Topic 1

Analytical Techniques
Analytical Techniques
Chromatography (paper chromatography)
Gas-Liquid Chromatography Apparatus

Topic 2
Mass Spectrum. Molecular Ion. M+1 peak

Topic 4
Infrared Frequencies. Infrared Absorption. Fingerprint region
Global warming (SAU 4 -1 part) video presentation

Topic 5
C-13 NMR. NMR Spectrum

Topic 6
Proton NMR
 Chromatography Mass spectrometry

Separation onto individual components Identification of the molecular formula

IR spectroscopy NMR spectroscopy

Structural analysis

Identification the functional groups Identification the structural formula

 Instrumental analysis
Advantages Disadvantages
Highly accurate and Expensive
sensitive
Relatively quick Specialist training
results required
Can analyse very Results can be
small samples interpreted only by
comparison with
known substances
 Chromatography (paper chromatography).
Gas liquid chromatography.
Skills: Knowledge and comprehension
LO: Assessment criteria:
11.1.4.28 understand the importance of  Explain and use the terms Rf value in
analytical techniques in modern chemistry
thin layer chromatography and
11.1.4.29 recall previous work on retention time in gas/liquid
chromatography and understand gas-liquid chromatography from chromatograms
chromatography as an extension of it
 Interpret gas/liquid chromatograms in
11.1.4.30 understand the importance of terms of the percentage composition
gas-liquid chromatography as a precursor of a mixture
process in many forms of analysis
 Describe the mechanism of working of
gas-liquid chromatography
 Invention of Chromatography
Mikhail Tswett
invented
chromatography in
1901 during his
research on plant
pigments.
He used the technique
to separate various
plant pigments such as Mikhail Tswett
chlorophylls, Russian Botanist
(1872-1919)
xanthophylls and
carotenoids.
 Paper chromatography
Let’s see the video and identify the main principals of
separated mechanism and purposes of using mobile and
stationary phases
https://www.youtube.com/watch?v=PvHvx7k7UPU
Use the bank phrase to explain to your group
Mobile phase/water, stationary phase/filter paper, Retention factor, solvent
front, colour bands, base line, solubility, adsorption, partition.
Descriptors:
- Describe the role of mobile and stationary phases
- Explain that chromatographic techniques involve adsorption on a stationary phase
- Explain how to calculate the retention factor (Rf) using experimental data for each component of the samples
Calculating Rf Values Example:

Type of chromatography

TLC
https://www.youtube.com/watch?v=rMGQavOMAmc
 Q1
The rose oil contains a mixture of compounds, some of which can
be separated by using thin-layer chromatography (TLC). The
chromatogram obtained is shown below.
i) Explain how TLC separates compounds in the mixture. In your
answer, you should use appropriate technical terms, spelled
correctly.
ii) The mixture of rose oil was spotted onto the base line. Explain
why the base line must be above the level of the solvent
iii) Describe how to calculate the Rf value of B.
Q2
Complete the following passage using words from the box to describe what affects how far the different
chemicals in a sample travel in paper chromatography.
Gas-liquid chromatography (GLC)
• Used to separate a mixture of http://www.youtube.com/watch?v=08Y
WhLTjlfo
compounds, like paper
chromatography, but the sample is Let’s see the video and identify the main principals
vaporised first of separated mechanism and purposes of using
mobile and stationary phases
• Can detect and identify compounds
in tiny amounts of a sample, e.g. can
be used to detect the presence of
banned drugs in samples from
athletes.

https://www.youtube.com/watch?v=iX25exzwKhI
 Fill the gaps
Stationary phase 1…………..
Mobile phase 2……………..
The components of the mixture leave the column in order of 3……………….
1st stage is 4…………………….
Sample mixture is 5………………….
A 6……………………………is used to move the vapour through a coiled column
The compounds in the sample will take different times (7………………) to move through the
column due to their attractions with the material inside the column and due to their
8………………………...
The separated compounds are recorded on a 9…………………………….. The retention times
can then be compared with results of a known substance for identification.

1. Column, packed with a solid or oil (high boiling point liquid) 5. Vaporized
2. Inert gas (Helium) 6. carrier gas
3. Volatility 7. retention times
4. Injection 8. volatility
9. chromatogram
 • The solvent used was ethanol. Which sample of
ink, A, B, or C, has the greatest relative solubility in
ethanol?
• Work out the Rf value of the inks.

A B C
Chromatogram interpretation

https://www.youtube.com/watch?v=03uDmX9ycIg

What is it used for?

 Pharmaceutical Industry
 Radioactive dating
 Drug testing
Three peaks this tells that the sample  Space Research
had three different compounds.   Forensic Science
 Q1
Analytical techniques are very important in modern chemistry.
a) Describe one use of gas-liquid chromatography in analytical chemistry
b) Describe how gas-liquid chromatography is used to separate organic compounds.

Q2
State which peak corresponds to the component that spends
the highest proportion of its time in the tube in the mobile
phase.

Q3
The diagram represent the output from gas/liquid chromatography carried out on a mixture. Determine
the percentage of each of the two components X and Y in the mixture.
 Mass Spectrum. Molecular Ion. M+1 peak
Skills: Knowledge and comprehension. Application
LO:
11.1.4.31 recall the block diagram and outline
operation of the mass spectrometer
11.1.4.35 be able to make sensible
suggestions about structure by inspection of
the fragmentation pattern
11.1.4.32 understand at an elementary level
the production of the molecular ion
and its subsequent fragmentation
11.1.4.33 know in outline the cause of the
M+1 peak
11.1.4.34 be able to find relative molecular
mass from a (simplified) mass spectrum
Assessment criteria:
 Describe the structure and working principle
of the mass spectrometer
 Use precise atomic masses and the precise
molecular mass to determine the molecular
formula of a compound
 Suggest the identity of molecules formed by
simple fragmentation in a given mass spectrum
 Deduce the molecular mass of an organic
molecule from the molecular ion peak in a mass
spectrum
 Deduce the number of carbon atoms in a
compound using the M+1 peak
Recall previous knowledge
For each of the statements below, indicate with the appropriate number (or vice
versa), the stage in the mass spectrometer at which that process occurs.
The first one has been done for you

3
Fragmentation Pattern Mass Spectrum
Cut the given molecule into
fragments and calculate their
masses
Pentan-2-one

Pentan-3-one
Match the suitable mass-spectrum to your
molecule

Pentan-3-one Pentan-2-one
More stable carbonations will be more abundant
the tallest line
base peak

Molecular ion
Parent ion

the presence of
[M+1] the  C isotope in
13

the molecule
1.1%
peak Possible group peak Possible group
15   27  
29   42  
43   43  
57   71  
 Calculation of C numbers using
M:M+1
Using this formula
Calculating the number of carbons
(a) An aromatic compound R has a mass spectrum in which the
M and M + 1 peaks are in the ratio 10:0.9. The M peak is at m/e 122.
Analysis of the compound gave the following composition by mass:
C, 78.7%; H, 8.2%; O, 13.1%.
Showing your working:
(i) use the data to determine the empirical and molecular formulae
of compound R (3 marks)
(ii) use the M and M + 1 data formula to confirm how many carbon atoms
are present in compound R. (2 marks)
 High resolution mass spectrometer
A gas was known to contain only elements
from the following list:
The gas had a molecular
ion peak at m/z = 28.0312
in a high resolution mass
spectrometer. What was
the gas?
C2H4
 Q1
Mass spectrometry is a machine, which determines the molecular weight of molecules
and atoms. Describe the role of the following parts of the mass spectrometer.
• Electric field
• Magnetic field

Q2
The mass spectrum of the element magnesium is shown on the
diagram.
From the mass spectrum, calculate the relative atomic mass, Ar, of
magnesium to two decimal places.

Q3
T is a saturated alcohol. It was analysed by mass spectroscopy. In the mass spectrum, the molecular ion
peak, M, was at an m/e value of 74 and the ratio of the heights of the M and M+1 peaks was 20.4 : 0.9.
i) Use the ratio of the heights of the M and M+1 peaks to calculate the number of carbon atoms in a
molecule of T. What is the molecular formula of T?
 O
Compound U have the formula CH 3CH 2 CH 2 C
Cl
Draw schematically the mass spectrum for the molecule U

Write down the molecular ions that correspond to the m/z peaks you represented on
your mass spectrum above
 Infrared Absorption. Fingerprint region
Global warming
Skills: Knowledge and comprehension. Application
LO: Assessment criteria:
11.1.4.36 understand the reasons for • Analyse an infrared spectrum of a simple
absorption of infrared frequencies by a molecule to identify functional groups
molecule • Use the Data Booklet to interpret the
11.1.4.37 understand and know the infrared spectrum
location of the ‘fingerprint region’ and be • Link between absorption of infrared
able to use it to identify compounds radiation by bonds in CO2, methane and
11.1.4.38 understand the implications of water vapor and global warming
infrared absorption for global warming
 Keywords:

Spectroscopy
Infra-Red
Using
infrared, radiation,
absorption, spectroscopy,
fingerprint region

Visible light is made up of a continuous

range of different electromagnetic
frequencies.....so is Infrared radiation
(the eye just can’t see it)
Each frequency of light (including infra-
red) has a certain energy.
 Principles of Infrared Spectroscopy
https://www.youtube.com/watch?v=BcOBMQlbFlk
 Different covalent bonds have different strengths due
to the masses of different atoms at either end of the
bond.
 As a result, the bonds vibrate at different frequencies
 The frequency of vibration can be found by detecting
when the molecules absorb electro-magnetic
radiation.
 Various types of vibration are possible.
 Examples include... STRETCHING and BENDING • a beam of infra red radiation is passed
through the sample
• a similar beam is passed through the
reference cell
• the frequency of radiation is varied
• bonds vibrating with a similar frequency
SYMMETRIC BENDING ASYMMETRIC
absorb the radiation
STRETCHING STRETCH
• the amount of radiation absorbed by the
Every bond vibrates at its own frequency, dependant upon sample is compared with the reference
bond strength, bond length and the mass of atoms • the results are collected, stored and plotted
involved in bond.
 INFRA RED SPECTRA - INTERPRETATION
Peak position depends on bond strength and masses of the atoms joined by the bond
strong bonds and light atoms absorb at lower wavenumbers
weak bonds and heavy atoms absorb at high wavenumbers

Vertical axis Absorbance the stronger the absorbance the larger the peak

Horizontal axis Frequency wavenumber (waves per centimetre) / cm-1

Wavelength microns (m); 1 micron = 1000 nanometres
FINGERPRINT REGION The region below 1500 cm , usually called the fingerprint region, has many
-1

peaks that are difficult to assign. The pattern of these peaks is unique to a
particular compound.

https://www.youtube.com/watch?v=Bxo_DQpFh1c
• organic molecules have a lot of C-C and C-H bonds within their structure
• spectra obtained will have peaks in the 1400 cm-1 to 800 cm-1 range
• this is referred to as the “fingerprint” region
• the pattern obtained is characteristic of a particular compound the frequency
of any absorption is also affected by adjoining atoms or groups.
Which functional groups are
present in this spectrum?
Which functional groups are present in this spectrum?
Which functional groups are present in this spectrum?
Alcohols

Carboxylic acids

Aldehydes and ketones

Put a tick of compounds have these bonds?
Stretch O-H C-H C=O C-O

Alcohols

Carbonyls

Carboxylics
 O-H C-H C=O C-O
Stretch
2500-3300 2850-3100 1840-1750 1000-1300

Alcohols   
3000-3300

Carbonyls  
Carboxylics    
Very Broad
 IR radiation and greenhouse gases
Nitrogen and oxygen, the most abundant gases in the atmosphere, do not absorb infrared
energy. This is because the vibrations caused would not change the dipole of the molecules.

• The Sun rays are reflected from

earth’s surface as an Infrared
radiation back into the
Sun
atmosphere.
• Some of these Infrared radiations
are absorbed by gases (such as
CO2, methane, nitrous oxide,
sulfur hexafluride, HFCs and Earth
PFCs) and re-radiated as heat,
back to earth.
• This is green house effect.
Global Warming
The three greenhouse gases most frequently found in nature are:
• Carbon dioxide (CO2) – a naturally occurring gas released as a by-product of fossil fuel combustion, selected
industrial processes and changes in the patterns of land-use, particularly deforestation. In terms of gross
volume of emissions, it is by far the most important greenhouse gas. Carbon dioxide is given the base global
warming potential value, 1.
• Methane (CH4) – a gas released in coal mining, landfill operations, livestock raising and natural gas/oil drilling
(among other processes). Methane has a global warming potential of 21 (in other words, it is 21 times more
potent in terms of global warming effect than carbon dioxide).
• Nitrous oxide (N2O) – a gas emitted during fertilizer manufacturing and fossil fuel combustion. The
transportation sector is usually a significant contributor to N2O emissions. N2O has a global warming potential
of 310. Human activity clearly contributes to the increased concentrations of CO2, CH4 and N 2O in the
atmosphere, but they can also be released through natural processes. It is expected that the vast majority of
CDM projects will involved these gases.
Engineered gases
Hydrofluorocarbons (HFCs) – a group of gasses emitted in selected
manufacturing processes and frequently used in refrigeration and air conditioning
equipment. HFC-23, HFC-12, HFC-134a and HFC 152a have global warming
potentials of 11,700, 2800, 1300 and 140 respectively.
 Using the fingerprint region
A substance may
be identified by
comparing the IR
spectrum to a
database of reference
spectra.

An exact match in
the fingerprint region
identifies a compound.
The infrared spectra shown are those of three compounds.
Compound A 1,4-dibromobutane
Compound B butane-1,4-diol
Compound C butanedioic acid
a) Identify the compound responsible for each spectrum by writing the correct letter, A, B or C, in the
box next to each spectrum

b) Infra-red spectra are helpful in the identification of functional groups present in a

molecule.
i) A molecule absorb IR at a wavenumber of 1720 cm -1. Which functional group could
account for this absorption?
ii) Cyclohexane and hex-1-ene are isomers. Suggest how you could use infrared
spectroscopy to distinguish between the two compounds.
iii) The in toximeter, used by police to test the alcohol levels in the breath of drivers,
measures the absorbance at 2900 cm-1. Identify the bond that causes ethanol to absorb
at this wavenumber.
iv) The infrared spectrum of hexane is shown below. The part of the spectrum below
1500 cm-1 is referred as the fingerprint region. Explain why fingerprint region is use as a
unique identifier?
 Nuclear Magnetic Resonance spectroscopy, C-NMR
Skills: Knowledge and comprehension. Application
LO:
11.1.4.39 understand an outline of C-13 NMR and
proton NMR and how spectra are obtained
11.1.4.40 understand the origin of, and be able to
interpret a simple C-13 NMR spectrum
Assessment criteria:
Analyse a carbon-13 NMR of a simple molecule to
deduce:
• the different environments of the carbon
atoms present
• the possible structures for the molecule
Predict the number of peaks in a carbon-13 NMR
spectrum for a given molecule
State the need for deuterated solvents, e.g. CDCl3,
when obtaining an NMR spectrum
Describe the use of tetramethylsilane, TMS, as the
standard for chemical shift measurements
 How NMR spectrometer works
https://www.youtube.com/watch?v=sfiQFQYgJuQ

magnetic field electromagnetic radiation orientation spin

orientation spin
Nuclear Magnetic Resonance- changing the …….. of nuclear ……. by
magnetic field electromagnetic radiation
placing in a ……………., caused by the absorption of ………………
NUCLEAR MAGNETIC RESONANCE (NMR)
is concerned with the magnetic properties of certain nuclei

C13 nuclei H1 nuclei

Protons number 6 1
Neutrons number 7 0
CAN BEHAVE AS A LITTLE MAGNET
Carbon-13 nmr spectroscopy investigates the absorption of radiation by nuclei of carbon-
13 atoms (13C) and is thus a technique for obtaining information about the number and
arrangement of carbon atoms in a molecule
Carbon-13 NMR spectra A typical carbon-13 NMR spectrum looks like this:
provide 2 pieces of information
about the structure of a
molecule:
1.Number of peaks present in
the spectrum = how many
different carbon
environments there are in
the molecule
2.Chemical shift of each peak
tells us about the nature of Chemical shift, δ, is a scale that compares the
the carbon environment frequency of an NMR absorption with the frequency
responsible for that peak. of the reference peak of TMS at δ=0 ppm.
Tetramethylsilane (TMS) is used as the standard for
chemical shift measurements because it is:
• non-toxic liquid - SAFE TO USE
• inert - DOESN’T REACT WITH COMPOUND
BEING ANALYSED
• has a low boiling point - CAN BE DISTILLED OFF
AND USED AGAIN
• all the hydrogen atoms are chemically equivalent -
PRODUCES A SINGLE PEAK
• twelve hydrogens so it produces an intense peak -
DON’T NEED TO USE MUCH
• given the chemical shift of σ = 0
• the position of all other signals is measured
relative to TMS

Analysis usually occurs in solution. Most organic solvents would produce an

NMR signal because they contain carbon-13 or hydrogen-1. Instead, we
use a deuterated solvent, which has had its hydrogen nuclei replaced by
deuterium. An example is CDCl3.
 From this spectrum, it is clear that there are
three peaks. This means that there are three
different types of carbon atom in the
molecule, each with a different chemical
shift.

* *
same
*
environments of
carbon atoms
 Pair Class Activity CARBON-13 NMR SPECTRA

How many carbon environment and hydrogen(proton) environment

are there in the following molecules?

• butane CH3CH2CH2CH3
• 2-methylpropane CH3CH(CH3)CH3
• butanal CH3CH2CH2CHO
• butanone CH3COCH2CH3
• pentan-2-one CH3COCH2CH2CH3
• pentan-3-one CH3CH2COCH2CH3
• cyclohexane C6H12
 CARBON-13 NMR SPECTRA -
Answers

How many peaks would you expect there to be in the carbon-13

spectrum of…check the environment.
• butane CH3CH2CH2CH3 2

• 2-methylpropane CH3CH(CH3)CH3 2

• butanal CH3CH2CH2CHO 4

• butanone CH3COCH2CH3 4

• pentan-2-one CH3COCH2CH2CH3 5

• pentan-3-one CH3CH2COCH2CH3 3

• cyclohexane C6H12 1
 Pair Activity
Instructions:
1.Draw three isomers of C4H8O
2.Identify the chemical environment of carbon in each isomer-hence the number of
possible peaks.
3.Match the isomers with the 13 CNMR provided.
4. Use the data booklet to identify each chemical shift.
5.Label it on the diagram (confirm if the number of carbons, Hydrogen and oxygen fit
into the spectra).
 Pair CARBON-13 NMR SPECTRA - QUESTIONS
Identify the isomers of C6H12
How could you tell from just a quick look at a C-13
NMR spectrum (and without worrying about
chemical shifts) whether you had propanone or
propanal (assuming those were the only options)?
Is the C-13 NMR spectrum for
ISOPROPYL PROPANOATE?
 Nuclear Magnetic Resonance spectroscopy, C-NMR
Skills: Knowledge and comprehension. Application
LO: Assessment criteria:
11.1.4.41 understand the origin of, and be • Analyse a proton NMR spectrum of simple molecule to deduce:
• the number of nonequivalent protons adjacent to a given proton
able to interpret a simple proton NMR
from the splitting pattern, using the n + 1 rule
spectrum
• the different types of proton present using chemical shift values -
11.1.4.43 understand that these the possible structures for the molecule
techniques can yield information both by • Predict the chemical shifts and splitting patterns of the protons in
empirical analysis of the spectral data or proton NMR spectrum for a given molecule
by comparison with the spectra of known • Describe the identification of O–H and N–H protons by proton
substances exchange using D2O
11.1.4.42 relate this to the use of • Describe the role of analytical methods
magnetic resonance imaging (MRT)
 How does an MRI machine work?
https://www.youtube.com/watch?v=nFkBh
UYynUw

MRI can be used to detect brain

tumors, traumatic brain injury,
developmental anomalies, multiple
sclerosis, stroke, dementia, infection,
and the causes of headache

Advantages
 Harmless as it uses low
energy radiofrequency
electromagnetic waves
Non invasive
Patient does not feel
anything
Good for soft tissue scans
Disadvantages
 Electromagnet is so strong
that anything magnetic flies
across the room which is
hazardous
Patients cannot have an
MRI scan if they have a
pacemaker or metal implant
(such as a pin in the leg)
High resolution proton-NMR provides one additional piece of information about the molecule. This
results from the interaction of protons with adjacent protons – this is known as spin-spin coupling
resulting in a splitting pattern. The splitting of a peak can be predicted using the n+1 rule.
Number of H on Peak with Splitting Multiplicity
adjacent carbon (n) pattern (n+1) This peak is due to
the R-CH protons in
0 0+1 = 1 Singlet
the CH3 group. This
1 1+1 = 2 Doublet peak is split into a
2 2+1 = 3 Triplet This peak is due to the –O- triplet which
3 3+1 = 4 Quartet H proton. The peak is a indicates that the
singlet, because the adjacent carbon
hydrogen atom is attached
The relative intensities of the lines can be determined by Pascal’s triangle.
to an oxygen atom (which must have 2 carbons.
i.e. a doublet has a relative intensity of 1:1
a triplet has an intensity of 1:2:1 has no other hydrogens
a quartet has an intensity of 1:3:3:1 attached).

This peak is due to the HC-

O proton. The peak is split
into a quartet (4 parts).
This means that the
neighbouring carbon must
contain 3 carbons. This is
correct – there are 3
hydrogens on the -CH3 group.
Differences between 13C NMR and 1H proton NMR
• 13C NMR has a scale ranging from 0-220
while 1H NMR scale ranges from 0-12.
• 13C NMR no splitting and integration
while 1H NMR splitting and integration of proton values
Which one produced the proton NMR spectrum below?