HOME PAGE * KS3 SCIENCES * GCSE BIOLOGY CHEMISTRY PHYSICS * ADVANCED LEVEL CHEMISTRY

Advanced Level Organic Chemistry: Amino acids: Structure, preparation and reactions

Part 6. The Chemistry of  Carboxylic Acids and their Derivatives

Doc Brown's Chemistry Advanced Level Pre-University Chemistry Revision Study Notes for UK KS5 A/AS GCE IB advanced level organic chemistry students US K12 grade 11 grade 12 organic chemistry

6.13 Amino acids - structure, preparation and reactions - two functional group chemistries including acidic and basic character

Sub-index for amino acids page

6.13.1 Molecular and ionic structure of amino acids, zwitterions and physical properties

6.13.2 A laboratory synthesis of amino acids from halogenated carboxylic acids

6.13.3 The multi-functional group chemistry of amino acids (focus on acidic and basic character)

6.13.4 The formation of polypeptides

6.13.5 Amino acids from proteins

6.13.5 Chromatography - a method of amino acid analysis

6.13.6 The multi-functional group chemistry of amino acids - other reactions of the carboxylic acid and amino (amine) functional groups (other than the acid-base chemistry described in section 6.13.3)

See also organonitrogen chemistry Section 8.8 Amino acids, peptides, polypeptides and types of proteins

and Amino acids as a case study of R/S isomerism

INDEX of all My carboxylic acids and derivatives notes

All My Advanced A Level Organic Chemistry Notes

Index of My GCSE/IGCSE basic organic chemistry notes

[SEARCH BOX] ignore ads at top



6.13.1 Molecular and ionic structure and physical properties of amino acids

Amino acid molecules have at least one amino/amino group (-NH2) and one carboxylic acid group (COOH).

zwitterions molecular structure of alpha amino acids aminoethanoic acid structural formula 2-aminopropanoic acid 3-aminopropanoic acid skeletal formula

The primary suffix name for an aliphatic carboxylic acid is based on the "longest carbon chain name *" for the -COOH bond system e.g.  ethanoic acid, propanoic  acid etc. The amino group -NH2, with its C-atom position number, is added as a prefix. [* without the end 'e']

They are usually colourless crystalline solids at room temperature.

Data on some of the simplest amino acids is quoted below.

(Mpt. = melting point;  dec. = decomposes;  sub. = sublimes)

IUPAC systematic name of amino acid (common trivial name) Molecular form and the isomeric zwitterion form (see notes below table) Mpt/oC Comments including solubility in water
Aminoethanoic acid

'glycine'

H2NCH2COOH

+H3NCH2COO-

dec. 232 Very soluble in water, the simplest amino acid
2-aminopropanoic acid

 'α-alanine'

CH3CH(NH2)COOH

CH3CH(NH3+)COO-

sub. 258 Moderately soluble in water, an α amino acid α & β alanine are structural positional isomers.
3-aminopropanoic acid

-alanine'

H2NCH2CH2COOH

+H3NCH2CH2COO-

200 Moderately soluble in water, , a β amino acid.  The simplest amino acid to exhibit R/S stereoisomerism.  The middle carbon is 'chiral' - 4 different groups attached.
2-amino-3-methylbutanoic acid

'Valine'

(CH3)2CHCH(NH2)COOH

(CH3)2CHCH(NH3+)COO-

298 Soluble in water.
2-amino-4-methylpentanoic acid

'Leucine'

(CH3)2CHCH2CH(NH2)COOH

(CH3)2CHCH2CH(NH3+)COO-

294 Soluble in water.
2-amino-3-sulfanylpropanoic acid

'Cysteine'

HSCH2CH(NH2)COOH

HSCH2CH(NH3+)COO-

dec. 240 Soluble in water. Note the presence of a sulfur based functional group HS-.

Notes on the data table

(i) For the size of the molecule (e.g. as measured in electrons) they have relatively high melting points, at which they sometimes thermally degrade and decompose - this indicates strong intermolecular bonding of some form between the amino acid molecules.

(ii) The intermolecular forces, apart from the 'usual' instantaneous dipole - induced dipole forces, are greatly increased by hydrogen bonding or ionic attraction between zwitterions (see below for their structure)..

(iii) The α and β (alpha and beta) refer to the second and third carbon atoms to which the amino group can be a substituent.

(iv) All the above have one -COOH group and one -NH2 group in the molecule, so the aqueous solution will tend to be neutral ~pH 7. See amino acid examples in section 6.13.3 which are not neutral.

 

There many amino acids essential to life and are found combined together in proteins e.g tissue, enzymes.

Amino acids are the building blocks for these substances form long chains called polypeptides.

Twenty amino acids are found in the structure of proteins in the human body.

Twelve can be synthesised from other amino acids, but eight cannot and are referred to as essential amino acids and must be found in our diet.

 

Amino acids and zwitterions

Amino-acids in aqueous solution, or in the crystalline state, exist as 'zwitterions' where the proton migrates from the acidic carboxylic -COOH group to the basic -NH2 amino group to form the ionic groups -NH3+ and -COO BUT within the same 'molecule'.

H2N-CHR-COOH +H3N-CHR-COO

For the functional groups present in the molecule, the carboxylic acid is weakly acidic and amino group is weakly basic.

This results in an equilibrium between the neutral molecular form and the zwitterion ionic forms (shown above).

For example, shown as structural formulae of the molecule and zwitterion AND the skeletal formulae of the molecule and zwitterion forms of ...

aminoethanoic acid (glycine) (c) doc b,   (c) doc b  AND   (c) doc b(c) doc b

2-aminopropanoic acid (alanine(c) doc b,   (c) doc b(c) doc b ,  (c) doc b,   (c) doc b

 

There is a strong electrical attraction between the oppositely charged ends of the zwitterions


6.13.2 A laboratory synthesis of amino acids from halogenated carboxylic acids

(not a biosynthesis method)

This is the nucleophilic substitution by ammonia of the halide atom in the carboxylic

e.g. chloroethanoic acid  + ammonia  ===> aminoethanoic acid  +  ammonium chloride

ClCH2COOH  +  2NH3  ===>  H2NCH2COOH  +  NH4Cl

 

A two stage synthesis of the amino acid alanine

2-aminopropanoic acid (the amino acid alanine) when extracted from broken down protein will show optical activity because it will consist of only one of the optical isomers, as it was produced, and used in protein formation, by stereospecific enzymes. It can be produced in the laboratory/industry by a two stage synthesis e.g.

(1) CH3CH2COOH  +  Cl2 ===>  CH3CHClCOOH  +  HCl

free radical chlorination of propanoic acid (no optical isomers)  with chlorine/uv gives 2-chloropropanoic acid which does exhibit optical isomerism (the reaction also forms isomeric 3-chloropropanoic acid).

(2) CH3CHClCOOH  +  2NH3 ===>  CH3CH(NH2)COOH  +  NH4+  +  Cl-  

treating 2-chloropropanoic acid with excess conc. ammonia gives 2-aminopropanoic acid, which again, can exhibit optical isomerism.

In practice this kind of laboratory synthesis yields a racemic mixture, a 50 : 50 mixture of the R and S stereoisomers.

In stage (1) the chlorine radical could abstract/substitute either of the two middle H's with equal probability and therefore a racemic mixture is likely to result.

OR if stage (2) went via a carbocation (with a trigonal planar bond arrangement, SN1 mechanism), substitution can take place by the NH3 molecule hitting either side of the carbocation 'centre' with equal probability.

Therefore either step could give an equimolar mixture of the possible optical isomers.

For more details on reaction (2) see carbocation mechanisms of haloalkane substitution reactions,.

 

Most synthetic amino acids are produced by complex biosynthetic pathways.


6.13.3 The multi-functional group chemistry of amino acids

The acidic and basic character of the -COOH and -NH2 functional groups

Amino acids are carboxylic acids (like ethanoic acid, with the -COOH group) but commonly, one of the hydrogen atoms of the 2nd carbon atom is substituted with an amino/amine group (a nitrogen + two hydrogens gives -NH2).

-COOH is a weakly acid carboxylic acid functional group in aqueous solution.

R-COOH(aq)  +  H2O(l)    R-COO-(aq)  +  H3O+(aq)

-NH2 is a weakly basic functional group in aqueous solution.

R-NH2(aq)  +  H2O(l)    R-NH3+(aq)  +  OH-(aq)

R is the 'rest of the molecule' and if the two functional groups are present in the same molecule, they can cancel each other out to give a ~neutral solution.

 

Another hydrogen on the same 2nd carbon can be substituted with other groups of atoms (R) to give a variety of amino acids.

or The simplest is aminoethanoic acid or 'Glycine'

and another amino acid called 2-aminopropanoic acid or 'Alanine'

All amino acids have the general structure H2N-CH(R)-COOH (see diagram by 5b heading).

R can vary, think of it as the 'Rest of the molecule!

R = H for Glycine, R = CH3 for Alanine.

Amino acids have 2 functional groups: -COOH carboxylic acid and -NH2 amino group.

BUT, there is the added complication of zwitterion form of the amino acid e.g. for alpha-amino acids.

H2N-CHR-COOH +H3N-CHR-COO

 

Amino acids can be classified into three groups depending on their acid-base molecular structure

Neutral amino acids

Here the amino acid has one amino group and one carboxylic acid group.

These will tend to cancel each other out and dissolves to give a neutral aqueous solution

e.g. CH3CH(NH2)COOH  2-aminopropanoic acid (Alanine)

Acidic amino acids

Here the amino acid typically has one amino group and two carboxylic acid groups.

The presence of the 2nd acid group gives an acidic aqueous solution.

e.g. HOOCCH2CH(NH2)COOH  2-aminobutane-1,4-dioic acid (Aspartic acid)

Basic (alkaline) amino acids

Here the amino acid typically has two amino groups and one carboxylic acid group.

The presence of the 2nd amino group gives an alkaline solution

e.g. H2NCH2CH2CH2CH2CH(NH2)COOH  2,6-daminohexanoic acid (Lysine)

 

A flavour of the dual chemistry - I'm giving the equations using both the 'molecular' and 'zwitterion' forms.

(i) Reaction with a stronger acid, amino acid acts as a base

RCH(NH2)COOH(aq)  +  H+(aq)    RCH(NH3+)COOH(aq)

The amine group is protonated giving an alkylammonium ion

RCH(NH3+)COO-(aq)  +  H+(aq)    RCH(NH3+)COOH(aq)

Here the -COO- group acts as the conjugate of the carboxylic acid -COOH.

 

You reverse the reaction by adding alkali

RCH(NH3+)COOH(aq)  +  OH-(aq) RCH(NH2)COOH(aq)   or   RCH(NH3+)COO-(aq)   +  H2O(l)

 

(ii) Reaction with a stronger base, amino acid acts as an acid

RCH(NH2)COOH(aq)  +  OH-(aq)    RCH(NH2)COO-(aq)   +  H2O(l)

The carboxylate anion is formed

RCH(NH3+)COO-(aq)  +  OH-(aq)    RCH(NH2)COO-(aq)   +  H2O(l)

Here the protonated zwitterion acts as a conjugate acid via the -NH3+ group.

(This is just like the ammonium ion, NH4+ which is the conjugate acid of the base ammonia NH3).

 

You reverse the reaction by adding acid

RCH(NH2)COO-(aq)   +  H+(aq) RCH(NH2)COOH(aq)  or  RCH(NH3+)COO-(aq)

 

(iii) Things get a bit more tricky when there are two -COOH or two -NH2 groups in the molecule e.g. several in the table below

Common name of amino acid IUPAC systematic name of amino acid (allowed alternative) Molecular form and the isomeric zwitterion form Comments
Aspartic acid 2-aminobutane-1,4-dioic acid

(2-aminobutanedioic acid)

HOOCCH2CH(NH2)COOH

HOOCCH2CH(NH3+)COO-

Will be weakly acidic in aqueous solution.
Glutamic acid 2-aminopentane-1,5-dioic acid

(2-aminopentanedioic acid)

HOOCCH2CH2CH(NH2)COOH

HOOCCH2CH2CH(NH3+)COO-

Will be weakly acidic in aqueous solution.
Lysine 2,6-diaminohexanoic acid H2NCH2CH2CH2CH2CH(NH2)COOH

H2NCH2CH2CH2CH2CH(NH3+)COO-

Will be weakly alkaline in aqueous solution.
       
       

If there is one -COOH group and one -NH2 group in the moleule, the aqueous solution will tend to be neutral ~pH 7.

If there are two -COOH groups and one -NH2 group, the aqueous solution will tend to be weakly acid with a pH <7

If there is one -COOH group and two -NH2 group, the aqueous solution will tend to be weakly alkaline with a pH >7.

 


6.13.4 The formation of polypeptides

Amino acids can polymerise together, by condensation polymerisation, forming polypeptides.

The peptide linkage is formed by elimination of water between two amino acids.

The simplest amino acid is glycine H2NCH2COOH and the polymerisation can be written as ...

n H2NCH2COOH  ===>  (-NHCH2COO-)n + nH2O,

where n can be quite a large number in the polymer.

These polymers are therefore 'polyamides' as well as 'polypeptides'.

In general the polymerisation to form a polypeptide/secondary amide link is ...

H2N-CH(R)-COOH + H2N-CH(R)-COOH ==> H2N-CH(R)-CO-HN-CH(R)-COOH + H2O ...

... to form one peptide linkage, so ...

n H2N-CH(R)-COOH ==> -NH-CO-CH(R)-NH-CO-CH(R)-NH-CO-CH(R)-NH-CO-CH(R)- etc. n units long

... where R is variable chemical group, as there over 20 known amino acids,

and so proteins are long chain polypeptides and are natural condensation polymers of amino acids.

Long chain polypeptides are what make the majority of the molecular structure of proteins.

Each polypeptide, protein, enzyme etc. has its own unique sequence of amino acids (all encoded for in an organism's DNA).

formation of a peptide link joining amino acids together aminoethane glycine dipeptide

Diagram showing the formation of the polyamide/polypeptide link as a water molecule is eliminated when the carboxylic acid of one amino acid, and the amino group condense together to give an polypeptide/amide link.

In this case two amino acids have a formed the simplest possible polypeptide - a simple dipeptide.

Note that at each end of the molecule, the amino/amine group (-NH2, on left) and the carboxylic acid group (-COOH, on right) can both form a bond with another amino acid molecule by further elimination of water molecules.

So, both functional groups are involved in the condensation to form the polymer.

So, if the process continues, as shown below), you build up a long chain polymer - known as a polyamide, polypeptide or a protein - they are all the same here.

formation of protein polypeptide by condensation of amino acids

Proteins have the same (amide) linkages as nylon but with different units.

Proteins are an important component of tissue structure and enzymes (powerful biological chemical catalysts) are also protein molecules.

Proteins tend to adopt a particular three dimensional shape (3D) which aids its function.

Apart from the structural proteins in you body e.g. muscle tissue, enzymes are protein molecules wrapped into a specific 3D shape to carry out their catalytic function.

For more detailed notes see Enzymes and Biotechnology

When proteins are heated with aqueous hydrochloric acid or sodium hydroxide solution they are hydrolysed to amino acids.

see chromatography below, about how amino acids are identified in proteins.

There are 20 amino acids that make up the proteins in the human body.

12 can be synthesised by us from other amino acids,

 


6.13.5 Amino acids from proteins and R/S stereoisomerism

All the alpha-amino acids obtained from proteins are optically active except glycine (aminoethanoic acid), that is they exhibit R/S stereoisomerism (pin-pointed in the diagram above) where you have two mirror image forms that cannot be superimposed on each other (as with your right and left hands!). R/S stereoisomerism page

 2-aminoethanoic acid, H2NCH2COOH, is not an optically active molecule because it has no chiral/asymmetric carbon atom because there is no carbon with 4 different groups attached, but all the other alpha-amino acids have four different groups attached to the 2nd carbon atom i.e.  R-CH(NH2)COOH

In aqueous solution, and in the solid state, they predominantly exist as zwitterions, the ionic form derived from proton transfer from the carboxylic group onto the amino group.

RCH(NH2)COOH (c) doc b RCH(+NH3)COO-

Even so, the zwitterions will also exhibit R/S stereoisomerism as well as the non-ionic molecular form.

structural formula diagram of R/S optical isomers of alpha-amino acids stereoisomers enantiomers stereoisomerism molecular structure R/S isomers of the non-ionic molecular form (R = CH3 for alanine)

skeletal diagram of R/S optical isomers of zwitterions of alpha-amino acids stereoisomers enantiomers stereoisomerism ionic structure R/S isomers of the ionic zwitterion form (R = CH3 for alanine)        

need zwitterion form too

Comparing 'natural' and 'laboratory' synthesised amino acids (other than biosynthetic routes)

When molecules capable of exhibiting optical isomerism are obtained from natural sources, they usually consist of one of the possible isomers (one of the enantiomers).

On extraction, purification and isolation, they show optical activity (that is rotating the plane of polarised light in a polarimeter tube).

This is due to the need for stereospecific structures from enzymes to proteins. The '3D' stereospecificity of enzyme sites is discussed in section 6.

However, when the same compound is synthesised in the laboratory by a non-biosynthetic route, it often consists of an equimolar mixture of the two optical isomers (R/S isomer enantiomers).

This is known as a racemic mixture and it is optically inactive due to one isomer cancelling out the optical effect of the other.

Warning: It is wrong to say that R/S optical isomers are not, or cannot be formed, in a laboratory synthesis!

 Its difficult, but no impossible, using very sophisticated synthesis techniques.

The most common explanation for the production of a racemic mixture lies in understanding the mechanisms of the laboratory synthesis reactions.

For example, if a carbocation is formed, which has three C-R bonds in a trigonal planer arrangement, the reagent molecule or ion (electron pair donor) can attack on either side with equal probability.

So when a possible chiral carbon molecule is formed in many a laboratory synthesis, it tends to be an equimolar mixture of the two spatial possibilities or enantiomers.

(see carbocation mechanisms of haloalkane substitution reactions, addition reactions of aldehydes/ketones.

However, since the 1990's the problem is being tackled by the use of chiral auxiliary molecules.

 


6.13.6 Chromatography - a method of amino acid analysis

Hydrolysis means breaking down a molecule with water to form two or more products.

Hydrolysis is accelerated if the substance is heated with acid or alkali solutions.

When proteins are heated with strong aqueous mineral acid they are hydrolysed to amino acids.

(1)chromatography at start  (2)chromatogram at the end  (3)chromatography (4)diagram of paper/thin layer chromatography at the end paper chromatography

Paper chromatography or thin layer chromatography (TLC) is used to separate coloured compounds (illustrated above).

Thin layer chromatography (TLC) uses a stationary phase of either silica gel or aluminium oxide immobilised on a flat inert surface that can be made of a glass or plastic plate.

In the procedure:

(1) Samples spotted onto start line of the paper/plate placed in solvent, but below start line of pencil.

(2) The solvent, the mobile phase called the eluent, rises up paper/plate.

(3) When solvent near top of paper, remove paper to dry.

However, amino acids are colourless, but can still be separated in this way, and made visible!

For colourless amino acids, you spray the paper with ninhydrin and gently heated, which gives a purple spot for each amino acid.

You measure the vertical distance travelled by each spot.

(4) You can then measure the Rf values to identify amino acids in the mixture.

To illustrate the method I've described the separation of coloured dye molecules to represent different amino acids.

1 to 5 represent five pure compounds, 6 is a mixture.

Red, brown and blue make up the mixture because its spots horizontally line up with the three known colours.

The substances (solutes) to be analysed must dissolve in the solvent, which is called the mobile phase because it moves.

The solvent may be water or other suitable solvent that the amino acids will dissolved in - can vary the pH of the eluent solvent - the liquid that moves up the paper or TLC plate.

The paper or thin layer of material on which the separation takes place is called the stationary or immobile phase because it doesn't move.

The distance a substance moves, compared to the distance the solvent front moves (top of grey area on diagram 2) is called the reference or Rf value (a simple ratio) and has a value of 0.0 (not moved - no good), to 1.0 (too soluble - no good either).

 Rf ratio values between 0.1 and 0.9 can be useful for analysis and identification of the amino acids.

Rf = distance moved by amino acid spot / distance moved by solvent

 

Thin layer or paper chromatography can still used to separate and identify the products of hydrolysis of proteins because you make them coloured by using another chemical reagent.

The hydrolysis can be done by boiling-refluxing the protein with hydrochloric acid.

The hydrolysed mixture is then 'spotted' onto the pencil base line of the chromatography paper or the TLC plate.

Known amino acids are also spotted onto the base line too.

The prepared paper is then placed vertically in a suitable solvent, which rises up the paper/TLC plate.

Since the products are colourless, the dried chromatogram is treated with another chemical to produce a coloured compound.

A reagent spray of Ninhydrin produces purple spots with amino acids

Using known amino acid samples, you can then tell which amino acids made up the protein that you hydrolysed.

The number of different spots tells you how many different amino acids made up the protein or peptide.

Spots which horizontally match the standard known molecule spots confirm identity their identity.

Theory of paper and thin layer chromatography


6.13.6 The multi-functional group chemistry of amino acids

Other reactions of the carboxylic acid and amino (amine) functional groups

The acidic and basic character of the -COOH and -NH2 groups has been dealt with in section 6.13.3

Other reactions of the carboxylic acid group

 

(a) Esterification

Amino acids can be reacted with alcohols to form esters using a strong acid catalyst like concentrated sulfuric acid e.g.

RCH(NH2)COOH  +  R'OH     RCH(NH2)COOR'  +  H2O

The strong acid ensures the amino acid is protonated and the carboxylic acid group is free to undergo esterification.

The zwitterion form of the amino acid cannot be esterified.

Technically it is the protonated form that is esterified and the ester freed on adding alkali

e.g. preparing ethyl 2-aminopropanoate in 2 stages

(i) Esterification under acid conditions

CH3CH(NH3+)COOH  +  CH3CH2OH     CH3CH(NH3+)COOCH2CH3 +  H2O

(ii) Addition of alkali

CH3CH(NH3+)COOH  +  OH-     CH3CH(NH2)COOCH2CH3 +  H2O

 

(b) -

 

Other reactions of the amine/amino acid group

(a) Acylation

Amino acids will react with acid chlorides and acid anhydrides to replace one of the hydrogens on the amino group with an acyl group (R-C=O, RCO) e.g.

RCH(NH2)COOH  +  R'COCl     RCH(NHOCR')COOH  +  H2O

The product is a secondary amide, but still with a carboxylic acid group.

 

(b) Acting as a nucleophile in nucleophilic substitution reactions

The zwitterion nature of amino acids means that the lone pair of electrons on the nitrogen is not available to allow the amino acid to act as a nucleophile unless the pH is much higher than 7 i.e. via reaction (i) below in alkaline conditions

(i)  RCH(NH3+)COO-  +  OH-    RCH(:NH2)COO-   +  H2O

The allows the amine group to acts as the 'front end' of an electron pair donating nucleophile (: in the right-hand formula).

The amino acid can then react with halogenoalkanes to yield a secondary amine, albeit with a carboxylic acid group retained.  (X = Cl, Br or I)

 

(ii)  2RCH(NH2)COO-  +  R'X  +  2H2O  ===>  RCH(NHR')COOH   +  RCH(NH3+)COOH  + X-   + 2OH-

-


Doc Brown's Advanced Level Chemistry Revision Notes

[SEARCH BOX] ignore ads at top

INDEX of all My carboxylic acids and derivatives notes

All My Advanced A Level Organic Chemistry Notes

Index of My GCSE/IGCSE Oil - useful products and basic organic chemistry notes

 

KS3 BIOLOGY QUIZZES ~US grades 6-8 KS3 CHEMISTRY QUIZZES ~US grades 6-8 KS3 PHYSICS QUIZZES ~US grades 6-8 HOMEPAGE of Doc Brown's Science Website EMAIL Doc Brown's Science Website
GCSE 9-1 BIOLOGY NOTES GCSE 9-1 CHEMISTRY NOTES and QUIZZES GCSE 9-1 PHYSICS NOTES GCSE 9-1 SCIENCES syllabus-specification help links for biology chemistry physics courses IGCSE & O Level SCIENCES syllabus-specification help links for biology chemistry physics courses
Advanced A/AS Level ORGANIC Chemistry Revision Notes US K12 ~grades 11-12 Advanced A/AS Level INORGANIC Chemistry Revision Notes US K12 ~grades 11-12 Advanced A/AS Level PHYSICAL-THEORETICAL Chemistry Revision Notes US K12 ~grades 11-12 Advanced A/AS Level CHEMISTRY syllabus-specificatio HELP LINKS of my site Doc Brown's Travel Pictures
Website content Dr Phil Brown 2000+. All copyrights reserved on revision notes, images, quizzes, worksheets etc. Copying of website material is NOT permitted. Exam revision summaries & references to science course specifications are unofficial.

 Doc Brown's Chemistry 

*

TOP OF PAGE