THE CHEMISTRY

OF AMINES
A guide for A level students

2008
SPECIFICATIONS
AMINES
CONTENTS
• Prior knowledge
• Structure and classification
• Nomenclature
• Physical properties
• Basic properties
• Nucleophilic properties
• Amino acids
• Peptides and proteins
• Amides
• Check list
 AMINES
Before you start it would be helpful to…

• know the functional groups found in organic chemistry

• know the arrangement of bonds around atoms
• recall and explain nucleophilic substitution reactions
What are amines?

• Amines is as near relatives of ammonia, NH3.

•Amines are organic compounds based on ammonia.

• In amines, the hydrogen atoms in the ammonia have been replaced one at a time

by hydrocarbon groups.

Example:
Amines
•Do not occur in free state in the living world.

•Amines are formed by action of bacteria on amino

acids when proteins break down.
• Some body odours detected from human who
do not wash frequently

Di and trimethylamine occur in

rotting fish – distinctive smell.
 STRUCTURE & CLASSIFICATION

Structure Contain the NH2 group

 STRUCTURE & CLASSIFICATION

Classification

H H

R N: R N:

H R
primary (1°) amines secondary (2°) amines

R R
R N: +
R N R
R R

tertiary (3°) amines quarternary (4°) ammonium salts

Aliphatic methylamine, ethylamine, dimethylamine

Aromatic NH2 group is attached directly to the

benzene ring (phenylamine)
Role of amine in life’s survival:

•involved in the creation of amino acids, the building blocks of

proteins in living beings.

•Many vitamins are also built from amino acids.

•Serotonin is an important amine that functions as one of the

primary neurotransmitters for the brain.

BE HAPPY
Role of amine in life’s survival:

- Controls the feelings of hunger

- Brain operating speed
- Affects the state of happiness
-Helps in regulating the sleeping and waking-up cycle of
the brain.
Some important applications of amines

water purification medication

development of crop protection •Amines are also used

chemicals. in many products of care.
 NOMENCLATURE

Nomenclature Named after the groups surrounding the nitrogen +

amine

C2H5NH2 ethylamine

(CH3)2NH dimethylamine

(CH3)3N trimethylamine

C6H5NH2 phenylamine (aniline)

The –NH2 group can be named by using suffix ‘amine’.

Eg: CH3NH2 :–

CH3 CH2NH2 :–

-NH2 group is named by prefix ‘amino’ along with locant if

there are isomers.

Eg: CH3 CH2 CH2NH2 :–

CH3 CH (NH2) CH3 :–

Naming amines can be quite confusing because there are so
many variations on the names.
For example, the simplest amine, CH3NH2,

amine

methyl

commonest name is methylamine

NH2 group is named by prefix ‘amino’ along
with the locant if there are isomers.
If the amine is not the principle functional group, the
prefix ‘amino’ is used.
Eg: NH2CH2 COOH :–

CH3 CH (NH2) COOH :–

Names of secondary and tertiary are written by using

di, tri, …if there are same alkyl groups
• If there are different alkyl groups, they should be named in
the alphabetical order.

• Eg.
 PHYSICAL PROPERTIES

The LONE PAIR on the nitrogen atom in 1°, 2° and 3° amines makes
them ...
H H

R N: R N:

H R

LEWIS BASES - they can be lone pair donors

BRØNSTED-LOWRY BASES - they can be proton acceptors

RNH2 + H+ ——> RNH3+

NUCLEOPHILES - provide a lone pair to attack an electron deficient

centre
 PHYSICAL PROPERTIES

Boiling point Boiling points increase with molecular mass

Amines have higher boiling

points than corresponding
alkanes because of their
intermolecular hydrogen bonding

Quarternary ammonium
salts are ionic and exist as salts
Boiling points of primary, secondary and tertiary amines

type formula boiling point (°C)

primary CH3NH2 -6.3

Hydrogen bonding and
primary CH3CH2NH2 16.6 Vander waals forces

primary CH3CH2CH2NH2 48.6

Hydrogen bonding and

secondary (CH3)2NH 7.4
Weak Vander waals forces
only
tertiary (CH3)3N 3.5
Weak Vander waals forces
 PHYSICAL PROPERTIES

Solubility Lower mass compounds are soluble in water due to

hydrogen bonding with the solvent.

Solubility decreases as the molecules get heavier.

Soluble in organic solvents.

 Preparation of
amines

Aliphatic amines Aromatic amines

Halogenoalkanes Reduction of Reduction of Reduction of

with conc NH3 nitrile amide nitroarenes
Amines can be prepared from halogenoalkanes

Reagent Excess, alcoholic ammonia

Conditions Reflux in excess, alcoholic solution under pressure

Product Amine (or its salt due to a reaction with the acid produced)

Nucleophile Ammonia (NH3)

Equation C2H5Br + NH3 (alc) ——> C2H5NH2 + HBr ( or C2H5NH3+Br¯ )

Ammonia attacks halogenoalkanes because it has a lone pair and is a

nucleophile.
C2H5Br + NH3 (alc)

The amine produced also has a lone pair C2H5NH2 so can also attack a
halogenoalkane; this leads to the formation of substituted amines.
WHY USE EXCESS AMMONIA?
Using excess ammonia ensures that all the halogenoalkane
molecules react with the ammonia before having the chance to
react with any amines produced.
HALOALKANES - NUCLEOPHILIC SUBSTITUTION
- Amines are also nucleophiles (lone pair on N) and
- can attack halogenoalkanes to produce a 2° amine.
- This too is a nucleophile and can react further producing a 3° amine
and, eventually an ionic quarternary ammonium salt.

C2H5Br + NH3 (alc) ——> C2H5NH2 + HBr

C2H5NH2 + C2H5Br ——> HBr + (C2H5)2NH diethylamine, 2° amine

(C2H5)2NH + C2H5Br ——> HBr + (C2H5)3N triethylamine, 3° amine

(C2H5)3N + C2H5Br ——> (C2H5)4N+ Br¯ tetraethylammonium bromide

a quaternary (4°) salt
Uses Quarternary ammonium salts with long chain
alkyl groups are used as cationic surfactants in fabric
softening.
e.g. [CH3(CH2)17]2N+(CH3)2 Cl¯
Reduction of nitrile and amide

Nitriles are compounds containing the -CΞN group

Reagent LiAlH4

Conditions Dry ethoxyethane (ether) followed by acid hydrolysis

Product primary amine

Nucleophile Ammonia (NH3)

Balanced equation

CH3CH2CΞN + 4[H] CH3CH2CH2NH2

Reduction of amide

Reagent LiAlH4

Conditions Dry ethoxyethane (ether) followed by acid hydrolysis, room temp.

Product primary amine

Nucleophile Ammonia (NH3)

Equation
Aromatic amines

Aromatic amines (phenylamines) are prepared by reducing nitroarene.

Reagent: Tin in concentrated Hydrochloric acid (Industrially Iron is

used instead of Tin)

Condition: Heat under reflux followed by excess NaOH

Type of reaction: Reduction

Example: reaction with nitrobenzene

Equation:

3H2
 This phenylamine forms the salt by reacting with hydrochloric acid and
it is soluble.

Getting the phenylamine back from its salt

To re- precipitate amine, dilute NaOH is added.

Reactions of amines
REACTIONS OF AMINES

Lone pair of the nitrogen atom of amine is responsible

for two main properties of amines

1. Base

2. Nucleophile
 Amines as Bases
- Electron donating groups, e.g: alkyl groups, increase the
basic strength.
- Electron withdrawing groups, e.g: C6H5, decrease the basic
strength.
Reaction with mineral acids

• Amines react with acids (dil. HCl or H2SO4) to form salts.

Observation:

Temperature increases (exothermic reaction) and

fishy amine smell is lost.

Example: Ethylamine reacts with hydrochloric acid to form

ethylammonium chloride

Equation:
Reaction with water (Solubility)

• A solution of an amine in water is alkaline.

Amines are quite strong bases- the nitrogen atom acts as a

proton acceptor.

Equation with ethylamine:

Complex ion formation with Copper(II) ions
Ammonia form complex ions with transition metal cations.
For example:
4NH3(aq) + [Cu(H2O)6]2+(aq) [Cu(NH3)4(H2O)2]2+(aq) + 4H2O(l)
Amines form complex ions in a similar way.
Equation with ethylamine:

Equation with phenylamine:

 Equation for the reaction of ethylamine and ethanoyl chloride

HCl

•Ethanoyl chloride reacts violently with a cold concentrated solution of

ethylamine.
•A white solid product is formed - N-ethylethanamide
(an N-substituted amide) and
ethylammonium chloride.
Making paracetamol
 Making paracetamol

Reagents Condition Reaction type

Electrophilic
Step 1 Dilute HNO3 Room temp. substitution

Step 2 Sn and Conc HCl Heat under reflux Reduction

Room temp Nucleophilic

Step 3 Acid chloride substitution
 Reaction with nitrous acid

Nitrous acid is a weak acid and so you get the reaction:

Primary amines and nitrous acid

Observation is a burst of colourless, odourless gas. Nitrogen is given off.

By measuring the amount of nitrogen produced, the amount of amine

present can be worked out
Aromatic arenes with nitrous acid

• When phenyl amine reacts with nitrous acid Diazonium salts are produced.
•They contain an -N2+ group.
•The "azo" bit of the name refers to nitrogen.
.

William Henry Perkin

(1860-1929)
The discoverer of azo dyes
•Azo coupling is widely used industrial
reaction in the production of dyes and
pigments.
•Aromatic diazonium ions acts as electrophiles
in coupling reactions with activated aromatics
such as anilines or phenols.
•The substitution normally occurs at the para position,
•if para position is already occupied, ortho position is favoured.

Methyl orange
Aromatic amine

Aromatic amines form a diazonium salt.

They are more stable than aliphatic diazonium salts, at

temperatures from 0-10 ºC.

Example with phenylamine

Equation:

WARNING: Diazonium salts are very unstable and tend to be explosive as solids.
• Above 10 ºC, this will decompose and give phenol
and N2. Below zero, the reaction is too slow.
Therefore, temperature should be carefully
controlled.

• Equation:
Benzene diazonium ion acts as an electrophile. It can undergo two
types of reactions

(a) Azocoupling reactions

(b) Nucleophilic Substitution.

(a) Azocoupling Reactions

Benzene diazonium ion acts as a strong electrophile

as it has a positive charge.

Benzene diazonium salts react with phenol, phenylamine and 2-napthol

to give highly coloured diazo-compounds.
Example with phenol (in alkaline condition)

Example with phenylamine

Example with 2-napthol (in alkaline condition)

(a) Azocoupling Reactions

Since two aromatic rings are joined together these reactions are known as
coupling reactions.

Unlike diazonium compounds from which they are made, diazo-compounds are
extremely stable and unreactive.
• These products can exhibit geometrical isomerism due
to restricted rotation in N=N
(b) Reaction with nucleophiles

Example: Reaction with steam

Reagent: H2O(g)

Condition: Heat under reflux

Type of reaction: Nucleophilic substitution

Equation:
Nitriles
What are nitriles?
Nitriles contain the -CN group, and used to be known as cyanides.

The smallest organic nitrile is ethanenitrile, CH3CN,

(old name: methyl cyanide or acetonitrile

- and sometimes now called ethanonitrile).
Suffix ‘nitrile’ is used if cyanide is the principle functional group

CH3CN ethanenitrile

CH3CH2CN propanenitrile

CH3CH2CH2CN butanenitrile

The prefix ‘cyano’ is used if it is a secondary functional group.

• NITRILES

• The suffix ‘nitrile’ is used if cyanide is the principle

functional group.

• The prefix ‘cyano’ is used if it is a secondary functional

group.

• Examples:
Ethane nitrile 2-hydroxybutane nitrile 2-cyanobutanoic acid
Some common pharmaceutical drugs - nitriles
 Preparation of nitriles - from amides

Nitriles can be made by dehydrating amides.

Reagent phosphorus(V) oxide, P4O10

Conditions Heat and distillation

Reaction type Dehydration

Equation + H 2O
Reactions of nitriles
1. Reduction

Reagents: LiAlH4
Condition: dry ethoxyethane followed by acid
(H+) hydrolysis

For example, reduction of ethanenitrile to ethylamine:

Nitriles can be reduced into primary amines

2. Hydrolysis
Acidic hydrolysis of nitriles
The nitrile is heated under reflux with dilute
hydrochloric acid.

For example,
with ethanenitrile and hydrochloric acid –
ethanoic acid and ammonium chloride.
 Alkaline hydrolysis of nitriles

For example, with ethanenitrile and sodium hydroxide

solution - sodium ethanoate and ammonia.

Acidify the final solution with a strong acid such as dilute

hydrochloric acid or dilute sulphuric acid.

CH3COONa(aq) + HCl(aq) ——> CH3COOH + NaCl

 AMIDES

Structure derivatives of carboxylic acids

amide group is -CONH2

Amides

• derivatives of carboxylic acid where the –OH group

in –COOH is replaced first by –Cl and then by –NH2.

• If the hydrogens of –NH2 group is replaced by alkyl

or aryl groups they are called as N-substituted
amides.
 AMIDES

Nomenclature White crystalline solids named from the corresponding acid

(remove oic acid, add amide)

CH3CONH2 ethanamide (acetamide)

C2H5CONHC6H5 N - phenyl propanamide - the N tells you the

substituent is on
the nitrogen

If the chain was branched, the carbon in the -CONH2 group counts as
the number 1 carbon atom. For example:

Nylons are examples of

polyamides
The three simplest amides are:

HCONH2 methanamide

CH3CONH2 ethanamide

CH3CH2CONH2 propanamide
The melting points of the amides are high for the size of the molecules because they
can form hydrogen bonds.
Amides form dimers (2 molecules joined together by hydrogen bonds
Amides can act as bases as the lone pair on nitrogen can accept a proton BUT
the lone pair is involved in delocalisation and is less available to accept a
proton.
Preparation of amides

Reagent: Acyl chloride + ammonia

Condition: Room temperature

Type of reaction: Nucleophilic substitution

CH3COCl + NH3 ——> CH3CONH2 + HCl

ethanoyl chloride ethanamide

 AMIDES - CHEMICAL PROPERTIES
Reagent Condition Reaction
type
-
Reaction 1 Br2(l) and Conc.NaOH Heat under reflux

LiAlH4 Dryethoxyethane Reduction

Reaction 2 (lithium followed by dilute
aluminium HCl
hydride)
 AMIDES - CHEMICAL PROPERTIES

Reagent Condition Reaction type

Reaction 3 P4O10(s) Heat and distillation Dedhydration

 AMIDES - CHEMICAL PROPERTIES

Identification Warming an amide with dilute sodium hydroxide solution

and testing for the evolution of ammonia using moist red
litmus paper is used as a simple test for amides.
 POLYMERISATION

General A process in which small molecules called monomers join

together into large molecules consisting of repeating units.

There are two basic types

ADDITION all the atoms in the monomer are used to form the polymer

CONDENSATION monomers join up the with expulsion of small molecules

not all the original atoms are present in the polymer
 POLYMERISATION

ADDITION
 CONDENSATION POLYMERS

• monomers join up the with expulsion of small molecules

• not all the original atoms are present in the polymer

Examples polyamides (nylon) (kevlar)

polyesters (terylene) (polylactic acid)
peptides
starch
 CONDENSATION POLYMERS

Synthesis

Reactions between;

• diprotic carboxylic acids and diols

• diprotic carboxylic acids and diamines

• amino acids

AMIDE LINK

ESTER LINK
Condenstation polymerisation for the formation of polypeptides

What are polyamides?

Polyamides are polymers where the repeating units are held together by
amide links.

An amide group has the formula - CONH2.

An amide link has this structure:
Nylon
•In nylon, the repeating units contain chains of carbon atoms.
•There are various different types of nylon depending on the nature
of those chains.

Nylon-6,6
Nylon-6,6 is made from two monomers each of which contain 6
carbon atoms - hence its name.
 POLYAMIDES - NYLON-6,6

Reagents hexanedioic acid hexane-1,6-diamine

HOOC(CH2)4COOH H2N(CH2)6NH2

Mechanism addition-elimination

Eliminated water

Equation

n HOOC(CH2)4COOH + n H2N(CH2)6NH2 ——> -[NH(CH2)6NHOC(CH2)4CO] n-

+ n H2O
 POLYAMIDES - NYLON-6,6

Repeat unit —[-NH(CH2)6NHOC(CH2)4CO-]n—

Product Nylon-6,6 two repeating units, each with 6 carbon atoms

 POLYAMIDES - NYLON-6,6

Properties contains a peptide (or amide) link

can be broken down by hydrolysis
the C-N bond breaks
behave as amides
biodegradable
can be spun into fibres for strength

Uses fibres and ropes

 NYLON-6

It is possible to get a polyamide from a single monomer.

Nylon-6 is made from a monomer called caprolactam.

This monomer already contains an amide link.

When this molecule polymerises, the ring opens,
and the molecules join up in a continuous chain.
 POLYAMIDES – KEVLAR

Reagents benzene-1,4-dicarboxylic acid 1,4-diaminobenzene

Repeat unit

Properties contains an amide link

Uses body armour

 Polyamides

• Kevlar (bullet-proof
vests) is also a
polyamide:
Uses body armour- tyres
 POLYESTERS - TERYLENE

Reagents terephthalic acid HOOC-C6H4-COOH

ethane-1,2-diol HOCH2CH2OH

Reaction esterification

Eliminated water

Equation

n HOCH2CH2OH + n HOOC-C6H4-COOH ——> -[OCH2CH2OOC(C6H4)CO] n-

+ n H2O
 POLYESTERS - TERYLENE

Repeat unit — [-OCH2CH2OOC(C6H4)CO-] n —

Product poly(ethylene terephthalate)

Properties contains an ester link
can be broken down by hydrolysis ‘Terylene’, ‘Dacron’
the C-O bond breaks
behaves as an ester
biodegradable

Uses fabrics

an ester link
 POLYMERISATION OF ALKENES
ADDITION POLYMERISATION

Process
• during polymerisation, an alkene undergoes an addition reaction
with itself
• all the atoms in the original alkenes are used – 100% atom economy
• long hydrocarbon chains formed
 POLYMERISATION OF ALKENES
ADDITION POLYMERISATION

The equation shows the original monomer and the repeating unit
in the polymer

n represents a
large number

ethene poly(ethene)
MONOMER POLYMER
 POLYMERISATION OF ALKENES
EXAMPLES OF ADDITION POLYMERISATION

ETHENE POLY(ETHENE)

PROPENE POLY(PROPENE)

CHLOROETHENE POLY(CHLOROETHENE)
POLYVINYLCHLORIDE PVC

TETRAFLUOROETHENE POLY(TETRAFLUOROETHENE)
PTFE “Teflon”
POLYMERISATION OF ALKENES
SPOTTING THE MONOMER
POLYMERISATION OF ALKENES
SPOTTING THE MONOMER
Water Attracting Addition Polymers
Addition polymers can be made from ethenol and propenamide.

CH2=CHOH

Ethenol also called acrylamide

CH2=CHCONH2

Propenamide also called poly (vinylalcohol)

Uses of poly(ethenol)

- disposable laundary bags in hospitals.

- liquid detergent capsules (liquitabs)
Produce interersting slimy material – a viscoelastic gel
 AMINO ACIDS

Structure Amino acids contain 2 functional groups

They all have a similar structure - the identity of R1 and R2 vary

H H

H2N C COOH H2N C COOH

H CH3
 AMINO ACIDS – OPTICAL ISOMERISM

Amino acids can exist as optical isomers

If they have different R1 and R2 groups H
α
H2N C COOH
Optical isomers exist when a molecule
Contains an asymmetric carbon atom CH3

Asymmetric carbon atoms have four

different atoms or groups attached
• This means they will rotate the plane of plane-polarised
monochromatic light.

• This can be demonstrated using a polarimeter.

H2N C COOH
Glycine doesn’t exhibit optical isomerism as
H
there are two H attached to the C atom
GLYCINE
2-aminoethanoic acid
Physical Properties of Amino Acids

• All the amino acids are solids at room temperature.

• High melting point.

• This results into positive and negative charges in the amino

acid molecule
 AMINO ACIDS - ZWITTERIONS

Zwitterion • a dipolar ion

• has a plus and a minus charge in its structure
• amino acids exist as zwitterions
• give increased inter-molecular forces
• melting and boiling points are higher
Why do amino acids have a high melting
point?

• Amino acids exist in solid state and they have high

melting points due to the strong electrostatic forces
of attraction between the oppositely charged ends of
these zwitterions.
What is isoelectric point?

• Each amino acid has a particular pH called the

isoelectric point at which the overall charge on the
amino acid is zero.

• Hence at isoelectronic point, they exist as zwitterions.

AMINO ACIDS - ACID-BASE PROPERTIES

• amino acids possess acidic and basic properties

• this is due to the two functional groups
• COOH gives acidic properties
• NH2 gives basic properties
• they form salts when treated with acids or alkalis.

R1

H2N C COOH

R2
 AMINO ACIDS - ACID-BASE PROPERTIES

Basic properties:

with OH¯ HOOCCH2NH2 + OH¯ ——> ¯OOCCH2NH2 + H2O

with NaOH HOOCCH2NH2 + NaOH ——> Na+ ¯OOCCH2NH2 + H2O

The pH of a solution of an amino acid is increased by adding

hydroxide ions, the hydrogen ion is removed from the -NH3+group.
 AMINO ACIDS - ACID-BASE PROPERTIES
Acidic properties:

with H+ HOOCCH2NH2 + H+ ——> HOOCCH2NH3+

with HCl HOOCCH2NH2 + HCl ——> HOOCCH2NH3+ Cl¯

When the pH of the solution is decreased by adding an acid to a

solution of an amino acid, the -COO- part of the zwitterion picks up a
hydrogen ion.
Evidence!
• The existence of the amino acid as either positive or
negative ion is shown using electrophoresis.
 • Amino acid is colourless but its position after a time
can be found by spraying it with ninhydrin.

When a solution of ninhydrin

is sprayed onto an amino acid
and warmed, a violet colour is
produced.
• Ninhydrin can be used to test for the presence of amino acids. It
is also used to detect finger prints since they leave behind lysine
(amine).
 PROTEINS

• are polypeptides with high molecular masses

• chains can be lined up with each other

• the C=O and N-H bonds are polar due to a difference in electronegativity

• hydrogen bonding exists between chains

dotted lines ---------- represent hydrogen bonding

 PEPTIDES - FORMATION & STRUCTURE

Amino acids can join together to form peptides via an amide or peptide link

2 amino acids joined dipeptide

3 amino acids joined tripeptide

many amino acids joined polypeptide

a dipeptide
 PEPTIDES - HYDROLYSIS

Peptides are broken down into their constituent amino acids by hydrolysis

• attack takes place at the slightly positive C of the C=O

• the C-N bond is broken
 PEPTIDES - HYDROLYSIS

Peptides are broken down into their constituent amino acids by hydrolysis

H H CH3

H2N C CO NH C CO NH C COOH

CH3 H CH3

H H CH3

H2N C COOH + H2N C COOH + H2N C COOH

CH3 H CH3
 PEPTIDES - HYDROLYSIS

• hydrolysis with water is very slow

• hydrolysis in alkaline/acid conditions is quicker
• hydrolysis in acid/alkaline conditions (e.g. NaOH) will produce salts
 PEPTIDES - HYDROLYSIS

Peptides are broken down into their constituent amino acids by hydrolysis

H H H

H2N C CO NH C CO NH C COOH

CH3 H CH3

Which amino acids are formed?

 PEPTIDES - HYDROLYSIS

Peptides are broken down into their constituent amino acids by hydrolysis

H H H

H2N C CO NH C CO NH C COOH

CH3 H CH3

H H

2x H2N C COOH + H2N C COOH

CH3 H
• 2. Peptide bonds occur between what two functional groups
of adjacent amino acids?

A. carboxyl group of one amino acid and amino group of the

other
B. hydroxyl group of one amino acid and the sulfhydryl group
of the other
C. carbonyl group of one amino acid and the carboxyl group
of the other
D. phosphate group of one amino acid and the hydroxyl
group of the other
• 1. Review the image and answer the following questions.

Choose the statement that is true about peptide bonds.

A. A peptide bond is an ionic bond occurring between adjacent amino acids.

B. A peptide bond is a covalent bond that occurs only between amino acids.
C. Peptide bonds occur in all organic macromolecules.
D. A peptide bond is a special type of hydrogen bond found only in large protein
molecules.
• A spot of a solution containing the
mixture of amino acids is placed about 2
cm from the bottom of a chromatographic
plate

• spots of solutions of known amino acids

placed alongside.

• bottom of the plate is placed in a suitable

liquid eluent in a beaker

• the beaker is covered to prevent the

solvent from evaporating.
Thin layer chromatography - TLC
Thin layer chromatography - TLC
• After 20 minutes, the plate is removed, dried, warmed
and sprayed with ninhydrin in a fume cupboard and
then dried it in a hot oven.
• amino acids will show up as violet/purple spots.

• If the height of one of the spots matches the height of

one of the known amino acids, that amino acid must
have been in the mixture.

• Rf value can be calculated and compared with known

amino acids.