MTY1107 1st Semester, AY 2020-2021

Biochemistry - Laboratory

Problem Set: Chromatography

Name: Turingan, Alliah Nicole T. Instructor: Paolo Robert Bueno

Section: 105

Answer the following questions. Show your solutions, as necessary.

1.) Five amino acids were subjected to paper chromatography, using a less polar solvent as the mobile
phase. After visualization using ninhydrin, the distance travelled by the amino acids were measured
as follows:
Distances:
Solvent - 7.5cm
A - 6.8cm
B - 5.7cm
C - 0.4cm
D - 1.2cm
E - 0.8cm

a.) What is the Rf value of each amino acid sample? (3pts)

A - 6.8/7.5 D - 1.2/7.5
= 0.90667 = 0.16
B - 5.7/7.5 E - 0.8/7.5
= 0.76 = 0.1066
C – 0.4/7.5
= 0.0583
b.) Arrange the amino acids from most polar to least polar. (2pts)
The most polar is C followed by E, D, B, and A.

empangan2020
MTY1107 1st Semester, AY 2020-2021
Biochemistry - Laboratory

2.) An unknown amino acid was subjected to paper chromatography with five amino acid standards
in order to identify it. After visualization using ninhydrin, the distances travelled were measured
as follows:
Distances:
Solvent - 9.3cm
A - 1.1cm
B - 4.7cm
C - 2.4cm
D - 7.6cm
E - 8.7cm
X - 1.6cm

a.) What is the Rf value of each amino acid sample? (3pts)

A = 0.1182 D = 0.817
B = 0.505 E = 0.935
C = 0.258 X = 0.172

b.) Arrange the amino acids from most non-polar to least non-polar. (2pts)
The most non-polar amino acid is E followed by D, B, C, X, and the least non-polar is A.

c.) What is the possible identity of the unknown amino acid? Answer can be up to two amino
acids. (1pt)
The value of X is 0.172. Therefore, X can be amino acid A or C.

empangan2020
MTY1107 1st Semester, AY 2020-2021
Biochemistry - Laboratory

3.) A mixture of amino acids was subjected to paper chromatography with six amino acid standards
in order to determine its composition. After visualization using ninhydrin, the measured
distances were as follows:
Distances:
Solvent - 10.3cm
A - 4.5cm
B - 1.0cm
C - 6.6cm
D - 3.9cm
E - 3.5cm
F - 9.1cm
G - 10.3cm
X1 - 0.7cm
X2 - 4.3cm
X3 - 7.1cm

a.) What is the Rf value of each amino acid observed? (5pts)

A = 0.4365 F = 0.8834
B = 0.0970 G=1
C = 0.6407 X1 = 0.0679
D = 0.3786 X2 = 0.4174
E = 0.3398 X3 = 0.6893

b.) What are the possible components of the amino acid mixture? Answer can be up to two amino
acids per component. (4pts)
X1 = amino acid B
X2 = amino acid A
X3 = amino acid C
Therefore, the mixture X is a mixture of amino acids B, A, and C.

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