Introduction to unit 2

Background to unit 2
Assessment –series of assignments
Keywords: Solvent, solute, solution.

Watch the video and write down what chromatography

is and how it works.
Extension – How can chromatography be used to test
pure or impure substances?
https://www.youtube.com/watch?v=PvHvx7k7UPU&t=123s
https://www.youtube.com/watch?v=PvHvx7k7UPU
https://www.youtube.com/watch?v=SnbXQTTHGs4
(15 min)
Mind map notes
Lesson 2
Overview for notes – PREZI – 1
lesson plus complete research for
homework
• https://prezi.com/ah1oxe_uj1yx/chromatography/
Chromatography key terms – copy
into notes
Paper chromatography
Practical lesson 3-4
WALT: How to interpret
chromatograms
• how chromatography can be used to distinguish pure substances from
impure substances
• how to interpret chromatograms
• how to determine Rf values from chromatograms.
 Food colours

Why did Nestle get rid of blue Smarties until 2006?

The artificial food colourings in Smarties was linked to hyperactivity.

Nestle had to separate the colours on the Smarties out to test which ones
caused hyperactivity.
 Separating the colours

How are food colours separated out?

Uses for paper chromatography

Forensics labs use it for DNA profiling.

It is used in the food industry to determine

what food colours are in a food.
Paper chromatography
• In paper chromatography, the stationary phase is a sheet of chromatography
paper. The mobile phase may either be an aqueous (water-based) liquid or a
non-aqueous (carbon-based) organic solvent.
• An example of an organic solvent is propanone - which is the main chemical in
nail varnish remover.
• For each chemical in the sample, there is a dynamic equilibrium between the
stationary phase and the mobile phase.
• The overall separation depends upon how strongly attracted the chemicals
are to the mobile and the stationary phases.
• This produces a chromatogram where different samples can be compared to a
reference material.
 Chromatography key words

Solvent – the substance that we dissolve the colours in. It is water in this
experiment.

Solute – the substances that are dissolved in the solvent.

Chromatogram – the piece of chromatography paper showing how the dyes

have separated.
Why we use a pencil for chromatography

We draw a pencil on the chromatogram because a pen mark will run up the
water.
Lesson content
• Investigation into the use of paper chromatography to separate and
identify a mixture of food colourings.

• You will use paper chromatography to separate the different colours

present in an unknown mixture of food colourings. You will then
measure the distance travelled by each colour and the solvents to
calculate Rf values.
• Risk assessment

• Safety goggles should be worn throughout

Draw the results table – use a pencil and ruler!

Distance travelled in mm
Food colouring Rf value
Solvent Spot

     
A

     
B

     
C

     
D
Method
• You are provided with the following:

• 250 cm3 beaker

• Glass rod
• A rectangle of chromatography paper
• Four known food colourings labelled A-D
• An unknown mixture of food colourings labelled U
• Glass capillary tubes.
Read these instructions carefully before you start work.
1. Use a ruler to draw a horizontal pencil line 2 cm from a short edge of the chromatography paper.
2. Mark five pencil spots at equal intervals across the line. Keep at least 1 cm away from each end.
3. Use a glass capillary tube to put a small spot of each of the known colourings on four of the pencil
spots. Then use the glass capillary tube to put a small spot of the unknown mixture on the 5th
pencil spot.
4. Try to make sure each spot is no more than 5 mm in diameter.
5. Label each spot in pencil.
6. Pour water into the beaker to a depth of no more than 1 cm.
7. Tape the edge of the chromatography paper to the glass rod. The paper needs to be taped at the
end furthest from the spots.
8. Rest the rod on the top edge of the beaker. The bottom edge of the paper should dip into the
water.

• Ensure that the:

• pencil line is above the water surface
• sides of the paper do not touch the beaker wall.
 Method continued  3 Hang the paper up to dry thoroughly.
4 Measure the distance in mm between the two pencil lines. This
1 Wait for the water solvent to travel at least three is the distance travelled by the water solvent.
quarters of the way up the paper. Do not disturb 5 Measure and record the same distance for each food colouring in
the beaker during this time. the table below.
2 Carefully remove the paper. Draw another pencil 6 For each of the four known colours, measure the distance in mm
line on the dry part of the paper as close to the wet from the bottom line to the centre of each spot. Write each
measurement in the table.
edge as possible.
7 Use the following equation to calculate the Rf value for each of
the known colours.
 

 
8 Write the calculated values in the table.
9 Match the spots in mixture U with those from A–D. Use the
colour and distance travelled to help you.
10 Which of colourings A–D are in mixture U?
11 Are there any other colourings in mixture U which do not match
A–D?
 Assignment
preparation
Title: Separate to
identify
Lesson 5-6
 The brief - C: Undertake chromatographic
techniques to identify components in mixtures.
• You are a newly appointed technical assistant at a
large chemical plant, Chemcalequip. As part of
your induction period and to progress in your
role, you have to demonstrate skills in a range of
practical procedures and techniques. The
company is often required to identify substances.
• You need to demonstrate that you can carry out
different chromatographic techniques to
separate and identify components in mixtures. In
a report of the techniques you have used, you
will need to explain the techniques, analyse and
evaluate your results and suggest improvements.
What do I need to hand in as
evidence?
• A report on the experiments you have carried out.

• Your teacher will also be assessing you:

• An observation report with a checklist, completed by the tutor,
identifying the level of independence and proficiency demonstrated
whilst carrying out each technique. Safe working practices must be
identified.
Criteria covered by this task:
Unit/Criteria
To achieve the criteria you must show that you are able to:
reference
Evaluate the chromatographic techniques used in relation
C.D3
to outcomes and suggest improvements.
Analyse own chromatograms and relate the factors that
C.M3 affect the separation of mixtures to the quality of results
obtained.
Correctly use chromatographic techniques to produce
C.P5
chromatograms.
Explain the use of chromatographic techniques to separate
C.P6
mixtures.
The assignment:
Evaluate the chromatographic techniques you
have used in relation to the outcomes.

• The Practicals (Teacher assessed and your results will be used in your
written evaluations)
• Demonstrate safe working practices and a high level of proficiency
when using the following chromatography techniques;
• paper chromatography of extracted plant pigments
• Thin layer chromatography, TLC, of extracted plant pigments
• paper chromatography of amino acids.
• Repeat any separations where you can justify a lack of satisfaction
with the quality of the separation obtained.
 Correctly use chromatographic
techniques to produce
How do I write up my practical? chromatograms.

• Practicals should be written up individually and each should follow the

format below:
• Title – written in full – this should describe your investigation
• Aim – what do you hope to achieve by doing this practical/ what is the
purpose of this?
• Equipment list / labelled diagram
• Full, bullet pointed method
• Risk assessment table (Hazard, risk, precaution, emergency action) – see
technicians for CLEAPSS information
• Results table with calculated Rf values
• Conclusion of your results (whole analysis can be completed in the report)
 Correctly use chromatographic
techniques to produce
chromatograms.

Pass: How do we display our results?

• Make suitable table for each practical that allows you to record:
• Distance solvent has moved
• Distance spot has moved
• Rf values
The report
See slides for guidance and links.
Produce a report to include (use this as a tick list,
additional information on following slides:
• An introduction to your report – what is chromatography and what types there are.
• an evaluation and analysis of the links between the chromatography techniques used
and the chromatograms produced.
• conclusions about the components and their effect on separation leading to the
results obtained.
• results from the paper chromatography and TLC of extracted plant pigments and from
paper chromatography of amino acids
• calculations of Rf values and consideration of the factors that influence separation.
• an explanation of the principles behind the chromatographic separations and their
suitability.
• justification of conclusions about the identification of components in the mixtures,
e.g. polarity of the solvents, effect of molecule size on its mobility.
• Suggestions and rationales for specific improvements to the chromatographic
procedures carried out and full justification of these suggestions.
 Explain the use of
chromatographic
techniques to

Introduction - a reminder! separate mixtures.

• Paper chromatography
• In paper chromatography, the stationary phase is a sheet of chromatography paper.
The mobile phase may either be an aqueous (water-based) liquid or a non-aqueous
(carbon-based) organic solvent.
• An example of an organic solvent is propanone - which is the main chemical in nail
varnish remover.
• For each chemical in the sample, there is a dynamic equilibrium between the
stationary phase and the mobile phase.
• The overall separation depends upon how strongly attracted the chemicals are to the
mobile and the stationary phases.
• This produces a chromatogram where different samples can be compared to a
reference material.
 Introduction – a reminder! Explain the use of
chromatographic
techniques to
separate mixtures.
• TLC Thin Layer Chromatography
• instead of paper, the stationary phase is a thin layer of an inert substance (eg silica) supported on a flat,
unreactive surface (eg a glass plate).
• TLC has some advantages over paper chromatography. For example:
• the mobile phase moves more quickly through the stationary phase
• the mobile phase moves more evenly through the stationary phase
• there is a range of absorbencies for the stationary phase
• TLC tends to produce more useful chromatograms than paper chromatography, which show greater
separation of the components in the mixture - and are therefore easier to analyse.
• The distance a sample travels can depend on the size or the polarity of the molecules involved. Larger
molecules take longer to move up the chromatography paper or TLC plate, whereas smaller molecules are
more mobile.
• Likewise, the polarity of the molecules can affect how far the spots travel, depending on the type of solvent
used. Polar molecules will be more strongly attracted to polar solvents, and so would move further if a
polar solvent was used as opposed to a non-polar solvent.
• The distance that spots move can be compared to the overall distance the solvent has moved and
comparisons and measurements made.
Pass (paragraph 1): an evaluation and analysis of the links between the
chromatography techniques used and the chromatograms produced.

• Look at the 3 methods you used.

• Which was the most appropriate method for each test and which
produced the most valid results close to given Rf values?
• Why do you think this happened?
Pass (paragraph 2): What effect do the components have on separation?
Explain the use of
chromatographic techniques
to separate mixtures.

• Components (chemicals) in a moving liquid mixture spread out because they

travel at different speeds over a stationary solid. The key thing to remember is
that chromatography is a surface effect.
• The distance a sample travels can depend on the size or the polarity of the
molecules involved. Larger molecules take longer to move up the
chromatography paper or TLC plate, whereas smaller molecules are more mobile.
• Likewise, the polarity of the molecules can affect how far the spots travel,
depending on the type of solvent used. Polar molecules will be more strongly
attracted to polar solvents, and so would move further if a polar solvent was used
as opposed to a non-polar solvent.
• The distance that spots move can be compared to the overall distance the solvent
has moved and comparisons and measurements made.
Pass: What are the factors that influence
Explain the use of
chromatographic
techniques to
separate mixtures.

separation?
• For chromatography to work effectively, the components of the
mobile phase need to separate out as much as possible as they move
past the stationary phase. That's why the stationary phase is often
something with a large surface area, such as a sheet of filter paper, a
solid made of finely divided particles, a liquid deposited on the
surface of a solid, or some other highly adsorbent material.
• Type of running solvent needs to be chosen well as this will also affect
the rate at which separation takes place and how affective the
separation is.
Pass: What are the principles behind the
Explain the use of
chromatographic
techniques to
separate mixtures.

chromatographic separations?
• Chromatography is used to separate mixtures of substances into their
components. All forms of chromatography work on the same
principle.
• They all have a stationary phase (a solid, or a liquid supported on a
solid) and a mobile phase (a liquid or a gas). The mobile phase flows
through the stationary phase and carries the components of the
mixture with it. Different components travel at different rates.
Merit: How can we justify the identification of Analyse own chromatograms
and relate the factors that affect
the separation of mixtures to

components in mixtures? the quality of results obtained.

• Rf values – describe what an Rf value is and how it is used. Use your calculated values
as examples.
• Polarity of solvents
• Polarity has a huge affect on how attracted a chemical is to other substances. Some
molecules have a positively charged side and a negatively charged side. For example,
the positive side is attracted to the negative side of another molecule (opposites
attract). The larger the charge difference, the more polar a molecule is.
• As polarity of the solvent is increased, all the components of the mixture move faster
during chromatography.
• https://socratic.org/questions/how-does-polarity-affect-chromatography
• Effect of molecule size on mobility – research this – Larger molecules take longer to
move up the chromatography paper or TLC plate, whereas smaller molecules are more
mobile. Use your results for reference (which would be larger and which are smaller
components of the substances? How can you tell? Can you back this up with research?
Distinction: Suggestions and rationales for specific improvements to the
chromatographic procedures carried out and full justification of these suggestions

• From the practical work and your research, you should be able to
come up with some specific improvements for the procedures carried
out.
• A justification needs to EXPLAIN WHY the suggested method (or part
of a method) is better than the method you have used and how it
would affect your results.
• You need to research this yourself!
Evaluate the
chromatographic
techniques used in
relation to outcomes and
suggest improvements.
References
• Some of the references you could use for your report alongside your own
research: (Web pages need address, title and date accessed)
• http://www.chromatography-online.org/Detection-and-Quantitatitation.php
• http://www.chemguide.co.uk/analysis/chromatography/paper.html
• http://www.explainthatstuff.com/chromatography.html
• http://www.chemguide.co.uk/analysis/chromatography/paper.html
• https://socratic.org/questions/how-does-the-solvent-effect-chromatography
• Use your methods for references
• Text book