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L1 Portail BECV LAST NAME : Group :

UE Biomolecules – BMOLsi
November 28, 2022 First name :

Mark : /20

CONTINUOUS ASSESSMENT
Duration : 45 min

Programmable calculators are allowed only if all memories are cleared before the test.
Documents are not allowed.
The exercises are independent of each other.

The scale is given as an indication only, it can eventually be slightly modified.

Exercise 1 : 7,5 points. Amino acids and study of an oligopeptide P (Recommended time: 15 min)

Q1. Give the name of the 4 amino acids below.

Name : Name : Name : Name :

Q2. Write the condensed formula of the tetrapeptide P at pH 7. The N-terminal amino acid of P contains
a sulphur atom and its radical is apolar. The 3 other amino acids are, in the following order: leucine,
asparagine and tyrosine.

Q3. Give the name of the N-terminal amino acid.

Name :

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Q4. Indicate the polarity of the radical of the 3 other amino acids.

Leu

Asn

Tyr

Q5. What is the overall charge of the tetrapeptide P at pH 7 ? Justify.

Q6. Using a spectrophotometer, the optical density of a solution that contains the tetrapeptide P is
measured at a wavelength of 280 nm. Why can we use this wavelength ?

Exercise 2 : 7,5 points. Determination of the structure of a lipid. (Recommended time: 15 min).

The lipid « Lip » was analysed by thin layer

chromatography (Figure 1). The solvent used
is a 80/19/1 mixture (petroleum
ether/ethyl ether/acetic acid). « Lip » was
analysed without treatment (deposit « Lip »),
after a soft alkalin hydrolysis (deposit « Lip* »)
or after a strong alkalin hydrolysis (deposit
« Lip** »). A mixture of standards (cholesterol,
cholesterol ester and triglycerids) was also
included (deposit « Std »).
Deposits
Std Lip Lip* Lip**
Q1. Annotate Figure 1 by identifying each of the spots observed for the mixture « Std » and rank these
standards by increasing hydrophobicity.

Q2. What types of molecules are 'theoretically' present in spots A, B, C and D? Give a schematic
structure for each of them.
Spot A Spot B

Spot C Spot D

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Q3. In order to obtain further details on « Lip » a
second, more resolutive thin layer
chromatography was performed (Figure 2). The
solvent used was a 70/12/1 mixture
(petroleum ether/ethyl ether/acetic acid). The
molecules present in the spot C of figure 1 were
analysed (deposit « C » in Figure 2). A mixture
of standard lipids (C18:0, C18:1 and C18:2) was also
included (deposit « Std* »).

Annotate Figure 2 by identifying each of the

spots observed for the mixture « Std* ».

Deposits

Std* C

Q4. Give the name of lipids that correspond to spots 1 and 2 knowing that these are common lipids and
that the lipid in spot 1 is not sensitive to oxidation by KMnO4.
Spot 1 : Spot 2 :

Justify :

Q5. Write the condensed formulas of molecules obtained after oxidation by KMnO4 of the lipid in spot 2.
Indicate their respective stoichiometry.

Q6. Knowing that « Lip » contains i) an omega 3 on carbon atom number 1 of the polyol, and ii) an
amino alcohol with 2 carbon atoms, give the condensed formula of « Lip ».

Q7. Give the name of « Lip » :

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Exercise 3 : 5 points. Determination of an unknown concentration (Recommended time: 10 min).

To determine the concentration of an unknown cholesterol solution « C » , you performed a standard curve
using a standard cholesterol solution at 25 mg.L-1. The equation of this standard curve (Optical density at
500 nm as a function of the quantity of cholesterol per tube in micrograms) was determined: y = 0,168 x.

Q1. The standard tube « G5 » must contain 5 microgram of cholesterol. What volume of the standard
solution (in µL) do you need to take to prepare this tube.

Q2. Prior to concentration determination, you first need to perform a 1 :50 dilution of solution « C ». You
then obtain the solution « Cdil ».
How do you perform this dilution using a 10 mL volumetric flask ?

Q3. The average optical density you obtain using 100 µL of « Cdil » is 0,420.
Determine the concentration of the unknown solution « C » in g.L-1.

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