th of August. 12 Lemons- picked on the 27th of August.

Times 2

0.100 mol/L Na2S2O3.5H2O (Standard Solution)

To make up 2 litres of Na2S2O3.5H2O (sodium thiosulfate): 1. Weigh out 49.64g of Na2S203 as- mass= 248.18 (molar mass) x 0.2 L 2. Dissolve in measured 2 litres of distilled water. 3. Store standard solution in dark bottle to prevent reaction with light. 0.0100 mol/L KIO3 1.0 mol/L H2S04 0.20 mol/L KI 1% starch solution 25 mL pipette 50 mL pipette Conical flask Clamp and stand Burette Gauze Beaker

PART A: Blank titration (to determine the total amount of I2 present.) 1. Pipette 25mL of 0.0100 mol/L of KIO3 into a conical flask. 2. Add 5mL of 1.0 mol/L H2S04. 3. Add 15 mL+ of 0.2 mol/L KI solution (solution will turn dark brown as iodine is formed). 4. Titrate solution in conical flask with standard solution of sodium thiosulfate until solution is pale yellow. 5. At this stage ad 1mL+ of starch indicator (solution will turn dark blue/black). 6. Continue titration with sodium thiosulfate until the blue colour of the starch disappears. 7. Repeat until concordant results are obtained. PART B: Determination of the Amount of Vitamin C present in juice. Pipette 25mL of 0.0100 mol/L of KIO3 into a conical flask. Add 5mL of 1.0 mol/L H2S04. DAY 1 3. Add 15 mL+ of 0.2 mol/L KI solution (solution will turn dark brown as iodine is formed). Titrations 4. Strain approximately 200mL of Simply Squeezed orange juice through the gauze to prevent pulp Carried out on th getting caught in the pipette. Store strained juice in a beaker, cover with lid to minimise oxidation. 27 August 5. Pipette 50mL of strained juice into the conical flask. 6. Titrate immediately against sodium thiosulfate solution until the brown colour of the iodine has lightened. 7. At this stage ad 1mL+ of starch indicator (solution will turn dark blue/black). 8. Continue titration with sodium thiosulfate until the blue colour of the starch disappears. 9. Repeat until concordant results are obtained, record results. 10. Repeat steps 1-9 using Simply Squeezed lemon juice. 11. Squeeze and strain the juice through gauze into beaker from 3 lemons and 3 oranges, repeat steps 19 on these also. Fridge storage- After initial titrations store Simply Squeezed Orange juice (used by date, 4th September 07)- brought on the 27th of August. Simply Squeezed Lemon juice (used by date, 23rd October 07)- brought on the 27th of August. 12 Oranges- picked on the 27th of August. 12 Lemons- picked on the 27th of August.

FRIDGE

Bench storage- After initial titrations store Simply Squeezed Orange juice (used by date, 4th September 07)- brought on the 27th of August. Simply Squeezed Lemon juice (used by date, 23rd October 07)- brought on the 27th of August. 12 Oranges- picked on the 27th of August. 12 Lemons- picked on the 27th of August. 1. 2. DAY 2 TITRATIONS: Juices have been in storage conditions for 24 hours. Repeat previous steps 1- 11 for all juices in both storage conditions. Carry out back titration (steps 1-11) on DAY 3 and DAY 4.

BENCH

Results: PART A: blank titration

titrated sodium thiosulfate 15.2 15.2

average 15.2

15.2

Calculations for Part A: PART A- gives the maximum amount of moles of iodine formed when no Vitamin C is present. Average volume of 0.100mol/L sodium thiosulfate used in blank titration (part A) = 15.2mL n(sodium thiosulfate) = 0.0152L x 0.100mol/L = 1.52x10-3 mol n(Iodine total) = n(sodium thiosulfate) = x 1.52x10-3 = 7.6x10-4 mol Results: PART B- determination of Vitamin C present in juice samples DAY 1Juice
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfate

11.3, 11.4, 11.2 10.4,10.4, 10.4 13.8,13.8, 13.8 12.8, 12.8, 12.8

titrated sodium thiosulfate average (mL)

11.3 10.4 13.8 12.8

DAY 2titrated sodium thiosulfate

11.3,11,3,11. 3 10.4,10.4, 10.4 13.8,13.8, 13.8 12.8, 12.8, 12.8

Fridge storage
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfat e average (mL)

11.3 10.4 13.8 12.8

Bench storage
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfate average (mL)

11.8,11.8,11.8,11. 8 12.5,12.5,12.5 14.1, 14.1, 14.1 14.9, 14.8, 14.8

titrated sodium thiosulfate average (mL)

11.8 12.5 14.1 14.87

DAY 3titrated sodium thiosulfate

11.8, 11.8, 11.8 10.6, 10.5, 10.6 14.2, 14.2, 14.2 13, 13.1, 13.1

Fridge storage
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfat e average (mL)

11.8 10.57 14.2 13.03

Bench storage
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfate average (mL)

12.4, 12.3, 12.2 14.5, 14.4, 14.3 14.7, 14.6. 14.7 15.1, 15.2, 15

titrated sodium thiosulfate average (mL)

12.3 14.4 14.67 15.1

DAY 4titrated sodium thiosulfate

12.2,12.2,12.2 10.7,10.7,10.6 14.6,14.8, 14.7 13.4, 13.3, 13.4

Fridge storage
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfate average (mL)

12.2 10.67 14.6 13.37

Bench storage
Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

titrated sodium thiosulfate average (mL)

13.5, 13.6, 13.5 15.1, 15.1, 15.2 15.1, 15.1, 15.2 15.1, 15.1, 15.1

titrated sodium thiosulfat e average (mL)

13.53 15.16 15.13 15.16

Calculations for Part B: Determination of Vitamin C present The amount of iodine present after titration can be found after it has reacted with the Vitamin C in the juice. n(iodine)total determined in part A = n(iodine)reacted with vitamin C + n(iodine)remaining to react with sodium thiosulfate in titration Therefore Vitamin C present in juice can be obtained. n(Vitamin C) = n(iodine)total determined in part A - n(iodine)remaining to react with sodium thiosulfate in titration

Example of Calculations: PART A n(iodine) total = 7.6x10-4 Vitamin C is a reducing agent so reacts with the iodine (I2) present in solution and reduces it to I-. The remaining I2 is determined by titration with sodium thiosulfate. Average titre volume for DAY 1- Simply squeezed orange juice is 11.3mL n(sodium thiosulfate) = 0.100mol/L x 0.0113L = 1.13x10-3 n(I2)remaining after reaction with Vitamin C = n(sodium thiosulfate) = x 1.13x10-3 = 5.65x10-4 n(iodine)reacted with vitamin C = n(iodine)total determined in part A - n(iodine)remaining to react with sodium thiosulfate in titration = 7.6x10-4mol 5.65x10-4mol = 1.95x10-4mol = n(Vitamin C present in 50mL of juice) Concentration (Vitamin C) = n/V = 1.95x10-4 0.050L = 3.9x10-4mol/L Converting concentration to mg/L = 3.9x10-4mol/L x M(Vitamin C)

= 3.9x10-4mol/L x 176g/mol = 0.6864 g/L = 686 mg/L

Validity of experiment: 1. Remake 1 L of standard solution (sodium thiosulfate). Mass (Na2S203) = 49.64/2 = 24.82g 2. Add 24.82g of Na2S203 crystals to 1 L of distilled water, dissolve. 3. Repeat part A. Results:
titrated sodium thiosulfate (mL) 15.2 15.2 15.2

Average (mL) 15.2

This is the same as the first titration; therefore PART A results must be valid. PART B- Using sample of Simply Squeezed orange juice (newly bought 3rd September) Results:
titrated sodium thiosulfate 11.5, 11.4, 11.3 titrated sodium thiosulfate average (mL) 11.43

Juice Simply Squeezed Orange

The average of 11.43 from newly bought orange juice is very similar to the original titration of 11.3mL. As results are concordant, the method must be valid for determining the amount of Vitamin C present. Results PART B- Vitamin C present in mg per litre

Comparison between Simply Squeezed orange and lemon Juice to freshly squeezed orange and lemon juice, and the comparison between found Vitamin C present and expected results:
Juices Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons Found Vitamin C present (mg/L)
686.4 884.8 246.4 422.4 400

Expected Vitamin C present (mg/L)

350 500

Comparison of Vitamin C in bottled 'Simply Squeezed' orange and lemon juice to freshly squeezed oranges and lemons
Amount of Vitamin C (mg/L) 1000 900 800 700 600 500 400 300 200 100 0 Simply Oranges Simply Lemons Squeezed Squeezed Orange Lemon Juice Samples

Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

This graph shows that previously bottling juice does have an effect on the amount of vitamin C present as freshly squeezed orange/lemon juice has much higher vitamin C content. This is partly due to air trapped within the bottle during processing which oxidises the Vitamin C slightly, despite preservatives, and due to air exposure before preservatives are added.
C p isonof fou dV inCinju sam les to om ar n itam ice p exp ectedr lts esu
Amount of Vitamin C (mg/L) 10 00 90 0 80 0 70 0 60 0 50 0 40 0 30 0 20 0 10 0 0 S ply im S queez ed O range O ranges S ply im S queez ed L on em L ons em

Series1 Series2

Found Vit. C Expecte d Vit. C

ju sa p ice m les

This graph compares the found amount of vitamin C in the juice samples with the expected results (those shown on the bottle under nutritional information and from http://en.wikipedia.org/wiki/Vitamin_C for oranges and lemons. Simply Squeezed lemon juice claimed to hold no Vitamin C however titration showed that 2246 mg/L was present. This is a very small amount of vitamin C, so was not included in nutritional information. The amount of vitamin C in lemons found was very close to the expected result with only a 22mg/L difference. The lemons were picked in August, so were mature in growth. Oranges gave results higher than the expected 500mg/L average for oranges. The results are most likely still accurate as the oranges were freshly picked and not yet ripe. Vitamin C degrades as fruit ripens, and as picked in August the fruit was not yet fully mature, resulting in a high Vitamin C content. Simply squeezed orange juice claimed to have 350 mg/L of Vitamin C, found results were much higher at 686.4 mg/L. Juice companies put significantly reduced figures under nutritional information for Vitamins as Vitamin C amounts vary between oranges, and so the lowest possible amount must be recorded. Vitamin C also degrades slowly within the juice, and so this is also compensated for in accordance with the used by date as the juice must still contain 350mg/L + by this deadline.

The comparison of degradation of Vitamin C over 4 days in Simply Squeezed orange and lemon juice and freshly squeezed orange and lemon juice in fridge storage and bench storage:
Vitamin C present mg/ L (FRIDGE STORAGE) Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons Vitamin C present mg/ L (BENCH STORAGE) Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons Day 1
686.4 884.8 246.4 422.4

Day 2
686.4 884.8 246.4 422.4

Day 3
598.4 814.88 176 381.92

Day 4
528 797.3 88 322.1

Day 1
686.4 884.8 246.4 422.4

Day 2
598.4 475.2 193.6 58.08

Day 3
510.4 140.8 93.28 17.6

Day 4
293.9 7.04 12.32 7.04

Degradation of Vitamin C over 4 days in juice samplesStored in Fridge

1000 900
Amount of Vitamin C (mg/L)

800 700 600 500 400 300 200 100 0 1 2 DAY 3 4

Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

Over a period of 4 days the amount of Vitamin C in the juice samples did not decrease greatly as colder temperatures preserves Vitamin C content for longer time periods. From the results a steeper loss of Vitamin C can be seen between Day 3-4; Simply squeezed orange juice loosing 70.4mg/L, orange loosing 17.58mg/L , Simply squeezed lemon juice loosing 88mg/L and lemons loosing 59.8mg/L. Though this is probable for the fruit, the bottled juice contains preservative and so this higher loss is not as likely. Error in titration or with chemical could have given these results, otherwise a longer exposure to air before titration which led to degradation. This is plausible as air is let into the bottle each time it is opened, trapped within the bottle when closed, it oxidised the Vitamin C.

Degradation of Vitamin C in juice samples over 4 days in juice samples- bench storage
1000 Amount of Vitamin C (mg/L) 900 800 700 600 500 400 300 200 100 0 1 2 DAY 3 4 Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

Juice samples were left on the bench in full sunlight for 4 days. Rapid degradation is shown in this time period as sunlight speeds up the process of oxidation, reducing the Vitamin C present.

C ma iso o V m Cd r d tio o er4d y inj ic o p r n f ita in eg a a n v as u e sa p b eenb ttleda dfr lysq eez j ic a d mles etw o n esh u ed u e n fr g a db c sto a e c n itio s id e n en h r g o d n
Amount of Vitamin C (mg/L) 10 00 90 0 80 0 70 0 60 0 50 0 40 0 30 0 20 0 10 0 0 1 2 DY A 3 4
S ply S u ezed im q e O e (frid e) rang g O es (frid e) rang g S ply S u ezed im q e L m (frid e) e on g L m (frid e) e ons g S ply S u ezed im q e O e (b rang ench) O es (b h) rang enc S ply S u ezed im q e L m (b e on ench) L m (b h) e ons enc

http://activin.com/FoodPres.htm

The orange line on this graph, found on http://activin.com/FoodPres.htm shows the degradation of originally 60 mg/Vitamin C (no preservatives present) over a period of 9 days. It is in accordance with my results as it shows similar deductions in the presence of Vitamin C, supporting results found.

Conclusion: The initial purpose of this investigation was to obtain whether or not previously bottling juice affected the amount of Vitamin C present. It was found that previously bottling juice did have an effect on Vitamin C present. Simply squeezed orange juice had a 198 mg/L less vitamin C to that of freshly squeezed juice. I also compared the degradation of bottled and freshly squeezed juices over time in fridge and bench storage conditions. In general there was an overall decrease in the amount of Vitamin C present over a time period of 4 days. Degradation of Vitamin C in fridge storage was much slower than that of juice left in the sun. While juice samples stored in the fridge only decreased by approximately 100mg/L, juice samples in the sun had a much greater loss with oranges and lemons reduced to 7.04mg/L over 4 days, an almost total loss of Vitamin C content. It was also apparent that bottled juice in sunlight did not lose as much vitamin C as the fruit due to added preservative 202 with orange juice decreasing from 686mglL to 293.4mg/L, a much high amount present compared to an almost complete loss in oranges and lemons. Discussion: Vitamin C is an essential vitamin for humans, and as it must be obtained on a daily basis it is important to ensure that degradation of this vitamin is prevented. This experiment shows the difference in degradation rates between juice kept on the bench, and juice in the fridge. This experiment supports the background information as the presence of sunlight caused rapid reduction in the amount of Vitamin C present. The loss of Vitamin C on bench stored juice was very rapid due to the damage of Vitamin C by radicals, or oxidants which are produced by sunlight in cells. Radicals are molecules with unpaired electrons. When an atom in a single bond receives one electron; this is known as the homolytic fusion of a covalent bond, it forms a radical. This unpaired electron (radical) is very reactive, producing a radical chain reaction. Radicals cause the homolytic fusion of non radical molecules- propagation. Two radicals can then react with each other (known as termination). This creates a rapid exothermic reaction, rapidly decreasing Vitamin C present as it is oxidised.
The total amount of vitamin C lost in Simply Squeezed orange juice after 4 days bench storage
Vitamin C lost Vitamin C retained

The total amount of vitamin C lost in oranges after 4 days bench storage

Vitamin C lost Vitamin C retained

After only one day of leaving the juice samples on the bench, Vitamin C had decreased by 409.6 mg/L, 54% in oranges. This was much greater than degradation in bottled orange juice which only decreased 88mg/L after one day. As found during background research, this is due to the presence of preservative 202 which prevents Vitamin C loss. The experiment supported the background information as Vitamin C was reduced very quickly in the presence of sunlight as oranges and lemons lost 95%+ of their Vitamin C present in only four days.

The total amount of Vitamin C lost in Simply Squeezed lemon juice after 4 days bench storage
Vitamin C lost Vitamin C retained

The amount of vitamin C present in oranges was significantly higher than the expected 500mg/L. As previously explained under graphs, this would be due to the fact that oranges picked were immature (not ripe) and so had a very high concentration of vitamin C The total amount of Vitamin C lost in present. Lemons however showed very close results to lemons after 4 days bench storage what was expected. This is because the lemons were picked at maturity.
Vitamin C lost

Vitamin C found in Simply squeezed orange juice was Vitamin C 198.4mg/L higher than the expected 350mg/L. This was retained possibly because Vitamin C present was titrated immediately after it was purchased and so degradation had no yet occurred. The used by date of the orange juice was for the 7th of September indicating that the juice had been bottled relatively recently. As previously mentioned in the background information it is known that lower than actual Vitamin C amounts do not fall below that which is recorded in the nutritional information. My first attempt at making a standard solution of Sodium thiosulfate did not work as the titrated results came out far too high at 22.8mL (the rest of the class was averaging around 15). This could have been due to contamination of the chemicals or equipment used. On repeating the procedure I obtained coordinate results of 15.2mL. Modifications were made to the original method in order to complete the procedure, and retain the validity of the experiment. I had to alter my original method by adding in a filtration of juices prior to titration. This was achieved by straining juice through fine gauze into a beaker. This was to prevent the problem of pulp getting caught in the pipette, and so made the titrations possible. Sources of error which could have influenced my results include parallax error (Observing measurements at eye level). To remove this error I ensured all measurements were taken at eye level with the meniscus. Juice bubbles in pipetting were sometimes present, which could potentially have altered the position of the meniscus. To reduce the error in each titration I repeated the titration until 3 concordant results (within a range of 0.2mL) were obtained. I then averaged these results to further reduce any error in titrations. If I was to do this experiment again I would add a dilution factor to my juice samples. By diluting juices (using distilled water) to 50% the results would be far more accurate as error in titrations would also be reduced. In repeating this experiment I would be far more careful with the time in which the lids of bottles were removed to get juice, and the time in which freshly squeezed juice was exposed to air. The validity of the experiment could have been affected as I did not have specific times set for how long bottles were opened and exposed to air, nor times set for how long juice samples were removed from storage conditions to carry out the experiment. Juices which were left open for longer periods of time than others would have more air trapped within the bottle. This would lead to increased oxidation of Vitamin C, degenerating the validity of the results/data obtained. In repeating this experiment I would ensure that times were set and recorded for all disturbances from opening and closing bottles, squeezing juice and times out of designated storage conditions. I would also like

to have carried the experiment on for a longer period of time to see the significance that preservatives have on retaining Vitamin C concentrations in Simply Squeezed juice. If further research proves that Vitamin C degrades immediately with exposure to air, set times of air exposure to juice samples would be very important to obtain Validity and consistency in the procedure. The oranges I used in titration were not ripe. A potential improvement to this experiment would be to titrate the juice of only ripe oranges as this would be in accordance with the expected amount of Vitamin C present. It would also be realistic to the experiment as people are more likely to consume ripe oranges. I would ensure the accuracy of the chemicals and would be more aware that these can contribute to error in titrations. There was contamination of these chemicals by students pouring used chemical back into the original containers. There were also problems with the KIO3 as it was exposed to air and oxidised at it turned yellow, this could have affected results. I would also make up 4 solutions of 0.0100 mol/L KIO3, 1.0mol/L H2S04, 0.20 mol/L KI before carrying out any titrations. This would greatly speed up the titration process as 50mL of juice would just have to be added before it was titrated. It would prevent the need to continuously wash out one beaker, and rough and concordant results could be obtained using the 4 prepared solutions. To increase the accuracy of the experiment I could increase the number of titrations repeated for each juice samples. This would limit random error and so give a more accurate average of the results. However due to the amount of time allocated this was not feasible. I could All equipment was washed consistently with distilled water before use and pipettes and burettes were cleaned with the solution they were to contain before use. This was necessary to prevent the contamination of other chemical which could of affected results. I tested the validity of the experiment by repeating it from the beginning, however only retesting one juice sample. The standard solution of sodium thiosulfate was remade, and as the titration result for PART A (15.2ml) was the same as my original titrations for PART A, the solution must have been valid. I bought new Simply Squeezed orange juice and obtained results very similar to those previously obtained in PART B with only a 0.13mL difference. This further supports the validity of the experiment. The degradation of fridge stored juices was unexpectedly high as it was expected that preservative 202 would retain Vitamin C. Bottled orange juice lost 23% of its vitamin C over 4 days, while lemon juice follows a similar degradation trend. Though this loss is very similar between the two juices there should possibly not be this much reduction. This could be due to the slightly oxidised KIO3 used (though it is not certain this has an effect), which could have affected the results. In general degradation of all samples in fridge storage follows a similar decreasing curve, with each loosing around 100mg/L over 4 days. The results for bench storage were expected with all juice samples loosing 95%+ Vitamin C, except Simply Squeezed orange juice (57%), after 4 days. The validity of this part of the experiment was consistent as over the 4 days it was continuously sunny so juices were exposed to the same conditions every day. This difference, between a 98% loss in Vitamin C in Simply Squeezed lemon juice and only 57% loss in Simply Squeezed orange juice is partly due to the originally high amount of Vitamin C present in Simply Squeezed
Degradation of Vitamin C in juice samples over 4 days in juice samples- bench storage
1000 Amount of Vitamin C (mg/L) 900 800 700 600 500 400 300 200 100 0 1 2 DAY 3 4 Simply Squeezed Orange Oranges Simply Squeezed Lemon Lemons

orange juice (686mg/L) before it was exposed to the sun. Bottled orange juice did in actual fact loose the same amount of Vitamin C as bottled lemon juice, with a total loss of 158mg/L. Bottled lemon juice had originally the lowest amount of Vitamin C present at only 246mg/L, and so reached a point where it had very little Vitamin C to loss. This is shown on the graph above as the deduction in the in amount of lemon juice present decreases as the gradient tends towards 0 after day 2. Similarly this occurred in Lemons and oranges also, as they had significantly higher amounts of vitamin C present originally, with the gradient tending towards 0 on day 3. Recent research from the University of Otago has found that Vitamin C is oxidised immediately on contact with air, significantly reducing the concentration of the Vitamin after only a few seconds of exposure. As bottled juices and the fruit juice I squeezed was exposed to the air for significantly longer than a few seconds, Vitamin C could already have reduced considerably before the titration was carried out. This recent research shows that the currently found amount of Vitamin C present in oranges (500mg/L) is actually much higher than expected as scientists are now able to test for Vitamin C without previous exposure to oxidation. I would be interested to follow up on this investigation, however unfortunately it is not possible to carry out this experiment with school facilities. Though there could have been marginal error my end results supported the background information showing a relationship between time and Vitamin C loss, as well as further supporting that the presence of preservatives greatly reduces degradation If I was able to do an experiment on Vitamin C content again I would be interested to test the effects that cooking vegetables has on Vitamin C concentrations, as this supposedly significantly reduces the Vitamins in which the vegetable is eaten to obtain. Overall the results for this experiment came out as expected according to background information. A very clear relationship between time and the amount of vitamin C present in juice was shown. True to the background information as they supported rapid degradation in sunlight, and reduced Vitamin C loss when samples were stored in the fridge. The importance of this experiment was to show just how quickly Vitamin C is lost when exposed to sunlight in comparison to fridge storage and this was very clearly shown in the results. In comparing bottled juice and fruit it was obtained that though fruit contains an originally much higher concentration of Vitamin C, it degrades much more quickly than in juice due to the addition of preservatives in Simply Squeezed products. Overall the results are valid to the extent that a clear relationship is shown and discussed. Though C ma iso o V m Cd r d tio o er4d y inj ic o p r n f ita in eg a a n v as u e Simply squeezed has be criticised sa p b eenb ttleda dfr lysq eez j ic a d mles etw o n esh u ed u e n for its brand name, confrontation fr g a db c sto a e c n itio s id e n en h r g o d n on the bases of health reason is S ply S u e im q e zed not supported as I found Simply 10 00 O ng (frid e ra e g) Squeezed orange juice to contain 90 0 O ng s (frid e) ra e g 80 0 a very high amount of vitamin C S ply S u e im q e zed 70 0 at 686.4mg/L. L on (frid e em g)
Amount of Vitamin C (mg/L) 60 0 50 0
L ons (frid e) em g

Calibration curves obtained S ply S u e im q e zed 40 0 O ng (b ) ra e ench showed the degradation of 30 0 O ng s (b h) ra e enc 20 0 Vitamin C over time, and the 10 0 S ply S u e im q e zed significance of this loss in the L on (b ) em ench 0 presence of sunlight. As shown on L ons (b h) em enc 1 2 3 4 the graph the calibration curves DY A for bench stored samples has a steeper graident- showing rapid degradation of vitamin C compared to the gradual decline of vitamin C when samples were stored in the fridge. In conjunction with background research this investigation enabled the purpose of this

experiment to be carried out, hence showing comparison of storage, bought juice and fruit and the impacts of time to Vitamin C. In comparing bottled and freshly squeezed juice it was obtained that more vitamin C is to be gained from eating fruit. Bottling orange and lemon juice reduced the Vitamin C significantly. Even in comparison to the expected 500mg/L I oranges, the expected 350mg/L is low. However added preservatives in juice retain Vitamin C efficiently, and so are seen as a better source of the vitamin if fruit is not eaten promptly, or is exposed to sunlight- As shown in the results sun exposure can reduce Vitamin C in oranges by 54% in only one day, so even a few hours in the sun would cause significant loss of this vitamin. Bibliography: Books: John Daintith, Oxford Dictionary of Chemistry, Oxford University Press, 2004 M. Clugston and R. Flemming. Advanced Cheminsty, Oxford University Press, 2000 Internet: http://actvin.com/FooodPres.html www.herbs2000.com/vitamins/v_c.htm http://en.wikipedia.org/wiki/Vitamin_C http://www.stuff.co.nz/thepress/4010259a19744.html http://www.stuff.co.nz/stuff/4179024a3600.html www.physicsforums.com/archive/index.php/t-117786.html Interview: National program interviewing Otago University Institution on the latest findings on Vitamin C degradation

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