Peptides and Amino Acids.docx

The first solution to prepare was solution 4 30 ml of

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The first solution to prepare was Solution 4. 30 mL of aspartame stock solution was added to a flask, along with 10 drops of 6M NaOH. The solution was then heated gently for 60 minutes. This was done first so that all other solutions could be prepared while solution 4 was heating. Solution 5, identified as Beverage A, was prepared by adding 5 mL of a diet soft drink to a test tube along with 10 drops of ethanol. Solution 6, identified as Beverage B, was prepared by
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Colton Aleman CHEM 238-516 TA: Jialuo Li adding 10 mL of the diet soft drink to a flask along with 10 drops of 6M HCl. This solution was stirred for 20 minutes, then half was poured into a test tube. 10 drops of ethanol were then added to the solution. Solution 7, identified as Beverage C, was prepared by adding 20 mL of the diet soft drink to a flask with 10 drops of 6M NaOH and heated for 15 minutes. After cooling, half the solution was poured into a test tube along with 10 drops of ethanol. The TLC plates were prepared by drawing a faint line in pencil approximately 1 cm from the bottom of the plate. This is where the solvent line was expected to begin. Each solution was then spotted onto the TLC plate. If a solution required multiple spots, the spot was dried with a blow drier before placing the subsequent spot. On the first TLC plate, Solution 1 was spotted onto the plate 3 times, Solution 2 was spotted once, Solution 3 was spotted once, and Solution 4 was spotted 12 times. On the second TLC plate, Solution 5 was spotted 12 times, Solution 6 was spotted 12 times, Solution 7 was spotted 12 times, and the fourth spot used Solution 1 again, spotting it 3 times. A table representation of the spotting is found in Table 1 and Table 2 . Once the spotting had been completed, the TLC plates were then inserted into the TLC chambers and allowed to absorb the solvents until the TA gave approval for their removal. The solvent line was marked and the TLC plates were then allowed to air dry. Upon drying completely, the plates were dipped in a ninhydrin solution, which acted as an indicator for the peptide bonds. The plates were then heated and dried using a heat gun to allow the spots to appear. The spots were then measured on their distance from the baseline in comparison to the length of the solvent front, providing the R f value of that particular spot. The second part of the experiment was the use of the KMnO 4 and Ceric Nitrate qualitative tests. For the KMnO 4 test, a ceramic spot plate was used to old the different samples.
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  • Fall '07
  • Staff
  • Soft drink, ninhydrin solution

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