Leighton Williams 1902815

Date: 07/10/22
Title of Experiment: Chromatography
Aim: To compare straight Aspartame before and after hydrolysis.
Method:
The following solutions were prepared: Aspartame/Equal, Hydrolyzed Aspartame solution,
Beverage A, Beverage B, Phenylalanine and Aspartic acid. Two spots were made up on TLC
plates, with five spots each as follows:
Plate 1: Aspartame, Hydrolyzed Aspartame solution, Beverage A, Beverage B, Phenylalanine
and Aspartic acid.
Plate 2: Aspartame, Hydrolyzed Aspartame solution, Beverage B, Phenylalanine and Aspartic
acid.
The TLCs were run in the solvent provided and the solvent was marked from the front and
allowed the plates to dry. The dry plates were treated with the Ninhydrin Reagent, warmed and
the Rf values for each spot were measured. The Rf values were recorded in the table below.
Results:
# Substance Distance spot Solvent Rf values
moved (cm)
Plate 1
1 Aspartame 4.65 6.1 0.76
2 Hydrolyzed Aspartame Spot 1: 1.50 6.1 Spot 1:0.24
Spot 2: 4.25 Spot 2: 0.69
3 Beverage A 4.65 6.1 0.76
4 Aspartic Acid 2.15 6.1 0.35
5 Phenylalanine 4.20 6.1 0.68
Plate 2
1 Aspartame 4.60 5.7 0.80
2 Hydrolyzed Aspartame Spot 1: 2.05 5.7 Spot 1:0.35
Spot 2: 4.45 Spot 2: 0.78
3 Beverage B 4.60 5.7 0.80
4 Phenylalanine 4.40 5.7 0.77
5 Aspartic Acid 2.60 5.7 0.45
Leighton Williams 1902815

# Solution KMnO4- pos/neg Ceric Nitrate

1 Aspartame Negative (Didn’t Negative
decolorize)
2 Hydrolyzed Aspartame Positive (Decolored) Negative
3 Beverage B Negative (Didn’t Negative
decolorize)
4 Methanol Negative (Didn’t Positive
decolorize)
5 Distilled Water Negative (Didn’t Negative
decolorize)

Discussion:
Aspartame is composed of two amino acid L-aspartic acid and L-phenylalanine. It is a white
crystalline, odorless intensively sweet powder. The density of Aspartame is 1.347g/cm3 with a
high melting point, between 246-247°C. Under strongly acidic or alkaline conditions, aspartame
may generate methanol by hydrolysis. Under more severe conditions, the peptide bonds are
also hydrolyzed, resulting in free amino acids.