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Bio1TC2Chromatography QA

1. Biological molecules can be separated by using


chromatographic techniques. The diagram beside shows the
separation of several spinach leaf pigments by paper
chromatography.
(a) Explain how paper chromatography can be used to
separate pigments based on their chemical and physical
properties.
[4m]
.
.
.
.
.
.
.

(b) Use the ruler shown above to determine the Rf value of


xanthophyll. Show your calculations.

[2m]

.
.
[AP 2010]

2 A dipeptide was heated with an acid for 20 minutes. Three different samples were taken and
loaded on to a piece of chromatography paper. These were:
I
the peptide only;
solvent
II the dipeptide after it had been heated with the acidfor
front
2 minutes;
III the dipeptide after it had been heated with the acidfor
spot X
20 minutes.
The resulting chromatogram is shown in the drawing.
(a) What type of reaction results in the conversion of a dipeptide
to amino acids?
[1 m]
_________________________________________________________________

(b) Explain why three spots were obtained when the dipeptide
was heated with the acid for 2 minutes.
[2 m]

II

III

_________________________________________________________________
II
(c) (i) Calculate the Rf value of spot X. Show your steps

(ii) Explain how you could use the Rf values to identify spot X.

[2 m]

[1 m]

__________________________________________________________________________________________________________
__________________________________________________________________________________________________________

3 The diagram shows a chromatogram of


some individual sugars and mixtures of
TAH

Bio1TC2Chromatography QA

sugars.
(i) The Rf value of maltose is 0.25.
Mark on the diagram the position of
the spot representing maltose.
[1 m]
(ii) Which of A, B, C or D represents the
complete hydrolysis of sucrose? [1 m]
.............................................................
(iii) The spots from mixture D were very
close together. What could you do to
a chromatogram of mixture D to
separate the spots further?
[2 m]
........................................................................................................................................................................................................................
........................................................................................................................................................................................................................
........................................................................................................................................................................................................................
AQA BYA1 Jan 06

4 (a)

The diagram shows apparatus used to separate


these sugars by chromatography.
A pencil line was ruled on the chromatography
paper to mark the origin. A concentrated spot
of the solution was produced at X.

(i) Explain why the origin should be marked


with a pencil and not in ink.
[1 m]

(ii) Describe how you would produce a


concentrated spot at X.

[2 m]

(b) Use the appropriate letters from A, B, C, D and E to write a simple equation to
show how the Rf value
of the sugar in spot S could be calculated.
[1 m]
AQA BYA1 Jan 04

Rf =

Answers
TAH

Bio1TC2Chromatography QA

1(a)

Description of chromatography protocol.


(b) Formula or description dpigment/dsolvent
Calculation 3.5/7.5 0.5
2 a) hydrolysis
b) some dipeptide plus two different amino acids
c) i) - R f recognized as involvimg distance moved by spot and solvent front
- measure to the center and leading edge of spot
3.9/4.8=0.813
ii) - Check with table of known values
- Compare with R f of known substance

(i)
Correct position indicated; 1
(ii) C; 1
(iii) Turn through 90/ two-way chromatography/2D;
Run in a different solvent; 2

4 (a) (i) Ink will dissolve in solvent/will obscure spots (as it rises)/ink will run;
(Look for principle of interfering with chromatogram) 1
(ii) Repeated placing of drops;
Allowing to dry between applications; 2
(b) C .
(B + C) ;

TAH