A-level Spectroscopy

An image of a human brain from a live patient recorded using magnetic resonance imaging - a 3D form of n.m.r. spectroscopy

Introduction
Spectroscopy is a collective name for the various techniques that use the interaction between molecules and electromagnetic radiation to elucidate the structure of molecules. Spectroscopic methods are fundamental to the study of Chemistry, Molecular Biology, Medicine and Astrophysics. This booklet covers the following techni ues!-

A" Infrared Spectroscopy - from

Medicine%#

!olymer "evolution# and $hat#s in a

&se relevant given data to interpret 'and ma(e predictions of) infrared spectra for organic compounds containing a limited range of functional groups 'hydro*yl, carbonyl and carbo*ylic acid groups). &nderstand that every compound has a distinctive fingerprint# in its infrared spectrum. &se information given in the +ata Sheet to interpret and predict infrared spectra for organic compounds, in terms of the functional group's) present, understand that specific frequencies of infrared radiation ma(e specific bonds vibrate more.

#" $ass Spectrometry

- from $hat#s in a Medicine%#

-nterpret and predict mass spectra. -dentify the M/ pea( and e*plain that it indicates the Mr 'synoptic), e*plain how the molecular formula can be wor(ed out from the high-resolution value of the M/ pea(, recall that other pea(s are due to positive ions from fragments and the mass differences between pea(s indicate the loss of groups of atoms, suggest the origins of pea(s e.g. pea(s at masses of 01 and 22 are usually due to the presence of the methyl and phenyl positive ions, loss of a methyl group would be indicated by a mass difference of 01.

%" &uclear $agnetic Spectroscopy

- from Medicines by +esign#

+escribe and e*plain how proton nuclear magnetic resonance spectroscopy '3M") can be used for the elucidation of molecular structure 'including splitting patterns up to quartets 4 using the n/0# rule, further e*planation of splitting not required, 5*plain how a combination of spectroscopic techniques 6MS, -"'synoptic) and 3M"7 can be used to elucidate the structure of organic molecules.

This work builds on AS topics of! -nteraction of radiation with matter ' 5lements of 8ife# and Atmosphere#), Mass spectrometry ' 5lements of 8ife#

A) Infrared 'i.r." spectroscopy

&sed to identify bonds 9 functional groups Can only identify the e*act molecule by comparison with library spectra

-nfrared radiation is passed simultaneously through the sample and a reference cell. :he reference ensures that pea(s due to water or carbon dio*ide in the air can be cancelled out. :he frequencies of i.r. radiation absorbed are determined by passing through a rotating prism to focus one frequency at a time onto the detector. -( :he spectrum shows the ;;;;;;;;;;;;;;;; 'cm ) on the * a*is 'which is 09) and the ;;;;;;;;;;;;;;;;;;;; on the y-a*is.

%alculations (" c ) *" +avenumber ) (, 'cm)

e.g. $hat wavenumber would appear on an i.r. spectrum if the frequency of radiation absorbed by a molecule was <.1 * 0=(3 >?%

Theory -" radiation corresponds to the energy required to ma(e chemical bonds vibrate more 9 move to a higher vibrational energy level. :herefore, energy of certain wavelengths is absorbed by molecules. :he actual energy depends on the mass of the atoms and the strength of the bond, so different bonds will absorb at different frequencies. An A3A8@AB between covalent bond and spring

c.f. spring oscillations

weight

Stronger bonds need more energy to ma(e them vibrate, so absorb a higher frequency of i.r. radiation 'higher wavenumber) e.g. hydrogen halides

Molecules with more than < atoms can vibrate in different ways e.g. sulphur dio*ide

So these spectra will contain more absorptions

Most organic molecules contain a number of types of bond, so characteristic absorptions will be seen for each bond. e.g. ethanol

:he following types of bond need to be recognised.#ond @4> @4> CE@ -unctional group Alcohols Carbo*ylic acids Aldehydes 9 (etones 9 carbo*ylic acids9 esters Alcohols 9 esters 9 ethers Al(anes 9 al(enes etc +ave number forAbsorbance 'cm-(" C<== 4 CD== 9 strong and broad. <1== 4 C<== 9 medium and very broad. 0DF= 4 021= 9 strong and sharp

C-@ C->

0=1= 4 0C== 9 medium <F1= 4 C0== 9 medium

GBroad due to >ydrogen Bonding between @-> groups i.r. bands.ppt

/0amples of infrared spectra

(" ethanol 'C>CC><@>)

displayed formula i.r. spectrum

#ond , '-unctional group"

Absorption , cm-(

*" ethanoic acid 'C>CC@@>)

displayed formula

i.r. spectrum

#ond , '-unctional group"

Absorption , cm-(

3" /thyl /thanoate 'C>CC@@C><C>C)

H H C H
i.r. spectrum

O C O

H C H

H C H H

#ond , '-unctional group"

Absorption , cm-( 021=

0<1=

C===

1" a" i.r. spectrum of an alcohol with molecular formula % 3234.

&#! :his Alcohol is o*idised to compound H)b) when heated under distillation with acidified potassium dichromate and H)c) when heated to reflu* with acidified potassium dichromate. Clue% #ond , '-unctional group" Absorption , cm-(

Displayed -ormula of 1a

1" b" i.r. spectrum of the compound with molecular formula % 3254 obtained by distilling compound 1"a" with acidified potassium dichromate

#ond , '-unctional group"

Absorption , cm-(

Displayed -ormula of compound 1b

10

1"c" i.r. spectrum of the compound with molecular formula % 3254* formed when compound 1a is heated to reflu0 with acidified potassium dichromate

#ond , '-unctional group"

Absorption , cm-(

Displayed -ormula of %ompound 1c

11

6"a" i.r. spectrum of an isomer of 1a which forms the same product 6"b" whether it is heated to distil or reflu0 with acidified potassium dichromate

#ond , '-unctional group"

Absorption , cm-(

Displayed -ormula of %ompound 6a

12

6"b" i.r. spectrum of the product of the reaction of 6a with acidified potassium dichromate when heated to reflu0 or distillation.

#ond , '-unctional group"

Absorption , cm-(

Displayed -ormula of %ompound 6b

13

5" Salicylic Acid '*-hydro0yben7oic acid - '>@CD>HC@@>)"

displayed formula

i.r. spectrum

#ond , '-unctional group"

Absorption , cm-(

14

8" Aspirin 'C>CC@@CD>HC@@>)

i.r. spectrum

#ond , '-unctional group"

Absorption , cm-( <I== v. broad

021=

02==

0<==

15

B) $ass Spectrometry
&se M9 'molecular ion) to measure Mr &se M/< isotope pea(s to identify Cl or Br &se fragmentation pattern to confirm structure of molecule

/0periment * : ;S% video

AS-level

<aporisation of atoms or molecules, Ionisation of atoms or molecules, Acceleration of ions, Time of -light Measurement of ions 'lighter ones quic(er), Detection of ions.

J.5. E Kmv<

:ime of Llight# E

16

A*-level
2igh ;esolution $ass Spectrometry
-n high resolution mass spectrometry, the ions are focused into a much smaller range of (inetic energies before entering the time of flight# measurement. Consequently. the mass to charge ratio of ions can be measured to a much higher level of precision, using accurate relative isotopic masses we can distinguish between molecules with the same Mr, this allows us to determine the molecular formula. -sotope 0 > 0< C 0H 3 0D @ Accurate relative isotopic mass 0.==2F 0<.==== 0H.==C0 01.IIHI

/0ample
C@ and 3< have the same Mr at low resolution '<F), but different at high resolution. % 0<.==== 4 01.IIHI %4 <2.IIHI & 0H.==C0 & 0H.==C0 &* <F.==D<

= !ropane, carbon dio*ide and ethanal all have a Mr of HH. A high resolution mass spectrum shows an accurate molecular ion pea( at HH.=<D0, which molecule is it%

17

-ragmentation
:he atoms or molecules are ionised by bombarding with high energy electrons.e 6C>CC@C>C7 M/ / / <e -

e.g. C>CC@C>C /

&sually, the resulting molecular ion has such high energy that it splits up into a smaller ion and an uncharged molecule 'fragmentation)

/ e.g. 6C>CC@C>C7 M/m9e E 1F or 6C>CC@C>C7 1F /

6C>CC@7

C>C

HC
C>CC@ / 6C>C7 01 /

&# :he first fragmentation route is more li(ely because fragments containing 9 the 6"-CE@7 group 'acylium cations) are particularly stable. :he following pea(s are often seen in the fragmentation patterns of mass spectra 4 the highlighted pea(s usually provide very useful clues in determining the structure of a molecule m,e (6 *> C= C0 13 HH H1 88 I0 (?6

fragment %23 %23%2* or %24 C><3>< C><@> %23%4 or %328 C@3>< C@@> %526 CD>1C>< %526%4

18

/0amples of fragmentation and the interpretation of mass spectra

(" @ropanone 'C>CC@C>C) displayed formula

mass spectrum

m,7 1F

-ormula

m,7 lost

Aroup lost

HC

01

19

*" @ropanal 'C>CC><C>@)

displayed formula

mass spectrum

m,7 1F

-ormula

m,7 lost

Aroup lost

12

<I

20

3" $ethyl #en7oate 'CD>1C@@C>C)

O C O

H C H H

mass spectrum

m,7 0CD

-ormula

m,7 lost

Aroup lost

0=1

22

21

1" /thyl /thanoate 'C>CC@@C><C>C)

H H C H

O C O

H C H

H C H H

mass spectrum

m,7 FF

-ormula

m,7 lost

Aroup lost

2C

HC

<I

01

22

6" Salicylic Acid '*-hydro0yben7oic acid - '>@CD>HC@@>)"

displayed formula

mass spectrum

m,7 0CF

-ormula

m,7 lost

Aroup lost

G 0<=

I<

. 3B C- or H- hydro*yben?oic acid isomers cannot eliminate water 4why not%

23

5" Aspirin '%23%44%521%442"

mass spectrum

m,7 0F=

-ormula

m,7 lost

Aroup lost

0CF

0<=

HC

24

8" ethanamide '%23%4&2*"

displayed formula

mass spectrum

m,7 1I

-ormula

m,7 lost

Aroup lost

HH

HC

25

3" paracetamol '1-hydro0yphenylethanamide" '>@CD>H3>C@C>C)

displayed formula

mass spectrum

m,7 010

-ormula

m,7 lost

Aroup lost

0=I

0=F

HC

26

C) &uclear $agnetic ;esonance 'n.m.r." spectroscopy

:he number of pea(s 4 number of proton types :he chemical shift 'M) 4 what are the proton types :he integration 4 how many protons of each type

5*periment C 4 "SC video

Sample is placed in a very strong magnetic field A pulse of radiofrequency radiation is applied "adiofrequency signal emitted from sample is detected

Theory

3uclei have a property called nuclear spin which generates a tiny magnetic field. :he nuclei therefore behave li(e tiny bar magnets.

$hen such nuclei are placed in a large magnetic field they will become aligned with or against the direction of the e*ternal field.

27

:he nuclei lined up with the field are slightly more stable 'lower energy) than those that oppose the e*ternal field.

:he energy gap between these two states corresponds to radiofrequency radiation.

-f the sample is irradiated with a pulse of radio waves, the nuclei in the lower energy state may be promoted to the higher energy state 'the tiny bar magnets flip# from being aligned with to against the e*ternal field). :he e*cited nuclei will then return to the ground state releasing fi*ed quanta of energy which will be detected.

28

:he energy gap depends on the chemical environment of the nuclei and can be used to deduce the e*act structure of the molecule.

5thanal has two proton types, so produces < signals in the n.m.r. spectrum .

:he important features of the spectrum are. :he number of pea(s 4 number of proton types :he integration 4 how many protons of each type :he chemical shift 'M) 4 what are the proton types :he following table can be used to lin( the chemical shift to the proton type 'chemical environment of > atom).type of proton "C>C 9 "C><" 'alkane" "C@C>C 'carbonylsC esters" "C><>al "@C>C 'estersC ethers" "@> 'alcohol" "CD>H> 'arenes" "CD>HC>C 'methylarene" "C@3>" 'amides" "C>@ 'aldehydes" "C@@> 'carbo0ylic acids" "CD>H@> 'phenols" "3>" 'amines" 6" represents an al(yl group7 chemical shift B , ppm =.F - 0.H 0.F - <.< C.< - H.D C.< - C.1 0.= - D.= D.= - I.= <.< - <.H 2.= - 0=.= I.2 - I.F I.= - 0<.= variable variable

/0amples of the interpretation of n.m.r spectra

29

(" propanone 'C>CC@C>C)

displayed formula

nmr spectrum

@roton >a

integration

inference

B , ppm

inference

*" ethanoic acid 'C>CC@@>)

30

nmr spectrum

@roton >a

integration

inference

B , ppm 00.H

inference

>b

<.0

3" propanal 'C>CC><C>@)

31

displayed formula

nmr spectrum

@roton >a

integration

inference

B , ppm I.2

inference

>b

<.H

>c

0.0

2igh ;esolution nmr Spectra

32

Most nmr spectra loo( more complicated than the first three e*amples. :he signal for each hydrogen atom may be split into a number of pea(s. :he pattern of the splitting tells us how many hydrogen atoms are bonded to the adNacent carbon atom. The n9( rule :he no. pea(s E the no. > atoms on the adNacent carbon / 0

e.g. ethanal

@ C >b

>a C >a >a

:he >a protons have one adNacent > atom '>b) :he signal will be split into ;;;; pea(s 4 a doublet.

:he >b proton has three adNacent > atoms '>a) :he signal will be split into ;;;; pea(s 4 a quartet.

2igh resolution spectra may be analysed as follows! :he number of pea(s 4 number of proton types :he integration 4 how many protons of each type :he chemical shift 'M) 4 what are the proton types :he splitting pattern 4 the number of > atoms on the adNacent C atom

&# :his level of analysis is now required for the A-level e*aminations, as the information from splitting patterns is e*tremely useful in wor(ing out the structure of comple* molecules.

+hy are the signals split by adDacent protonsE

33

5ach > nucleus generates its own tiny magnetic field, which may be aligned with or against the e*ternal magnetic field. :his will affect the magnetic environment e*perienced by > nuclei bonded to adNacent C atoms.

e.g. ethanal

@ C >b

>a C >a >a

:he >a protons have one adNacent > atom '>b) >b may be aligned with or against the field

( !

:his means that there are two possible environments for the >a protons, of equal probability.

:he signal for >a will be split into < pea(s 4 a 0.0 doublet.

@ C 34 >b

>a C >a >a

:he situation is slightly more complicated for the >b proton, which has three adNacent > atoms '>a). 5ach of the three >a protons may be aligned with or against the field. :his means that there are four possible orientations of the >a nuclei.a. b. c. d. All nuclei are aligned with the field '0), :wo nuclei with and one against the field 'C), @ne nucleus with and two nuclei against the field 'C), All nuclei aligned against the e*ternal magnetic field '0).

( ! 3 ! 3 !

:his means that there are four possible environments for the >b proton, with a relative probability of 0.C.C.0

:he signal will be split into H pea(s 4 a 0.C.C.0 quartet.

1" /thanal 'C>CC>@)

35

@ C >b

>a C >a >a

>igh "esolution nmr Spectrum of ethanal 3

>a 1 >b

0=

9 ppm

<

@roton >a

integration

inference

B , ppm <.0

inference

splitting

inference

>b

I.2

6" ethyl ethanoate 'C>CC@@C><C>C)

36

Ha Ha C Ha
nmr spectrum

O C O

Hb C Hb

Hc C Hc Hc

@roton >a

integration

inference

B , ppm <.0

inference

splitting

inference

>b

H.0

>c

0.<

>a >c C >c @ C >b >c C >c >c

5) propan-*-ol 'C>CC>'@>)C>C)

>c 37

nmr spectrum

@roton >a

integration

inference

B , ppm <.0

inference

splitting singlet

inference

>b

C.I

septet

>c

0.<

doublet

8" Salicylic Acid '*-hydro0yben7oic acid - '24%521%442""

38

displayed formula

nmr spectrum

@roton >a

integration

inference

B , ppm F.=

inference

>b

2.D

>c

2.=

$here are the @-> groups%

39

3" Aspirin 'C>CC@@CD>HC@@>)

H H

H H

nmr spectrum

@roton >a

integration 0

inference

B , ppm 00.C

inference

>b

H*0

2-F

>c

<.0

40

>" $ystery compound : O$hy are there no aspirin in the Nungle%P

n.m.r. spectrum

@roton >a

integration 0

inference

B , ppm I.2

inference

>b

I.0

>c

<

2.H

>d

<

D.2

>e

<.=

Structure

41

%ombined Spectral Techni ues (" @redict the irC nmr and mass spectra of propanoic acid

a" I; spectroscopy #ond , '-unctional group" Absorption , cm-(

b" &mr spectroscopy @roton >a integration inference B , ppm inference splitting inference

>b

>c

c" $ass Spectrometry

m,7 -ormula m,7 lost Aroup lost

42

*" Deduce the structure of the molecule from these spectra a. ir spectrum

#ond , '-unctional group"

Absorption , cm-(

43

b" nmr spectrum

@roton >a

integration

inference

B , ppm <.<

inference

splitting singlet

inference

>b

C.D

triplet

>c

0.1

overlappin g quartet of triplets

>d

=.I

triplet

44

c" mass spectrum

C0

D=

m,7 D=

-ormula

m,7 lost

Aroup lost

C0

Structure of &n(nown Molecule

3ame

45