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Lab MICRO 1
From Juniata College SIM
Vitamin C is a water soluble vitamin that is required daily since it is not synthesized by humans and cannot be stored by the body. Vitamin C is required for cellular respiration, for enzyme function, as a component in collagen formation, and has been reported to increase iron absorption within the body. Dr. Linus Pauling has found that Vitamin C dosages in excess of the recommended daily allowance (RDA) appears to reduce the severity and frequency of the common cold. The RDA of vitamin C in adults is 60 mg per day. The vitamin is nontoxic even at high dosage levels. A dietary deficiency in vitamin C may result in scurvy which is characterized by fatigue, shortness of breath, muscle degeneration, gum bleeding, tooth loss, and even death. Just 10 mg of vitamin C is enough to cure scurvy. Vitamin C, also known as ascorbic acid, is a weak acid found mostly in citrus fruits and leafy green vegetables (broccoli, parsley, and spinach). Apples, grapes, and peaches are low in vitamin C. The method of measuring the amount of vitamin C in fruit juices in this lab utilizes the ease at which vitamin C can be oxidized (forced to lose electrons). Iodine oxidizes + vitamin C. (C6H8O6 + I2 --> 2H + 2I + C6H6O6) After the reaction, excess I2 remaining is detected by a starch indicator solution that turns blue - black in the presence of iodine. Vitamin C has a long storage life; however, it rapidly decomposes when in solution because it is oxidized by atmospheric oxygen. The decomposition is sped up by heat, light, bases, oxidative enzymes, and traces of iron and copper. In the diet, vitamin C concentrations are greatly reduced by cooking foods in water and then draining off the water.
The amount of vitamin C in various citrus juices is measured using an I2 / KI solution and a starch indicator.
Gannon University SIM
so gloves and aprons are recommended. Dangerous chemicals may dissolve into the juices after exposure. Record the number of drops of iodine used for each trial in the table on the data sheet. Repeat step 4 for as many juices as your instructor requests. Gannon University SIM Micro 1-2 . Iodine stains hands and clothes.Vitamin C in Citrus Juices EQUIPMENT/MATERIALS 1% starch solution iodine solution fruit juices 24 well spot plate 1. 2. Add two drops of the starch solution to each well containing the vitamin C solution.0 mg/mL Vitamin C solution medicine droppers for each juice & solution SAFETY • • • Always wear safety glasses in the lab. After each drop is added. Next. Using a medicine dropper. 5. drop iodine solution into each well until the solution turns a purple to dark blue color for at least 15 seconds. 3. Place 5 drops of a fruit juice in place of the vitamin C standard in each well of the next row (6 wells) and follow steps 2 and 3. Each well used will act as a different trial. place 5 drops of the standard vitamin C solution in 6 wells (one row) of the spot plate. gently shake the spot plate to mix the iodine solution with the solution. PROCEDURE 1. 4. Do not drink the fruit juices after they have entered the lab.
Fill in the appropriate spaces on the table. Conc Vit C Standard Juice 1 Juice 2 Juice 3 Juice 4 CALCULATIONS 1.Vitamin C in Citrus Juices Name_______________________________ Name_______________________________ Period______________________________ Date________________________________ MICROANALYSIS OF VITAMIN C IN CITRIC JUICES DATA TABLE Number of drops of iodine solution used in each trial 1 2 3 4 5 6 Ave. Calculate the average number of drops of iodine solution required to react with the Vitamin C standard. Gannon University SIM Micro 1-3 . Repeat for each of the fruit juices.
List two nutritional sources of vitamin C other than citrus fruits. 3. Which fruit juice contained the largest concentration of vitamin C? 2. QUESTIONS 1. 4. How many ounces of each sample provide the RDA for Vitamin C? Gannon University SIM Micro 1-4 .Vitamin C in Citrus Juices 2. The concentration of the vitamin C standard is 1. Calculate the concentration of vitamin C in each of the fruit juices by comparing the amount of iodine required for vitamin C and the amount needed for the fruit juice. Using the RDA for vitamin C listed in the introduction section of the experiment.0 mg / mL. calculate the mL of each sample required to supply the RDA for vitamin C.
Then.6 g KI in 250 mL distilled H2O. V: The van can supply materials. 1% starch indicator: Add 1 g soluble starch in 100 mL distilled H2O. Gannon University SIM Micro 1-5 . Prepare fresh weekly.Vitamin C in Citrus Juices MICROANALYSIS OF VITAMIN C IN CITRUS JUICES TEACHER NOTES Time: 30.6 g I2 in 25 mL ethanol. add solution A to solution B and dilute to 500 mL with deionized water. Iodine solution: First make solution A by dissolving 0. T: Prepare the following solutions: Standard Vitamin C: To make the 1 mg/mL standard. make solution B by dissolving 0. including solutions. dissolve 100 mg of vitamin C in 100 mL of water (or use one 100 mg tablet of vitamin C). 1 mL of laundry starch diluted to 100 mL with water can be substituted if soluble starch is not available. as requested. Next.45 minutes Preparations: Time: 30 minutes T: Make available a set of the equipment and materials for each lab group. heat to dissolve and filter. This solution should be freshly prepared and refigerated.
Considerations: The use of another indicator. List two nutritional sources of vitamin C other than citrus fruits. throughout the lab. The starch endpoint color fades with time in the fruit juices. if possible. Using the RDA for vitamin C listed in the introduction section of the experiment. is discussed in Chemtrek by Steven Thompson. Also. fruit juices should be bought on day of lab or 1 or 2 days before and refrigerated. Allyn and Bacon. Variations of the lab can include comparing temperature effects on vitamin C concentration.Vitamin C in Citrus Juices Answers to Questions: 1. Gannon University SIM Micro 1-6 . etc. 1990. storage time vs. Indophenol. Drops were easier to control using medicine droppers. The amount of the iodine solution needed to produce an endpoint decreased when running 8 samples of a juice (or Vitamin C) suggesting the Vitamin C was being oxidized by the air. The vitamin C solution should be freshly prepared and refrigerated. Answers will vary but may include fruit juices green leafy vegetables. Answers will vary depending on data collected. No difference in endpoints was observed using either the 1% soluble starch solution or a 1% laundry starch solution. calculate the mL of juice 1 required to supply the RDA for vitamin C. since the vitamin C concentration will significantly change throughout the day if kept at room temperature. rather than Beral pipets. Which fruit juice contained the largest concentration of vitamin C? Answers will vary depending on data collected. the color is stable with the Vitamin C solution. 3. The juices should be refrigerated. 2. vitamin C concentration.
21 13 9 Concentration of Vit C in OJ = # drops iodine used for OJ X Concentration of Standard # drops iodine used for Stand.0 mg/mL 21 drops Concentration of Vit C in OJ = 0.Vitamin C in Citrus Juices Sample Data/Calculations: DROPS IODINE SOLUTION Sample Vit C Orange Juice Grapefruit Juice 1 24 14 10 2 22 14 9 3 20 15 10 4 19 14 9 5 20 13 8 6 19 12 8 7 21 12 9 8 24 12 9 Ave.62 mg/mL Microscale Techniques Gannon University SIM Micro 1-7 . Concentration of Vit C in OJ = 13 drops X 1.
masking tape ** Note .500mL bottle of deionized water to dilute ascorbic acid 1-wash bottle of DDI water to dilute the ascorbic acid 2. 1 Demo 9-15mL glass dropper bottles labeled starch solution. *** • Styrofoam cups to put juice samples in (2/type of juice). Then. make solution B by dissolving 0. Pack 20. The size of the flask sent will depend on the number of classes and class sizes. Do not dilute to volume until ready to use.Vitamin C in Citrus Juices Microanalysis of Vitamin C in Citrus Juices 12/4/02 For class of 24 students. 8 groups. This means that a volumetric flask containing the proper amount of vitamin C must be taped shut and the mobile educator will add the water at the school. Next. add solution A to solution B and dilute to 500 mL with deionized water. Preparation of solutions: Standard Vitamin C: To make the 1 mg/mL standard. Cover flask with lid.6 g I2 in 25 mL ethanol.Sharpie pen for labeling pipets and Styrofoam cups • 1. This solution should be freshly prepared the morning of the lab and refrigerated if possible.6 g KI in 250 mL distilled H2O.The vitamin C and starch solutions must be prepped the same day as the lab.250 mL vol flasks with lids into which 250 mg of ascorbic acid per flask has been massed. • 9-spot well plates • 10-plastic droppers labeled for juices (done by Mobile Educator or Teacher borrowing experiment • 1. Iodine solution: First make solution A by dissolving 0. filled 9-15mL glass dropper bottles of iodine solution. 1% starch indicator: • • • • • • • • • Gannon University SIM Micro 1-8 . plus several extra.25 grams) of vitamin C in a 250 mL volumetric flask. filled 9-Beral pipets labeled iodine 9-plastic bottles labeled vitamin C solution 9-Beral pipets labeled vitamin C solution funnel to fill bottles with vitamin C standard 1. Pack enough DI water to dilute to volume and include a wash bottle of DI water. mass 250 mg (0.
heat to dissolve and filter.Vitamin C in Citrus Juices Add 2. Prepare fresh weekly Gannon University SIM Micro 1-9 .5 g soluble starch in 250 mL distilled H2O. 1 mL of laundry starch diluted to 100 mL with water can be substituted if soluble starch is not available.
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