37 Analytical techniques

ANALYSIS - CIE A-LEVEL CHEMISTRY

New syllabus for CIE
This content is for exams in 2022.

AS
◦ If you’re starting in September 2020 with Year 12, then you’ll need to use the old syllabus for the AS in
May/June 2021.
◦ Only start this syllabus with AS in September 2020 if your students will take ALL the exams (AS & A2) at the
end of two years (May/June 2022).

A2
◦ Year 13 September 2020 old syllabus examined in 2021
◦ Year 13 September 2021 new (this) syllabus examined in 2022
Mastery learning information for the teacher:
There are increasingly more mastery learning definitions however they are all centred on the idea that students must achieve a defined proficiency
in a given unit of work before proceeding.

Repetition and practice are critical to mastery of different concepts as students proceed through the course. We need also to be mindful of retention
of the information to maintain mastery as a student works through the course over the 8 or 9 months before the examinations.

Mastery learning develops a growth mindset as the implication is that all students will understand all of the work. The variable is the time it takes
to master the different concepts.

In these A2 slide decks, mastery is encouraged by different styles of questioning, practice questions and relies of constant formative assessment by
the teacher. Only move on once everyone in the class is getting over 80% of the problems correct.

There are some more detailed suggestions on the website of how to use these presentations but arguably one of the most effective and the intended
approach is to use the flipped classroom model. It is advised to prepare additional practice as you progress through your own worksheets,
homeworks and extension activities, these can then be made available for those who work more quickly.

Retention is enhanced through spaced repetition in the starter activities and the allocation of complete A2 past paper questions.

The notes section of each slide provides links back to mastery learning ideas to provide the teacher with a rationale of what this slide is aiming to
achieve or test. It also includes answers and/or explanation of problems and questions for the teacher to consider as their students progress.

For additional support of questions related to mastery learning please email me at: content@chemcatalyst.co.uk
Starter 1: Retrieval Questions
What did you learn last lesson?
What did you learn last week?
What did you learn through an activity you organised yourself?
◦ independent reading, additional questions, research?
 Starter 2: Amino acids

Glycine, Alanine, Valine and Leucine are all amino acids

1. Which functional groups common to all amino acids react to form amide groups (peptides in Biology)?

2. Draw a diagram to show the structure of glycine at pH 3

3. Draw a diagram to show the structure of alanine at pH 11

Starter 3: Nitrogen compounds
What are the names of these compounds?
Which one of these compounds is the strongest base?
37 Analytical techniques
ANALYSIS - CIE A-LEVEL CHEMISTRY
Subject content
Learning outcomes 37.1 from the syllabus
Subject content
Learning outcomes 37.2 from the syllabus

Learning outcomes 37.3 from the syllabus

Subject content
Learning outcomes 37.4 from the syllabus
Expected learning gains
Describe and understand different separation techniques to be able to analyse different chemical
mixtures
Describe and understand different analytical techniques to be able to analyse the different results they
produce
◦ Use the MS data to determine the relative molecular mass of the unknown substance and the presence of
certain functional groups
◦ Use the 13C-NMR to determine the number of carbon environments
◦ Use the 1H-NMR to determine the number of hydrogen environments

Combine analytical data to identify organic molecules

TLC
37.1 THIN-LAYER CHROMATOGRAPHY
Chromatography
Chromatography is a separation technique
A mixture of different substances is dissolved in a fluid called the mobile phase, which carries the
substance/s through another material called the stationary phase
Different substances in the mixture will travel at different speeds depending on their size and how
much they interact with the stationary phase
These differences in speed cause them to separate
High Performance Liquid Chromatography HPLC

LINK
Thin Layer Chromatography
TLC is similar to paper chromatography which you
have almost certainly done earlier at school
Rather than using chromatography/filter paper the
stationary phase is a thin uniform layer (usually
silica gel) coated on a piece of plastic, metal or glass
The mobile phase is a liquid which acts as a solvent
for the substances you are trying to separate or
analyse

paper chromatography
Thin Layer Chromatography 1
1. A base line is drawn in pencil across the plate and dots of the substances are placed along the
linepeated application is used to increase the concentration
2. The plate is placed in a beaker of the solvent such that the level of the solvent is below the line and
the solvent moves up the plate.
3. A lid is placed to ensures that the atmosphere in the tank is saturated with the often volatile solvent
which climbs up the plate causing the mixture to separate

1 2 3
Thin Layer Chromatography 2

4 5 6

4. As the solvent front moves up the plate the purple mixture separates into a
fast moving red spot and a slower moving blue spot
5. The solvent reaches near to the top of the plate and the plate is removed
6. The solvent front (4) is noted and the solvent is left to evaporate
Rotating the sheet
This method can be developed into two dimensional chromatography
The sheet is rotated through 90° and a second solvent is used to further split the different components if
they initially overlap

rotated
through
90°
Calculating Rf
Assuming the following distances
◦ Base line to solvent front: 6 cm
◦ Base line to blue chemical: 2 cm
◦ Base line to red chemical: 5 cm

distance moved by the compound

Rf 
distance moved by the solvent

Rf = 2/6 = 0.33
Rf = 5/6 = 0.83

Providing the temperature and solvent is the same these R f values will remain the same if the experiment is
repeated
Calculating Rf
Once you have calculated the Rf value you can compare to a data source and identify which protein or
chemical substance you are dealing with

Amino acid Rf (solvent X)

Alanine 0.51
Tyrosine 0.68
Arginine 0.16
TLC demonstration

LINK
GLC
37.2 GAS / LIQUID CHROMATOGRAPHY
Gas-liquid
chromatography
A gas chromatography oven,
open to show a capillary
column
Gas-liquid chromatography 1
In GLC a long column is often filled with a polysiloxane liquid compound
In thinner tubes, the stationary phase is coated to the inside surface of the tube
The sample is vaporised and pushed through the column by an inert carrier gas (most often helium)
Gas-liquid chromatography 2
Depending on the solubility of the different compounds in the stationary liquid phase, their boiling
points and the temperature of the column, the different compounds will travel through the GLC column
at different speeds
The length of time a compound remains in the column is known as the retention time
It is calculated from the time the sample is injected until the maximum peak height for that
compound is observed on the display/chromatogram
Gas-liquid chromatography

LINK
Example results
In this example gas/liquid chromatogram we can see 3
peaks
Each peak corresponds to a different compound in the
sample and we can observe that they arrive at the detector in
sequence
They have different retention times
Proportion of components
C Remember the area of a triangle
The area under the peaks ◦ = ½ x base x height
can be used to calculate • Peak C
the composition of the Peak A
◦ Height: 17mm • Height: 110mm
mixture • Width: 5mm
◦ Width: 9mm
◦ Area: 76.5mm2 • Area: 225mm2

Peak B
B ◦ Height: 50mm
◦ Width: 4mm
A
◦ Area: 100mm2

Total area = 401.5mm2

Peak A = 76.5/401.5 x 100% = 19%
Peak B = 100/401.5 x 100% = 25%
Peak C = 225/401.5 x 100% = 56%
Retention Time Practice
Retention Time (s) Compound
16.9 Phenol
33.0 Octan-1-ol
54.7 Ethanoic acid
23.0 Tetradecane
60.6 Ethandioic acid

96mm2 92mm2

80mm2 What percentage of

50mm2 this mixture is
Ethanoic acid?
sample injected
32mm2

time
NMR Spectroscopy
37.3 CARBON-13 NMR SPECTROSCOPY
NMR Spectroscopy
The nucleus’ of carbon and hydrogen atoms have magnetic
properties
Chemists use this property to determine information about
the structure and chemical environment of molecules
which contain the carbon and hydrogen atoms
NMR stands for
nuclear
magnetic
resonance

NMR spectrometers can be

small benchtop sized devices
or much larger
Nuclear magnetic resonance

Nuclear magnetic resonance is a physical phenomenon

where the nucleons of an atom absorb electromagnetic
radiation which changes their quantum property of spin

We have seen this spin idea before with electrons when

drawing diagrams for electronic configuration
Nuclear magnetic resonance
An external magnetic field is applied to the sample and a specific radio wave
frequency in the electromagnetic spectrum causes the protons in a nucleus to absorb
energy and ‘flip’ to their resonance condition
This flipping is measuring by the device and is seen as a peak on the NMR spectra
The peak can be analysed to tell us information about the structure of the molecule
Splitting of nuclei spin states

Nuclei behave like they have their own magnetic moments

In an external magnetic field there is a high-energy state and a low-energy state
depending on the relative orientations of the magnet to the external field
The surrounding environment 1
The electrons of other atoms in the surrounding environment of
the atom reduce the affect of the external magnetic field
The bonding electrons are further or closer to the carbon
depending on relative electronegativities
Hence an isolated proton for example would need a different
strength of magnetic field to cause the proton to flip into its
resonance condition than a similar proton bonded to another atom
The surrounding environment 2
For the nucleon to flip there must be exactly the right
combination of external magnetic field and radio frequency
for the resonance condition
Historically all NMR machines varied the magnetic field and
kept the frequency of the radio waves constant but some
newer models fix the magnetic field and vary the frequency of
the radio
Carbon-13 NMR
1% of carbon atoms exist as carbon-13 atoms
The odd number of nucleons means that a resonance condition can exist
◦ The small percentage is irrelevant as any sample will contains millions (if not trillions) of individual molecules

The other atoms attached to a carbon-13 atom mean that a slightly different magnetic field will need to
be applied to cause the resonance condition to occur in the carbon-13 nucleus
This can be used to work out the carbon atom's environment in the molecule
13
C-NMR Propan-1-ol
• In this 13C-NMR we can
see three peaks
• Is this what we would
expect?
• The structure of propan-
1-ol is:

1 2 3

180 160 140 120 100 80 60 40 20 0

δ ppm

• Carbon ‘1’, ‘2’ and ‘3’ are all in different environments when you look at the atoms they are
attached to and also the atoms that are attached to them
 13
C-NMR Propan-1-ol

• The value at around 63 corresponds to the carbon with the alcohol group
• The value at 11 is the leftmost carbon
• The value at 26 is the middle carbon which is shifted further downfield as it
is nearer the electronegative oxygen
The scale
The x-axis represents the chemical shift, δ
It is the frequency of the resonance relative to a standard compound which sets the scale at 0 ppm

downfield upfield

180 160 140 120 100 80 60 40 20 0

δ ppm
 13
C-NMR Cyclobutanol
• In this 13C-NMR we can
see three peaks
• Is this what we would
expect?
• The structure of
cyclobutanol is:
5
OH
4 1

3 2
180 160 140 120 100 80 60 40 20 0
δ ppm

• Carbon ‘2’ and ‘4’ are all in the same environment when you look at the atoms they are attached
to and also the atoms that are attached to them
• Carbon ‘1’ and ‘3’ are in different environments
Calculating the number of peaks
How many peaks would a molecule of heptane have?

1 2 3 4 5 6 7

Peak 1: Carbons 1 and 7 have identical environments

Peak 2: Carbons 2 and 6 have identical environments
Peak 3: Carbons 3 and 5 have identical environments
Peak 4: Carbon 4 has its own unique environment
Calculating the number of peaks
A 13C-NMR Spectra has 5 peaks, does it belong to
◦ 2,2-dimethylpentan-3-ol or

◦ 3,3-dimethylpentan-2-ol?
NMR Spectroscopy
37.4 PROTON ( 1 H) NMR SPECTROSCOPY
1
H-NMR
In the same way as a nucleus of carbon-13, hydrogen nuclei (a single proton) are affected by a
magnetic field
The magnetic spin can align with a magnetic field or against it in a higher energy state
The difference in energy between the two states once again corresponds to the energy of a specific
radio frequency
The size of energy gap between the two alignments depends on the environment of the hydrogen
atom
H-NMR Propan-1-ol
1

• In this 1H-NMR we can see four peaks

(2) (3) • Is this what we would expect?
• The structure of propan-1-ol is:
A low resolution spectra

(1)
(2)

9 8 7 6 5 4 3 2 1 0
δ ppm

• Hydrogen nuclei exist in four separate environments

H-NMR Propan-1-ol
1

• In this low resolution spectra the

(2) (3) area under the peak can be used to
tell you how many hydrogen nuclei
A low resolution spectra exist in a given environment

(1)
(2)

9 8 7 6 5 4 3 2 1 0
δ ppm

• (2), (1), (2) and (3) are relative areas and again are in line with our structure
• areas not to scale
H-NMR Propan-1-ol
1

• Data section values help to confirm

the analysis
H-NMR Propanal and Propanone
1

If these two molecules were examined independently how

many peaks would they each have and what would be
the ratio of those peaks in each individual molecule?
H-NMR Propan-1-ol
1

(2) (3) • At a higher resolution we can observe that the

peaks are split
A high resolution spectra
• The amount of splitting of each signal peak
depends on the number of hydrogens attached
to the carbon atom/s next in the chain

• Hydrogen atoms on the same carbon are

(1) equivalent and don’t affect each other
(2)
• Splitting occurs because of spin-spin coupling
• This is university chemistry but essentially
slightly different magnetic environments exist
for that specific proton due to interaction with
other neighbouring protons

9 8 7 6 5 4 3 2 1 0
δ ppm
Naming split signals
We classify the number of peaks a signal has with the following names:
Singlet
◦ 0 hydrogen neighbours = 1 peak

Doublet
◦ 1 hydrogen neighbour = 2 peaks

Triplet
◦ 2 hydrogen neighbours = 3 peaks

Quartet
◦ 3 hydrogen neighbours = 4 peaks

}
Quintet (Pentet)
◦ 4 hydrogen neighbours = 5 peaks

Sextet multiplets
◦ 5 hydrogen neighbours = 6 peaks

Septet
◦ 6 hydrogen neighbours = 7 peaks
1
H-NMR Propan-1-ol
If a proton has n neighbours
(protons attached to adjacent carbons) le t
It will split into n+1 lines , t rip
=3
n+1
2 ,
n= plet
t ri
= 3,
2 , n+1
n =

-OH
gro
ups
are
al way
s sing
l et s

n = 5, n+ 1 = 6,
sextet
-OH and –NH bonds
The hydrogen on the -OH or –NH always forms a singlet
They do not interact with other hydrogens to produce any splitting in their peak or the peaks of
neighbouring hydrogens
D2O is used to eliminate the peak from –OH and –NH bonds and allow more accurate analysis
Introducing D2O causes hydrogens on the –OH and -NH in the compound to be replaced by deuterium
due to proton exchange in the alcohol and amino groups
Peaks created by –OD and –ND bonds appear in a different region of the spectra and are not
displayed
Spectra Examples

Follow the link for collections of

spectra for different molecules

Examples
Creating a baseline
In order to compare peaks tetramethylsilane is used as an internal standard for
calibrating chemical shift for 1H and 13C NMR

All four carbons atoms and all twelve hydrogen atoms (relative to each other) in a
tetramethylsilane molecule are equivalent so a strong peak is created
Creating a baseline
The peak appears to the far right of the spectra due to the silicon atom
◦ The electrons in the C-Si bond are closest here to the carbon atom and hence shield it the most effectively
from the magnetic field meaning a very large magnetic field (hence far right) is needed to create the resonance
condition in the carbon atoms in 13C-NMR

◦ For 1H-NMR twelve hydrogens creates a strong singlet which can be easily identified

All other chemical shifts are determined relative to this peak

 More chemical shift detail
• The peak created by TMS is often omitted
from spectra
• The horizontal scale is the chemical
shift measured in parts per million
• A peak to the left of the TMS peak
is downfield 
• 100ppm is more downfield than 40ppm

180 160 140 120 100 80 60 40 20 0

δ ppm

• Assuming the device varies magnetic field, a peak at 80 means that the carbon atoms causing the peak need a
magnetic field 80 millionths less than the field needed by the TMS to create a resonance condition
Solvents
Solid molecules need to be dissolved into solution for the NMR to be run
Deuterated solvents such as CDCl3 are used
The hydrogen atom in this chloroform molecule has been replaced with an atom of deuterium, an
isotope of hydrogen
As such it doesn’t have any hydrogen to interfere with a 1H-NMR spectra
It does have carbon atoms but CDCl3 creates an easily identifiable peak which can be easily discounted
when analysing the spectra for 13C-NMR
Summary: Proton Nuclear Magnetic Resonance (NMR)

LINK
Homework task
Create a worksheet for another member of the
class
It should focus on one particular area of the
course
You should have an answer sheet so you can
assist and your colleague and mark their work
 Paper Session Year Question

Past Paper
43 October 2017 6
42 October 2017 3

Questions
42 October 2017 8
43 May 2017 6
42 May 2017 2

Complete the entire question 42 February 2017 7

42 February 2017 9
42 October 2018 1
42 October 2018 5
41 October 2018 1
41 October 2018 6
42 Summer 2018 8
41 Summer 2018 7
42 February 2018 9
42 October 2019 8
42 October 2019 9
41 October 2019 4
41 October 2019 9
42 May 2019 7
41 May 2019 8
42 March 2019 6
Write a paragraph explaining a concept from this unit using the ‘wordcloud’ above to give you ideas and remind
you of keywords
Some people will then be asked to read their paragraphs out to the class.
Which of these ideas has been difficult?
Mass Spectrometry
Chromatography
H-NMR Spectroscopy
1

C-13 NMR Spectroscopy

Teacher self-evaluation
Changes to be made to PowerPoint, teaching, resources?
◦ was the pace too fast or too slow for your students?
◦ more/less slides for next year?
◦ any of your current resources you could add?

Ask the students for feedback