Laboratory Manual for Practical Exercises Properties of organic compounds

Properties of organic compounds

Contents
Properties of organic compounds ........................................................................................................1
Exercise: Detection of ethanol by iodoform reaction........................................................................3
Exercise: Oxidation of ethanol by KMnO4 in solution with alkaline, neutral and acidic pH .............4
Exercise: Color reactions of phenols................................................................................................6
Exercise: Reduction of Fehling reagent aldehydes ...........................................................................8
Exercise: Detection of acetone by Lestradet reagent ........................................................................9
Exercise: Reaction of oxalic acid with Ca 2+ ................................................................................... 12
Exercise: Detection of lactic acid .................................................................................................. 13
Exercise: Detection and determination of salicylic acid with FeCl 3 ................................................ 14
Exercise: Hydrolysis of acetylsalicylic acid ................................................................................... 15
Exercise: Preparation of esters of carboxylic acids......................................................................... 16
Control questions .......................................................................................................................... 17

Chemical properties of organic compounds depend on the size and the arrangement of molecule
and the presence of the characteristic functional groups. Based on the functional groups the organic
compounds can be divided into several groups. The most important include hydroxyderivates, oxo-
compounds and carboxylic acids. Nomenclature overview of organic compounds derivates is in the next
table:
NOMENCLATURE OF HYDROCARBONS DERIVATES

Name of derivate Formula Group name

chlorethane CH3CH2Cl halogen - hal

aminomethane CH3NH2 amino - NH2

dimethylamine CH3 N HCH3 imino - NH
CH3CHCH3
2-nitropropane nitro - NO2
NO2

methanethiol CH3SH tiol - SH

Benzenesulfonic acid SO 3H sulfo group - SO3H

ethanol CH3 CH2OH

hydroxylic - OH
phenol OH

Dimethylether metoxymethane CH3 OCH3 ether -O-

Dimethyltioether dimethylsulfid CH3 SCH3 tioether -S-

O O
Ethanol acetaldehyde CH3 C aldehyd C
H H

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Laboratory Manual for Practical Exercises Properties of organic compounds

Propanone dimethylketone CH3COCH3 keton C O

O O
Ethanoic acid / Acetic acid CH3 C carboxylic C
OH OH

Hydroxyl derivatives contain in the molecule one or more hydroxyl (-OH) groups. If is the hydroxyl
group bonded to the aliphatic chain, compounds are called alcohols, when linked to the aromatic ring,
they are phenols.
Oxo compounds have in their molecules carbonyl C = O functional group, which can be an
aldehyde (when binding a hydrocarbon radical and one H) or ketone (binds the two hydrocarbon
radicals). If the carbonyl groups are bound by a hydrocarbon radical and one-OH group, together form
the carboxylic acid.
Referred groups of substances provided on the basis of characteristic functional group specific reactions,
which can be used in the analysis of their practical proof or quantitative determination.

Reactions of alcohols
Alcohols are classified according to the number of hydroxyl groups to the mono-,
di-, polyhydroxylic. According to the location of the hydroxyl group in the chain alcohols are divided
into primary (functional group-CH2-OH), secondary (-CH-OH), and tertiary (group
-C-OH).
Properties of hydroxyl group results from polarity of covalent bond between the atoms O and H.
Biologically important reaction is the oxidation (dehydrogenation) of alcohols. Primary alcohols there
give the aldehydes, which can be oxidized to the carboxylic acid. Secondary alcohols are oxidized to
ketones, tertiary alcohols are oxidized to strong oxidizing agents, and gives a mixture of different
oxidation products, one of which is usually ketone.

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Detection of ethanol by iodoform reaction

Ethanol is oxidized by concentrated iodine in Lugol solution to acetaldehyde, which react

with iodine and form triiodoacetaldehyde according:
CH3-CH2-OH + I2 CH3-CHO + 2HI
CH3-CHO + 3I2 CI3-CHO + 3HI
After addition of NaOH iodoform is formed with characteristic odor:

CI3-CHO + NaOH CH3I + HCOONa

Materials
Test tubes, methanol solution, ethanol solution, Lugol solution, NaOH solution c = 1 mol/l

Procedure
1. Prepare and label two test tubes. Into first test tube pipette 0,5 ml of methanol.
2. Into second test tube pipette 0,5 ml of ethanol. Add 2 ml of Lugol solution into both test tubes and
mix the solutions by shaking.
3. Then slowly drip the solution of NaOH until the brown color of solution will be changed to yellow.
4. Content of both test tubes heat at 60 C°.
5. Cool the test tubes under tap water.
6. In second test tube with ethanol observe creation of yellow crystals of iodoform with
characteristic odor. First test tube with methanol will become colorless.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Oxidation of ethanol by KMnO4 in solution with alkaline, neutral and acidic
pH

During oxidation of alcohols is permanganate reduced into different oxidation numbers according
pH of solution where reaction proceeds. In the test tube with alkaline pH, ethanol will cause the
reduction from permanganate (VII) to manganate (VI) with green-brown color:

Oxidation in other test tubes proceeds slower. After additional adding of ethanol to test tube with
neutral pH the manganese (IV) dioxide is formed. Reaction proceeds without any visible color
change according:

In the test tube with acidic pH the solution of permanganate after addition of etanol becomes
orange transparent solution. The reaction proceeds according:

Material
–3 –3
Test tubes, ethanol, KMnO4 solution c = 0,01 mol∙dm , H2SO4 solution c = 6 mol∙dm , NaOH
solution c = 6 mol∙dm–3, distilled water

Procedure
1. Prepare three labelled test tubes.
2. Pipete 1 ml of KMnO4 into all test tubes.
3. Add 1 ml of distilled water to the first, 1 ml of H2SO4 to the second and 1 ml of NaOH into third
test tube.
4. To all test tubes slowly add 1 ml of ethanol, vortex the solutions and observe color changes. Reaction
in first test tube proceeds slower and needs additional 1 ml of ethanol.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Reactions of phenols
Phenols have a hydroxyl group bonded directly to the aromatic ring. Similarly
as alcohols, they are classified according the number of functional groups on mono-,
di- and polyhydroxylic. The names use prefix hydroxy- (hydroxybenzene,
1,2-dihydroxybenzene), but typically they are mainly using trivial names
(phenol, catechol, resorcinol). Alcohols and phenols exhibit due to the presence of-OH group a number
of similar reactions. They differ in terms of acidity, reactivity with oxidizing agents as well as the ability
to form complexes. The fact that the hydroxyl group in the aquatic environment release hydrogen ion is
due to the influence of the aromatic ring. Therefore, phenols act as weak acids and in the reactions with
inorganic hydroxides provide phenolates.
Polyhydroxylic phenols have a great affinity for certain metal ions (Fe3 +), which generate during
reciprocal reactions significantly colored salt complexes - chelates. These reactions are usually used as
a proof of phenols, as well as evidence of complexing metals. Coloration or color stability depends on
the relative positions of the hydroxyl groups, respectively with the groups like -COOH, -COH or -SO3H.
Rising phenolate complex has a structure:

The condition of color formation with Fe3+ is unavoidable presence of at least

one free hydroxyl on the aromatic ring. Colors of the complexes of aromatic alcohols are in the following
table:

Compound Colour of Fe3+ complex species

Phenol violet

Resorcinol violet

Pyrocatechol blue-green

Hydroquinone blue-green (unstable)

Naphtol violet-brown

Salicylic acid violet

p-aminosalicylic acid (PAS) red

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Color reactions of phenols

Phenols are getting into the body by inhalation of vapors, skin absorption, or contact with mucous
membranes. After rapid absorption is part of its molecules oxidized to pyrocatechol and hydroquinone,
an the resting part is completely oxidized. The organism is able to excrete total quantity of absorbed
phenol within 16h. 80% is excreted through the kidneys in unchanged or conjugated form with
glucuronic acid and the remaining part is excreted through breathing.
High concentrations of phenols may affect the ability of blood to transport oxygen, causing
headache, nausea, bluing limbs and lips. Prolonged exposition may lead to breathing difficulties,
collapse and subsequent death. High repeated exposure may cause damage to the liver, kidneys and
central nervous system. Phenolic compounds are mutagenic and can also cause irregular heartbeat
(cardiac arrhythmia).
Practical medical importance of these reactions is that they can be applied to urine tests for
phenylketonuria, or for the control of treatment with p - aminosalicylic acid (PAS).

Materials
Test tubes, pipettes.
Phenol solution (c = 0.05 mol/L), pyrocatechol solution (c = 0.05 mol/L), hydroquinone solution (c =
0.05 mol/L), -naphtol solution in ethanol (c = 0.05 mol/L), FeCl3 solution (c = 0.1 mol/L).

Procedure
1. Into individual test tubes pipette 1 mL of phenol, or pyrocatechol, or hydroquinone, or -naphtol
solution.
2. Add few drops of FeCl3 solution into each test tube.
3. Observe and record the colours of products.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Reactions of carbonyl compounds - aldehydes and ketones

Aldehydes and ketones contain a carbonyl functional group C = O, which is bound in the aldehydes
at the end of the chain and in the middle in ketones. The names are derived from basic hydrocarbons:
for aldehydes adding the suffix -al (methanal, ethanal), ketones have added suffix -one (propanone,
butanone). However, commonly are used trivial names (formaldehyde, acetaldehyde, acetone).
Reactions based on the presence of carbonyl groups are the common for aldehydes and ketones.
Reactions could be additions and condensation. However, only the aldehydes undergoes redox reactions.
They are easily oxidized to carboxylic acids.
From the addition reactions is very important addition of water. First, are produced relatively
unstable aldehydehydrates or ketonehydrates, which can be stabilized by reaction with an alcohol. This
reaction usually results in hemiacetals or acetales. From ketones arise usually hemiketals and ketals.
Formation of hemiacetals is important during the formation of internal hemiacetals in carbohydrates.
From the group of condensation reactions is significant reaction with hydrazine and its derivatives, in
which arise aldehydehydrazones and ketonehydrazones:
R1-CHO + H2N-NH-R2 → R1CH = N-NH-R2 + H2O
The greatest importance, however, have reactions with primary amines. The reaction is based on
the interaction between the aliphatic amine and aromatic aldehyde, or between
aromatic amines and aliphatic aldehydes. Similarly, this reaction also proceeds well with ketones. The
final product of this condensation is called Schiff base.
Ar-CHO + H2N-R → Ar-CH = N-R + H2O
R-CHO + H2N-Ar → R-CH = N-Ar + H2O
This reaction is applicable to a wide variety of biochemical processes, including transamination, a key
reaction in the conversion of amino acids in living organisms. The aldehydes include acetaldehyde,
which is in the body by the action of aldehyde dehydrogenase oxidized to acetyl coenzyme A. Has been
shown that acetaldehyde is able to interact with proteins and affects their structure and functions. This
creates harmful substances mainly in the liver, muscle, heart, brain and gastrointestinal tract. The most
toxic is for muscles. The immune system perceives these pollutants as a direct attack and responds by
inflammatory processes. Arises rheumatoid arthritis, Alzheimer's disease or heart attack. Acetaldehyde
attacks not only proteins but also DNA. Mutations and chromosomal changes are made. Therefore,
acetaldehyde is classified as a dangerous carcinogen.

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Reduction of Fehling reagent aldehydes

The toxic aldehydes include formaldehyde, which enters the body mainly through inhalation and
ingestion. In the lungs and digestive tract is formaldehyde easily absorbed. Formaldehyde undergoes
very rapid metabolism, so inhalation of lower concentrations will not increase its blood levels. Its
metabolites (formic acid and carbon dioxide) enter into macromolecules through one-carbon metabolic
pathway or are eliminated through the lungs and kidneys. Formaldehyde, which wasn’t metabolized,
can directly in a place of entry into the body cause formation of cross-linking in macromolecules and
DNA. Therefore, the reactions of aldehydes are important for the detection of intoxication.
The Fehling reagent serves generally for detection of aldehyde group. The aldehyde group is
oxidized to carboxylic and Cu2+ present in the reagent is reduced to Cu+ (Cu2O).

2 Cu2+ + H-CHO 2 Cu+ + H-COOH

Material
Pipettes, test tube.
Fehling I reagent (CuSO4.5 H2O), Fehling II reagent (sodium potassium tartrate and NaOH ).

Procedure

1. Pipette 1 mL of formaldehyde to 2 mL of freshly prepared Fehling reagent (mix 1 mL of the Fehling

I reagent and 1 mL of Fehling II reagent) in a test tube.
2. Boil the solution in the test tube.
3. On the bottom red Cu2O precipitates.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Detection of acetone by Lestradet reagent

Acetone in the human body occurs in higher amount during disorders of fat metabolism. Excessive
ketone bodies are substances, such as acetoacetic acid, hydroxybutyric and acetone. They are formed
also during fasting. Especially in children aged 3-7 years may occur vomiting problems accompanied
by the presence of ketones in the urine (ketonuria). Ketones in the blood originates from the increased
production of acetoacetic and hydroxybutyric acid in the liver, which is well metabolized in muscles
and kidney. In the liver, there is an active enzyme system, which catalyzes the formation of ketone
bodies. These enzymes have, however, very low activity. The heart muscle gain energy also through
oxidation of acetoacetic acid. This acid is largely excreted in the urine and its transformation arises
acetone. The characteristic odor of acetone is typically in some metabolic diseases, especially in
diabetics, but also due to the so-called acetonemic vomiting. The amount of acetone in the urine is
usually small (up to 60 mg/24 h) and undetectable by common reactions.
A positive finding (ketonuria) is a sign of impaired lipid metabolism without
adequate supply of carbohydrates, as is fasting, or strenuous physical exertion in
diabetes (diabetes mellitus) and other pathological conditions. With proven ketonuria is
always necessary to search for the primary cause.
Acetone proof is based on a color reaction with sodium nitroprusside (Na2[Fe(CN)5NO]). Agent is
unstable in solution, so the reaction is adjusted for the addition of sodium nitroprusside in the solid state.

Material
A watch glass, a pipette.
Acetone, Lestradet reagent.

Procedure
1. Put a bit of the Lestradet reagent on a watch glass.
2. Moisture with a drop of water.
3. Add a drop of acetone solution. The reagent turns to violet after reaction of nitroprusside with oxo
group. Alkaline medium is given by presence of ammonium sulfate and sodium carbonate.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Reactions of a carboxylic acid

Physical properties of carboxylic acids are expressed from mutual competition between
hydrocarbon and carboxylic groups. The members of the series of carboxylic acids with short chain
predominate characteristics of the carboxyl group, above the physical characteristics of hydrocarbons.
Characteristic chemical properties of the carboxylic acid results from the reactivity of the carboxyl
group, which is composed of two formally known carbonyl and hydroxyl groups.
+
The essential characteristic of carboxylic acids is the ability of dissociates of H ions and form salts.
Carboxyl functional group also comprises functional derivatives, such as: anhydrides, esters, amides.
Other substitution derivatives of carboxylic acids have in addition to general characteristics and
properties added specified substituent like hydroxy acids react as alcohols, or phenols respectively.
Reactivity of carboxylic acids can be illustrated as follows:

Derivates of carboxylic acids

Derivate Formula Example

O
Functional R C
X
derivate

O O
Halogenide R C CH3 C Acetylchloride
Cl Cl

O O
Amide R C CH3 C Acetamide
NH2 NH2

O O
R C CH3 C
Anhydride O O Acetanhydride
R C CH3 C
O O

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Laboratory Manual for Practical Exercises Properties of organic compounds

O O
Ester R C CH3 C Methylester of acetic acid
O R O CH3

O O
Thioester R C CH3 C Methylthioester of acetic acid
S R S CH3

Acetonitril
Nitril R CN CH3 C N

Nitril of acetic acid

R CH COOH
Substitutional
X
derivate

CH3 CH COOH
R CH COOH Cl
Halogenacid Cl
2–chlor propanoic acid

CH3 CH COOH
R CH COOH NH2
Aminoacid 2–aminopropanoic acid
NH2

CH 3 C COOH
R C COOH NH
Iminoacid 2–iminopropanoic acid
NH

CH3 CH COOH
R CH COOH OH
Hydroxyacid OH
2-hydroxypropanoic acid

R C COOH CH3 C COOH

Oxoacid O
2–oxopropanoic acid
O

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Reaction of oxalic acid with Ca2+

Oxalic acid is a dicarboxylic acid, which forms normal and acidic salts. From its salt quite
significant is calcium oxalate (forming kidney stones), which is insoluble in water but soluble in mineral
acids.

Material
Test tubes, pipettes, pH paper strips, a glass rod.
Oxalic acid (solid), concentrated ammonia, HCl solution (10 %), calcium chloride (20 g /L).

Procedure
1. Dissolve a small amount of oxalic acid in water in a test tube.
2. Neutralize the solution with ammonia.
3. Add approximatelly 2 mL CaCl2 solution. A white precipitate forms.
4. The precipitate is dissolved by the addition of diluted HCl.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Detection of lactic acid

Lactic acid similarly to other hydroxy carboxylic acid forms complex species with metal ions.
Binding of Fe3+ to lactic acid is much stronger than to phenol. Therefore lactic acid displaces phenol
from its complex with Fe3 (Uffelman reagent).

The reaction can be used for detection of lactic acid in the gastric juice. The reaction is not very specific
because of other hydroxy acids react similarly.

Material
Test tubes, pipettes.
Concentrated lactic acid, Uffelman reagent (0.5 % phenol solutioon, 2 % FeCl3 solution).

Procedure
Add few drops of lactic acid to 1-2 mL of Uffelman reagent in a test tube and shake. Violet colour will
change to yellow or green-yellow due to the formation of iron(III) lactate complex.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Detection and determination of salicylic acid with FeCl3

Salicylic (2-hydroxybenzoic) acid is a colourless crystalline substance with low solubility in hot
water. During heating it sublimes and decomposes to phenol and CO2. It reacts with acids as a phenol,
but with alcohols and phenols it reacts as an acid. It forms iron(III)-salicylate complex species with
FeCl3.

Materials
Test tubes, a spectrophotometer, pipettes.
Salicylic acid (crystalline), FeCl3 solution (c = 0.12 mol/L).

Procedure
a) Detection of salicylic acid:
1. Put few grains of salicylic acid into a test tube.
2. Add 5 drops of dist. water and 1-2 drops of FeCl3 solution. The violet colour forms due to iron(III)-
salicylate complex. The reaction proves the presence of phenolic hydroxy group.

b) Determination of salicylic acid:

1. Pipette 2 mL of salicylic acid solution (unknown sample - cx) into a test tube.
2. Pipette 0.1 mL of FeCl3 solution into the test tube.
3. Mix well and measure the absorbance Ax in a 1 cm cell at 510 nm against dist. water as blank.
4. Repeat the steps 1-3 with the standard solution of salicylic acid (cs = 0.36 mmol/L).
5. Calculate the unknown concentration c x of salicylic acid by the formula:

Aa
ca = As
. cs
where As is the absorbance of the standard solution of salicylic acid.

Observation and calculation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Hydrolysis of acetylsalicylic acid

Acetylsalicylic acid is a white crystalline compound, slightly soluble in water. It is easily

hydrolyzed to salicylic acid and acetic acid. It has wide application in medicine (acetylsalicylic acid) as
an antipyretic. Nature of the functions of aspirin is irreversible inhibition of the enzyme cyclooxygenase
(COX), which is in the organism responsible for the synthesis of prostaglandins and thromboxanes.
Irreversible inhibition is due to the transfer of acetyl groups from the molecule of acetylsalicylic acid on
coenzyme, other anti-inflammatory drugs are able to inhibit the enzyme reversibly.

Materials
Test tubes, pipettes, funnel, filter paper.
Acetylsalicylic acid (crystalline), dist. water, 2 % FeCl3 solution.

Procedure
1. Dissolve a few crystals of acetylsalicylic acid in 2-3 mL dist. water in two test tubes.
2. Test the presence of salicylic acid by the addition of the FeCl 3 solution into one of them. The test
should be negative when acetylsalicylic acid was not hydrolyzed.
3. Boil the content of the second test tube to hydrolyze of acetylsalicylic acid.
4. On cooling the crystals of salicylic acid are excluded.
5. Filter the solution, in the filtrate remains acetic acid (odour).
6. Dissolve crystals in hot water and test with FeCl 3 solution (positive test).

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Exercise: Preparation of esters of carboxylic acids

Esters of lower carboxylic acid are mostly volatile compounds with pleasent (frequently fruit)
odour.

Materials
Test tubes, a scoop, pipettes, a 600 mL beaker, a thermometer.
Glacial acetic acid, salicylic acid, benzoic acid, ethanol, methanol, amyl alcohol, concentrated H 2SO4.

Procedure
1. Prepare a water bath from a 600 mL beaker and water. Heat up to 85 C.
2. Label by numbers 4 test tubes with cork stoppers.
3. Add the substances into four test tubes by the table below:

Test tube No. Acid Amount Alcohol mL H2SO4

1 glacial acetic 0.4 mL ethanol 0.5 10 drops
2 salicylic 1/4 scoop methanol 0.5 4 drops
3 glacial acetic 0.4 mL amyl 0.5 5 drops
4 benzoic 1/4 scoop ethanol 0.5 10 drops

4. Mix the content of the test tubes and close with stoppers.
5. Put the test tubes into water bath (85 C).
6. After 10 min check the odour in the test tubes. You should feel banana, rum, nail polish, mint
odours, respectively.

Observation

Conclusion

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Laboratory Manual for Practical Exercises Properties of organic compounds

Control questions

1. Write the formula of two medically and toxicologically important halogen and hydroxy derivatives
of hydrocarbons and define their use.

2. Name and write the formula of four glycol oxidation products

3. Write the equation of dehydrogenation of tioetanolu and name the product.

4. Write the reaction of formation of glycerol trinitrate and its use in practice.

5. Write the formula of four unsaturated fatty acids and name them.

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Laboratory Manual for Practical Exercises Properties of organic compounds

6. Write the formula of aromatic carboxylic acid and its sodium salt which is used as a preservative
in food and write the detoxification reaction of this salt in the body.

7. Write the formula of EDTA, chemical name and the importance of this compound.

8. Define what is Seigert`s salt.

9. Write the reaction of production of malic acid.

10. Define phosgene and write the formula of its reaction with water.

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