Report Sheet

Experiment # 6
Chromatographic Analysis of Amino Acids

DATA
A. Spotting the Chromatography paper

Spot No. Color of the Spot Distance Distance Rf Value

travelled by the Travelled by the
Spot Solvent
1 Light Brown 45 cm 0.85
2 Reddish Pink 46 cm 0.87
3 Blueish Purple 53 cm 1.00
53 cm
4 Yellowish 53 cm 1.00
5 Brownish 6 cm 0.11
Orange

Guide Questions
 1. Why should you avoid getting your fingertips on the chromatography paper? How will it
interfere?
Touching the chromatography paper can contaminate it with dirt, oils and proteins. Oils from
fingertips can interfere with migration of pigments up the paper.

2. Comment on the amino acid content of the diet soda amino acid based on your chromatography
results.
The chromatography result of the diest soda shows that the Rf value of the said substance is 0.11,
meaning that the substance is polar and based on research, Histidine amino acid have the same
Rf value as the given substance and the substance can be conjenctured to have histidine amino
acid.

3. Comment on the amino acid content of your oregano extract based on your chromatography
results.
The chromatography result of oregano shows that its Rf value is 1 and the same result with the
ink. The result can mean two things, one of which is that the substance is very non-polar to have
a high Rf value or that chromatography result is unreliable since Rf values, by principle, should
be less than one.

4. Enumerate and describe the reasons behind the differences in the RF value of different amino
acids.
Affinity to the solvent- when the hydrophobic solvent moves over to the location of the
biomolecule , the rate at which the biomolecule moves up the paper is related to its relative
affinity for the solvent. Hydrophobic molecules (glycine, etc) will move faster because they are
more attracted to the solvent.
Affinity to the paper- paper is hydrophilic and the amino acids that are hydrophilic tends to be
attracted to the paper and moves slower.

5. Enumerate the sources of error in the experiment and their respective impact.
 Touching the chromatography paper which affected the movement of pigments.
 Insufficient or wrong spotting which caused pigments to not manifest.
 Miscalibration of other compounds (i.e. phenylalanine) that resulted to it becoming diluted or
concentrated.
 Lack of time for proper execution leading to the solvent not reaching the top and giving
incorrect result.

References:
Paper Chromatography (2012). Retrieved from: https://msu.edu