Name: PARREÑO – PACULANANG – PEROCHO Date Performed: 11/29/2021

Section: C-LAB Date Submitted: 12/6/2021

Experiment No. 9
INTRODUCTION TO CHROMATOGRAPHY

I. Data
1. Chromatographic Separation of Leaf Extract

Distance travelled by solvent: 10 cm

Distance travelled by component A: 7.5

B: 0.4

C:

D:

2. Chromatographic Separation of Ink

Distance travelled by solvent: 11 cm

Distance travelled by component A: 9.6 cm

B: 7.7 cm

C: 5.5 cm

D: 3.2 cm

3. Chromatographic Identification of an Unknown Amino Acid

Distance travelled by solvent: 6.0 cm

Distance travelled by P: 0.83

T: 0.58

U: 0.58
II. Questions

1. What would be the effect of the following errors on chromatographic work:

a) the solvent level in the developing chamber is higher than the spotted sample
 In a chromatographic work, if the solvent level in the developing chamber is higher than
the spotted sample, then it will cause the spotted sample to dissolve and it will not be
carried along with the solvent. This will contaminate the solvent and the work will
become inaccurate.

b) too much sample is applied to the paper

 If we are going to have too much sample applied to the paper, then this will result to
tailing or streaking, meaning the samples will overlap, because of too much
concentration. This will also make the distance traveled difficult to measure.

c) the paper remained in the chamber after the solvent front reached the top of the paper.
 If the paper will remain in the chamber after the solvent front reached the top of the
paper, then this will make the spots difficult to see or may cause the spots to run into
each other, and the Rf values will not be accurately measured. The paper should be
removed before the solvent front reaches the top in case the solvent will keep on
running even after removing it.

2. Why is it necessary to cover the developing chamber tightly during the

development of the chromatogram?
 It should be securely covered during chromatogram to make sure that the chamber's
atmosphere is saturated with solvent vapor, keeping the solvent from evaporating as it
rises and separates the ink in the process.

3. What is the identity of your unknown amino acid? Explain clearly how you arrived at this
conclusion.
 After the paper is developed, the distance that the solvent traveled and the distance
that each amino acid has traveled are measured. This is called the retention factor, Rf,
and has a different value for each amino acid is then calculated. By comparing Rf values
of unknown amino acids to those of known values, the unknowns can be identified.
After calculating, the phenylalanine has an Rf of 0.83. The glutamic acid and the
unknown sample have the same Rf value which is 0.58, therefore glutamic acid is the
unknown amino acid.
4. Can paper chromatography be used to separate and identify a very volatile substance?
Explain your answer
 No, paper chromatography cannot be used to separate or identify highly volatile
substances because volatile molecules evaporate fast, causing the solvent to dry out
before the chromatography experiment can be completed. As a result, highly volatile
substances cannot be separated or identified using paper chromatography.

5. Why are you required to handle the chromatographic paper only at its corners in Part C?
 It is necessary to handle the chromatographic paper only at its corners in order to avoid
leaving fingerprints as these may hinder the elution process