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INVESTIGATION OF FACTORS AFFECTING VITAMIN C IN LIQUID
Vitamin C is important for the human body because it is needed in the production of collagen to make connective tissue. Vitamin C also helps the body to absorb iron, helps wounds to heal, helps red blood cell formation and helps to fight infections. Some studies say that vitamin C prevents cancer. A lack of vitamin C can cause a disease called scurvy, iron deficiency and poor wound healing. The healthy diet should include high amounts of vitamin C because the human body cannot make it’s own vitamin C. Oranges are an excellent source of vitamin C. (Gordon, 1995-2005, p.1)(Larsen,1997-2005, p. 1)(General Health Encyclopedia,1998, p. 1)(Royston, 2003, p.19) Orange juice and drinks are common in our diet and some people think they are receiving vitamins in any type of orange drink. I conducted two experiments to compare the content of vitamin C in different kinds of orange juice and drinks.
My hypothesis is that freshly squeezed orange juice will have the most vitamin C compared to the other types of orange juice and drinks, because the fruit is picked fresh, and it is not stored, preserved or exposed to oxygen.
Vitamins help to speed up important reactions in the body. Vitamin C, also known as ascorbic acid, is water soluble, meaning it will dissolve in water and is not stored in the body.
We need to get vitamin C from the foods we eat. (Gordon, 1995-2005, p.1) (Silverstein, 1992, p.8) Vitamin C is found in fruits such as oranges, limes, and grapefruit, and vegetables including tomatoes, green peppers, and potatoes. The recommended amount of vitamin C is 60 to 90 milligrams per day. (Gordon, 1995-2005, p. 2 ) People who smoke need more vitamin C in their diet, because they lose 25 mg. of vitamin C every time they smoke a cigarette. People who are stressed, have infections, take antibiotics, drink lots of alcohol or have been injured need more vitamin C in their diet. (Sullivan, 1997, p. 30) Factors that Affect Vitamin C in Orange Juice Vitamin C is sensitive to light, heat and air. (Alpert, 1997-2004, p. 1) Vitamin C is the most easily destroyed vitamin and it is easily harmed during food preparation. This can happen during chopping, exposure to air, cooking, and boiling. (Joanne Larsen, 1995-2003, p. 1and 2)
There are many factors that will affect how much vitamin C is in orange juice. Townsend (1999) summarized some of these factors. For example, freezing preserves vitamin C. Exposure to oxygen will destroy vitamin C. Oranges will have more vitamin C if they are picked when they are less ripe, if they are early maturing like Navel oranges, if they are grown in soil with low levels of nitrogen or if they are exposed to cooler temperatures while growing. How the juice is made will also affect the amount of vitamin C in it. For instance, frozen orange juice is made from a mix of early ripening and late ripening oranges, so it tends to have higher concentrations of vitamin C. The type of container that the juice is stored in can affect vitamin C levels. For instance, enamel lined cans lose more vitamin C than tin cans. Orange juice in glass jars or cardboard cartons have less. Today cartons have oxygen and light barriers to protect the vitamin C and frozen juices are stored in cardboard cans lined with foil to help keep vitamin C. For the best vitamin C levels, orange juice must be stored at cool temperatures with oxygen barriers. (Townsend, 1999, p. 1 and 2)
When all of the ascorbic acid is used up, that is the end point of the reaction. If there is so much vitamin C that all of the iodine in the starch solution is used up, the solution will become clear because of the dehydroascorbic acid. If there is very little vitamin C, less iodine will be used up and more blue colour will remain. (Thomas, 1999, p. 1)
• Vitamin C Indicator Solution (2% Iodine solution, cornstarch and water)
• Medicine droppers, Test tubes, Test tube holder
• Beverages -12 different kinds of orange juices and drinks:
Indicator Solution Mix one tablespoon of cornstarch and water to make a paste, add 250 millilitres of water and boil. Add 10 drops of this solution to 75 millilitres of hot water, stirring. To this solution, add 12 drops of 2% iodine solution. (USDA Agricultural Research Service, 2005, p. 1)
- Experiment One
Add 10 drops of one juice to 5 ml. of indicator solution in a test tube. Stir. Repeat for each beverage. Line the tubes up from lightest to darkest. Record the rank of each beverage from ‘12’ being the lightest (with the most vitamin C) to ‘1’ being the darkest colour (with the least vitamin C). (USDA Agricultural Research Service, 2005, p. 1)
- Experiment Two
Add juice one drop at a time to 5 ml. of indicator solution in the test tube until the indicator solution changes from blue to colourless or until the solution no longer changes colour. This is the endpoint. Repeat for each beverage. Observe and count the numbers of drops of orange beverage added to the indicator to cause it to lose its colour or to stop changing colour. Record the number of drops. (Dipaolo, 2002, p. 1)
Ascorbic acid is one of the important water soluble vitamins. It is essential for collagen, carnitine and neurotransmitters biosynthesis.  Most plants and animals synthesize ascorbic acid for their own requirement. However, apes and humans can not synthesize ascorbic acid due to lack of an enzyme gulonolactone oxidase. Hence, ascorbic acid has to be supplemented mainly through fruits, vegetables and tablets. . The current US recommended daily allowance (RDA) for ascorbic acid ranges between 100–120 mg/ per day for adults. Many health benefits have been attributed to ascorbic acid such as antioxidant, antiatherogenic, anti-carcinogenic, immunomodulator and prevents cold etc. Thus, though ascorbic acid was discovered in 17th century, the exact role of this vitamin/nutraceutical in human biology and health is still a mystery in view of many beneficial claims and controversies . Ascorbic acid is a labile molecule; it may be lost from foods during cooking/processing even though it has the ability to preserve foods by virtue of its reducing property. Syntethis ascorbic acid is available in a wide variety of
supplements, tablets, capsules, chewable tablets, crystalline powder, effervescent tablets and liquid form. L-ascorbic acid (C6H8O6) is the trivial name of Vitamin C. The chemical name is 2-oxo-L-threo-hexono-1, 4-lactone-2, 3-endiol, Fig. 1 . Ascorbic acid being a water soluble compound is easily absorbed but it is not stored in the body. The major metabolites of ascorbic acid in human are dehydroascorbic acid, 2, 3-diketogluconic acid and oxalic acid, Fig. 2. The main route of elimination of ascorbic acid and its metabolites is through urine. It is excreted unchanged when high doses of ascorbic acid are consumed. Ascorbic acid is generally non-toxic but at high doses (2-6g/day) it can cause gastrointestinal disturbances or diarrhea [5, 6].
Keeping in view its importance, the analysis of food products and pharmaceuticals containing this vitamin assumes significance. Such attempts to quantify ascorbic acid in these samples have resulted in a large number of methods: titrimetry, voltametry, fluorometry, potentiometry, kinetic-based chemiluminiscence (CL), flow injection analyses and chromatography [7–9].
For determination of ascorbic acid were used two methods: a titrimetric method with potassium brommat-bromide solution in the acid medium  and a conductometric method  based by the calibration curve which was plotted under the fallowing operative conditions: 4°C, 18°C, 30°C and 40°C, the linearity range is 0,008–0,1N ascorbic acid. The methods have been applied to many samples to determine the Vitamin C quantity at different temperature .
All reagents were of analytical-reagent grade and all solutions were prepared using distilled-deionized water. A stock standard aqueous ascorbic acid solution (0,1N) was prepared from L-ascorbic acid (Merck). Other standard ascorbic acid solutions were obtained by appropriate dilutions of the stock solution. For the titrimetric method, the reagents used have been: Na2S2O3 0,1N, KBrO3-KBr 0,05N, K2Cr2O7 0,1N, H2SO4 1N, H2SO4 1:2, KI, starch indicator 1%.
For conductometric procedure was used a Conductivity Meter WTW, LF 340-A/SET, made in Germany.
- CONDUCTOMETRY PROCEDURE
First was plotted the calibration graph (Fig. 3) of different ascorbic acid concentrations (0,008N; 0,01N; 0,03N; 0,05N; 0,08N respectively 0,1N) at the room temperature by reading it at the conductivity meter (ìS/cm).
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