Workbook "Chemistry of Biomolecules and Nanosystems"

for the 1st year students of medical faculty

RNRMU, 2018, 82 pages
This workbook contains common terms and definitions, homework exercises,
problems for self-study and class lessons, typical questions for test control and unit
tests, guidelines for laboratory works and various supplementary materials.
The workbook "Chemistry of Biomolecules and Nanosystems" meets the
requirements of FSES–3+.
This workbook was prepared by Prof. I. Yu. Belavin with essential contributions
from Prof. I. Yu. Baukov, Prof. V. N. Sergeev, Dr. N. A. Anisimova, Dr. L. P. Butba,
Dr. S. Yu. Bylikin, Dr. E. V. Gromova, Dr. N. A. Kalashnikova, Dr. S. A. Tarasenko,
Dr. I. V. Yankovich and V. P. Sergeeva.
Edited by Professor Vad. V. Negrebetsky
Additional information is available on the department website at the following
address: http://rsmu.ru/coboch_lf.html.
Literature for self-study and reading assignments
1. Glinka N. L. General Chemistry. Vol. 1 & 2. Moscow, 1990 (university library).
2. Richard Zsigmondy (Author), Ellwood B. Spear (Contributor). The Chemistry
of Colloids: Part I. Leopold Classic Library (April 1, 2016); ASIN B01DP1DFPM.
3. John E. McMurry, David S. Ballantine. Fundamentals of General, Organic and
Biological Chemistry, ISBN 978-1292123462.
4. Zurabyan S. E. Fundamentals of Bioorganic Chemistry. Moscow, GEOTAR-Media,
2003.
5. https://www.westminsterpublicschools.org/cms/lib/CO01001133/Centricity/
Domain/398/AP%20Chemistry%20Zumdahl%20Textbook.pdf
Additional literature for Russian-speaking students
1. Ленский А. С. , Белавин И. Ю. , Быликин С. Ю. Биофизическая и бионеоргани-
ческая химия. Учебник для студентов медицинских ВУЗов. М.: Медицинское
информационное агентство, 2008.
2. Попков В. А., Пузаков С. А. Общая химия. М.: ГЭОТАР-Медиа, 2010 (2007).
3. Сергеев В. Н. Курс коллоидной химии для медицинских ВУЗов. Учебное
пособие для студентов медицинских ВУЗов, обучающихся по специальности
фармация. М.: Медицинское информационное агентство, 2008.
4. Тюкавкина Н. А., Бауков Ю. И., Зурабян С. Э. Биоорганическая химия. М.:
ГЭОТАР-Медиа, 2014 (2010–2013).
5. Тюкавкина Н. А., Бауков Ю. И. Биоорганическая химия. М.: Дрофа, 2010
(2004–2009).
6. Руководство к лабораторным занятиям по биоорганической химии. Под ред.
Тюкавкиной Н. А. М.: Дрофа, 2010.
Contents
The program of the second semester includes two units.
Unit 1 — Bioorganic chemistry.
Unit 2 — Chemistry of organic nanosystems and biopolymers.
Detailed contents is given on page 82.
2
 Safety instructions
General rules
1. Students should never stay alone in a laboratory or chemical storage area. Any
experimental work should never be carried out without supervision of an instructor
or laboratory assistant.
2. Students should wear laboratory coats at all times. Eye goggles are highly
recommended. Long hair should be confined or bound.
3. Usage of unauthorized chemicals or equipment is strictly prohibited.
4. Unknown or unauthorized electrical equipment should never be plugged into
electrical outlets or turned on.
5. Before moving an electrical device, switch it off and unplug from the outlet.
6. No food or beverages should be stored or consumed in the laboratory. No chemicals
or equipment should be ever brought out of the laboratory.
7. Any safety issues arising in the course of experimental work should be immediately
reported to the instructor.
Before you start
1. Read the description of the experimental work you are going to carry out. Discuss all
unclear topics with your instructor or other students.
2. Write down the description of any planned practical work into your laboratory
journal in advance. All initial calculations for a practical work should be performed
before the work begins.
3. Make sure that all the equipment is in good order and the glassware is clean.
Impurities may affect chemical reactions.
Operating procedures
1. Acids, bases, heavy metals and most of other common chemicals (such as bromine,
phenol, etc.) are serious health hazards. Take care to prevent contact of chemicals
with your skin and eyes. Never taste chemicals or inhale deeply their vapours.
2. All operations with volatile, toxic or otherwise hazardous chemicals must be
performed under the fume hood. Some routine operations may be carried out at the
student's workplace when approved by the instructor.
3. Alcohols and other organic solvents are fire hazards. Their quantities should be kept
at minimum. Open flames must never be used to heat a flammable liquid.
4. When heating or boiling a liquid, use a special holder and make sure that the open
end of a flask or a test tube is never pointed towards you or other people.
5. When transporting glassware with hot liquids, hold it with two hands: one hand at
the neck, the other below the bottom. Always use a towel or gloves to protect your
skin from heat.
6. Always use funnel to transfer liquids. The funnel should be supported with a metal
ring fixed above the target flask.
5. Never pipette by mouth. Always use a bulb or other device for suction.
After the work is finished
1. Place all chemicals under the fume hood.
2. Switch off and unplug all electrical equipment.
3. Clean your workspace. Glassware should be washed immediately after use.

3
In the case of emergency
1. Seek immediate medical assistance whenever signs or symptoms of exposure to
hazardous chemicals are registered.
2. Promptly wash away with plenty of water any hazardous chemical that has contacted
your skin. After that, wash damaged skin with dilute (1–3%) solutions of acetic acid
(in the case of contact with bases) or sodium hydrogencarbonate (in the case of
contact with acids).
3. Whenever potentially harmful chemicals enter the eye(s), the eye(s) should be
immediately flushed with water for a prolonged time. After that, a diluted solution of
boric acid should be used for further washing. Medical assistance is required.
4. Minor thermal skin burns should be treated with ethanol and covered with a bandage.
Severe burns may require medical assistance.
5. If a fire occurs in a small area, isolate it from air by covering the affected area with
fire blankets. Remove any flammable materials nearby to avoid spread of the fire.
Switch off all electrical equipment.
6. If a fire occurs over a larger area, all students must evacuate the laboratory at once
and the fire department must be called.
Students who do not comply with safety instructions will be removed from the
laboratory and may be subject to civil penalties. Any violation of the safety rules will
be reported to the University officials.
Graphical techniques
A graph illustrates the relationship between two variables – in other words, it is a
"picture" of the numerical data. Many practical works require the representation of
experimental results in the form of graphs, so the students must be familiar with basic
rules of such representation.
1. Always use graph paper when drawing a graph.
2. The axes must be drawn along the bold lines of the graph paper.
3. In a graph of Y versus X, the independent variable (i.e., the cause) is plotted on the
x-axis and the dependent variable (i.e., the effect) is plotted on the y-axis.
4. Drawn graphs have labelled and scaled axes, and are used in quantitative
measurements. Drawn graphs always display the appropriate units for all variables.
5. The scale on each axis must match the range of the variable and make the most
effective use of the graph paper. The data should be critically examined to establish
whether it is necessary to start the scale(s) at zero.
6. When plotting a linear relationship, you may find that not all the data points lie
exactly on the same line. In such case, you should draw a line of best fit. This line
does not have to contain all experimental data points.
7. Fractional values plotted on the graph must be rounded to the nearest fine line on the
graph paper.

4
UNIT 1. BIOORGANIC CHEMISTRY

This unit covers the structure and reactivity of the main types of poly- and hetero-
functional compounds: metabolites, bioregulators and structural components of
biopolymers. The basic principles of biological oxidation and reduction, as well as the
chemical behaviour of compounds with several functional groups, are considered.
Biologically important transformations of monosaccharides, the structure and properties
of biologically important heterocyclic systems, nucleosides and nucleotides are studied
in detail. The structures of important biopolymers, polysaccharides and nucleic acids are
considered. The material studied in this unit allows to predict the behaviour of various
organic substances in the human body. The main goal of the section is to prepare
students for other medicine-related disciplines, including biochemistry, pharmacology.

Topic 1. Redox reactions of organic compounds.

Oxidation and reduction of different classes of organic compounds. Dehydrogenation
and hydrogenation. Oxidation of alcohols, amines and aldehydes. Peroxide oxidation.
Oxidative decarboxylation. Oxidation of thiols. Specific products of oxidation:
hydroperoxides, epoxides, quinones. Enzymatic hydroxylation. Biological redox
systems: FAD / FADH2, NAD+ / NADH, lipoic / dihydrolipoic acid, disulfide / thiol,
quinone / hydroquinone.

Concepts and terms

Reduction is a chemical reaction that involves the gain of electrons. In organic
chemistry, it is often accompanied by the formation of new carbon–hydrogen bonds.
Oxidation is the loss of electrons. Oxidation of organic compounds often involves the
loss of hydrogen atoms and the formation of either C=C or C–X bonds, where X is a
highly electronegative atom (O, N, S or halogen).
Active forms of oxygen (AFO) are active species that form in the respiratory chain or
can be generated in other redox processes: O2•– (superoxide radical), HOO•
(hydroperoxide radical), HOO– (hydroperoxide anion), H2O2 (hydrogen peroxide), HO•
(hydroxyl radical).
Hydroxylation is a replacement of a hydrogen atom with a hydroxyl group (–OH).
Aliphatic hydroxylation introduces a hydroxyl group (–OH) at an sp3-hybrid carbon
atom.
Aromatic hydroxylation introduces a hydroxyl group (–OH) into a benzene ring.
Hydroquinone is a common reducing agent with the formula HO OH .

Decarboxylation is a chemical reaction that removes a carboxyl group and releases

carbon dioxide. 2-Oxoacids undergo decarboxylation readily and form ketones:
O
R C CH2 C R C CH3 + CO2
O OH O

5
Oxidative decarboxylation is a typical reaction of 1-oxoacids that produces carbon
dioxide and a carboxylic acid:
O [O] O
R C C R C + CO2
OH OH
O
Peroxide oxidation, or hydroperoxidation is the replacement of a hydrogen atom with
a hydroperoxido group (–OOH).
Peroxide oxidation of lipids is a process leading to destruction of cell membranes and
programmed (apoptosis) or pathological cell death (for example, radiation sickness).
Respiratory chain (electron transport system) is a series of electron transporters
embedded in the inner mitochondrial membrane that shuttles electrons from NADH and
FADH2 to molecular oxygen, which is reduced to water. The energy released is used for
the ATP synthesis and maintaining the body temperature.
Oxidoreductase is a class of enzymes involved in the redox processes in the body.
Cofactors are transition metal cations (Cu2+, Fe2+, Fe3+, Mn2+, etc.) that form complex
compounds with proteins or enzymes.
Coenzyme is a nonprotein part of enzyme. Coenzymes of oxidoreductases can exist in
oxidized and reduced forms.
Xenobiotic is a foreign chemical substance found within an organism.
Oxidative stress is a disturbance in the balance between the production of reactive
oxygen species (free radicals) and antioxidant defences. Oxygen-free radicals damage
biological molecules such as lipids, proteins or DNA.
OH
Pyrocatechin (catechol) is a common reducing agent with the formula . The
OH
fragment of pyrocatechin is present in the molecules of catecholamines (important
bioregulators).
Epoxides are organic compounds containing a three-membered ring with two carbon
atoms and one oxygen atom (epoxy, or oxirane ring): С С .
О
Epoxidation is the oxidation of unsaturated compounds leading to formation of an
epoxide ring.
Glutathione (GSH) is an important antioxidant in plants, animals, fungi, and some
bacteria and archaea. Glutathione is capable of preventing damage to important cellular
components caused by reactive oxygen species such as free radicals, peroxides, lipid
peroxides and heavy metals. In all these processes glutathione is converted to its
oxidized form, glutathione disulfide (GSSG).

6
 Biologically important coenzymes participating in redox processes
Oxidized form Reduced form
Pyridine nucleotides — compounds containing the fragment of pyridine
O H H O
C C
NH2 NH2
+
N N
R R
+
NAD (nicotinamide adenine dinucleotide) NADH
+
NADP (Nicotinamide adenine dinucleotide
NADPH
phosphate) (phosphorylated NAD+)
Flavin nucleotides — compounds containing a flavin fragment
O H O
N N
N H N H

N N O N N O
R R H

FMN — Flavin adenine nucleotide FMNH2

FAD — flavin adenine dinucleotide FADH2
Thiol/disulfide systems contain readily oxidizable –SH group
Lipoic acid Dihydrolipoic acid
S S H S S H
(CH2)4 COOH (CH 2)4 COOH

CH2 CH COOH
S NH2
CH2 CH COOH
Cystine S Cysteine
SH NH2
CH2 CH COOH
NH2

Ubiquinones — derivatives of para-benzoquinone*

O OH
CH3O C R CH3O C R

CH3O C H CH3O C H
O OH

Ubiquinone or coenzyme Q (CoQ) Ubiquinol CoQH2

*Quinones are six-membered cyclic unsaturated diketones (oxidising agents):
О
ortho-quinone (ortho-benzoquinone)
О

para-quinone (para-benzoqinone) О О.

OH

Hydroquinone ( НО ОН ) and pyrocatechol ( OH ) are diatomic phenols

that act as reducing agents.

7
 Important biogenic carboxylic acids
Formula Trivial name Salts and esters
Dicarboxylic acids
HOOC–COOH Oxalic Oxalates
HOOC–CH2–COOH Malonic Malonates
HOOC–(CH2)2–COOH Succinic acid Succinates
HOOC–(CH2)3–COOH Glutaric acid Glutarates
Hydroxyacids
H O CH 2 COOH Glycolic Glycolates
СН3 СН СООН
Lactic Lactates
ОН
O
CH2 CH C Glyceric Glycerates
OH OH OH

HOOC CHCH2COOH
Malic Malates
OH
HOOC CH CH COOH
Tartaric Tartrates
OH OH
COOH
HOOC CH 2 C CH 2 COOH Citric Citrates
OH
O O
C CH CH CH2 C Isocitric Isocitrates
HO OH
OH COOH
Oxoacids (keto acids)
O
C COOH Glyoxylic Glyoxylates
H
СН3 С СООН
О
Pyruvic Pyruvates
CH 3 C CH 2 COOH
O
Acetoacetic Acetoacetates
HOOC C CH 2 COOH
Oxalacetic Oxalacetates
O
HOOCCH2CH2CCOOH 2-Oxoglutaric 2-Oxoglutarates
O (α-Ketoglutaric) (α-Ketoglutarates)
O O
C C CH CH2 C Oxalosuccinic Oxalosuccinates
HO OH
O COOH

8
 Unsaturated acids
H H
Maleic
Maleates
HOOC COOH (cis-butenedioic)
Н COOH Fumaric
С C
H Fumarates
HOOC (trans-butenedioic)
O O Aconitic
C CH C CH2 C (an intermediate in the Aconitates
HO OH
COOH Krebs cycle)
Biologically important derivatives of ethanamine
Name Formula Function
Colamine An important metabolite, a part of
HOCH2CH2NH2
(2-animoethanol) the molecules of phospholipids
+ Precursor of acetylcholine, a part of
Choline HOCH2CH2N(CH3)3
the molecules of phospholipids
O Neurotransmitter (participates in the
Acetylcholine +
CH 3C OCH CH N(CH )
2 2 3 3 transmission of nerve impulses)
Cysteamine A part of the coenzyme A (HSCoA),
HSCH2CH2NH2
(2-aminoethanethiol) the precursor of taurine
O
Taurine (2-amino- H2NCH 2CH 2 S OH A part of the molecules of bile acids
ethanesulfonic acid)
O
Cystamine (SCH2CH2NH2)2 Radioprotector

In-class exercises
1. Complete the following oxidation reactions, name the reactants and products:
O
1) CH3 CH C + NAD+ ; 2) CH3CH2CH2COOH + FAD ;
OH OH
O
3) CH3CH(NH2)COOH + NAD +
; 4) CH2 CH C + NADP+
OH OH H
O O
5) C CH2 CH2 C + O O ;
HO OH
O
6) CH2 CH C + G-S-S-G ; 7) CH2OOH + G-S-H ;
SH NH2 OH

H S S H + O O O2, NADP+
8) + NAD ; 9) C CH CH C ;
(CH 2)4 COOH
HO OH epoxidation

9
 О2, NADPH/H+
10) CH2 CH COOH aromatic hydroxylation ;
NH2
O2
11) HO CH2 CH COOH ;
oxidation of
NH2 pyrocatechol fragment
HO
O2, vit.С
12) HO CH2 CH2 NH3+ ;
aliphatic hydroxylation
HO
O2, HO
13) CH2 CH3 ;
peroxide oxidation
O2, HO
14) CH2 CH CH2 COOH ;
peroxide oxidation
FAD H2O
15) HOOCCH2CH2CHCOOH A ;
NH2
NAD+ NAD+
16) (CH3)3NCH2CH2OH A.
3. Complete the following reduction reactions, name the reactants and products::
O O О H+
1) С CH CH C + FADH2 ; 2) СН3 С С + NADH ;
HO OH ОН
О
CH2OH
H3CO OCH3
H OH
O reduction
3) O O + FADH2 ; 4) O ;
R O O
O
S S H+ ; H+
5) + NADPH 6) CH3 C C + NADH ;
OH
(CH2)4COOH NCH3
CH2CHCOOH
7) S NH2 + GSH
S NH2
CH2CHCOOH
3. Write reactions of decarboxylation and oxidative decarboxylation for: a) oxalacetic
acid (2-oxobutanedioic acid); b) oxalosuccinic acid.
4. The conversion of lactic acid into pyruvic acid in the body occurs in the presence of
the enzyme lactate dehydrogenase, the coenzyme of which is NAD+. Write the
corresponding equation, calculate the Emf and determine whether the reaction is
spontaneous under standard biological conditions. [E°'(CH3C(O)COO– + 2H+ + 2ē
→ CH3CH(OH)COO–) = –0.185 V; E°' (NAD+ + H+ + 2ē → NADH) = – 0.32 V).
Which factors will favour the formation of pyruvate ions?

10
 Online test "Redox reactions of organic compounds" (typical questions)
1. State the class name of the following compound: СН3 С СООН
О
a) hydroxyacid; b) dibasic acid; c) keto acid; d) alcohol.
2. What is the name for the following compound: НООССН2ССООН?
О
a) α-ketogluraric acid; b) oxalic acid; c) oxalacetic acid; d) glutaric acid.
3. Which of the following compounds can act only as oxidants in biological redox
processes?
a) thiol; b) FAD; c) O2; d) pyrocatechol.
4. The reaction cysteine (2R–SH) – 2ē → cystine (R–S–S–R) + 2H+ is:
a) reduction process;
b) neither reduction nor oxidation;
c) oxidation.
5. How will an increase in pH affect the value of the reduction potential for the system
NAD+ + H+ + 2ē → NADH?
a) potential will increase; b) potential will decrease; c) potential will not change.
6. Determine whether the reaction
–
OOCCH2C(O)COO– + NADH + H+ → –OOCCH2CH(OH)COO– + NAD+
is spontaneous under standard biological conditions if
E°'(oxalacetate, 2H+/malate) = – 0.17 V and E°'(NAD+, H+/NADH) = – 0.32 V.
a) spontaneous; b) impossible to say; c) nonspontaneous.
7. Classify the following reaction:
HO
vit.C
CH2CHCOOH + O2 CH2CHCOOH
N N NH2
NH2
H H
a) aromatic hydroxylation; b) hydration;
c) aliphatic hydroxylation; d) hydrogenation.
8. Classify the following reaction:
Х
СН3(СН2)7СН=СНСН2(СН2)6СООН + О2 СН3(СН2)7СН=СНСН(СН2)6СООН
ООН
a) reduction; b) neither reduction nor oxidation; c) oxidation.
9. Classify the product of the reaction:
СН2 СН СООН + Н2О СН2 СН2 СООН
ОН
a) α-hydroxy acid; b) α,β-unsaturated acid; c) β-hydroxy acid; d) oxoacid.
10. Name the product of the following reaction:
O O + FADH2 HO OH + FAD

a) para-benzoquinone; b) pyrocatechol; c) hydroquinone; d) ortho-benzoquinone.

11
Topic 2. Poly- and heterofunctional compounds.
Classes and reactivity of poly- and heterofunctional compounds in living organisms.
Decarboxylation and oxidative decarboxylation, elimination, intra- and intermolecular
cyclisation, alkylation, acylation, phosphorylation. Chelate complexes. Keto-enol,
lactim-lactam and ring-chain tautomerism.
Homework exercises
1. "Phenibut" is a contracted name of 4-amino-3-phenylbutyric acid, which is a central
depressant and analogue of the inhibitory neurotransmitter γ-aminobutyric acid
(GABA). Which reaction occurs when phenibut is heated up? Write the equation for
this reaction.
2. Which amino acid produces 2-butenoic acid upon heating? Write the equation for
this reaction.
3. Write the decarboxylation reaction for oxalacetic (oxobutanedioic) acid.
4. Write the reaction that produces sodium salicylate from the corresponding acid.
Which reagent (sodium hydroxide or sodium carbonate) should be used? Why?
5. Write the reaction of alkaline hydrolysis for novocaine [2-(diethylamino)ethyl-4-
aminobenzoate].
6. Match the class names (1 or 2) with the formulae (a)–(f):
1) lactam 2) lactone
CH3 CH3
NH O
a) b) c) O O d) O e) O NH2 f) O O
O O O O O

7. Match the chemical names (1 or 2) with the formulae (a)–(f):

1) p-aminosalicylic acid 2) salicylamide
COOH
COOH COOH CONH2 COOH CONH2
a) b) c) d) e) f)
H2N OH HO NH2 H2N OH NH2 OH
OH
8. Which reaction (intermolecular or intramolecular esterification) can occur for each
of the following compounds upon heating?
a) CH3CH(OH)CH(CH3)COOH b) HOOCCH2CH(OH)CH(CH3)COOH
c) (CH3)2C(OH)COOH d) HOOCCH(CH3)CH(OH)CH(CH3)COOH
e) CH3CH(OH)CH2CH(CH3)COOH
9. Which product (lactam or lactone) will form upon heating of the following
compounds?
a) 4-hydroxy-2-methoxyhexanoic acid b) 3-hydroxycyclohexanecarboxylic acid
c) 2-hydroxy-4-methoxypentanoic acid d) 2-amino-3-hydroxypentanoic acid
e) 3-hydroxypentanedioic acid
10. The elimination reaction proceeds readily upon heating of:
a) CH3CH(OH)CH2CH2COOH b) CH3CH(NH2)CH2COOH c) (CH3)2CHCH(OH)COOH
d) HOOCCH2CH(OH)COOH e) CH3CH(OH)CH2COOH
11. Which of the following compounds can be hydrolysed in the presence of acid?
NH NH
a) b) O O c) d) HN O e) O O f) O
O O HN O O O

12
12. Which product will form a) on heating of lactic acid in the presence of H2SO4(conc);
b) in the reaction of lactic acid with ethanol in the presence of H2SO4(conc)?
a) CH3C(O)COOH b) CH2=CHCOOH + H2O c) CH3CH(OH)COOC2H5 + H2O
d) CH3CH=O + HCOOH e) CH3CH(OC2H5)COOC2H5 + H2O

13. The hydrolysis of novocaine hydrochloride H2N .

COOCH2CH2N(C2H5)2 HCl in the
presence of HCl produces:
a) H2N–C6H4–COOH + 2C2H5OH + [HOCH2CH2NH3]+Cl–
b) [H3N–C6H4–COOH]+Cl– + [HOCH2CH2NH(C2H5)2]+Cl–
c) H2N–C6H4–COOH + HOCH2CH2N(C2H5)2
d) [H3N–C6H4–COOH]+Cl– + HOCH2CH2N(C2H5)2
e) C6H5NH2 + HOCH2CH2N(C2H5)2 + CO2
14. The hydrolysis of aspirin (acetylsalicylic acid) in the presence of aqueous NaOH
produces:
COONa COONa COOH
a) + CH3COONa b) + CH3COONa c) + CH3COONa
OH ONa OH
COOH
d) + CH3COONa e) C6H5ONa + CH3COONa + Na2CO3
ONa

15. The hydrolysis of phenacetin CH3CONH COOC2H5 in the presence of aqueous

NaOH produces:
a) H2N–C6H4–OC2H5 + CH3COOH b) H2N–C6H4–OH + CH3COONa + C2H5OH
c) HO–C6H4–OC2H5 + NH3 + CH3COOH d) H2N–C6H4–OC2H5 + CH3COONa
e) H2N–C6H4–ONa + CH3COONa + C2H5OH
16. The compound that undergoes decarboxylation most readily is:
a) HOOCCH2C(O)CH2COOH b) H2NCH2CH2COOH c) HOOCC(O)CH2CH2COOH
d) HOOCCH2CH2COOH e) CH3CH(OH)CH2COOH
17. Which of the following compounds can form chelate complexes with copper(II)
hydroxide?
a) HOCH2CH2OH b) HOCH2CH(OH)CH2OH c) HOOCCH(OH)CH(OH)COOH
d) H2NCH2COOH e) H2NCH2CH2OH
18. Which of the following compounds is the enol form of 3-methyl-2,4-pentanedion?
a) CH3CH2C(OH)=CHCH3 b) CH3C(O)CH2C(OH)=CH2 c) CH3C(OH)=CHCH2CH3
d) CH2=C(OH)CH2COCH3 e) CH3C(O)C(CH3)=C(OH)CH3
19. Match the enol forms (1 or 2) with oxo-forms (a)–(e) of the compounds:
1) HOOCC(OH)=CHCOOH 2) CH3C(O)CH=C(OH)CH3
a) CH3C(O)CH2C(O)CH3 b) CH3C(O)CH2COOCH3 c) CH3C(O)CH2CH(CH3)COOH
d) HOOCCH2C(O)COOH e) HOOCC(O)CH2CH(COOH)COCH3

Concepts and terms

Lactams are cyclic amides derived from γ-, δ- and ε-aminocarboxylic acids by the
intramolecular elimination of water from the amino and carboxyl groups.
O O
C C
(CH2)n OH
(CH2)n
H
N - H 2O N
H
H
n=2-4

13
Lactims are tautomeric form of lactams:
O
C C OH
(CH2)n (CH2)n
N H N
lactam lactim
Lactones are cyclic esters derived from γ-, δ- and ε-hydroxycarboxylic acids by the
intramolecular elimination of water from the hydroxyl and carboxyl groups.
O O
C + H+ C
(CH2)n OH
(CH2)n
H O
O - H2O
n=2-4
Cyclic hemiacetals are cyclic tautomers of hydroxyaldehydes or hydroxyketones:
O
C C OH
(CH2)n R (CH2)n R
O H O

Tautomerism is the dynamic equilibrium between two or more isomers.

Chelation is the formation of chelate complexes of poly- and heterofunctional
molecules with metal ions.
Cyclisation (intermolecular or intramolecular) is the formation of cyclic compounds.
Five- and six-membered rings are the most stable.
Phosphorylation is the addition of a phosphate group to an organic compound.
Common types of prototropic tautomerism
Scheme and name Example
Keto-enol
H O O OH H3C O H3C O
+ H+ - + H+
C C C C C CH2 C C CH C
C C
O SCoA HO SCoA
oxo-form enolate enol form
Enamine-imine
.. H .. H
N H N H
N N H H
H H C C
C C C C C
C
H COOH H COOH
H
enamine imine
Lactim-lactam
O O
O O H
C C
(CH 2)n (CH 2)n N OH N
N O
N H
H
lactam lactim
Ring-chain
O OH O OH
C CH 2 C CH 2 C
C H
H H H
(CH 2)n (CH 2)n CH 2 CH 2
O H O CH 2 O
CH 2 O H

oxo-form cyclic hemiacetal

14
 Chelate complexes of metals with poly- and heterofunctional ligands
a) with glycols: b) with amino alcohols: c) with diamines:
H2C CH2 2– H2C CH2 H2C CH2 2+

O O H2N O H2N NH2

Cu Cu Cu
O O O NH2 H2N NH2
H2C CH2 H2C CH2 H2C CH2

d) with amino acids: e) with oxalic acid: f) with 8-hydroxyquinoline:

H 2–
O O O
R C C C C
N
H2N O O O
O
Cu Cu Co
O
O NH2 O O
N
C C R C C
O H O O

Hydrolytically unstable bonds

Halides (X = Cl, Br, I) Derivatives of aldehydes and ketones
O O a) Acetals and ketals
C C + H2O C C + HX
OR H+
X OH C + H2O C O + 2 ROH
OR
C X + H2O C OH + HX
b) Imines
+
C C C X + H2O C C C OH + HX H
C N R + H2O C O + R NH2

Derivatives of carboxylic acids

a) Esters and lactones
O H+ O
'
Esters of phosphoric acids
R C + H2O R C + ROH O O
OR' OH
R O P X + H2O ROH + HO P X
b) Amides and lactams
OH OH
O O R'
R C R' + H2O R C + H N
N OH R''
R''

In-class exercises
1. Replace chemical names with formulae and complete the alkylation reactions:
a) colamine + CH3CH2Cl b) glycine + (CH3)2CHBr
H+
c) Choline + CH3 N d) N COOH + [CH3SR2]Cl
SAM
(S-adenosilmethionine)
e) H2N O
C + (C2H5)2NCH2CH2Cl
OH
PABA
2. Replace chemical names with formulae and complete the acylation reactions:
a) colamine + CH3C(O)SCoA b) lactic acid + CH3C(O)SCoA
c) aminoacetic acid + CH3C(O)OPO3H2

15
3. Replace chemical names with formulae and complete the phosphorylation reactions:
a) 2-hydroxybutanoic acid + ATP ADP + …
b) glycerol + H3PO4 2-glycerophosphate + …
4. Write cyclisation reactions for the following compounds:
CH3
O CH O O
a) H2N(CH2)4C b) 2 5 CH(CH2)3C c) HO C (CH2)2C
OH HO OH H
CH3
5. Draw the structures of chelate complexes:
Cu(OH) /NaOH
2 2 Cu(OH) /NaOH
a) Glycerol b) 2-aminopropanoic acid
6. Write hydrolysis reactions under acidic and basic conditions for the following
compounds:
O O O O
a) C2H5O NHCCH3 ; b) H2NCOCH2 CH2OCNH2 ; c) O2N O
CH N NHCNH2 .
C
CH3 (CH2)2CH3
phenacetin meprotan furacilin
7. Complete the following elimination reactions:
a) HSCH2CH2COOH b) C6H5CH(NH2)CH2COOH
c) CH3CH(OH)CH2COOH
8. Draw tautomeric forms for the following compounds:
O O O NH
a) C CH2 C b) C CH2 C
H3C SCoA H3C COOH
9. Write reactions of decarboxylation and oxidative decarboxylation for oxalacetic acid
(2-oxobutanedioic acid).
10. Write reactions of carboxylation for pyruvic acid and acetyl-CoA
11. Replace letters with formulae and write chemical and/or class names for the
products of the following reactions:
FAD H2O/H+ NAD+ to
1) Butyric acid A B C – CO D
2
elimination tautomerism hydrolysis oxidative
2) Cysteine A B C decarboxylation D
decarboxylation HSCoA choline
3) Malonic acid A B C
condensation NADH/H+ hydrolysis elimination
4) 2 Acetyl SCoA – HSKoA
A B C D
NAD+ H3PO4 elimination
5) Glyceraldehyde A B (2-phosphate) C
hydrolysis tautomerism
C D E
+
5) 2 AcetylCoA condensation A NADH/H B hydrolysis C elimination D
–HSKoA

6) a-Oxoglutaric NH3 NADPH/H+ B HSCoA 3 SAM

A C D
acid esterification
+
7) Malic acid NADP A decarboxylation B oxidative
C
decarboxylation
16
 carboxylation + FADH2
8) Pyruvic acid A NADH/H B elimination C

Online test "Poly- and heterofunctional compounds" (typical questions)

CH3

1. Identify the class of the compound O N :

H
a) lactone; b) cyclic hemiacetal; c) amine; d) lactam.
2. The intramolecular dehydration of the following compound CH3CH2CHCH2CH2COOH
NH2
produces:
a) cyclic hemiacetal; b) lactam; c) lactone; d) anhydride.
3. What is the chemical name of the compound HOOC–COOH?
a) malonic acid; b) oxalic acid; c) acrylic acid; d) acetoacetic acid.
4. Identify the class of the compound CH3–C(O)–CH2–COOH:
a) hydroxyacid; b) diatomic alcohol; c) oxoacid; d) hydroxyacid.
H3C C2H5
5. Which type of tautomerism is possible for the compound ?
O O
a) lactim-lactam; b) keto-enol; c) ring-chain; d) none.
6. The product of dehydration of β-hydroxybutyric acid is:
a) γ-hydroxybutyric acid (GHB); b) crotonic acid;
c) γ-aminobutyric acid (GABA); d) 3-butenoic acid.
H
H3C N O
7. Identify the type of the following reaction: CH3CHCOOH + CH3CHCOOH
_ 2H O
NH2 NH2 2 O N CH3
H
a) hydrogenation; b) hydrolysis; c) cyclisation; d) lactim-lactam tautomerism.
8. Crotonic acid is the product of:
a) elimination of γ-aminobutyric acid; b) hydration of 3-butenoic acid;
c) elimination of β-aminobutyric acid; d) dehydrogenation of γ-aminobutyric acid.
O C3H7 O
9. The products of the hydrolysis of meprotan (H2N C OCH2CCH2O C NH2) in excess
CH3
aqueous NaOH are:
CH2OH CH2ONa
a) H3C C C3H7 + NH3 + Na2CO3; b) H3C C C3H7 + NH3 + Na2CO3;
CH2OH CH2ONa
CH2OCOOH CH2OCOONa
c) H3C C C3H7 + NH4 OH; d) H3C C C3H7 + NH3
CH2OCOOH CH2OCOONa
10. Decide whether the following statements are true or false:
a) α-hydroxyacids do not form lactones on heating;
b) the decarboxylation of acetoacetic acid proceeds more readily than that of
propionic acid;
c) colamine is weaker as a base than ethylamine;
d) para-aminobenzoic acid is stronger than salicylic acid.
17
Additional problems
Part I
1) H2N OH + HCl 2) H2NCNH2 + HCl
O

3) N CONH2 + C2H5I 4) CH3OOC NH2 + C2H5I

+
H O
5) CH3 N + CH2 CH2 6) H2N C + (C2H5)2NCH2CH2Cl
OH
OH N(CH3)3 PABA
+
COOC2H5

7) + CH3SCH2CH2Cl 8) N COOH + [CH3S(R)2]Cl

NH2
O
9) HOCH2CH2NH2 + 3 [CH3S(R)2]Cl 10) CH3CHCOOH + CH3C
OPO3H2
OH
O O
11) H2NCH2COOH + CH3C 12) HOOC NH2 + CH3C
OPO3H2 OPO3H2
O O
13) HOCH2CH2NH2 + CH3C 14) HOCH2CH2NH2 + C
OPO3H2 SCoA
+ O O
15) HOCH2CH2N(CH3)3 + CH3 C 16) HOCH2CH2NH2 + CH3 C
SCoA SCoA
ATP
17) CH2 CH CH2 + H3PO4 18) CH3CHCH2COOH_
ADP
OH OH OH OH
elimination elimination
19) CH3CHCH2COOH 20) CH2CHCOOH
NH2 SH NH2
CH3
elimination
21) HOOCCHCHCOOH 22) CH2CHCOOH elimination
SH OH NH2
CH3
23) CH3CHCHCOOH elimination 24) CH3CHCH2COOH elimination
OH OH
elimination elimination
25) CHCH2COOH 26) HOOCCHCH2COOH
OH OH

H2O / H+ H2O / H+
27) C2H5OOC NH2 28) H2NCNHCCH3
O O

H2O / H+ H2O / H+
29) H3CCNH OC2H5 30) H2NCOC2H5
O O

18
 H2O / H+ H2O / H+
31) CH3OCCH2CNHCH3 32) CH3CCH2CNH2
O O NH O

H2O / H+ H2O / H+
33) H3COCNHC 34) H3CCNH CHOCH3
O O O OCH3

O C2H5
H2O / H+ H2O / H+
35) H3C O 36) O
N
CH3 H
NaOH
37) HOCH2CH2NH2 + Cu(OH)2 NaOH 38) CH3CHCOOH + Cu(OH)2 chelation
chelation
OH
NaOH
39) CH2 CH2 + Cu(OH)2 NaOH 40) CH2 CH CH2 + Cu(OH)2 chelation
chelation
OH OH OH OH OH
tautomerism O tautomerism
41) CH3CCH2COOC2H5 42) HOCH2CHC
H
O OH
tautomerism tautomerism
43) CH3CCOOH 44) CH3CCH2COOCH3
O O
tautomerism tautomerism
45) CH3CCH2CCH3 46) CH2CCOOH
O O O
tautomerism O cyclization
47) CH3CH2CCOOH 48) HOCH2CHCH2C
CH3 H
NH
CH3
O cyclization cyclization
49) CH3CHCH2CH2C 50) CH2CH2CHCHCOOH
H NH2 CH3
OH
oxidative
51) C2H5CHCH2CHCOOH cyclization 52) CH2CCOOH decarboxylation
NH2 CH3 O
oxidative oxidative
53) CH3CCOOH decarboxylation 54) HOOCCCH2CH2COOH decarboxylation
O O
oxidative FADH2
55) HOOCCCH2COOH decarboxylation 56) HOOCCH CHCOOH
O
CH3 O
FADH2 FADH2
57) O O 58) O CH3

+ +
59) CH3CH2CH2NH2 NAD 60) CH3CH NCH3 NADH/H
19
 NADH/H+ +
61) CH3CCH2COOH 62) CH2CH2COOH NAD
O OH
CH2CH3 NAD+
HO
63) HO 64) CH3CH CHCOOH FADH2

FAD FAD
65) CH3CH2CH2COOH 66) CH2NH2

CH3
O NADPH/H+ NADPH/H+
67) N CH + 2
aromatic 68) CH2COOH + O2
3 aromatic
hydroxylation hydroxylation
NADPH/H+
69) CH2CH CH2 + O2 epoxidation

Part II
+ FADH2 2 HSCoA
dehydration
1) HOOC C CH2COOH NADH/H A B C D
O
2) HOOC CH2 COOH
decarboxylation
A
HSCoA B HOCH2CH2NH2 C NAD+ D
O H2O / H + FAD B hydration NAD
+
3) CH3CH2CH2C A C D
SCoA hydrolysis
+ +
O H2O / H CH3OH CH3OH / H NAD+
4) CH3CH CHC A B C D
H + hydrolysis
B decarboxylation C HSCoA
NADP oxidative
5) CH3 CH CH COOH A _
NH3
CH3 NH2
elimination tautomerism hydrolysis NADH/H+
6) CH2 CH COOH A B _ C D
NH3
SH NH2
O NADH/H+ dehydration FADH2 carboxylation
7) CH3CCH2C A B C D
SCoA
O
_ NH
3 H2O / H + 2 HSCoA NAD+
8) HOOC CH CH2COOH A B C D
NH2
+ + + +
9) CH3CO(CH2)2N(CH3)3 H2O / H A NAD B NAD C HSCoA D
hydrolysis
O
NAD+ oxidative FAD HCl
10) C2H5 CH COOH A decarboxylation B C D
OH
O ester
condensation H2O / H + decarboxylation CH3NH2
11) 2 CH3C SCoA A B C D
hydrolysis
+ ester
12) 3
CH CH COOH
NAD
A
oxidative HSCoA condensation
D
decarboxylation B C
OH
dehydration tautomerism oxidative HSCoA D
13) HOOC CH CH COOH A B decarboxylation C
OH OH

20
 NAD+
14) CH3CH2CH2COOH HSCoA A FAD B hydration C D
+ _ O
3 [(CH3)3S] I CH3C SCoA +
H2O / H
15) HOCH2CH2NH2 _ A B C
2 HI hydrolysis
dehydration FADH2 HSCoA carboxylation
16) CH3 CH CH2 COOH A B C D
OH
+
NAD oxidative FAD
17) CH3CH2 CH COOH A decarboxylation B C HCl D
OH
ester
decarboxylation HSCoA condensation +
18) HOOCCH2COOH A B C NADH/H D
decarboxylation HOCH2CH2NH2 NAD+
19) HOOC CH2 COOH A HSCoA B C D
O H2O / H + retroaldol cleavage CH3NH2 FADH2
20) CH3CH CHC A B C D
H

Test "Poly- and heterofunctional compounds" (typical questions)

1. Complete the following equations:
Cu(OH)2/NaOH
a) HO (CH 2)2 NH 2 + CH 3CH 2Cl ; b) 2 СH3CHCOOH ;
NH2
O
c) H 2NСH 2COOH + CH 3 C OPO 3H 2 ; d) СH3CHCH2COOH −H O
OH 2

2. Replace the letters below with formulae:

O ester
condensation H2O / H + decarboxylation CH3NH2
2 CH3C SCoA A B C D
hydrolysis

Topic 3. Spatial structure of organic compounds.

Stereoisomerism of organic compounds. Chirality, asymmetrical atoms. Projection
(stereochemical) formula. Fischer projections. Configurational isomers: cis-, trans- and
D, L-conventions.

For self-study
1. Conformations of organic molecules.
2. R,S- and Z,E-nomenclature.
Homework exercises
1. Maleic (hydroxybutanedioic) acid and fumaric (trans-butenedioic) acid are involved
in the Krebs cycle. Which of these acids can exist as a pair of enantiomers?
2. Can adrenaline HO CH(OH)CH2NHCH3 exist as a pair of enantiomers?
HO

21
3. How many asymmetrical carbon atoms are there in the molecules of menthol and
camphor?
CH3 CH3 CH3

OH
CH(CH3)2 CH3 O
menthol camphor
4. Which of the following is the projection of L-alanine (L-2-aminopropanoic acid)?
COOH H
a) H3C H b) H3C COOH
NH2 NH2

5. Draw the Fischer projections of the two enantiomers of 2-hydroxybutanoic acid.

6. Which of following compounds can exist as cis/trans isomers: 2-methyl-3-hexene or
2-methyl-2-hexene?
7. Which of following compounds can exist as π-diastereomers: propenoic acid or
butenedioic acid?
8. Which of the following formulae represent the gauche-conformation of 1-butanol?
H OH H HH OH H
HO H H H H OH
1) H ; 2) ; 3) H5C2 ; 4) ; 5) H C2H5
H C2H5 H C2H5 HO H H H
H H
H C2H5 H
9. Which of the following is a) anti-conformation of 3-hydroxypropanoic acid;
b) constitutional isomer of 3-hydroxypropanoic acid?
H COOH OH HH H H OH
H H H H
1) H ; 2) H H
; 3) H ; 4) H OH
; 5) H
H H HO COOH H COOH
OH H H
COOH COOH
10. Arrange the following conformations in order of increasing energy:
CH(CH3)2 C2H5 CH3
H3 C CH(CH3)2
1) ; 2) CH3 ; 3) H C ; 4)
5 2
CH3

11. Which of following compounds can exist as enantiomers?

1) HOCH2CH(OH)CH2OH; 2) HOCH2CH(CH3)COOH; 3) CH3CH2CH(OH)CH3;
4) CH3CH2CH(OH)CH2COOH; 5) HOOCCOCH2CH2COOH
12. Which of following molecules does not contain a chiral centre?
1) (CH3)2CHCH2CH(OH)CH3; 2) CH3CH(NH2)COOH; 3) CH3CH2CHBrCH3;
4) HOOCCH2CH(OH)COOH; 5) HOOCCH2CH(OH)CH2COOH
13. Identify the pairs of enantiomers:
CH3 OH COOH OH
1) H OH and H CH3 ; 2) H OH and HO H ;
CH2OH CH2OH CH3 COOH

CH3 OH COOH CH2COOH

and
3) H C and H3C C ; 4) H2N C C COOH
C2H5 C2H5 CH2COOH H
OH H H NH2

22
14. Which of the following are D-isomers?
COOH COOH COOH CHO COOH
1) H OH ; 2) HO H ; 3) H NH2 ; 4) H OH ; 5) H NH2
CH3 C2H5 CH2C6H5 CH2OH CH3
15. Fill in the gaps:
The maximum possible number of isomers is calculated by the formula ,
where n is .
Diastereomers are isomers, which are not .
16. Which of the following are Fischer projections of meso-tartaric acid?
COOH COOH COOH COOH OH
H OH HO H HO H H OH H COOH
1) HO H
; 2) H OH ; 3) HO H ; 4) H OH ; 5) H COOH
COOH COOH COOH COOH OH

17. Which of the following are cis-isomers?

H3C CH3 H5C2 CH3 I Cl
1) C C ; 2) C C ; 3) C C ;
H COOH H H H H
HOOC H H3 C CH2Cl
C C C C
4) H COOH ; 5) F CH2CHO
18. The number of configurational isomers for compound C2H5CH(CH3)CH(NH2)COOH is:
1) 3; 2) 4; 3) 8; 4) 2; 5) 16.
Concepts and terms
Stereochemical formulae are used to represent on paper the three-dimensional spatial
arrangements of the atoms in the molecule.
Conformations are the different spatial arrangements of the atoms produced by rotation
about a single bond.
Newman projections are two-dimensional projections of molecules where the carbon
in front is represented by the point at which three bonds intersect, and the carbon at the
back is represented by a circle. A viewer is looking down the length of a particular
carbon–carbon bond. The three lines coming from each of the carbon atoms represent
their other three bonds.
Eclipsed conformations are the highest energy (least stable) conformations in which
two substituents eclipse each other.
Staggered conformations (gauche- and anti-conformations) are the lowest energy
conformations in which each sigma bond on one carbon atom is fixed at a 60 degree
angle from a sigma bond on the adjacent atom.
CH3
CH3
Gauche-conformation (for example, H ) is a staggered conformation where
H H
H
two substituents are staggered next to each other.
CH3
H H
Anti-conformation (for example, ) is the staggered conformation where two
H H
CH3
substituents at the adjacent carbon atoms lie as far apart from each other as possible.

23
Configurational isomers are stereoisomers that can cannot be converted into one
another by rotation around a single bond.
Asymmetric carbon atom (chiral carbon) is a carbon atom that is attached to four
different types or groups of atoms.
Chirality is the property of an object to be distinguishable from its mirror image.
Enantiomers are chiral molecules that are mirror images of one another.
Fischer Projection is a two dimensional projection of a molecule with chiral carbon
atoms. The Fischer projection represents every stereocentre as a cross. The horizontal
line represents bonds extending out of the plane of the page, whereas the vertical line
represents bonds extending into the plane of the page.
O
Glyceraldehyde (CH2 CH C )
H
is chosen as the standard for defining stereochemical
OH OH
configuration. Glyceraldehyde has two configurational isomers:
COOH COOH

L-glyceraldehyde HO H and D-glyceraldehyde H OH

CH2OH CH2OH
Diastereomers are configurational isomers that are not mirror images of each other.
π-Diastereomers are stereoisomers that differ in arrangement of substituents with
respect to a π-bond.
σ-Diastereomers are stereoisomers that differ in arrangement of substituents with
respect to the plane of the ring or several chiral centres.
Isomerism of organic compounds
isomerism of the principal chain (carbon skeleton)
Constitutional
isomerism of a multiple bond or a functional group position
isomers
isomerism of a functional group
enantiomers D- and L-
Configurational π-diastereomers cis- and trans- (relative to the π-bond plane)
isomers diastereomers cis- and trans- (relative to the ring plane)
σ-diastereomers
compounds with more than one chiral atom
eclipsed
Conformations* gauche- (skewed)
staggered
anti- (trans-)
*)
Usually undergo a fast interconversion and thus are not considered as isomers
In-class exercises
1. Determine the conformation type and draw the Newman projections for the most
and the least stable conformations of: a) butane (along C1–C2 and C2–C3 bonds),
b) 1,2-dibromoethane, c) ethylene glycol (consider the possibility of an
intramolecular H-bond formation).
2. Draw molecular structures for cis- and trans-isomers of 1-bromo-2-chloroethene
and 1,3-dimethylcyclobutane. Which type of configurational isomers do these
substances belong to?
3. Draw the molecular structure of 2-aminopropanoic acid (α-alanine). Label the
asymmetrical carbon atom with a asterisk. Underline four different substituents at
this atom. Assemble the "ball-and-stick" model for this molecule.

24
4. Use the model from the previous exercise to determine configurations (D- or L-) for the
following isomers of α-alanine:
CH3 COOH H NH2
COOH
a) H2N C COOH b) C NH2 c) C d) C
HOOC H
H3C H2N CH3 CH3
H H
5. Transform the projections below in accordance with the Fischer-Rosanoff
convention and determine the isomer configurations (D- or L-):
a) CH2COOH b) H c) NH2 d) NH2
O
H OH C OH H CH2SH H COOH
H
COOH CH2OH COOH CH2OH
Note that an exchange of any two substituents or rotation of the whole structure by
90° changes the isomer configuration from D- to L- or vice versa.
6. Draw Fischer projections for all possible isomers of 2,3-dihydroxybutanoic acid.
Indicate asymmetrical carbon atoms. Determine configurations (D- or L-) for the
isomers and find all pairs of enantiomers and diastereomers.
Online test "Isomerism of organic compounds" (typical questions)
1. Which types of constitutional isomers can be found in the mixture of the following
compounds: propanol, ethyl acetate, isopropyl alcohol, butyric acid?
a) isomers of carbon skeleton; b) isomers of a multiple bond position;
c) isomers of a functional group; d) isomers of a functional group position.
CH3
H CH3
2. Name the following conformation:
H H
H
a) eclipsed; b) gauche-conformation;
c) staggered; d) anti-conformation.
3. Select molecules with chiral centres:
a) lactic acid; b) glycerol;
c) pyruvic acid; d) maleic acid.
4. The number of asymmetric carbon atoms in the molecule below is:

a) 1; b) 2; c) 3; d) 4.
5. Which of the following molecules can exist as cis-/trans-isomers?
a) CH3CH CHCH3 ; b) ; c) ; d)
H3C CH3

6. Among the following Fischer projections, the formulae of D-isomers are:

COOH COOH H CH3

a) H NH2; b) H2N H ; c) HOOC NH2; d) H2N H .

CH3 CH3 CH3 COOH

25
7. Which types of stereoisomers are possible for the following molecule?

a) enantiomers; b) π-diastereomers;
c) σ-diastereomers; d) no stereoisomers are possible.
8. The existence of π-diastereomers is possible for:
a) 2-butenoic acid; b) 3-butenoic acid;
c) 2,3-dichlorobutanoic acid; d) 1,4-dimethylcyclohexane.
9. Which of the following molecules can exist as σ-diastereomers?
a) СH2 CH CHCOOH; b) Br ;
OH Cl Cl H3C
c) ; d) СH2 CH CH2COOH.
H3C Br OH Cl
10. Decide whether the following statements are true or false:
a) asymmetric carbon atom is attached to four different substituents;
b) the most stable conformation for 1,2-dibromoethane is gauche-conformation;
c) a molecule is chiral if it can not be superimposed with its mirror image;
d) diastereomers have the same physical properties.
Test "Isomerism of organic compounds" (typical questions)
1. Which of the following substances can exist as diastereomers? Draw molecular
structures of all possible diastereomers.
a) CH3CH2NH2; b) 1,2-dibromocyclohexane;
c) (CH3)2C=C(CH3)2; d) C2H5C(Cl)=CHCl.
2. Draw the structure of 2-aminopropanoic acid. Indicate the asymmetrical carbon
atom. Draw the Fischer projection for its D- and L-isomers.
CH3
3. Determine the configuration (D- or L-) of the following enantiomer: H2N COOH
H .

Topic 4. Carbohydrates (two lessons)

Classification and stereoisomerism of carbohydrates. Ring-chain tautomerism of
monosaccharides. Reactivity of monosaccharides: alkylation, acylation, epimerization,
reduction, and oxidation. Glycosides (pyranosides and furanosides). Decarboxylation of
uronic acids. Reducing and nonreducing disaccharides. Polysaccharides.
Homework exercises
1. Draw the formulae of enantiomers for mannose and fructose and name them.
2 Draw Fischer projections for the C-3 epimer of D-xylose and the C-4 epimer of
D-galactose. Name these compounds.
3. Draw the scheme of open-chain tautomerism for D-mannose in solution.
4. Classify the type (enantiomers, epimers, anomers) of isomers in each pair of
compounds:
a) D-glucose and D-mannose; b) D-arabinose and L-arabinose;
c) D-ribose and D-xylose; d) D-glucose and D-fructose;
e) α-D-glucopyranose and β-D-fructofuranose.

26
5. Write the reaction of hydrolysis for glycoside arbutine, specify the reaction
conditions and name the products.
Arbutin is a glycosylated hydroquinone. It inhibits tyrosinase and thus prevents the formation of melanin.
Arbutin is therefore used as a skin-lightening agent. Bearberry, which contains arbutin, is a traditional
treatment for urinary tract infections. CH2OH
O O OH
OH
OH
OH
6. Draw the formula of β-D-glucopyranosyl-(1→4)-α-D-glucopyranose. What is the
trivial name of this disaccharide? Draw the scheme of ring-chain tautomerism for
this compound.
7. Complete the reactions, write chemical names for the products:
+
2 H O/H H2
sucrose hydrolysis A + B cat.
8. Match the formulae (1 or 2) with the chemical names (a)–(f):
CH2OH CH2OH CH2OH CH2OH
HO O O OH O O OH
1) OH O OH 2) OH
O
OH
HO
OH OH OH OH
a) β-D-glucopyranosyl-(1→4)-α-D-glucopyranose;
b) α-D-glucopyranosyl-(1→4)-β-D-glucopyranose;
c) β-D-glucopyranosyl-(1→4)-β-D-glucopyranoside;
d) β-D-galactopyranosyl-(1→4)-α-D-glucopyranose;
e) α-D-galactopyranosyl-(1→4)-α-D-glucopyranose;
f) β-D-galactopyranosyl-(1→4)-β-D-glucopyranose.
9. Which statements are true for: a) amylose; b) amylopectin?
1) it consists of the residues of α-D-glucopyranose;
2) it belongs to branched polysaccharides;
3) it is soluble in water;
4) it forms methyl ethers;
5) it undergoes acid hydrolysis;
6) it forms coloured compound with iodine.

Concepts and terms

Carbohydrates (sugars) are biological molecules with the empirical formula Cm(H2O)n
(where m could be either the same as n or different from n).
Aldose is a monosaccharide that contains one aldehyde group and two or more hydroxyl
groups.
Ketose is a monosaccharide containing one ketone group (usually at the second carbon
atom) per molecule.
Trioses, tetroses, pentoses, hexoses are monosaccharides containing three, four, five
and six carbon atoms, respectively.
Epimers are diastereomers that differ in configuration at only one chiral centre.
D and L series for carbohydrates: L-isomers have the hydroxyl group attached to the
left side of the asymmetric carbon furthest from the carbonyl, while D-isomers have the
hydroxyl group on the right side. Naturally occurring sugars are D-isomers.

27
 The family of D-aldoses
H O
C
H OH
CH2OH
D-glyceraldehyde
H O H O
C C
H OH HO H
H OH H OH
CH2OH CH2OH
D-erythrose D-threose

H O H O H O H O
C C C C
H OH HO H H OH HO H
H OH H OH HO H HO H
H OH H OH H OH H OH
CH2OH CH2OH CH2OH CH2OH
D-ribose D-arabinose D-xylose D-lyxose

H O H O H O H O H O H O H O H O
C C C C C C C C
H OH HO H H OH HO H H OH HO H H OH HO H
H OH H OH HO H HO H H OH H OH HO H HO H
H OH H OH H OH H OH HO H HO H HO H HO H
H OH H OH H OH H OH H OH H OH H OH H OH
CH2OH CH2OH CH2OH CH2OH CH2OH CH2OH CH2OH CH2OH
D-allose D-altrose D-glucose D-mannose D-gulose D-idose D-galactose D-talose

Other monosaccharides and their derivatives

CH2OH CH2OH
H OH
H OH
HO H
HO H H OH
H OH H OH
CH2OH CH2OH
xylitol D-glucitol (sorbitol)
H O CH2OH CH2OH CH2OH
C
C O
H H C O C O
CH2OH HO H
H OH HO H H OH H OH
H OH C O H OH H OH H OH
CH2OH CH2OH CH2OH CH2OH CH2OH
D-deoxyribose dihydroxyacetone D-xylulose D-ribulose D-fructose
H O H O H O
C C C COOH H O
C COOH
H NH2 H2N H H NH2 H OH H OH H OH
HO H HO H HO H HO H HO H
HO H
H OH H OH HO H H OH H OH H OH
H OH H OH H OH H OH H OH H OH
CH2OH CH2OH CH2OH CH2OH COOH COOH
D-glucosamine D-mannosamine D-galactosamine D-gluconic D-glucuronic D-glucaric
acid acid acid

28
 Di- and polysaccharides
CH2OH CH2OH CH2OH CH2OH
O O OH O O OH
OH OH OH O OH
HO O HO
OH OH OH OH
β-maltose β-cellobiose
CH2OH CH2OH CH2OH CH2OH
HO O O OH O O
OH O OH ... OH O OH O ...

OH OH OH OH
β-lactose cellulose
CH2OH CH2OH CH2OH
O O O
OH OH OH
... O
HO
OH OH OH
O
HOCH2 CH2OH CH2O CH2OH
O O O O
HO OH OH OH
... O O O ...
CH2OH
OH OH OH OH
sucrose amylopectin (a component of starch)

Concepts and terms (continued)

Ring-chain tautomerism of carbohydrates is the dynamic equilibrium between cyclic
and oxo forms of carbohydrates.
Mutarotation is a gradual change in the optical rotation of freshly prepared solutions of
sugars due to the formation of different tautomeric forms.
Glycosidic hydroxyl group is the hydroxyl group derived from the carbonyl group due
to the formation of cyclic forms of monosaccharide.
Furanose is a five-membered cyclic form of monosaccharide containing four carbon
atoms and one oxygen atom in the ring.
Pyranose is a six-membered cyclic form of monosaccharide containing five carbon
atoms and one oxygen atom in the ring.
Haworth projection is a common way of drawing structural formulae to represent the
cyclic structures of monosaccharides with a simple three-dimensional perspective.
Anomer is a cyclic saccharide and an epimer that differs in configuration, specifically
at the hemiacetal/acetal carbon (also called the anomeric carbon).
Monosaccharide is a sugar (such as glucose) that cannot be hydrolysed to give a
simpler sugar.
Dihydroxyacetone (DHA) is a simple carbohydrate (a triose); an intermediate product
of fructose metabolism. DHA is primarily used as an ingredient in tanning products.
Dihydroxyacetone phosphate (DHAP) is one of the two products of the breakdown of
fructose 1,6-bisphosphate, along with glyceraldehyde 3-phosphate.
Ribose is an aldopentose that has all the hydroxyl groups on the same side in the
Fischer projection. A cyclic form of ribose, β-D-ribofuranose, is part of the RNA
backbone.
29
Ribulose is a ketopentose. As the 1,5-bisphosphate, it combines with CO2 at the start of
the photosynthetic process in green plants.
Deoxyribose (D-deoxyribose or 2-D-deoxyribose) is a pentose that is a key component
of deoxyribonucleic acids (DNA).
D-xylose participates in carbohydrate metabolism. Reduction of xylose by catalytic
hydrogenation produces the sugar substitute xylitol.
Xylulose is a ketopentose. D-xylulose 5-phosphate (D-xylulose-5-P) is an intermediate
in the pentose phosphate pathway.
Glucose is a monosaccharide containing six carbon atoms and an aldehyde group and is
therefore referred to as an aldohexose. It is the most important simple sugar in human
metabolism.
Glucosamine is an amino sugar and a constituent part of the structure of the
polysaccharides chitosan and chitin.
Galactose is a monosaccharide that is less sweet than glucose and fructose. The residue
of D-galactose is a part of lactose.
Galactosamine is the 2-amino-2-deoxy derivative of galactose, in which the 2-OH
group is replaced with an NH2 group; the D-isomer occurs in various
mucopolysaccharides, notably of chondroitin sulfate.
Mannose is an aldohexose. Mannose is important in human metabolism, especially in
the glycosylation of certain proteins.
D-Mannosamine (2-amino-2-deoxymannose) is a hexosamine derivative of mannose
and a part of different heteropolysaccharides.
Fructose (fruit sugar) is a ketohexose participating in carbohydrate metabolism.
Ascorbic acid (also known as L-ascorbic acid and vitamin C) has the formula
СН2ОН
Н ОН
О
О
and participates in biologically important redox reactions.
ОН ОН
СН2ОН
Н ОН
О
О
Dehydroascorbic acid ( ) is the oxidized form of ascorbic acid.
О О
Alditol is a polyol derived from aldose or ketose by reduction.
Sorbitol (glucitol) is a polyol produced by the reduction of glucose.
Aldaric acid is a carbohydrate in which both terminal groups (hydroxyl and aldehyde)
have been oxidized to carboxyl groups.
Aldonic acid is any of a family of sugar acids obtained by oxidation of the aldehyde
functional group of an aldose to form a carboxyl functional group.
Uronic acid is the compound obtained by replacing the hydroxymethyl group (CH2OH)
in the molecule of an aldose with a carboxyl group. A laboratory synthesis of an uronic
acid from an aldose requires protecting the aldehyde group from oxidation, for example
by its conversion to a glycoside.
Epimerization of carbohydrates is the interconversion of epimers of
monosaccharides.

30
Enediol is an unsaturated compound with two hydroxyl group attached to both carbon
atoms of the double bond ( С С ). Enediols are intermediates in the
ОН ОН
interconversion of monosaccharides under alkaline conditions.
Glycoside is the product of substitution of glycosidic (hemiacetal) hydroxyl group.
Glycosidic bond is the covalent bond between an anomeric carbon atom and an
aglycone molecule.
Glycoside-glycosidic bond is the glycosidic bond formed between the two anomeric
carbon atoms.
Aglycone (genin) is the non-sugar part of a glycoside.
Disaccharide is the sugar formed when two monosaccharides are joined together by a
glycosidic linkage.
Lactose (milk sugar) is a reducing disaccharide derived from the condensation of
galactose and glucose, which form a β-(1→4) glycosidic linkage.
Maltose (malt sugar) is a reducing disaccharide derived from the condensation of two
molecules of glucose, which form a α-(1→4) glycosidic linkage.
Cellobiose is a reducing sugar consisting of two glucose units linked by a β-(1→4)
bond, the product of the incomplete hydrolysis of cellulose.
Sucrose (beet or cane sugar) is a nonreducing disaccharide composed of glucose and
fructose linked via their anomeric carbon atoms.
Reducing disaccharide has a free hemiacetal group and thus can undergo a ring-chain
tautomeric transformation (for example, cellobiose, lactose and maltose).
Nonreducing disaccharide is a sugar that cannot undergo the ring-chain tautomeric
transformation and thus cannot act as a reducing agent (for example, sucrose).
Bionic acid is the product of oxidation of a reducing disaccharide.
Polysaccharides are polymeric carbohydrates composed of long chains of
monosaccharide units bound together by glycosidic linkages. The complete hydrolysis
of a polysaccharide gives the constituent monosaccharides.
Heteropolysaccharides (heteroglycans) are polysaccharides containing two or more
different monosaccharide units.
Homopolysaccharides are polysaccharides composed of a single type of sugar
monomer.
Starch is a homopolysaccharide consisting of amylose and amylopectin. This
polysaccharide is produced by most green plants and used as an energy storage.
Amylose is a linear (unbranched) homopolysaccharide made of α-D-glucose units,
linked to each other through α-(1→4)-glycosidic bonds.
Amylopectin is a branched homopolysaccharide made of α-D-glucose units, linked to
each other through α-(1→4) and α-(1→6)-glycosidic bonds.
Glycogen is a branched biopolymer consisting of linear chains of glucose residues
connected by α-(1→4)-glycosidic bonds with further chains branching through
α-(1→6)-glycosidic bonds.
Oligosaccharide is a saccharide polymer containing a small number (typically two to
ten) of monosaccharide units. The complete hydrolysis of an oligosaccharide produces
monosaccharides.

31
Dextrins are a group of low-molecular-weight polymers produced by incomplete
hydrolysis of starch or glycogen.
Cellulose is an unbranched homopolysaccharide made up of glucose monomers
connected via β-(1→4)-glycosidic linkages.
Heparin is a water-soluble heteropolysaccharide that consists of repeating disaccharide
units of D-glucosamine and uronic acid linked by (1→4)-glycosidic bond. It is used as
an anticoagulant (blood thinner).
In-class exercises
1. Draw Haworth formulae of α- and β-anomers for the following carbohydrates:
D-xylofuranose, D-galactopyranose, and D-fructofuranose.
2. Modify the formulae from the previous exercise to draw 2-deoxy-α-D-xylofuranose,
2-amino-2-deoxy-β-D-galactopyranose and N-acetyl-2-amino-2-deoxy-β-D-galacto-
pyranose.
3. Draw the scheme of formation of O-glycosides (α- and β-anomers) from
D-galactose and methanol. Name the reaction products.
4. Write the hydrolysis reaction for O-methyl-α-D-fructofuranoside. Specify the
reaction conditions and name the products.
5. Write reactions of formation and hydrolysis for lactose and sucrose. Is it possible for
these disaccharides to undergo a ring-chain tautomerism and mutarotation? If yes,
draw the corresponding reaction schemes.
6. Draw the chemical structure of glycogen, amylose and amylopectin fragments.
Indicate the α-(1→4)- and α-(1→6)-glycosidic bonds. What is the difference
between glycogen and amylopectin?
7. Draw the chemical structure of a cellulose fragment. Indicate the β-(1→4) and
glycosidic bonds. What is the difference between amylose and cellulose structures?
8. Complete the reactions and state the chemical names for the products:
NADH/H+ CH3I/KOH H2O/H+
a) D-xylose b) β-D-ribofuranose (excess) A B
NAD+ CH3C(O)SCoA
c) D-mannose d) 2-amino-2-deoxy-D-glucopyranose
HNO3 (diluted) (CH3CO)2O
e) D-galactose f) α-D-glucopyranose (excess)
+ ATP
g) α-D-fructofuranose-6-phosphate – ADP α-D-fructofuranose-1,6-diphosphate
aldol cleavage
h) D-fructose-2,6-diphosphate
aldol condensation
i) glyceraldehyde-2-phosphate + dihydroxyacetone phosphate
9. Replace letters with chemical formulae, name the product:
C2H5OH/HCl 2 NAD+ hydrolysis decarboxylation
D-glucose A B C D
11. Draw the isomerisation scheme for D-mannose and D-ribose under alkaline
conditions. Indicate the enediol intermediate and name the isomerisation products.

32
 Online test "Structures of carbohydrates" (typical questions)
1. Which of the following compounds is aldopentose?
a) D-ribulose; b) D-mannose; c) D-xylose; d) D-fructose.
H O
C
HO H
2. Name the following compound: HO H
H OH
H OH
CH 2 O H

a) D-xylose; b) D-mannose; c) D-galactose; d) D-glucose.

CH2OH
O
3. Name the following compound: OH OH
OH OH
a) β-D-mannopyranose; b) α-D-mannofuranose;
c) β-D-mannofuranose; d) α-D-mannopyranose.
H O
C
H NH2
4. Name the following compound: H OH
H OH
C H 2O H
a) D-galactosamine; b) D-ribosamine; c) D-mannosamine; d) L-mannose.
5. The number of asymmetrical carbon atoms in the molecule of D-glucose is:
a) 1; b) 2; c) 3; d) 4.
CH2OH
O OH
6. The compound OH NH2 is the cyclic form of:
OH
a) 2-amino-2-deoxy-L-mannose; b) 2-amino-2-deoxy-L-glucose;
c) 2-amino-2-deoxy-D-glucose; d) 2-amino-2-deoxy-D-mannose.
CH2OH CH2OH
HO O O OH
7. The compound is: OH O OH

OH OH
a) α-lactose; b) nonreducing disaccharide;
c) β-lactose; d) sucrose.
8. D-xylose and L-xylose are:
a) tautomers; b) anomers at C-2;
c) enantiomers; d) isomers of a functional group position
9. Which of the following compounds is a disaccharide?
a) cellulose; b) glycogen; c) starch; d) cellobiose.
10. Choose the true statements about glycogen:
a) it is a branched polysaccharide; b) it is a linear polysaccharide;
c) it contains β-D-glucopyranose units; d) it is a reducing disaccharide.

33
 Online test "Chemical properties of carbohydrates" (typical questions)
1. The reaction of a monosaccharide with an alcohol under acid conditions produces:
a) glycosides; b) polyol;
c) uronic acids; d) aldonic acids.
2. Name the reaction product(s) of D-glucose and methanol:
a) O-methyl-α-D-galactopyranoside; b) O-methyl-α-D-glucopyranoside;
c) a mixture of O-methyl-α-D-glucopyranoside and O-methyl-β-D-glucopyranoside;
d) O-methyl-β-D-glucopyranoside
3. The reduction of monosaccharides gives:
a) glycosides; b) disaccharides; c) alditols; d) aldonic acids.
4. The reduction of D-xylose produces:
a) D-mannitol; b) xylitol; c) sorbitol; d) D-glucitol.
5. The mild oxidation of monosaccharides produces:
a) lactams; b) aldonic acids; c) glycosides; d) aldaric acids.
6. Oxidation of D-glucose with bromine water gives:
a) sorbitol; b) D-glucaric acid; c) D-gluconic acid; d) galactaric acid.
7. Oxidation of aldoses with a strong oxidant gives:
a) aldaric acids; b) aldonic acids; c) glycosides; d) disaccharides.
8. Oxidation of D-glucose with HNO3 produces:
a) sorbitol; b) D-glucopyranose; c) D-xylonic acid; d) D-glucaric acid.
9. Oxidation of a primary hydroxyl group in aldoses produces:
a) uronic acids; b) aldaric acids; c) aldonic acids; d) glycosides.
10. The equilibrium mixture that forms after isomerisation of D-glucose under alkaline
conditions consists of:
a) D-mannose and D-fructose; b) D-glucose, D-mannose and D-fructose;
c) D-glucose and D-mannose; d) D-glucose and D-fructose.
Test "Carbohydrates" (typical questions)
1. Draw the scheme of ring-chain tautomerism for a monosaccharide. Indicate α- and
β-anomers, glycosidic hydroxyl group. Name the tautomers.
2. Write the hydrolysis reaction for a disaccharide or polysaccharide. Indicate the
glycosidic hydroxyl group and name the products.
3. Write the following reactions:
a) acylation of D-galactosamine with acetyl coenzyme A;
b) hydrolysis of 5-phosphato-β-D-ribofuranosyl-1-diphosphate.
c) reduction of D-fructose.
Specify the reaction conditions and name the products.
4. Draw the isomerisation scheme for a monosaccharide under alkaline conditions.
Indicate the enediol intermediate and name the carbohydrates.

Topic 5. Heterocyclic compounds. Nucleosides, nucleotides and nucleic acids

Five-membered heterocycles with one heteroatom (pyrrole, furan, thiophene) and two
heteroatoms (pyrazole, imidazole, oxazole, thiazole). Six-membered heterocycles
(pyridine, pyrimidine, tetrahydropyran). Bicyclic heterocycles (indole, quinoline,
purine). Pyrimidine and purine nucleic bases. Nucleosides and nucleotides.
34
Polynucleotide chains. RNA and DNA. Primary and secondary structures of nucleic
acids. Complementary pairs of nucleic bases. Nucleoside polyphosphates. Nicotinamide
nucleotides and flavin adenine dinucleotide as redox cofactors.
Homework exercises
1. Write the reactions of nitration for pyrrole and thiophene. Which of these
heterocycles is acidophobic? Which nitration reagents are used for it?
2. Dibasol is a drug with vasodilating, spasmolytic, hypotensive and moderate
immunomodulatory actions. From the chemical point of view, it is 2-benzyl-
benzimidazole hydrochloride: . Which of the two nitrogen

atoms in the imidazole ring is protonated readily in this compound?

3. Write the reactions of nitrous acid with cytosine, adenine and guanine. Name the
reaction products and draw their tautomeric forms. Name all types of tautomeric
conversions.
4. Match the names of heterocycles (a)–(e) with the names of their biologically active
derivatives (1 or 2):
CH2CH2NH2
N HO
1) CH2CH2NH2 2)
N N
H H
histamine serotonin
a) pyrazole; b) imidazole; c) indole; d) pyridine; e) quinoline.
5. Arrange the following heterocycles in order of decreasing basicity:
a) pyrrole; b) imidazole; c) pyrimidine; d) piperidine.
6. The reaction product of pyridine with H2SO4 at 20 °C is:
SO3H
SO3H HO3S SO3H _
a) ; b) ; c) ; d) ; e) + HSO4 .
N SO3H N N N N
H
7. Match the formulae of the reagents (1 or 2) with the products of their reactions with
pyridine (a)–(e):
1) CH3Br 2) Cl2, AlCl3
CH3
CH3 _ Cl
a) ; b) ; c) ; d) + Br ; e) .
N Cl N N N N
CH3
8. Match the reaction types (1)–(3) with the reaction schemes (a)–(e):
1) SN2 at sp3 hybrid carbon; 2) SN at sp2 hybrid carbon; 3) acid-base reaction
_ COOC2H5 CONH2
CH3I NH3
a) + I ; b) ;
N N N N
CH3
KMnO4, H2O, t o _
H2SO4, 20 C HSO4 .
c) ; d) +
N CH3 N COOH N N
H
35
9. Arrange the following heterocycles in order of decreasing reactivity in electrophilic
substitution reactions:
a) pyrrole; b) benzene; c) pyridine; d) pyrimidine.
10. Match the type of isomerism (1 or 2) with its scheme (a)–(e):
1) keto-enol; 2) lactim-lactam.
O O NH2 NH O O
NH NH N NH NH NH
a) ; b) ; c) .
N O N OH N O N O O N O HO N O
H H H H H
O OH O O
H3C H3C H5C2 H5C2
NH N NH H5C2 NH
d) ; e) H5C2 ;
N O N OH O N O O N OH
H H

Heterocyclic compounds

N N N N O S
H H H H
aziridine pyrrole pyrroline pyrrolidine furan thiophene
N N N
N
N N O S
H H
pyrazole imidazole oxazole thiazole
N
N N N O N N
H tetrahydro-
pyridine pyrimidine quinoline H
piperidine pyran indole
O O O
N N HN N HN N HN NH

N N N N O N N O N N O
H H H H H H
purine hypoxanthine xanthine uric acid

Nucleic (nitrogenous) bases

O O NH2 NH2 O
H3C N N HN N
NH NH N
N O N O N O N N H2N N N
H H H H H
uracil thymine cytosine adenine guanine
(Ura) (Thy) (Cyt) (Ade) (Gua)

36
Concepts and terms
Heterocyclic compound is a cyclic compound that has atoms of at least two different
elements as members of its ring(s).
Azoles are a class of five-membered heterocyclic compounds containing a nitrogen
atom and at least one other non-carbon atom (i.e. nitrogen, sulfur, or oxygen) as part of
the ring.
Pyrimidine nucleic bases: uracil, thymine and cytosine
Purine nucleic bases: adenine and guanine.
Hypoxanthine is the product of the deamination of adenine.
Xanthine is the product of the deamination of guanine.
Uric acid is the final product of metabolism of purine nucleic bases.
Urates are salts or esters of uric acid.
Tautomeric forms of nucleic bases: the four bases of DNA can exist in at least two
tautomeric forms each. Adenine and cytosine can exist in either amino or imino forms,
and guanine, thymine and uracil (which are cyclic amides) can exist in either lactam or
lactim forms. The tautomeric forms of each base exist in equilibrium but the amino and
lactam tautomers are more stable under physiological conditions.
"Pyridine-like" nitrogen is an sp2-hybrid nitrogen atom with the lone pair located in
an sp2 orbital and thus is not involved in the aromatic conjugation.
"Pyrrole-like" nitrogen is sp2-hybrid nitrogen atom with the lone pair located in the
p-orbital and thus is involved in the aromatic conjugation.
Tautomerism of azoles is the proton transfer between "pyrrole-like" and "pyridine-
like" nitrogen atoms in azoles.
Tautomerism of purine is the proton transfer between the 7th and 9th atoms in the five-
membered ring.
N-glycosidic bond is a covalent bond between a nitrogen atom of a nucleic base and the
anomeric carbon atom of ribose or deoxyribose.
Nucleosides are N-glycosides in which a nitrogen atom of a nucleic base is attached to
the anomeric carbon atom of ribo- or deoxyribofuranose via a glycosidic bond.
Nucleotides are the phosphate esters of nucleosides.
Ester bonds in nucleotides are covalent bonds P–O between the residue of phosphoric
acid and a cyclic form of pentose.
Nucleoside polyphosphate is a nucleotide with several residues of phosphoric acid.
Numbering and naming conventions: the purine nucleotides end in "-sine" (adenosine
and guanosine), the pyrimidine nucleotides end in "-dine" (cytidine, uridine,
deoxythymidine). The pentose rings are numbered with the prime symbol (').

37
 O
H
N
Uridine (U) HOCH2 N O
ribonucleotide containing the residue of uracil. O

OH OH
O
CH3 H
Thymidine (T) N

deoxyribonucleotide containing the residue of HOCH2

O
N O
thymine.
OH
NH2
H
N
Cytidine (C)
HOCH2 N O
ribonucleotide containing the residue of cytosine. O

OH OH
NH2
N N
Adenosine (A) HOCH2 N N
ribonucleotide containing the residue of adenine. O

OH OH
O
H
N N
Guanosine (G) HOCH2 N
O N NH2
ribonucleotide containing the residue of guanine.
OH OH
NH2
N N
O
AMP (adenosine-5'-monophosphate) НO P OCH2 N N
ribonucleotide. OH
O

OH OH
NH2
N
ADP (adenosine-5'-diphosphate) O O N
ribonucleotide with one macroergic bond (see НO P O P OCH2
O N N
OH OH
below).
OH OH
NH2
ATP (adenosine-5'-triphosphate) N N
O O O
ribonucleotide containing two phsophoanhydride HO P O P O P OCH2 N N
bonds (macroergic bonds) that are responsible OH OH OH
O

for high energy content of this molecule.

OH OH

38
Anhydride (phosphoanhydride) bonds in nucleoside polyphosphates are covalent
bonds between the residues of phosphoric acid.
Macroergic bond is a high-energy bond in the sense that free energy is released when it
is hydrolyzed (ΔG0298 > 30 kJ/mol).
O
C
NH2
O
HO P OCH2 N
O
+
NAD (nicotinamide adenine dinucleotide) O
NH2

is a coenzyme in redox reactions. OH OH N N

HO P OCH2 N
O N
O

OH OH
O
C
NH2
O
HO P OCH2 N
O
+
NADP (nicotinamide adenine dinucleotide O
NH2

phosphate) is a coenzyme in redox reactions. OH OH N N

HO P OCH2 N
O N
O

OH OPO3H2

Nucleoside cyclophosphate is a nucleotide in which the residue of phosphoric acid is

attached to two oxygen atoms of ribose.
NH2

cAMP (Adenosine 3',5'-cyclic monophosphate) N N

is a second messenger involved in numerous O CH2
O N N
cellular signalling processes. O P
HO O OH
NH2
S-Adenosylmethionine (SAMe) is a naturally N N
CH3
occurring compound found in almost every tissue SCH2 N N
and fluid in the body; a methyl donating HOOCCHCH2CH2 O
NH2
compound in metabolic reactions.
OH OH
NH2
N
Coenzyme A O O CH3 O O N
(CoA or HSCoA) is HSCH2CH2NH CCH2CH2 NH CCH C CH2 O P O P OCH2
O N N
OH CH3 OH OH
the coenzyme in the O
acylation reaction. HO P O OH
O

Nucleic acid are biopolymers consisting of nucleosides units linked through

(5'→3')-phosphodiester bonds.
O
Phosphodiester bond O Р O is a phosphorus atom involved in two ester bonds
OH
that link two mononucleotide units in nucleic acid.
39
Primary structure of a nucleic acid is the sequence of nucleotides in a polynucleotide
chain.
DNA is deoxyribonucleic acid.
RNA is ribonucleic acid.
Complementary pairs of nucleic bases are the pairs of bases of two DNA strands
linked together by hydrogen bonds. The base pairing of complementary nucleotides
leads to the formation of the secondary structure of nucleic acids.
In-class exercises
1. for the compounds given below:
NH2 OH
N
NH N N N
N N O N N N
H H H HO N OH HO N N OH
H
a) identify aromatic heterocycles;
b) determine the number of atoms participating in each cyclic conjugate system;
c) indicate acidic and basic centres;
d) name the compounds.
2. Write the reactions between: a) uric acid and NaOH; b) pyrrole and Na metal;
c) pyridine and H2O; d) pyridine and H2SO4.
3. Draw the following schemes: a) hydrogen bonds between two molecules of
imidazole; b) formation of a 8-hydroxyquinoline complex with Cu2+ ions;
c) tautomeric forms of imidazole, cytosine and 5-pyrazolone.
4. Indicate hydrolytically labile bonds in the following compounds and write the
appropriate hydrolysis reactions:
O H O
C
HO CH2 O P OH
O
OH O2N O CH N NH C
NH2
H3C N
Pyridoxal-5-phosphate (PLP, vitamine B6) Furacilin
5. Draw tautomeric forms of uracil and guanine. Which tautomeric form of guanine
will produce xanthine upon hydrolysis? Write the corresponding equation.
6. Which nucleic bases can react with nitrous acid? Write the corresponding equations.
7. Draw the structures of adenosine and thymidine. Indicate N-glycosidic bonds and
numbering of the atoms in both heterocyclic and monosaccharide fragments.
8. Draw the structural formula for adenosine-5'-triphosphate. Indicate the N-glycosidic
and ester bonds. Write the hydrolysis reaction for this nucleotide.
9. Draw the structural formula for adenosine 3',5'-cyclic monophosphate (cAMP).
Indicate N-glycosidic and ester bonds. Write the hydrolysis reaction for this
nucleotide.
10. Draw the structure of a DNA fragment of the following composition: -dT-dC-dA-.
Indicate 3'-5'-phosphodiester bonds, N-glycosidic bond, 5'- and 3'- terminals.

40
11. The fragment of DNA strand contains the following sequence of nucleotides:
TAAGTCAGAGATC. Deduce the sequence of nucleotides in the complementary
fragment of DNA.
12. Calculate the free Gibbs energy change for the following reaction:
pyruvatekinase
H2C C COO– + ADP2– + H+ CH3 C COO– + ATP3–
OPO2–
3 O
phosphoenolpyruvate (PEP) pyruvate
The ΔG°' values for the hydrolysis reactions of PEP and ATP are –62 and –30.5
kJ/mol, respectively. [Answer: –31.5 kJ/mol]
13. Write the reaction of formation of uridine 5'-phosphate from 5-phospho-α-D-ribo-
furanosyl 1-diphosphate and the appropriate nucleic base.

Trivial names of some heterocyclic compounds

N N N N O S
H H H H
aziridine pyrrole pyrroline pyrrolidine furan thiophene
N N N
N
N N O S
H H
pyrazole imidazole oxazole thiazole
N
N N N O N N
H tetrahydro- quinoline H
pyridine pyrimidine
piperidine pyran indole

O O O
N N HN N HN N HN NH
N N N N O N N O N N O
H H H H H H
purine hypoxanthine xanthine uric acid

Nucleic (nitrogenous) bases

O O NH2 NH2 O
H3C N N HN N
NH NH N
N O N O N O N N H2N N N
H H H H H
uracil thymine cytosine adenine guanine
(Ura) (Thy) (Cyt) (Ade) (Gua)

41
 Online test "Heterocyclic compounds" (typical questions)
1. Match the names of heterocycles (1–4) with their formulae (a–d):
1) pyridine; 2) furan; 3) guanine; 4) pyrazole
O
HN N
a) N; b) ; c) ; d) .
N O N
H2N N N
H
H
2. Chose the correct statement about the properties of indole:
a) it is a strong acid; b) it is a strong base;
c) it is aromatic compound; d) it is a weak acid.
3. Which of the following compounds are aromatic?
N
a) ; b) ; c) N; d) .
O N N N
OH H H
4. Identify the ester bond:
O
1 N H
N
O
НO P OCH2 N N NH2
O
OH
2 3
OH OH
5. Which of the following compounds have acidic properties?
COOH
N
a) ; b) ; c) ; d) N.
S N N N
H
6. Which of the following compounds show only basic properties?
O
N N N NH
a) ; b) ; c) ; d) .
N N N N O N
H H H
7. Which of the following compounds are aromatic?
NH2
N N
a) ; b) ; c) ; d) .
N N O S N
H H H
8. The fragment of adenine in RNA or DNA is:
NH2 O NH2 O
N N N H3C
HN N NH
a) ; b) ; c) ; d) .
N O H2N N N N N N O

42
9. What are the structural units of uridine?
a) deoxyribose and uracil;
b) deoxyribose, phosphoric acid and uracil;
c) ribose, phosphoric acid and uracil;
d) ribose and uracil.
10. Which of the following compounds will undergo deamination in the reaction with
nitrous acid?
NH2 O NH2
N N NH N
a) ; b) ; c) ; d) .
N N N N O N O
H H H

Test "Heterocyclic compounds" (typical questions)

1. Draw the structural formula of uridine-5'-diphosphate. Indicate N-glycosidic and
ester bonds.
2. Write the reaction of hydrolysis for this compounds.
3. Which nucleic base is the product of this reaction? Will it react with nitrous acid? If
yes, write the reaction and draw at least three tautomeric forms of the reaction
product. Otherwise, draw at least three tautomeric forms of the nucleic base itself.

Unit test No. 1

1) Online unit test (contains 30 questions randomly chosen from the previous tests).
2) Paper test (typical questions).
1. Complete the following reactions and name the reaction products:
+ oxidative
1) HO
NAD O
OH 4)CH3C COOH decarboxylation
O cyclisation
2) HO(CH2)3C
H NH tautomerism
NH 2 HNO 5) CH 3C
3)CH3CH COOH 2 CH2COOH
2. Draw the scheme of the formation of α- and β-anomers for D-glucose. Name the
tautomers and indicate the glycosidic OH groups.
3. Write the reaction of complete hydrolysis for deoxycitidin-5'-diphosphate. Indicate
ester and N-glycosidic fragments. Draw the tautomeric forms of the nucleic base
formed in this reaction.
3) Laboratory works. Copy the description of the following lab works into your lab
journal and write the equations for all reactions. After the experiments are complete,
record the results observed.
Chemical properties of tartaric acid
1. Formation of potassium hydrotartrate
Work procedure
Put one drop of a 15% tartaric acid solution into a test tube, add two drops of a 5%
potassium hydroxide solution and shake. A white crystalline precipitate of a slightly
43
 soluble in water potassium hydrotartrate gradually begins to form. If the precipitate
does not form, cool down the tube with cold water and rub the inner wall of the tube
with a glass rod.
2. Formation of complex compounds
Work procedure
Put two drops of a 2% solution of copper(II) sulfate and a 10% sodium hydroxide
solution into two test tubes. Add the solution of potassium-sodium tartrate obtained
in the previous experiment into the first test tube. The precipitate of copper(II)
hydroxide dissolves. The resulting solution has a blue colour. This solution is called
Feling's reagent, which is used for detecting glucose in urine.
Heat the liquids in both test tubes to boil. In the first test tube, the colour of the
solution will not change, in the second test tube the blue precipitate of copper(II)
hydroxide is converted to black copper(II) oxide.
Formation of chelate complex compounds of α-amino acids
Work procedure
Put one spatula of dry copper(II) carbonate into a test tube and add 1 mL of a 1%
glycine solution. Heat the test tube, note the colour of the solution.
Pour carefully the resulting solution from the precipitate into another test tube and
add 1–2 drops of a 10% sodium hydroxide solution.
Deamination of α-amino acids
Work procedure
Mix 0.5 mL of a 1% glycine solution with equal volume of a 5% sodium nitrite
solution in a test tube. Add two drops of concentrated hydrochloric acid. Shake the
mixture and observe the formation of gas bubbles.
Chemical properties of glucose
1. The presence of hydroxyl groups in glucose
Work procedure
Place one drop of a 0.5% solution of D-glucose and six drops of a 10% sodium
hydroxide solution into a test tube. Add one drop of a 2% solution of copper(II)
sulfate. The precipitate of copper(II) hydroxide formed dissolves rapidly. The
resulting solution has a blue colour. Save this solution for the next experiment.
2. The reduction of copper(II) hydroxide with glucose under alkaline conditions
Work procedure
This reaction is called Trommer's test and is used to detect glucose in urine.
To the solution obtained in the previous experiment, add a few drops of water, until
the height of the liquid becomes 18–20 mm. Heat the tube above the burner flame,
keeping it tilted so that only the upper part of the solution is heated, and the lower
one remains cold for comparison. Heat the solution until it just begins to boil (do not
boil). The blue colour of the solution at the top of the tube changes to orange.

44
 Chemical properties of sucrose
1. Formation of chelate compounds
Work procedure
Put one drop of a 1% sucrose solution and six drops of a 10% sodium hydroxide
solution into a test tube. Add 5–6 drops of water for dilution (the height of the liquid
layer should be 18–20 mm). Add one drop of a 2% solution of copper(II) sulfate.
The resulting solution has a blue colour. Use the solution in the next experiment.
2. Sucrose as a nonreducing sugar
Work procedure
Heat the tube above the burner flame, keeping it tilted so that only the upper part of
the solution is heated, and the lower one remains cold for comparison. Heat the
solution until it just begins to boil (do not boil). The solution colour does not
change. Remember that in the similar experiment with D-glucose the colour of the
upper part of the solution changed from blue to orange.
The reducing properties of lactose
Work procedure
Put one drop of a 1% lactose solution and 5–7 drops of a 10% sodium hydroxide
solution into a test tube. Add one drop of a 2% solution of copper(II) sulfate. The
blue precipitate of copper(II) hydroxide dissolves when the tube is shaken, forming
a blue solution of the complex copper(II) salt with lactose. Add a few drops of water
to dilute the mixture until the height of the liquid becomes 18–20 mm. Heat the tube
above the burner flame, keeping it tilted so that only the upper part of the solution is
heated, and the lower one remains cold for comparison. Heat the solution until it just
begins to boil (do not boil). The colour of the upper part of the solution changes
from blue to orange. Compare this result from those of similar experiments with
D-glucose and sucrose.

45
UNIT 2. CHEMISTRY OF ORGANIC NANOSYSTEMS AND BIOPOLYMERS
The second unit covers: 1) organic compounds that can form organic heterogeneous
nanosystems (colloidal surfactants), and their structural components (higher fatty acids,
simple and complex lipids); 2) amino acids, peptides and proteins; 3) physico-chemistry
of disperse systems and solutions of macromolecules. The study of this section is
necessary for understanding the structure and properties of biological membranes, as
many liquids and dense tissues of the human body are disperse systems. In the same
section, the techniques of chromatography and electrophoresis, widely used in modern
medicine, are discussed. The chemistry of macromolecules will help a physician to
understand various phenomena occurring in a living organism. A medical professional
must have clear understanding of the laws governing the processes of dissolution of
biopolymers, salting out, gelation and viscous flow.
Topic 6. Lipids
Classification and structural units of hydrolysable lipids: glycerol, sphingosine, fatty
acids, 2-aminoethanol (colamine), choline and serine. Waxes, fats and oils.
Glycerophospholipids: phosphatides, phosphatidic acids, phosphatidylserines and
phosphatidylcholines. Sphingolipids: ceramides and sphingomyelins. Glycolipids:
cerebrosides and gangliosides. Reactions of hydrolysable lipids: hydrolysis, peroxide
oxidation and β-oxidation of fatty acids.
Homework exercises
1. When treated with nitrogen oxides, oleic acid (cis-isomer) converts into elaidic acid
(trans-isomer). Draw the formulae of these π-diastereomers.
2. Draw the structural formula for 2-oleoyl-2-palmitoyl phosphatidylserine and
indicate ester bonds in the molecule of this phospholipid.
3. Draw the structural formula of the phosphatidylcholine that contains residues of
palmitic and linolenic acids. Write the reaction of hydrolysis for this lipid under
acidic and basic conditions.
4. Draw the structural formula for 2-linoleoyl-2-stearoylphosphatidylethanolamine.
Which products will be formed by hydrolysis of this lipid under acidic conditions?
5. Choose the correct statements about unsaturated fatty acids that are components of
hydrolysable lipids:
a) their molecules contain even number of carbon atoms;
b) their double bonds commonly have cis-configuration;
c) their double bonds are conjugated;
d) saturated parts of their hydrocarbon chains typically have zigzag conformations;
e) the multiple bond is typically located between C-9 and C-10 atoms.
6. Match the names of fatty acids (1–5) with their compositions (a–f):
1) palmitic; 2) linoleic; 3) linolenic; 4) oleic; 5) arachidonic.
a) 20:4 5,8,11,14; b) 18:1 9; c) 18:0; d) 18:2 9,12; e) 16:0; f) 18:3 9,12,15.
7. Match the types of acid (1–3) with their names (a–d):
1) ω-3; 2) ω-6; 3) ω-9.
a) palmitic; b) linoleic; c) linolenic; d) oleic.

46
8. Match the structural formulae (1 or 2) with the names of lipids (a–e):
CH2O C(O)C17H33
a) 2-linolenoyl-1-oleoyl-3-palmitoylglycerol;
1) CHO C(O)C17H31
CH2O C(O)C15H31
b) 2-linolenoyl-3-palmitoyl-1-stearoylglycerol;
CH2O C(O)C17H35
c) 2-linolenoyl-3-oleoyl-1-stearoylglycerol;
2) CHO C(O)C17H29 d) 2-linoleoyl-1-oleoyl-3-palmitoylglycerol;
CH2O C(O)C15H31 e) 1-linolenoyl-2-linoleoyl-3-palmitoylglycerol.
8. Which of the following triglycerides are likely to be oils?
a) 1,2,3-tripalmitoylglycerol; b) 2-linolenoyl-1-linoleoyl-3-stearoylglycerol;
c) 1,3-dilinoleoyl-3-palmitoylglycerol; d) 1,2-dioleoyl-3-stearoylglycerol;
e) 1,2,3-tristearoylglycerol; f) 2-linoleoyl-1-palmitoyl-3-stearoylglycerol.
9. Which of the following compounds can be hydrolysed under basic conditions?
CH2O COC17H33 CH2O COC17H35 CH2O C17H35
a) CHO COC17H31 b) CHO COC17H29 c) CHO C17H29 d) CH3(CH2)14COOC30H61
+
CH2O COC15H31 CH2O P(O)OCH2CH2NH3 CH2O C15H31
_
O
10. Match the name of the process (1 or 2) with the equations (a)–(e):
1) saponification; 2) hydrogenation.
CH2O C17H35 CH2O C17H35 CH2O C17H35
+
a) CHO C17H29 3 KOH ; b) CHO C17H29 H2O / H ; c) CHO C17H29 H2, Ni ;
CH2O C15H31 CH2O C15H31 CH2O C15H31
d) C17H35COOH + NaOH ; e) C17H33COOH + KMnO4 .
11. Choose the major products of the peroxide oxidation of oleic acid:
COOH
a)
OOH
COOH
b)
OOH
COOH
c)
OOH
OOH
COOH
d)
OOH
COOH
e)
12. Which of the following acids will undergo peroxide oxidation most readily?
a) stearic b) palmitic c) oleic d) arachidonic
13. Match the class name of lipids (1 or 2) with structures (a)–(e):
1) lecithin; 2) cephalin
CH2O COC17H35 CH2O COC17H35 CH2O COC17H35
a) CHO COC17H33 b) CHO COC17H29 c) CHO COC17H29
+
CH2O P(O)OCH2CH2NH3 CH2O P(O)OCH2CH3 CH2O PO3H2
_
O OH
CH2O COC15H31 CH2O COC17H35
d) CHO COC17H29 e) CHO COC17H29
+
CH2O P(O)OCH2CHCOOH CH2O P(O)OCH2CH2N(CH3)3
_ _
O NH3 O
+

47
14. Match the class name of lipids (1 or 2) with structures (a)–(e):
1) sphingolipid; 2) glycerophospholipid
CH2O COC17H35 CH2O COC15H31 CH2O COC17H35
a) CHO COC17H33 b) CHO COC17H33 c) CHO COC17H33
+ +
CH2O P(O)OCH2CH2NH3 CH2O P(O)OCH2CH2N(CH3)3 CH2O COC15H31
_ _
O O
CH CH(CH2)12CH3
CH2OH
CHOH
d) CHOH e)
+ CHNH COC17H33
CH2O P(O)OCH2CH2NH3 +
CH2O P(O)OCH2CH2N(CH3)3
OH
_
O
15. Choose the correct statements about phospholipids:
a) in glycerophospholipids, fatty acids are linked by ester bonds to carbon atoms 1
and 2 of glycerol;
b) the residue of glycerol is linked by ester bonds to one residue of fatty acid and
two residues of phosphoric acid;
c) the residue of aminoalcohol is linked by an ester bond to the residue of
phosphoric acid;
d) phospholipids are derivatives of L-phosphatidic acids;
e) glycerophospholipids exist in bipolar form.
16. Choose the correct statements about the following compound:
CH2O (CH2)nCH3
CHO COCH3
+
CH2O P(O)OCH2CH2N(CH3)3
_
O
a) it contains four ester bonds; b) it belongs to phospholipids;
c) it contains one ether bond; d) it contains the residues of fatty acids;
e) this is a derivative of an L-phosphatidic acid.
17. Choose the correct statements about the following compound:
CH CH(CH2)12CH3
CHOH
CHNH COC17H35
CH2O
CH2OH
O
O
OH
OH
OH
a) it contains one ester bond; b) it is a sphingolipid;
c) it contains β-glycosidic bond; d) it contains two residues of fatty acids;
e) this is a derivative of an L-phosphatidic acid.

48
 Common fatty acids
Chain length Formula Common name
Saturated carboxylic acids
C12 : 0 C11H23COOH Lauric
C14 : 0 C13H27COOH Myristic
C16 : 0 C15H31COOH Palmitic
C18 : 0 C17H35COOH Stearic
C20 : 0 C19H39COOH Arachidic
Unsaturated carboxylic acids
C18 : 1 CH3(CH2)7CH=CH(CH2)7COOH Oleic
C18 : 2 CH3(CH2)3(CH2CH=CH)2(CH2)7COOH Linoleic
C18 : 3 CH3(CH2CH=CH)3(CH2)7COOH Linolenic
C20 : 4 CH3(CH2)3(CH2CH=CH)4(CH2)3COOH Arachidonic

Concepts and terms

Lipids are a large and diverse group of naturally occurring organic compounds that are
related by their solubility in nonpolar organic solvents (e.g. ether, chloroform, acetone
and benzene) and general insolubility in water. Hydrolysable lipids can be cleaved by
hydrolysis into smaller molecules (fatty acids or their salts, alcohols or aminoalcohols,
phosphoric acid, and so on).
Fatty acid is a carboxylic acid with a long aliphatic chain, which is either saturated or
unsaturated. Most naturally occurring fatty acids have an unbranched chain of an even
number of carbon atoms, typically from 4 to 28.
Soaps are water-soluble sodium or potassium salts of fatty acids.
Arachidonic acid (C19H31COOH) is involved in cellular signalling. This acid is a key
inflammatory intermediate, and can also act as a vasodilator.
Simple lipids are esters of fatty acids with various alcohols (waxes, triglycerides).
O
Waxes (R C
O R'
) are esters of fatty acids and long-chain alcohols. They belong to the
class of simple lipids.
Glycerolipids are derivatives of glycerol.
Fats and oils (triacylglycerols or triglycerides) are simple lipids derived from glycerol
and three fatty acids.
Complex lipids are phospholipids and sphingolipids.
Phosphatidic acids are the simplest diacylglycerophospholipids, the precursors of other
glycerophospholipids.
Glycerophospholipids are the derivatives of glycero-3-phosphoric acid.
Phosphatidylcolamines (colaminecephalins) are glycerophospholipids containing the
residue of colamine (ethanolamine, monoethanolamine, or 2-aminoethanol).
Phosphatidylserines (serinecephalins) are glycerophospholipids containing the
residue of serine. They are components of cell membranes and participate in cell cycle
signalling, including apoptosis.
Phosphatidylcholines (lecithins) are glycerophospholipids containing the residue of
choline. They are major constituents of cell membranes.

49
 CH3 (CH2)12 H
C C CH OH
H
Sphingosine ( CH NH2
) is an aminoalcohol with an unsaturated hydro-
CH2 OH
carbon chain. It is the primary constituent of sphingolipids.
Sphingolipids are derivatives of sphingosine.
Ceramides are N-acylated derivatives of sphingosine. Making up to 50% of our skin's
natural protective barrier, ceramides retain moisture levels in the skin and help regulate
cell activity.
Cerebrosides are glycolipids that consist of a ceramide with a single sugar residue
(typically glucose or galactose) at the 1-hydroxyl moiety. Galactocerebrosides are often
found in neural tissue while glucocerebrosides are found in other tissues.
Sphingomyelins are sphingolipids found in animal cell membranes, especially in the
membranous myelin sheath that surrounds some nerve cell axons.
Glycolipids are sphingolipids with a carbohydrate attached by a glycosidic bond to the
residue of sphingosine.
Gangliosides are acidic glycosphingolipids containing one or more residues of sialic
acid (N-acetylneuraminic or N-glycolylneuraminic acid) in their carbohydrate moiety.
β-Oxidation of fatty acids is a catabolic process that breaks down fatty acid molecules
and generates acetyl-CoA.
Peroxide oxidation of lipids is the oxidative degradation of polyunsaturated fatty acids
residues in lipids resulting in cell damage. This process occurs by a free radical chain
reaction mechanism.
Common hydrolysable lipids
Glycerolipids:
O
CH2 O C R phosphatidylcolamines phosphatidylcholines
fats phosphatidic phosphatidylserines
O and oils acids (serinecephalins) (colaminecephalins) (lecithins)
CH O C R'
O O O O
O P OH P O CH2 P O CH2 P O CH2
CH2 O C R"
OH O- CH O- CH2 O- CH2
+
H3N COOH H3N+ (CH3)3N+

Sphingolipids:
H
CH3(CH2)12 C CH OH
sphingomyelins cerebrosides gangliosides
C O ceramides
CH NH C R
H O CH2OH
CH2 O H P O CH2 O
O- CH2 OH oligosaccharide
(CH3)3N+ OH
OH
In-class exercises
1. Draw the formula of a triacylglycerol that contains residues of stearic, linoleic, and
linolenic acids. Indicate the configuration of double bonds in the last acid. Write
reactions of saponification, hydrogenation and iodination for this lipid.
50
2. Write the reaction of saponification for cetyl palmitate (n-hexadecyl hexadecanoate
CH3(CH2)14COOC16H33). State the class of this lipid.
3. An L-phosphatidic acid contains residues of palmitic and oleic acids. Draw a
scheme of formation of this glycerophospholipid. Indicate the ester bonds.
4. Complete the reactions and write the chemical names for their products:
decarboxylation exhaustive saponification
a) Phosphatidylserine A N-methylation B C+…
HSCoA
FAD hydration NAD+ cleavage
b) StearoylCoA A B C D+E
5. Write the saponification reaction for a ceramide that contains a residue of
arachidonic acid.
6. Write the saponification reaction for a sphingolipid that contains residues of
palmitic acid, phosphoric acid and choline. State the class of this lipid.
7. Draw a scheme of the peroxide oxidation for a lipid that contains a residue of oleic
acid.
Online test "Lipids" (typical questions)
1. Match the chemical names with the structures given below:
1) choline; 2) oleic acid; 3) stearic acid; 4) glycerol.
a) C17H35COOH; b) C17H33COOH;
CH2 CH CH2
c) HOCH2CH2N(CH3)3 ; d) .
OH OH OH
2. State the number of double carbon–carbon bonds in the molecules of the following
compounds:
1) arachidonic acid; 2) 1-palmitoyl-2,3-dilinolenoylglycerol;
3) trioleoylglycerol; 4) tristearoylglycerol.
a) 0; b) 3; c) 4; d) 6.
3. The molecule of 1-oleoyl-2-palmitoyl-stearoylglycerol contains the residues of:
a) different fatty acids; b) the same fatty acids;
c) glycerol; d) sphingosine.
2
CH2O C(O)R
CHO C(O)R
4. Identify the ester bond in the following molecule: 1
O
CH2O P OH
OH 3
CH2O C(O)R
CHO C(O)R
5. The hydrolysis of O produces:
CH2O P OH
OH
a) aldehydes; b) carboxylic acids; c) glycerol; d) phosphoric acid.
6. The hydrolysis of a lipid produced glycerol, 2-aminoethanol, palmitic, linoleic and
phosphoric acids. State the type of the initial lipid:
a) colaminecephaline; b) ceramide; c) cerebroside; d) glycerophospholipid.

51
7. One mole of a glycerolipid can add four moles of hydrogen. The name of initial
glycerolipid is:
a) 1-oleoyl-2-palmitoyl-3-arachidonoylglycerol;
b) 1-linolenoyl-2-palmitoyl-3-stearoylglycerol;
c) 1-oleoyl-2-linolenoyl-3-palmitoylglycerol;
d) 1-oleoyl-2-palmitoyl-3-linoleoylglycerol.
8. Which compounds will decolourise bromine water?
a) stearic acid; b) palmitic acid;
c) oleodistearin; d) arachidonic acid.
9. The mass of iodine (M = 254 g/mol) that can react with one mole of 1-oleoyl-
2-palmitoyl-3-stearoylglycerol is:
a) 127 g; b) 254 g; c) 381 g; d) 508 g.
10. Which statements are true?
a) triacylglycerols derived from animal tissues are usually liquid compounds;
b) the molecule of a phosphatidylserine contains the residue of an amino acid;
c) soaps are sodium or potassium salts of fatty acids;
d) the hydrolysis of complex lipids produces only fatty acids and alcohols.
Test "Lipids" (typical questions)
1. Write the hydrogenation reaction for 1-oleoyldistearoylglycerol. State the
configuration of the double bond in the unsaturated acid residue.
2. Hydrolysis of a lipid produces palmitic acid and cetyl alcohol (1-hexadecanol,
C16H33OH). Determine the lipid type and write the its saponification reaction.
3. Draw the formula of a sphingomyelin containing the residue of arachidonic acid.
Indicate amide and ester bonds. Which products may be formed by the hydrolysis of
this lipid? Write the appropriate reactions, indicate the reaction conditions.

Topic 7. Lyophilic sols. Microheterogeneous and coarse dispersed systems.

Lyophilic colloids. Anionic, cationic zwitterionic and nonionic surfactants. Critical
micelle concentration (CMC). Solubilisation. Emulsion types: direct (o/w) and reverse
(w/o). Emulsifying agents. Hydrophilic-lipophilic balance (HLB). Phase inversion of
emulsions. Foams, foam multiplicity. Aerosols. Suspensions. Factors of stability.
Homework exercises
1. List the factors of stability for o/w type of emulsion stabilised by sodium oleate.
2. Which type of emulsion is formed by shaking vegetable oil (trioleoylglycerol) in the
presence of aqueous NaOH? Draw the structure of the particle of this emulsion.
3. Which of the following compounds can act as emulsifying agents in the system
toluene/water: calcium chloride, sodium stearate, glucose, bile, calcium oleate?
4. Which compounds can be used to produce foam in the system water/air: sodium
chloride, gelatine, sodium palmitate, sulfuric acid, benzene, bile, sucrose? List the
factors of stability for foams.
5. What will happen to the emulsion stabilized with potassium oleate if: a) an
aluminium sulfate solution is added to the emulsion; b) the emulsion is filtered
through a hydrophilic filter (such as paper)?

52
6. How the following interconversions can be achieved?
aerosol powder suspension
7. Explain why the most effective way to destroy aerosols is the condensation of water
vapour in the aerosol phase.
8. The drug is an inverse emulsion (w/o). What is the most likely route of its
administration (topical or oral)?
9. Pulmonary oedema is the accumulation of fluid in the lungs, which can be
accompanied by the formation of foam. How this foam can be destroyed?
10. Which of the following compounds are colloidal surfactants? a) C6H5CH2OH;
b) CH3CH2COOH; c) C16H33NH3+Cl–; d) CH3(CH2)2COOH; e) C18H37N(CH3)3+Cl–;
f) C17H31COONa; g) C10H21OSO3Na; h) CH3(CH2)4NH2.
11. Draw the structure of micelle for C12H25OSO3Na in aqueous solution. Indicate the
core, granule, diffuse layer, E- and ϛ-potentials. Which of the following compounds
can be included into the core of this micelle: toluene, KI, C6H13OH, C18H37NH2?
What is the name of this process?
12. The experimental values of CMC for two surfactants are 2 × 10–6 mol/L and
3 × 10–4 mol/L. Compare a) the lengths of hydrophobic tails, and b) the degrees of
aggregation for these surfactants.
13. Draw the scheme of solubilisation of C18H33OH and C12H25NH2 in an ionic micelle.
In which case the relative solubilisation will be greater?
14. List all types of disperse systems that can be obtained using water, toluene and
sodium oleate. Explain the terms "CMC" and "solubilisation".
Concepts and terms
Colloidal surfactants are long-chain surfactants that can form colloidal solutions. They
can be classified as ionogenic (anionic, cationic and zwitterionic) and nonionic
(nonelectrolytes).
Anionic colloidal surfactants contain alkylsulfate, alkylsulfonate and carboxylate
anions. The dissociation of such surfactants produces negatively charged surface active
anions.
Cationic colloidal surfactants are usually salts of primary, secondary, or tertiary
amines that produce positively charged surface active cations upon dissociation.
Zwitterionic (amphoteric) surfactants have both cationic and anionic centres attached
to the same molecule. The most common biological zwitterionic surfactants have a
phosphate anion with an amino or ammonium group, such as the phospholipids
phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine, and sphingo-
myelins.
Nonionic colloidal surfactants are nonelectrolytes and do not produce ions. Some
examples are fatty alcohols, fatty acids, amines, phenols, aminoalcohols,
polyethyleneglycol alkyl ethers and carbohydrates esters.
Micelles of colloidal surfactants are aggregates of amphipathic (having both hydrophilic
and hydrophobic parts) molecules or ions. Micelles in solution exist in equilibrium with
molecules or ions of a surfactant.
Critical micelle concentration (CMC) is the concentration of a surfactant above which
micelles begin to form.

53
Solubilisation is a spontaneous transfer of a compound insoluble in the continuous
phase into the solution due to incorporation into the surfactant micelles.
Emulsions are microheterogeneous systems containing two immiscible liquids, one of
which is dispersed as globules (dispersed phase) in the other liquid (continuous phase).
In direct emulsions (o/w), the dispersed phase is "oil" (organic compound) and the
continuous phase is water; in reverse emulsions (w/o), the dispersed phase is water and
the continuous phase is "oil".
Emulsification is the breakdown of fat globules into tiny droplets. Emulsification
provides a larger surface area on which the enzyme pancreatic lipase can act to digest
the fats into fatty acids and glycerol.
Bile salts and bile acids are surfactants that facilitate emulsification of fats and stabilize
the emulsion formed.
Hydrophilic-lipophilic balance (HLB) of a surfactant is a measure of the degree to
which it is hydrophilic or lipophilic.
Emulsion lifetime is the time it takes for the emulsion to be destroyed .
Foams are highly concentrated dispersed systems with a gaseous dispersed phase and a
liquid or solid continuous phase.
Foam multiplicity is the ratio of the foam volume and volume of the liquid from which
the foam was formed.
Foam stability (the foam half-life) is the time required for half the original foam
volume to settle.
Aerosols are microheterogeneous or ultramicroheterogeneous systems with a liquid
(clouds, fog) or solid (smoke) dispersed phase and a gaseous continuous phase.
Powders are microheterogeneous and coarse systems with a solid dispersed phase and a
gaseous continuous phase.
Suspensions are microheterogeneous and coarse systems with a solid dispersed phase
and a liquid continuous phase.
In-class exercises
1. Which of the following compounds are colloidal surfactants? a) CH3(CH2)3OH;
b) C15H31COONa; c) CH3(CH2)2NH2; d) C12H25OSO3Na; e) C8H17C6H4SO3Na;
f) [C18H37NH3]+Cl–; g) CH3COOH; h) C6H5OH.
2. Classify the following compounds as anionic, cationic zwitterionic or nonionic
surfactants: a) C17H33COOK; b) [C18H37NH3]+Br–; c) NH2(CH2)9COOH;
+ -
d) C12H25OSO3Na; e) [C16H33N ] Cl .
3. Why do lyophilic colloidal systems form spontaneously? What are the signs of ΔH
and ΔS for such processes?
4. Draw the isotherms of surface tension for colloidal and truly soluble surfactants and
explain the difference between these isotherms.
5. Draw the structure of micelle for C17H33COOK in aqueous solution. Indicate the
core, granule, diffuse layer, E- and ϛ-potentials. Which of the following compounds
can be included into the core of this micelle: benzene, KCl, CH3OH, C18H37NH2?
What is the name of this process?
6. Explain why the increase in concentration of a surfactant affects the shape of
micelle but not its diameter.
54
7. Which type of emulsion will form a) from toluene and water in the presence of soot
powder; b) from benzene and water in the presence of sodium oleate? Draw
schematically the structure of the second emulsion. What will happen to the
emulsion from question (b) if calcium chloride is added? What is the name of this
process?
8. Explain the fact that sodium oleate stabilises o/w emulsions while calcium oleate
stabilises w/o emulsions.
9. Which types of emulsions will form in the presence of sodium palmitate and
calcium oleate if their HLB values are 19 and 4.3, respectively?
10. What will happen to an emulsion or foam stabilised by a surfactant with HLB = 18
if: a) an alcohol is added; b) an inorganic acid is added; c) the system is centrifuged;
d) the system is heated up; e) the system is sonicated?
11. The height of the emulsion column (h) is 45 cm. Determine the lifetime of the
emulsion (τ) if the phase separation rate (U) is 1.5 cm/s. [Answer: 30 s]
12. A foam was formed from 180 mL of a protein solution, which together with the
remaining liquid (40 mL) occupied a volume of 400 mL. Determine the stability of
the foam (τ1/2) and its multiplicity (β) if the phase separation rate was 1.5 mL / s.
[Answer: τ1/2 = 120 s, β = 2.57]

Laboratory work "Determination of CMC by stalagmometric method"

Objective
To determining CMC using the graph of surface tension vs. the logarithm of
surfactant concentration.
Background
The critical micelle concentration (CMC) is defined as the concentration above
which micelles form. At low surfactant concentration, the surfactant molecules are
adsorbed on the surface. When more surfactant is added, the surface tension of the
solution decreases sharply at first, as more and more surfactant molecules migrate to the
surface. When the surface becomes saturated (fully covered with the surfactant
molecules), further addition of the surfactant will lead to the formation of micelles. This
concentration point is called the CMC. To see a sharp slope change at the CMC, you
will need to plot the surface tension against the logarithm of surfactant concentration.
Work procedure
1. For one of the following variants (assigned by your instructor), calculate the surface
tension for each solution of the colloidal surfactant using the formula
σx = σH2O·nH2O/nx, where nH2O is the number of drops of water, nx is the number of
drops of the surfactant solution, and σH2O is 72.8 × 10–3 J/m2.
2. Fill in the table and plot the experimental data of the surface tension (σ) versus the
logarithm of the surfactant concentration. Determine the CMC from the graph.

55
 Variant 1: nH2O = 77.
No. csurfactant, M nx σx, J/m2 × 10–3 lg c
1 0.000125 81
2 0.00025 85
3 0.0005 92
4 0.001 108
5 0.002 159
6 0.004 164
7 0.008 164
CMC = _____ mol/L
Variant 2: nH2O = 68.
No. csurfactant, M nx σx, J/m2 × 10–3 lg c
1 0.000125 75
2 0.00025 82
3 0.0005 90
4 0.001 109
5 0.002 154
6 0.004 155
7 0.008 155
CMC = _____ mol/L
Variant 3: nH2O = 68.
No. csurfactant, M nx σx, J/m2 × 10–3 lg c
1 0.000125 78
2 0.00025 83
3 0.0005 91
4 0.001 108
5 0.002 153
6 0.004 158
7 0.008 158
CMC = _____ mol/L
Online test "Microheterogeneous and coarse dispersed systems" (typical questions)
1. Which of the following compounds are colloidal surfactants?
a) CH3(CH2)3OH; b) C15H31COONa; c) CH3(CH2)2NH2; d) C12H25OSO3Na;
e) CH3COOH; f) C8H17C6H4SO3Na; g) [C18H37NH3]+Cl–; h) C6H5OH.
2. The critical micelle concentration (CMC) is the surfactant concentration in the
solution where:
a) surfactant molecules begin to distribute between the boundary surface and the
bulk of the solution;
b) the surface tension value of the solution reaches its maximum;
c) the surfactant begin to precipitate;
d) micelles of the surfactant begin to form.
56
3. Indicate the compounds that will be solubilised in direct micelles formed in the
following aqueous solutions:
a) hydrocarbons; b) inorganic salts;
c) water insoluble vitamins; d) water soluble dyes.
4. Microheterogeneous systems with a gaseous continuous phase and a solid dispersed
phase are called:
a) suspensions; b) aerosols; c) emulsions; d) foams.
5. Which of the following compounds will stabilize a reverse emulsion?
a) sodium chloride; b) sodium oleate (HLB = 20);
c) calcium stearate (HLB = 4.3); d) oleic acid (HLB = 1).
6. Which property of a disperse system suggests that it is a direct emulsion (o/w)?
a) low electroconductivity;
b) colouring of dispersed phase droplets with water-soluble dye;
c) ability to wet a hydrophobic surface;
d) the system is miscible with polar liquids.
7. Choose the correct statement about foams:
a) they are systems with liquid dispersed phase and liquid continuous phase;
b) they are heterogeneous systems;
c) they are aggregatively stable systems;
d) they form without stabilizers.
8. The quantitative characteristic of the stability of foams is the time required to:
a) decrease the foam volume by 50%;
b) destroy the foam completely;
c) retain the initial volume;
d) decrease the foam volume three times.
9. Choose the correct statement about aerosols:
a) they are systems with high kinetic (sedimentation) stability;
b) they are homogeneous systems;
c) they are l/g or s/g systems;
d) they are systems with high aggregative stability.
10. Decide whether the following statements true or false.
1. The driving force for the formation of micelles in solutions of colloidal
surfactants are hydrophobic interactions.
2. Foams are polydisperse g/g systems.
a) both are true; b) only the first is true; c) only the second is true; d) both are false.

57
Topic 8. α-Amino acids, peptides and proteins.
α-Amino acids. Classification, stereoisomerism, acid-base properties, isoelectric point
(pI). Derivatives of α-amino acids: esters, N-acylates, imines (Schiff's bases). Reactions
of α-amino acids: transamination, decarboxylation, elimination, retro-aldol reaction,
oxidative deamination, hydroxylation. Peptides, peptide bonds. Hydrolysis of peptides.
Proteins, their primary and secondary structures (α-helix, β-sheets).
Homework exercises
1. Draw the structural formula of cysteine and indicate the acidic centres in the
molecule. Assign the pKa values (pKa1 = 2.0, pKa2 = 8.2, pKa3 = 10.3) to the
appropriate functional groups. Calculate the pI for cysteine. Which forms of
cysteine exist in strongly acidic and strongly alkaline solutions?
2. Indicate the acidic and basic centres in lysine. Assign the pKa values (pKa1 = 2.2,
pKa2 = 9.0 and pKa3 = 10.45) to the appropriate functional groups. Calculate the pI
for lysine. Which forms of cysteine exist in strongly acidic and strongly alkaline
solutions, at pH= pI and pH = 7? For pH = 7, calculate the ratio of these forms.
3. Write the reaction of alanine with benzaldehyde and state the nucleophilic reagent.
4. The excess of ammonia, which is highly toxic for brain cells, reacts with glutamic
acid and removes from blood. Which α-oxoacid is the precursor of glutamic acid?
5. Write the reaction of L-aspartic acid with nitrous acid and name the products.
6. Draw the structural formula for dipeptide Val–Phe. Write the hydrolysis reaction for
this peptide under acidic conditions. Draw the electronic and spatial structure for the
peptide group.
7. Which compounds will form after the enzymatic hydrolysis of the tetrapeptide
Ala–Lys–Asp–Val? Note that the enzyme trypsin cleaves peptide bonds formed by
carboxyl group of lysine or arginine.
8. Indicate
_
the correct_structure for_ the zwitterion _of glutamic acid: _
1) OOC(CH2)2CHCOO ; 2) OOC(CH2)2CHCOO ; 3) HOOC(CH2)2CHCOO ;
+ NH + NH
NH2 3 3
_
4) OOC(CH2)2CHCOOH ; 5) HOOC(CH2)2CHCOOH .
+ NH + NH
3 3
9. In which form(s) does glutamic acid exist in: a) strongly acidic solutions; b) strongly
alkaline
_
solutions? _ _ _
1) OOC(CH2)2CHCOO ; 2) OOC(CH2)2CHCOOH ; 3) HOOC(CH2)2CHCOO ;
NH2 NH2 + NH
_ 3
4) HOOC(CH2)2CHCOO ; 5) HOOC(CH2)2CHCOOH .
+ NH
NH2 3
10. Which acid is the product of the transamination reaction for: a) alanine; b) aspartic
acid?
1) CH3CH2CCOOH ; 2) HOOCCH2CCOOH ; 3) HOOC(CH2)2CCOOH ;
O O O
4) CH3CCOOH ; 5) CH3CCH2COOH .
O O

58
 O O
11. Choose the correct name for the peptide H2NCHCNHCH2CNHCHCOOH :
CH3 CH2C6H5
1) alanylglycinephenylalanine; 2) alanylglycylphenylalanine;
3) phenylalanineglycinealanine; 4) alanylglycylphenylalanyl.
12. Select all correct statements about the structure and properties of a peptide bond:
1) the hybridisation state of C, O and N atoms is sp2;
2) the peptide bond can be hydrolyzed by both acids and bases;
3) the C=O bonds in peptides are longer than those in amino acids;
4) the peptide fragment is a three-centre p,π-conjugated system;
5) the peptide fragment is planar.
Concepts and terms
α-Amino acids are heterofunctional compounds whose molecules contain amine and
carboxyl group attached to the same carbon atom:
R CH COOH
NH2
There are about 100 naturally occurring α-amino acids. Of these, 20 are encoded by the
universal genetic code.
Ionogenic α-amino acids are amino acids with ionisable side chains.
Nonionogenic α-amino acids are amino acids with non-ionisable side chains.
Polar α-amino acids contain polar side chain.
Nonpolar α-amino acids contain nonpolar side chain.
Acidic α-amino acids have acidic side chains (such amino acids contain two carboxyl
groups and one amino group in the whole molecule).
Basic α-amino acids have basic side chains (such amino acids contain one carboxyl
group and two or more basic functional groups in the whole molecule).
Neutral α-amino acids are amino acids that contain one carboxyl and one amino group.
Ornithine NH2 CH2 CH2 CH2 CH COOH is a non-proteinogenic amino acid that
NH2
plays a role in the urea cycle.
CH2 CH COOH
S NH2
Cystine (Cys-S-S-Cys) S
is a nonessential amino acid, the oxidized
CH2 CH COOH
NH2
dimeric form of the amino acid cysteine.
Isoelectric point (isoionic point, pI) is the pH at which the amino acid has zero net
ionic charge (i.e., the zwitterion form is dominant).
Dipolar ion (zwitterion) is a neutral species with both positive and negative electrical
charges. α-Amino acids exist in their dipolar forms at pH = pI.
Anionic form of α-amino acid is a negatively charged form of its molecule (anion) that
exists at pH > pI.
Cationic form of α-amino acid is a positively charged form of its molecule (cation)
that exists at pH < pI.
59
Elimination reaction for α-amino acids can involve the elimination of ammonia (non-
oxidative deamination), water (from hydroxyamino acids) or H2S (from cysteine). In the
last two cases, the amino acids are converted into α-oxoacids. The reaction involves
vitamin B6.
Hydrolytic deamination of α-amino acids is the substitution of the amino group with a
hydroxyl group. Hydrolytic deamination in vivo takes place in bacteria and fungi;
in vitro it can be carried out by the reaction with nitrous acid.
Non-oxidative deamination of α-amino acids is the elimination of ammonia
accompanied by the formation of a double carbon-carbon bond. It is typical for aspartic
acid and histidine.
Transamination is a chemical reaction that transfers an amino group to a oxoacid to
form a new amino acid and a new oxoacid. This pathway is typical for most amino acids
and occurs in the presence of vitamin B6.
Aldol cleavage of α-amino acids is the cleavage of serine and threonine into glycine
and corresponding aldehyde (formaldehyde or acetaldehyde) catalyzed by vitamin B6 .
Decarboxylation of α-amino acids is a removal of the carboxyl group and the
formation of biogenic amines and CO2. It involves vitamin B6.
N
Histamine CH2 CH2 NH2 is a biogenic amine, produced by the decarboxylation of
N
H
histidine. Histamine plays a major role in many allergic reactions.
Tryptamine CH2 CH2 NH2 is a common precursor to many hormones and

N
H
neurotransmitters. It can be obtained by the decarboxylation of tryptophan.
Aliphatic hydroxylation of α-amino acids is the introduction of a hydroxyl group into
a molecule of lysine and proline in the synthesis of connective tissue proteins. It
involves vitamin C.
5-Hydroxylysine (5-OH-Lys, Hyl) NH2 CH2 CH CH2 CH2 CH COOH is a hydroxylated
OH NH2
derivative of the amino acid lysine that is present in certain collagens.
HO

4-Hydroxyproline (4-OH-Pro, Hyp) COOH is a hydroxylated derivative of

N
H
proline, a major component of the protein collagen.
Aromatic hydroxylation of α-amino acids is the introduction of a hydroxyl group into
the aromatic ring. Aromatic hydroxylation converts phenylalanine into tyrosine,
tyrosine into catecholamines, and tryptophan into serotonin.
HO
Serotonin or 5-hydroxytryptamine (5-HT) CH2 CH2 NH2 is a neuro-

N
H
transmitter, the product of the aromatic hydroxylation of tryptophan. It is thought to
contribute to the feelings of well-being and happiness. .

60
Dihydroxyphenylalanine (DOPA) HO CH2 CH COOH is the product of the
NH2
HO
hydroxylation of tyrosine.
Dopamine (DA) HO CH2 CH2 NH2 is the product of DOPA decarboxylation.
HO
I

Diiodotyrosine (DIT) HO CH2CHCOOH is a precursor in the production of thyroid

NH2
I
hormone, and is formed by the aromatic iodination of monoiodotyrosine.
I I

Thyroxine (T4) HO O CH2 CH COOH is the thyroid hormone, the

NH2
I I
prohormone of triiodothyronine (T3), produced by the thyroid gland from 3,5-
diiodothyronine.
3-Iodotyrosine (mono-iodotyrosine) HO CH2 CH COOH is the product of
NH2
I
tyrosine iodination and a thyroid hormone that can be further iodinated by thyroid
peroxidase to form di- and triiodo forms.
Oxidative deamination of α-amino acids is the conversion of amino acids into
α-oxoacids that involves the oxidation of the amino acid into imino acid and subsequent
hydrolysis with the release of ammonia.
Reductive amination of α-oxoacids is the conversion of a carbonyl group into an
amino group via an intermediate imine (a reverse reaction with respect to oxidative
deamination). Reductive amination is a step in the biosynthesis of many α-amino acids.
Peptide is the condensation product of two or more amino acids, in which a carboxyl
group of one is linked with an amino group of another. Formally, it can be represented
as a product of intermolecular dehydration of several amino acids in a certain sequence.
Peptide group is the –CO–NH– linkage formed between the carboxyl group of one
amino acid and the amino group of another; it is an amide linkage joining amino acids
to form peptides.
O
Peptide bond is the amide bond C N in peptides and proteins.
H
Oligopeptide is a peptide with a relatively small number (2–10) of amino acid residues.
Polypeptides contain more than 10 residues of amino acids.
Proteins are large biomolecules, or macromolecules, consisting of one or more long
chains of amino acid residues.
C-terminal is the end of a protein or polypeptide with a free carboxyl group (–COOH).
N-terminal is the end of a protein or polypeptide with a free amino group (–NH2)
Primary structure of proteins and peptides is the linear sequence of amino acids.
Secondary structure of proteins is the three-dimensional shape of local segments of
proteins. The most common types of secondary structures are the α-helix and the

61
β-pleated sheet. Both structures are stabilized by hydrogen bonds, which form between
the fragments of peptide bonds: C=O HN .
Tertiary structure of proteins is the overall three-dimensional structure of the
polypeptide chain supported by interactions between the functional groups in the side
chains of amino acid units.
Quaternary structure of proteins is the three-dimensional arrangement of two or more
polypeptide chains (protein subunits).
Protection of functional group in peptide synthesis is the first step in peptide
synthesis that allows to reduce the reactivity of a functional group in order to obtain
chemoselectivity in subsequent chemical reactions. Amino groups are usually protected
by acylation, and carboxyl groups by esterification.
Activation of carboxyl group in peptide synthesis is the increase in reactivity of the
carboxyl group, which is typically achieved by converting it into a mixed anhydride.
The removal of protecting group is the final step in peptide synthesis.
Simple protein is a protein that yields only amino acids and no other major products
when hydrolyzed.
O O
R CH C
Aminoacyl adenylates O P O CH2 Ade are intermediates in protein
NH3 O
O

OH OH
biosynthesis.
Glycoproteins are proteins that contain oligosaccharide chains (glycans) covalently
attached to polypeptide side-chains.
Glutathione (GSH) is a tripeptide γ-glutamylcysteinylglycine (γ-Glu-Cys-Gly) with a
gamma peptide linkage between the carboxyl group of the glutamate side chain and the
amino group of cysteine. The carboxyl group of cysteine is attached by normal peptide
linkage to the amino group of glycine. GSH is an important antioxidant in plants,
animals, fungi and some bacteria.
Nucleoproteins are any proteins that are structurally associated with nucleic acids,
either DNA or RNA.
Chromoproteins are proteins (such as hemoglobin or rhodopsin) containing a pigment
non-protein group (such as heme, riboflavin or retinal).
Essential (indispensable) α-amino acids (Val, Leu, Ile, Met, Phe, Trp, Lys and Thr)
cannot be synthesized in the human body and must be obtained from food.
Non-essential (dispensable) α-amino acids (Ala, Asp, Asn, Glu, Gln, Gly, Pro and
Ser) can be synthesized by the human body and so are not essential for the human diet.
Semi-essential amino acids (Cys and Tyr) are the amino acids that must be consumed
in adequate amounts to prevent the use of essential amino acids for their synthesis. Cys
and Tyr are synthesised from the essential amino acids Met and Phe, respectively.
Conditionally essential α-amino acids (His and Arg) are usually not essential, except
at times of illness and stress.

62
 Proteinogenic α-amino acids
Neutral amino acids with nonpolar side chains (hydrophobic except glycine)
H CH COOH
NH2 N COOH
Glycine (Gly or G) H
CH3 CH COOH Proline (Pro or P)
NH2
CH2 CH COOH
Alanine (Ala or A)
NH2
CH3
CH CH COOH Phenylalanine (Phe or F)
CH3
NH2
CH3S (CH2)2 CH COOH
Valine (Val or V) NH2
CH3
CH CH2 CH COOH Methionine (Met or M)
CH3
NH2 CH2 CH COOH
Leucine (Leu or L) NH2
C2H5 N
CH CH COOH
CH3 H
NH2
Tryptophan (Trp or W)
Isoleucine (Ile or I)
Neutral amino acids with polar side chains (hydrophilic)
HO CH2 CH COOH HS CH2 CH COOH
NH2 NH2
Serine (Ser or S) Cysteine (Cys or C)
CH3
CH CH COOH HO CH2 CH COOH
HO
NH2 NH2
Threonine (Thr or T) Tyrosine (Tyr or Y)
H2N(O)C CH2 CH COOH H2N(O)C (CH2)2 CH COOH
NH2 NH2
Asparagine (Asn or N) Glutamine (Gln or Q)
Amino acids with acidic side chains (hydrophilic)
HOOC CH2 CH COOH HOOC (CH2)2 CH COOH
NH2 NH2
Aspartic acid (Asp or D) Glutamic acid (Glu or E)
Amino acids with basic side chains (hydrophilic)
H2N (CH2)4 CH COOH CH2 CH COOH H2N C NH (CH2)3 CH COOH
N
NH2 NH2 NH NH2
N
Lysine (Lys or K) H Arginine (Arg or R)
Histidine (His or H)

63
In-class exercises
1. Complete the following reactions:
1) Reactions involving the carboxyl group:
2 NH
3
a) Ala + C2H5OH + HCl A –NH Cl B
4
NH3 /ATP
b) Glu Gln
2) Reactions involving the amino group:
a) Lys + HNO2 (excess) b) Gly + acetylphosphate
c) Met + ethanal
3) Biologically important reactions:
a) Trp (decarboxylation) b) Phe (aromatic hydroxylation)
c) Cys (oxidation) d) Asp (non-oxidative deamination)
+
NAD /H2O transamination
e) Ala oxidative deamination f) Glu + pyruvic acid
elimination tautomerism hydrolysis
g) Ser A B C
–H2O
2. Draw the structural formula for the tetrapeptide Ala–Pro–Glu–Lys. Indicate peptide
bonds, C-terminal and N-terminal.
3. Draw the structural formula for the dipeptide Asp–Lys. Write the hydrolysis
reactions for this peptide under acidic and basic conditions.
4. Draw the structure of asparagine and indicate acidic centres in its molecule. Assign
the pKa values (pKa1 = 2.0, pKa2 = 8.8) to the appropriate functional groups.
Calculate the pI for asparagine. Which forms of asparagine are present in solution at
pH = 7.8? Calculate the ratio for these forms.
5*. For lysine pKa1 = 2.2, pKa2 = 9.0, pKa3 = 10.45 (NH3+ in the side chain). Calculate
the pI for lysine. Which forms of this amino acid are present in solution at pH = 8.0?
Calculate the ratio for these forms.
Online test "Amino acids" (typical questions)
1. Match the names of amino acids with their formulae:
1) glycine; 2) valine; 3) asparagine; 4) proline
CH3
a) CHCHCOOH; b) COOH; c) H2N C CH2CHCOOH; d) H2NCH2COOH.
CH3 N
NH2 O NH2
H
2. For glycine pI = 6.0. In which forms does it exist at pH = 4.3?
a) cationic and dipolar forms; b) anionic and dipolar forms;
c) cationic and anionic forms; d) dipolar form only.
3. Choose the pH range for a buffer system of glycine (pKa1 = 2.34, pKa2 = 9.60)
consisting of the cationic and dipolar forms:
a) 1.34–3.34; b) 2.34–4.34; c) 7.60–9.60; d) 2.34–9.60.
4. What is the common three-letter abbreviation for the amino acid below?
CH2CHCOOH
NH2

64
5. Match the names of peptides with their formulae:
1) glycylvaline; 2) alanylglycyl; 3) valylglycyl; 4) leucylvaline.
O O
a) H2NCHC NHCH2COOH b) H2NCHC NHCHCOOH;
CH(CH3)2 CH2 CH(CH3)2
CH(CH3)2
O O
c) H2NCH2C NHCHCOOH; d) H2NCHC NHCH2COOH.
CH(CH3)2 CH3
6. Which of the following is the dipeptide Cys–Thr?
O CH3 O
a) H2NCHC NHCHCOOH; b) H2NCHC NHCHCOOH;
CH2COOH CH2SH CH OH
CH3
O O CH3
c) H2NCHC NH(CH2)4CHCOOH; d) H2NCHC NHCHCOOH.
CH(CH3)2 NH2 (CH2)3NH C NH2
NH
7. The hydrolysis of the dipeptide Leu–Val under acidic conditions produces:
+ + + +
a) H3N CHCOOH ;b) H3N CHCOOH; c) H3N CHCOOH; d) H3N CHCOOH .
CH2CH(CH3)2 CH3 CH(CH3)2 H3C CH CH2CH3
8. Which of the following is the product of decarboxylation for leucine?
a) tert-butylamine; b) isopentylamine;
c) 3-methylpentanoic acid; d) 2-methylbutanoic acid
9. Which of the following is the product of oxidative deamination for alanine?
a) lactic acid; b) pyruvic acid; c) malic acid; d) ethylamine.
10. The reaction of isoleucine with 2-propanol in the presence of hydrogen chloride
taken in excess gives:
a) isoleucine isopropyl ester hydrochloride;
b) isoleucine isopropyl ester;
c) isopropyl ester of 2-aminohexanoic acid;
d) isoleucine hydrochloride.

Test "Amino acids" (typical questions)

1. For glycine pKa1 = 2.3, pKa2 = 9.6. Calculate the pI. Which forms of this amino acid
are present in solution at pH = 8.6? Calculate the ratio for these forms.
2. Draw the formula for the following tripeptide Ala–Trp–Lys. Indicate peptide bonds,
N- and C-terminals.
3. Complete the following reactions:
a) Val + acetyl SCoA b) Phe (hydroxylation)
c) Ser (elimination) d) Trp (decarboxylation)

65
Topic 9. Polymers. Acid-base properties of biopolymers. Solutions of polymers:
formation, properties and stability.
Classification of polymers (natural, synthetic, linear, branched and cross-linked
polymers, non-electrolytes, polyelectrolytes, polyampholytes). Acid-base properties of
proteins, ionic forms of protein molecules, isoelectric state and isoelectric point (pI) of
proteins. Formation of polymers solutions, swelling, factors affecting the degree of
swelling, thermodynamics of swelling.

Homework exercises
1. Which phenomena (from listed below) are typical for: a) the first stage of swelling
b) the second stage of swelling?
1) insignificant increase in mass and volume of HMW; 2) significant increase in
mass and volume of HMC; 3) swelling pressure; 4) hydration of macromolecules;
5) contraction; 6) diffusion of macromolecules into the solvent; 7) diffusion of the
solvent into HMC.
2. Which changes (listed below) of the thermodynamic parameters in the system
HMW/solvent are observed at: 1) the first stage of swelling; 2) the second stage of
swelling?
1) ΔH > 0; 2) ΔH < 0; 3) ΔS > 0; 4) ΔS < 0; 5) ΔG > 0; 6) ΔG < 0?
3. Compare a) the degree of swelling in water, and b) the solubility in water at 25 °C
for albumin (pI = 4.7) and hemoglobin (pI = 6.8).
4. Compare the degree of swelling for gelatine (pI = 4.7) in pure water and in the
presence of a small amounts of: a) CH3COOH; b) CH3COONa; c) NaCl; d) KCNS.
Explain your answer.
5. The swelling of natural rubber is limited in ethanol and unlimited in toluene while
the swelling of vulcanized rubber is limited in both solvents. Explain this difference.
6. Calculate the degree of swelling for natural rubber if 2 g of the rubber absorbs 5 mL
of benzene with ρ = 0.86 g/mL. [Answer: α = 215%]
7. The pI values for polymers A and B are 6.4 and 4.7, respectively. Draw the graph of
the swelling degree against pH for these polymers. In which pH range the
competitive swelling of these polymers can be observed? Which polymer will
dehydrate and which one will swell if pH increases from 4.7 to 6.4?
8. What is the difference between the properties of "bound water" and "free water"?
What is the biological role of "bound water"?

Concepts and terms

High molecular weight (HMW) compounds are natural or synthetic polymers
(Mr > 10 000–15 000) that are composed of simpler chemical units called monomers.
Biopolymers are polymers produced by living organisms (polymeric biomolecules).
Polyelectrolytes are polymers that contain ionisable (ionogenic) groups in their
repeating units.
Polyampholytes are amphoteric polyelectrolytes containing either acidic or basic
ionisable groups (for example, –COOH and –NH2).

66
Isoelectric state of protein is the state of the protein in solution when the number of
ionized carboxyl groups is equal to the number of protonated amino groups, i.e., the
total number of negative charges is equal to the total number of positive charges, and
the total charge of the macromolecule is zero.
Isoelectric point (pI) of a protein is the pH at which the protein has zero net charge
(i.e., the protein exists in its isoelectric state).
Unfolded form of a polypeptide molecule exists in a solution at pH >> pI or pH << pI.
Random coil is the conformation of a protein or other polymer where the monomer
subunits have random orientations.
Globule is a spherical ("globe-like") macromolecule.
Swelling is the increase in volume and mass of a polymer due to the absorption of
solvent.
Contraction is the decrease in the total volume of the system polymer/solvent at the
first stage of swelling.
Bound water is the water molecules that form thin hydration layers tightly bound to the
molecules of polymer.
Free water is the bulk of the solvent in an aqueous solution.
Swelling degree is the extent of swelling of polymers that can be determined via the
m − m0 V −V
changes in mass or volume: swelling = s × 100% or swelling = s 0 × 100%,
m0 V0
where m0 or V0 is the mass or volume of the original polymer and ms or Vs is the mass or
volume of the swollen polymer.
Limited swelling occurs when the macromolecules are bonded to one another strongly,
and swelling stops after having reached a certain limit. The swelled polymer retains its
shape and a distinct boundary with the liquid phase.
Unlimited swelling leads to the gradual disappearance of the interphase boundary
between the polymer and liquid, and eventually to complete dissolution of the polymer.
Competitive swelling is the swelling of one HMW compound due to the dehydration of
another.
Electrophoresis is the movement of charged particles in a fluid or gel under the
influence of an electric field.

In-class exercises
1. Classify the following polymers by a) source, b) structure, c) acid-base properties:
cellobiose, polyvinyl acetate, rubber, albumin, heparin, DNA.
2. Determine the type (polymer, polyelectrolyte, polyampholyte) of the following
compounds: a) polyalanine, b) polyacrylic acid, c) acrylic acid, d) rubber, e) nucleic
acid, f) cellobiose, g) polyethylene.
3. Decide whether the following statements are true or false.
1) Molecules of HMC differ from colloidal particles by their: a) size; b) shape;
c) flexibility.
2) Solutions of biopolymers form spontaneously and are thermodynamically stable.
3) Solutions of biopolymers are hydrophobic systems.
4) Solutions of biopolymers are true solutions.

67
4. The process of ionization of a peptide molecule can be represented as follows:
(+NH3)nPt(COOH)m (+NH3)nPt(COO–)m + mH+ (NH2)nPt(COO–)m + nH+
pH < pI m = n, pH = pI pH > pI
Which forms (cationic, zwitterionic, anionic) of peptide molecules can produce
buffer systems at: a) pH < pI; b) pH > pI?
5. The pI of an albumin is 4.7. What is the ratio of acidic and basic functional groups
in its molecule? In which form (zwitterionic, cationic or anionic) does this molecule
predominantly exist at pH = 7.4?
6. The pH of a solution containing β-lactoglobulin (pI = 5.2) and γ-globulin (pI = 6.6)
is 5.0. What are the charges of these proteins? Which electrode (electrodes) will
they move to? Which of the proteins will move faster (assume that the radii of their
solvated ions are identical)?
7. The pI range of blood plasma peptides is 4.9–7.0. Which types of buffer systems do
these peptides form in the blood (pH = 7.4)?
8. Calculate the number of water molecules bound with: a) one molecule of albumin,
b) one amino acid residue in this protein if 1 g of albumin binds with 0.3 g of water,
M(albumin) = 68 000 g/mol, and one molecule of the protein contains 515 amino
acid residues. [Answer: 1133; 2.2]
9. Which of the following polymers will swell and/or dissolve in: a) water; b) nonpolar
solvents (such as benzene): globular proteins, rubber, vulcanized rubber, fibrous
proteins, starch?
10. Which of the following statements are true for: a) the first stage of unlimited
swelling; b) the second stage of unlimited swelling?
1) the solvent molecules diffuse into the polymer to produce a swollen gel;
2) the solvated macromolecules diffuse into the solvent;
3) ΔH < 0;
4) ΔH ≈ 0;
5) ΔS < 0;
6) ΔS > 0;
7) ΔG < 0?
11. Compare the degree of swelling for gelatine (pI = 4.7) in pure water and in the
presence of a small amount of: a) HCl; b) NaOH; c) Na2SO4; d) C2H5OH. Explain
your answer.
12. Calculate the degree of swelling for starch if 1 g of the polymer absorbs 0.3 mL of a
solution with ρ = 1.05 g/mL. [Answer: α = 31.5%]
13. The pI values for polymers A and B are 5.4 and 3.7, respectively. In which pH range
the competitive swelling of these polymers can be observed?
14. The pI value of a protein is 6.8. How will the following properties
a) stability; b) posm; c) viscosity
change if the pH of the protein solution increases from 4 to 8?
15. How will the osmotic pressure of hemoglobin (pI = 6.8) in water change (decrease,
increase remain the same) if a small amount of a) HCl; b) NaOH; c) C2H5OH is
added to the solution? Explain your answer.

68
 Test "Acid-base properties of biopolymers, swelling" (typical questions)
1. At which of the given pH values the protein myoglobin (pI = 8.2) will:
1) have the maximum degree of swelling;
2) move to the cathode during electrophoresis;
3) have the lowest solubility in water?
a) pH = 4.2; b) pH = 6.9; c) pH = 8.3.
2. Which forms of a protein with pI = 6.9 will produce a buffer solution at pH = 8.5?
Draw these forms schematically.
3. Calculate the degree of swelling for gelatine if 2 g of the polymer absorbs 5 mL of a
solution with ρ = 1.05 g/mL. [Answer: α = 26.25%]

Topic 10. Properties of solutions of HMW compounds. Factors affecting the

stability of protein solutions. Aggregation in solutions of HMW compounds.
Properties of protein solutions that differ from those of solutions of low molecular
weight compounds. Salting out, denaturation and gelation.
Homework exercises
1. When a 0.1% agar-agar solution was cooled from 45 to 10 °C, a gradual increase in
the solution viscosity and the loss of fluidity were observed. Which process could
cause these phenomena?
2. In which of the two sols of iron(III) oxide (2% or 3%) the process of gelation will
begin earlier? Explain your answer.
3. A 2% solution of gelatine turns into gel at room temperature while a 0.5% solution
of gelatine does not. How the 0.5% solution can be gelated at the same temperature?
4. The addition of K2SO4 causes the gelation of a 5% gelatine solution in 25 minutes,
the addition of KI in 200 minutes, and KCNS does not cause gelation at all. How
can you explain it?
5. Consider three sols with the same mass fraction of the dispersed phase but different
shapes of dispersed phase particles: the first colloid contains rod-shaped particles,
the second contains leafy particles, and the third contains spherical particles. Which
of the sols will convert into gel first?
6. How does the temperature affect the gelation process? What can happen to a gel
when its temperature increases?
7. When the blood clot is heated to 35–40 °C, a slightly coloured, almost transparent
serum is separated. Name this phenomenon.
8. At the chemistry exam, the student gave the following definition of syneresis: "This
is when the gel is crying when aging." Do you think that the examiner could accept
this answer?
9. A suspension of bentonite clay with a mass fraction of the dispersed phase of 10% is
a semi-solid material that has plasticity but not fluidity. However, it can be liquefied
by shaking and then easily poured out of the vessel. After a while, the liquid
suspension solidifies again. What is the name of the phenomena described?
10. What kind of structural change (condensation of coagulation) is possible in a
thixotropic system?

69
Concepts and terms
Association is the interaction of identical or closely related molecules in solutions or
pure liquids with the formation of relatively unstable groups (associates), in which the
molecules are held together by non-covalent (such as van der Waals') forces.
Relaxation time is the time required for an exponentially decreasing variable to drop
from its initial value to 1/e or 0.368 of that value (where e is the base of natural
logarithms). Relaxation time depends on the type of the system and can vary from
10–13 s to millions of years.
Coacervation is the phenomenon of phase separation and formation of a very
concentrated colloidal phase ("coacervate") that coexists with a dilute colloidal phase.
Coacervate phase can remain in the medium as droplets or form a continuous layer.
Flocculation is the formation of aggregates (flocs) from coacervate droplets.
Salting out is the precipitation of proteins from their solutions containing salts at high
concentrations.
Complex coacervation is the coacervation caused by the interaction of two oppositely
charged colloids.
Native state (of a protein or nucleic acid) is the state in which the biopolymer is
properly folded and/or assembled and thus is operative and functional.
Denaturation is the loss of native structure (quaternary, tertiary and secondary) by proteins or
nucleic acids as a result of external stress or added chemicals.
Gelation is the formation of a gel from a sol.
Gel is a dispersion of a liquid within a solid where the solid is a cross-linked system
(continuous phase) and the liquid is the discontinuous phase.
Coagulation structures are elastic structures formed at the beginning of the gelation
process, when the interlayers of the dispersion medium still exist at the sites of particle
contact.
Condensation structures are brittle structures formed from coagulation structures by
extruding medium interlayers and increasing the areas and strength of the contacts
between particles.
Thixotropy is the ability of some gels to become liquid when stirred or shaken.
Syneresis is the contraction of a gel accompanied by the separation of a liquid.
In-class exercises
1. Match the definitions (1–5) with the process names (a–g).
1) The gradual addition of ammonium sulfate to blood serum caused the sequential
precipitation of two protein fractions (globulins and albumins).
2) A solution of the enzyme pepsin, inactivated by heating, regains its original
activity after cooling down to ambient temperature.
3) When a 5% solution of gelatine is added to a 5% solution of starch, small droplets
are formed, and after a while the liquid separates spontaneously into two layers with
a clear boundary between them.
4) The addition of copper(II) sulfate solution to a solution of egg protein produces a
pale blue precipitate that is insoluble in water.
5) When a 5% solution of gelatine is cooled down, an increase in the solution
viscosity and the loss of fluidity are observed.

70
 a) salting out; b) reversible denaturation; c) irreversible denaturation;
d) coacervation; e) complex coacervation; f) gelation; g) syneresis.
2. Which factor (a–f) will initiate the following processes in a protein solution:
1) salting out; 2) gelation; 3) denaturation.
a) addition of guanidine; b) addition of ethanol; c) decrease in temperature;
d) addition of Pb(NO3)2; e) addition of ammonium sulfate; f) pH ≈ pI.
3. Explain how sodium chloride can be used to separate albumin (M ≈ 68 000,
pI ≈ 4.9) and globulin (M ≈ 160 000, pI ≈ 6.4) from their mixture. Which other
electrolytes can be used for the separation of these proteins? Is it possible to use the
Cohn method for this purpose?
4. What is the role of denaturation in biological systems? Can denaturation be used to
isolate proteins for medical purposes?
5. Why egg white can be used as the first aid in the treatment of poisoning with soluble
salts of heavy metals?
6. Explain the fact that the structuring of proteins is favoured at pH ≈ pI.
7. Which phenomena cause the aging of protein structures in the body?
8. What is the role of thixotropy in biological systems?
9. Which types of intermolecular interactions are possible in protein solutions?
10. What is the main difference between properties of solutions of HMW and LMW
compounds?
11. Why the solutions of HMW compounds are thermodynamically stable?
12. Using the table below, describe a technique that can be used for isolating fibrinogen
and albumin from their mixture in a solution. You are allowed to use the following
reagents: a solution of an acid, a solution of a base and either a solid salt or a
solution of a salt.
Selected proteins and enzymes
Number of amino acid residues
Protein Type Mr pI
total basic acidic neutral
insulin globular 8000 50 6 4 40 8.2
pepsin globular 22000 150 4 71 75 1.0
albumin globular 62000 520 99 126 295 4.7
hemoglobin globular 66000 570 94 52 430 6.8
collagen fibrillar 135000 960 85 124 750 6.0
fibrinogen fibrillar 360000 2000 400 400 1200 6.0
13. Is it possible to use denaturing reagents for isolating proteins in their native states?
Explain your answer.
14. What are the roles of complex coacervation and denaturation of proteins in
biological systems?

71
 Laboratory work "Salting out of proteins"
Objective
Salting out of albumins (M ≈ 68 000, pI ≈ 4.9) and globulins (M ≈ 160 000,
pI ≈ 6.4) by addition of NaCl at different pH and (NH4)2SO4 of different concentrations.
Background
Proteins are precipitated from aqueous solutions when the salt concentration exceeds a
certain critical level. This process is known as salting out, because all water in the
solution is "bound" to the salts and therefore is not available for hydrating the proteins.
Ammonium sulfate, (NH4)2SO4, is commonly used because it is readily soluble in water
and forms slightly acidic solutions. Other salts, such as NaCl or KCl, may also be used.
The isoelectric point (pI) of a protein is the pH where the net charge on the protein is
zero. Proteins tend to aggregate and precipitate at their pI due to the lack of electrostatic
repulsion between their macromolecules. Proteins have different isoelectric points
because of their different amino acid composition (i.e., relative numbers of anionic and
cationic groups) and thus they can be separated by adjusting the solution pH. When the
pH becomes equal to the pI of a particular protein, that protein precipitates while other
proteins remain in the solution.
Salting out of proteins by NaCl at different pH
Work procedure
1. Place approximately 5 mL of the protein solution into a test tube and add powdered
sodium chloride in small portions until no more salt can be dissolved (complete
saturation).
2. Let the solution stand for 10–15 min, then filter off the precipitate.
3. Check the filtrate for the absence of proteins by using the biuret test.
The biuret test is a chemical test used for detecting the presence of peptide bonds.
Peptides form violet-coloured complexes with copper(II) ions in alkaline solutions.
The test is named so because it also gives a positive reaction to the peptide-like
bonds in the biuret molecule, (H2NCO)2NH:
O
C NH2 H2N C O O C NH2
2 HN + Cu(OH)2 N Cu N
C NH2 −2 H2O C O O C
H2N NH2
O
Transfer 1 mL of the filtrate into another test tube, add 5–7 drops of a 10% NaOH
solution and 2 drops of a 1% CuSO4 solution. Stir the mixture. Record the solution
colour.
4. Add 1.5 mL of a 1% acetic acid solution to the filtrate obtained in step 2. Let the
solution stand for 10–15 min, then filter of the precipitate.
5. Check the filtrate for the absence of protein by using the biuret test.
Results and evaluation
Which proteins were precipitated in a saturated NaCl solution? Which proteins were
precipitated in the NaCl solution after acetic acid was added? Explain your answer.

72
 Salting out of proteins by (NH4)2SO4 of different concentrations
Work procedure
1. Place approximately 5 mL of the protein solution into a test tube, add 2.5 mL of a
semi-saturated (ω ≈ 22%) solution of (NH4)2SO4 solution and stir the mixture.
2. Let the solution stand for 5 min, then filter off the precipitate, transfer 1 mL of the
filtrate into another test tube and check the absence of proteins by the biuret test.
3. Add powdered ammonium sulfate in small portions to the remaining filtrate until no
more salt can be dissolved and let the mixture stand for 5 min.
4. Filter off the precipitate and check the filtrate for the absence of protein by using the
biuret test.
Results and evaluation
1. Which proteins were precipitated in semi-saturated (NH4)2SO4 solution? Which
proteins were precipitated in saturated (NH4)2SO4 solution? Explain your answer.
Laboratory work "Denaturation of proteins"
Objective
Explore different methods of denaturing proteins.
Background
Denaturation is the disruption of secondary, tertiary, and quaternary structures of
proteins, which leads to the loss of their biological activity. Denaturation can be
caused by heat, heavy metal ions, pH change and the addition of organic solvents.
Work procedure
1. Place 5 drops of the protein solution into a test tube and add 1–2 drops of a 10%
CuSO4 solution. Record your observations.
2. Place 5 drops of the protein solution into a test tube. Tilt the tube slightly and
carefully place 5 drops of concentrated nitric acid on the side of the tube so as to
form two layers. Record your observations.
3. Place 5 drops of the protein solution into a test tube and add 2 drops of a 20%
trichloroacetic acid solution. Record your observations.
4. Place 5 drops of the protein solution into a test tube, add 15–20 drops of acetone and
1 drop of a saturated NaCl solution. Record your observations.
Results and evaluation
1. Explain the mechanism of action for the denaturing reagents you have used.

Online test "Solutions of HMW compounds" (typical questions)

1. Which of the following HMW compounds are proteins?
a) synthetic organic; b) natural inorganic;
c) synthetic inorganic; d) natural organic.
2. Proteins consist of:
a) α-amino acid residues; b) α,β-diamino acid residues;
c) β-amino acid residues; d) γ-amino acid residues.

73
3. If the isoelectric point of a protein is less than 7, it consists of:
a) approximately equal numbers of acidic and basic amino acid residues;
b) mainly acidic amino acid residues;
c) mainly basic amino acid residues;
d) mainly aromatic amino acid residues.
4. At which pH the electrophoretic mobility for albumin (pI = 4.7) will be the lowest?
a) 11.0; b) 8.6; c) 5.3; d) 4.6.
5. What will happen to the degree of swelling of gelatine (pI = 4.7) in water if a small
amount of HCl is added?
a) it will decrease first, then increase; b) it will increase;
c) it will not change; d) it will decrease.
6. Which of the following phenomena is typical for the first stage of swelling?
a) absorption of 70–80% of water (relative to dry polymer weight);
b) a slight increase in volume of the HMW compound;
c) the diffusion of the HMW compound into the solvent;
d) a significant increase in the volume of the HMW compound.
7. Salting out is:
a) the formation of spatial structures in sols;
b) the reversible conversions gel–sol or gel–solution of HMW compounds;
c) the formation of spatial structures in solutions of HMW compounds;
d) precipitation of proteins caused by the addition of a salt to the solution.
8. The addition of copper(II) chloride to a protein solution causes:
a) salting out of protein; b) complex coacervation;
c) syneresis; d) denaturation.
9. Gelation is:
a) the reversible conversions gel–sol or gel–solution of HMW compounds;
b) the structuring of protein molecules in a solution;
c) the precipitation of proteins caused by the addition of salts or organic solvents;
d) the precipitation of sol particles from a sol.
10. When a solution of an HMW compound loses its fluidity, it turns into:
a) slime; b) gel; c) glue.

Topic 11. Chromatography

Chromatography, mobile and stationary phases; column, paper and thin-layer
chromatography (TLC); adsorption, size-exclusion, ion-exchange, partition and affinity
chromatography; the retention time and the retention factor.
Homework exercises
1. Assign each type of chromatography to its mechanism:
1) adsorption chromatography; 2) ion-exchange chromatography; 3) size-exclusion
chromatography; 4) partition chromatography; 5) affinity chromatography.
a) selective interaction between analytes and specific molecules of stationary phase;
b) different abilities of analytes to enter the pores of the stationary phase;
c) different ratios of analyte concentrations in two immiscible liquids;

74
 d) different adsorption abilities of analytes;
e) migration of cations and/or anions between analytes and the stationary phase;
f) different solubility of the reaction products of analytes with the adsorbent.
2. Suggest the most effective chromatographic method for:
a) purification of proteins from low-molecular organic compounds and inorganic
electrolytes;
b) quick test for the identity and purity of a drug;
c) isolation of basic amino acids from the products of a protein hydrolysis;
d) isolation of antibodies to a specific antigen.
e) quick determination of the concentration and type of phosphorganic insecticide.
3. Can the following mixtures be separated by ion-exchange chromatography?
a) phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine;
b) glucose, lactose and sucrose;
c) phenol, o-nitrophenol and p-methylphenol;
d) xylitol, sorbitol and mannitol.
4. Typical heats of adsorption (Qads, kJ mol–1) for carboxylic acids and their derivatives
on a certain adsorbent are given below.
Compounds Qads
aliphatic carboxylic acids 7.6
aromatic carboxylic acids 6.1
esters of aliphatic carboxylic acids 5.3
amides of aliphatic carboxylic acids 9.6
nitriles of aromatic carboxylic acids 3.3
In which order the compounds will elute from the column if the following mixture is
separated by adsorption column chromatography:
butanamide; ethyl hexanoate, benzoic acid, p-methylbenzonitrile; pentanoic acid.
5. A mixture of nitrogenous bases and nucleosides was analysed by TLC, producing
the following data: the distance travelled by the solvent front l0 = 257 mm, the
distances travelled by the centres of spots l1 = 77 mm, l2 = 94 mm, l3 = 205 mm,
l4 = 184 mm. Using the table below, identify the compounds in the mixture.
Compound adenine adenosine guanine guanosine cytosine uracil uridine
Rf 0.3 0.53 0.37 0.58 0.8 0.72 0.81
6. The retention times of ethyl acetate, methyl formate and isopropyl acetate are 2.00,
0.55 and 3.65 min, respectively. Sketch the GC chromatogram of a sample that
contains equal amounts of these three esters.

Concepts and terms

Chromatography is a method of separation of mixtures based on different partitioning
of components between the mobile and stationary phases.
Mobile phase (eluent) is a liquid or gas that flows through the system, moving the
mixture components over the stationary phase at different rates.

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Gas chromatography (GC) is a type of chromatography used for volatile compounds.
The mobile phase (carrier gas) is usually an unreactive gas, such as helium, hydrogen or
nitrogen.
Gas-solid chromatography (GSC) is a type of gas chromatography with solid
stationary phase and gaseous mobile phase.
Gas-liquid chromatography (GLC) is a type of gas chromatography with liquid
stationary phase and gaseous mobile phase.
Liquid chromatography: the mobile phase is a liquid.
Liquid-solid chromatography is a type of chromatography with liquid stationary
phase.
Liquid-liquid chromatography is a type of chromatography where both the mobile
and stationary phases are immiscible liquids.
Column chromatography uses columns, which are glass or metal tubes filled with an
adsorbent or solid support, depending on the type of chromatography.
Planar chromatography is a separation technique where the stationary phase has a
planar shape. The stationary phase can be a piece of paper, serving as such or
impregnated with a liquid (paper chromatography) or a layer of solid particles spread
on a flat support such as a glass plate (thin-layer chromatography, or TLC).
Adsorption chromatography is a type of chromatography where the separation
depends on the different affinities of liquid or dissolved analytes to the solid stationary
phase.
Ion-exchange chromatography involves an exchange of ions of the same sign. It uses
a charged stationary phase to separate charged compounds, such as anions, cations,
amino acids, peptides and proteins.
Size-exclusion chromatography (gel permeation chromatography, or gel filtration
chromatography) separates molecules by their size.
Affinity chromatography is a method of separating biochemical mixtures based on a
highly specific interaction between antigens and antibodies, enzymes and substrates, or
receptors and ligands.
Partition chromatography is based on the partition of the solutes between two liquid
phases (a mobile solvent and an immobile film of another solvent on an inert support).
Partition coefficient (P) or distribution coefficient (D) is the ratio of concentrations of
a compound in a mixture of two immiscible phases at equilibrium.
Retention time is the time taken for a particular compound to travel through the column
to the detector. It is measured from the time at which the sample is injected to the point
at which the display shows the maximum peak height for that compound.
Retention factor (Rf) is the ratio of the distance travelled by the centre of the spot
produced by an analyte to the distance travelled by the solvent front.

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In-class exercises
1. Assign each type of chromatography (adsorption chromatography, ion-exchange
chromatography, size-exclusion chromatography) to its separation mechanism:
a) migration of cations and/or anions between the mobile and the stationary phases;
b) solubility of analytes in polar and nonpolar solvents;
c) ability of analytes to enter the pores of the stationary phase;
d) variation in the retention factors of analytes;
e) affinity of analytes to the stationary phase.
2. Which type of chromatography (TLC, paper chromatography or column
chromatography) is commonly used for:
a) isolation of compounds and quantitative analysis of their mixtures;
b) rapid detection and identification of impurities in a drug?
3. Which type of chromatography (in terms of the chromatographic bed shape,
physical state of the mobile and stationary phases, and the separation mechanism)
should be used to separate proteins (such as albumin) from low-molecular
compounds (NaCl, KCl, NaHCO3, urea, etc.)?
4. Which type of chromatography (in terms of the chromatographic bed shape,
physical state of the mobile and stationary phases, and the separation mechanism)
should be used to separate lysine (pI = 8.5) and aspartic acid (pI = 3.5)?
5. Decide whether the following mixtures can be separated by ion-exchange
chromatography:
a) 1-hexanol, 1-pentanol and 2-methyl-2-hexanol;
b) D-glucosamine, D-glucuronic acid and D-mannaric acid?
Explain your answer.
6. The mixture of lysine (pI = 9.8), glutamic acid (pI = 3.2) and alanine (pI = 6.0) is
separated by ion-exchange chromatography using a mobile phase with decreasing
pH (from 11 to 2.5). In which order the amino acids will elute from the column?
7. The mixture of fatty acids produced by the hydrolysis of a glycerolipid was
analyzed by TLC. The chromatogram showed three spots with Rf of 0.43, 0.59 and
0.75. Draw a possible structure of the initial lipid if the Rf values for stearic, oleic,
linoleic, linolenic and arachidonic acids obtained under the same experimental
conditions are 0.76, 0.60, 0.51, 0.42 and 0.34, respectively.
8. A mixture of lipids was analysed by TLC, producing the following data: the
distance travelled by the solvent front l0 = 200 mm, the distances travelled by the
centres of spots l1 = 132 mm, l2 = 88 mm, l3 = 31 mm. Using the table below,
identify the lipids in the mixture.
lipid distearate dioleoate cholesterol ceramide lecithin sphigomyelin
Rf 0.73 0.70 0.67 0.43 0.15 0.11

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9. The chromatographic separation of a mixture of penicillins was carried out in the
presence of three penicillins with known Rf. The chromatogram is shown below.

Two other Rf values for the same experimental conditions were found in literature:
0.60 for phenoxymethylpenicillin and 0.71 for 6-aminopenicillanic acid. Which
compounds are present in the sample if l0 = 120 mm, lA = 113 mm and lB = 85 mm?
Which component of the sample cannot be identified?
10. The heats of adsorption (kJ mol–1) for organic halides (RX) are given below.
Adsorbent RF RCl RBr RI
alumina (Al2O3) 6.89 7.64 8.40 8.40
silica gel (SiO2) 5.46 5.54 5.54 5.38
a) Which adsorbent should be used for the column chromatography of a mixture
containing 1-fluorohexane, 1-bromohexane and 1-iodinehexane?
b) In which order the compounds will elute from the column?
11. The retention times of methanol, ethanol and propanol are 0.45, 0.62 and 0.90 min,
respectively. Sketch the GC chromatogram of a sample that contains equal amounts
of these three alcohols.
Laboratory work "Identification of anesthesin and novocaine by TLC"
Background
Chromatography is used to separate mixtures of substances into their components.
The mobile phase flows through the stationary phase and carries the components of
the mixture. Different components travel at different speeds. In TLC, the stationary
phase is a thin layer of silica gel fixed on a flat aluminium plate. Substances can be
identified by the distances they travel on the TLC plate by calculating their retention
factors (Rf). The Rf value is a constant for a given substance under certain
experimental conditions. The Rf can be calculated as follows:
distance from the start line to the center of the spot
Rf =
distance from the start line to the solvent front

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 The Rf value is unitless. To identify an unknown substance, it is often necessary to
run a chromatogram of a known substance simultaneously with the unknown.
Work procedure
1. Place about 2 mL of the eluent into the beaker and cover it with a Petri dish or a
piece of glass to allow the solvent vapours to saturate the air inside the beaker.
2. Using a pencil (not a pen!), draw a very light and thin line across the silica gel plate
at about 1.5 cm from the bottom edge to mark the start line. Draw three marks on
the start line and label them.
3. Dip a clean capillary tube in one of the solutions and quickly touch the plate at the
start line with the capillary tip. Hold the capillary tube at right angle to the plate. Do
not scrape off the adsorbent with the capillary tube. Dry the spot and repeat the
application two more times at exactly the same spot.
4. Repeat step 3 for the second and the third solutions.
5. Carefully lower the plate into the beaker, trying not to splash or disturb the eluent.
Cover the beaker immediately with a Petri dish or a piece of glass.
6. Let the solvent rise to about 0.5 cm from the top of the plate, then remove the plate
from the beaker, mark the solvent front with a pencil and leave the plate to dry.
7. Place the plate for a few seconds into a desiccator filled with iodine vapour to reveal
the spots.
Results and evaluation
1. Calculate the retention factors (Rf) for all visible spots. Does the mixture contain
anesthesin, novocaine or both of these compounds?
Online test "Chromatography" (typical questions)
1. Chromatography is:
a) any method of separation of compounds;
b) purification of a solid by recrystallisation;
c) method of identification of substances based on the synthesis of their coloured
derivatives;
d) method of separation of compounds based on their different partitioning between
the mobile and stationary phases.
2. Size-exclusion chromatography is based on:
a) the process of exchanging ions with the charges of the same sign;
b) the different partition coefficients of compounds between mobile and stationary
phases;
c) highly specific interaction between analytes and the stationary phase;
d) different abilities of analytes to enter the pores of the stationary phase.
3. Gas-liquid chromatography can be used for:
a) separation of biopolymers;
b) detection of methanol impurity in ethanol;
c) separation of disaccharides;
d) analysis of minute quantities of amino acids.

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4. Which mixtures can be separated by ion-exchange chromatography?
a) 1-hexanol, 2-pentanol, 2-methylhexan-2-ol;
b) D-glucosamine, D-glucuronic acid, D-mannaric acid;
c) pentanal, hexanal, butanal;
d) propanone, pentan-2-one, butanone.
5. Which mixtures can be separated by size-exclusion chromatography?
a) heptane, hexane, decane;
b) maltose, lactose, sucrose;
c) benzene, toluene, xylene;
d) proteins with significantly different molar masses.
6. The TLC method can be used for:
a) separation of ethanol and acetone;
b) detection of acetaldehyde impurity in ethyl alcohol;
c) analysis of minute quantities of amino acids;
d) isolation of a large quantity of glucose from a mixture of sugars.
7. Affinity chromatography can be used for:
a) isolation of a certain enzyme from protein mixture;
b) detection of diethyl ether impurity in ethyl alcohol;
c) separation of amines;
d) separation of alcohols with low molecular masses.
8. The most effective chromatographic method to purify a protein solution from low
molecular weight impurities is:
a) ion-exchange chromatography; b) high performance liquid chromatography;
c) size-exclusion chromatography; d) thin-layer chromatography.
9. The mixture of 1) lysine (pI = 9.8), 2) glutamic acid (pI = 3.2) and 3) alanine
(pI = 6.0) is separated by ion-exchange chromatography using the mobile phase with
increasing pH (from 2.5 to 11). In which order the amino acids will elute from the
column?
a) 1–3–2; b) 2–3–1; c) 2–1–3; d) 3–2–1.
10. Are the following statements true or false?
1) Ion-exchange chromatography involves the exchange of ions between a solution
and an adsorbent.
2) Partition chromatography is based on different ability of analytes to enter the
pores of the stationary phase
a) only 1 is true; b) only 2 is true; c) both are true; d) both are false.

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 Unit test No. 2
1) Online unit test (contains 30 questions randomly chosen from the previous tests).
2) Paper test (typical questions).
1. The hydrolysis of a lipid produces sphingosine, stearic acid and galactose. Draw the
formula of this lipid.
2. Draw the formula of a micelle of sodium dodecyl sulfate (C12H25SO3Na) with the
aggregation number n. Draw the scheme of octanol incorporation into the micelle.
3. Draw the formula of the tripeptide Gln–Lys–Phe and the reaction of its hydrolysis
under acidic conditions.
4. Consider a peptide with pI = 5.8. Choose the pH value (pH ≈ 5.8; > 5.8; < 5.8) at
which this peptide is:
a) least soluble; b) has the lowest electrophoretic mobility;
c) negatively charged; d) has the lowest degree of swelling.
5. The mixture of fatty acids produced by the hydrolysis of a lipid was analyzed by
TLC. The chromatogram showed three spots with Rf of 0.43, 0.59 and 0.75. The
reference retention factors obtained under the same experimental conditions for
stearic, oleic, linoleic, linolenic and arachidonic acids are 0.76, 0.60, 0.51, 0.42 and
0.34, respectively. Identify the acids present in the original sample.

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 Contents

Literature for self-study and reading assignments ...................................................... 2

Safety instructions ....................................................................................................... 3
Graphical techniques ................................................................................................... 4
Unit 1. Bioorganic chemistry .................................................................................... 5
Topic 1. Redox reactions of organic compounds ........................................................ 5
Topic 2. Poly- and heterofunctional compounds ...................................................... 12
Topic 3. Spatial structure of organic compounds ...................................................... 21
Topic 4. Carbohydrates ............................................................................................. 26
Topic 5. Heterocyclic compounds. Nucleosides, nucleotides and nucleic acids ...... 34
Unit test No. 1 ............................................................................................................ 43
Unit 2. Chemistry of organic nanosystems and biopolymers .............................. 46
Topic 6. Lipids ........................................................................................................... 46
Topic 7. Lyophilic sols. Microheterogeneous and coarse dispersed systems ........... 52
Topic 8. α-Amino acids, peptides and proteins ......................................................... 58
Topic 9. Polymers. Acid-base properties of biopolymers. Solutions of polymers:
formation, properties and stability ................................................................................. 66
Topic 10. Properties of solutions of HMW compounds. Factors affecting the
stability of protein solutions. Aggregation in solutions of HMW compounds ......... 69
Topic 11. Chromatography ........................................................................................ 74
Unit test No. 2 ............................................................................................................ 81

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