Tutorial class

Exercise 1:
The absorbance of a fixed concentration of phenylephrine at 292 nm is found to be 1.224 in
0.1 M NaOH and 0.02 in 0.1 M HCl. Its absorbance in buffer at pH 8.5 is found to be 0.349.
Calculate the pKa value of its acidic phenolic hydroxyl group.

Exercise 2:

The determination of pyrethrins (an insecticidal agent) in a food sample gives a concentration
of 10 μg/kg. Express it in ppb and as a percentage.

Exercise 3: Which of the following molecules absorbs at longer wavelength?

a.

b.

c.

Exercise 4.
Phosphoric acid H3PO4 is a weak acid with 3 acidic functions : (pKa1=2.1, pKa2 =7.2,
pKa3 = 12.4).
a- Give the ladder diagram as a function of the pH
b- We want to prepare 1L of a 0.1M phosphate buffer solution at pH 7.2 . How would
we do it ? (Molecular weight of KH2PO4 : potassium dihydrogen phosphate= 136,09,
molecular weight of sodium hydrogen phosphate heptahydrate Na2HPO4 7H2O = 288,07)
 A B C

Exercise 5:

Phenolphthalein is used as a pH indicator. This reagent exists in 3 forms (A, B, C)

.
a) Which form appears at pH <8.2, pH 10-13, pH>13?
b) Which form is colored and why?

Exercise 6.

Zolpidem (figure above) is a common hypnotic with the chemical formula C 19H21N3O.
Stilnox - the brand name of zolpidem in Vietnam- is the hemitartratre of zolpidem: meaning
that the salt contains 1 molecule of tartaric acid for 2 molecules of zolpidem : (C19H21N3O)2
C4H6O6 . The molar mass (molecular weight) of zolpidem is 307.39 g/mol while the
molecular weight of the hemitartratre salt is 764.87 g/mol.
a- We want to prepare 100mL of a 1mM zolpidem solution. How much zolpidem base
should we weigh?
b- How much should we weigh if we start with the hemitartrate salt?
c- The pKa of zolpidem is 6.2, Is zolpidem mostly ionized or nonionized in plasma?

Exercise 7
Write the ladder diagram of para aminophenol A (pK OH = 9).
A and B are in plasma, are they extracted in chloroform
a) at pH = 7.4 A B
b) at pH = 12

Exercise 8
Write the ladder diagram of cysteine (pK COOH = 2.0, pK NH2 = 10, pK SH = 8.2 )
Write the structure in plasma.
Exercise 9
We extract a weak base at pH = pKa +3 with twice the volume of chloroform. The
distribution ratio of the unionized form is equal to 10. What is the % of the total amount in
the organic phase ?

Exercise 10
Green tea – a very common drink in South East Asia - is a hot water extract of the dried
leaves of Camellia sinensis. This extract contains hundreds of compounds; among those are
polyphenols, caffeine, amino acids, ….. Polyphenols are being extensively studied because
in-vitro they have antioxidant, antineoplastic and many other biological properties. These
therapeutic claims have not always been substantiated in vivo.
In the leaves of C. sinensis (and in green tea beverage) the most abundant class of
polyphenols are catechins, which represent about 20-30 % of the weight of the dried leaves.
The 4 major catechins are derived from epicatechin (-)-EC, and there are minor ones derived
from (+) catechin: (+) C.

Total polyphenols can be quantitated with the Folin-Ciocalteu reagent and the absorbance is
read in the visible range at 750-760 nm. This is the basis for the International ISO 14502 Part
1 procedure . The quantitation is done
by first having a calibration curve with
a standard and the results are
expressed as equivalents of that
standard. Here we have a linear
calibration curve between 10 and 100
µM of catechin (C15H14O6, molar
mass= 290.27 g/mol) with a regression
line A = 0.007 x - 0.0324

a) What are the limits of the

calibration curve expressed in ng/mL?
b) A stock solution of catechin was prepared for the calibration curve by dissolving 2.9 ±
0.1 mg of catechin in 10 ± 0.025 mL of methanol. What is the uncertainty (Relative
deviation standard) in each measurement (the mass of catechin and the volume of
methanol) and also in the concentration of the stock solution?
c) What volume of this stock solution do we need to prepare 1 mL of the 100 µM
calibration point.
d) Is the uncertainty of the concentration coming mainly from the mass or the volume
measurement?
 e) What is the concentration when A = 0.5? Can we calculate the concentration if A =
0.8?
1. The stability of catechins depends on the pH, the neutral molecule is more stable than
the ionized one. The lowest pKa is about 6.6. (it does not matter which phenol ionizes
first). We assume that the degradation starts to accelerate when the ratio
ionized/unionized is > 10 % . What is that pH value.

Question 11:

An experiment was realized 10 times. Given the data (in ng/ml) of this experiment as follows:
9.8; 10.5; 9.3; 8.9; 9.4; 10.5; 9.1; 10.5; 10.7; 9.6
a. Calculate the mean, standard deviation and coefficient of variation of this experiment.
b. We obtained a result of 11.6 ng/ml. Is this data an outlier?

Question 12: Theobromine has formula C7H8N4O2

a. How many mg of theobromine should we weigh to make 100mL of 5 mM solution?
b. How can we prepare 200mL of a 10-fold diluted theobromine solution?

Question 13:
HCl is a strong acid while benzoic acid is a weak one with a dissociation constant Ka = 6.5 x
10-5 M. Glutamic acid (the figure below) is an amino acid with 2 carboxylic groups. The pKa
values of the alpha carboxylic group, the amino group and the side chain carboxylic group are
respectively 2.2, 9.7 and 4.2.
a- What is the pH value of a 0.01 M HCl solution (in water)
b- What is the pH value of a 0.01M benzoic acid solution in water
c- Give the ladder diagram of glutamic acid as a function of pH
d- In which form will be glutamic acid in HCl 0.01M
e- In which form will be glutamic acid in a 0.01M benzoic acid

Question 14:

Fill the following blank with “water” or “fat”. Explain your answer
Question 15:
𝛼-Mangostin (C24H26O6, MW = 410.5 g/mol) is the major xanthone in both the arils and
pericarp of mangosteen (quả măng cụt), it has several peaks of absorption at 246 and 317 nm.
The molar absorption coefficient at 246 nm is quite high: 49 987 [Link]-1
a) Using the Beer lambert Law in a 1 cm cell, what is the absorbance at 246 nm of a 2
µM 𝛼-mangostin solution ?
b) In the above conditions what is the percentage of light absorbed?

Question 16
A stock solution of 𝛼-mangostin was prepared by weighing 10 mg dissolved in 10 mL of
methanol, we could measure the weight at ± 0.1 mg, and the volume in the 10 mL
measuring flask at ± 0.025 mL
a) What is the concentration of the stock solution?
b) What is the uncertainty in the concentration of this stock solution?
c) We use that stock solution to prepare 10 mL solutions for the calibration curve. What
is the dilution factor to get the 10 µg/mL solution?
d) What is the volume of the stock solution we need to get at the end 10 mL of the 10
µg/mL solution?

Question 17
Predict the order of elution, from first to last, of the following steroids from an ODS column
with acetonitrile/water (80:20) as the mobile phase.