PURIFICATION & ANALYSIS

Chemistry OLevels SHR - Scholastica

Topics
Pure Substances & Mixtures
Parameters of Purification
Techniques of Purification
Importance of Analysis
Pure Substances
A Pure Substance contains
Only one substance
One type of atom or molecule
No contaminating impurities
Mixtures
Mixtures are
formed by a physical change
Separated by methods of purification

Parameters of Purification
A pure substance has a fixed and definite
melting and boiling point.
Precise & predictable
Can be used to test the purity of a sample
A pure substance has a fixed solubility
Solubility curve
Chromatogram
A pure substance has physical properties
which are EXACT, PRECISE, DEFINITE &
PREDICTABLE
Parameters of Purification
The presence of an impurity in a substance
Lowers the melting point
Raises the boiling point of a substance
Changes will be spread over a range of
temperatures
Sharpness at a particular point will not be
observed
Methods of Purification
Mixture Type Method
Solid + Solid Soluble &
Insoluble
Filtration & Decantation
Solid + Solid Soluble Chromatography
Solid + Liquid Insoluble Solid Filtration
Centrifugation
Decantation
Solid + Liquid Soluble Solid Evaporation
Crystalization
Distillation
Liquid + Liquid Immiscible Using a Separating
Funnel
Liquid + Liquid Miscible Fractional Distillation
Separating Mixtures
Decantation solid insoluble particles from a liquid (set, particles settle and
pour off liquid)



Separating Mixtures
Filtration solid insoluble particles from a liquid



Filtration
Used to separate heterogeneous
mixtures composed of solids and
liquids
Uses a porous barrier to separate
the solid from the liquid
Liquid passes through leaving
the solid in the filter paper
Filtration
Gravity Filtration
This is the simplest kind of filtration when the
solution to be filtered is poured through a
filter paper in a filter funnel. The filtration of
hot solutions through a heated funnel and
fluted filter paper is often carried out as part
of a recrystallisation. Gravity filtration is
generally carried out to remove impurities
rather than to isolate solids.
Buchner Filtration
When a solid needs to be
isolated from a solution it
is normally done at a
reduced pressure using a
Buchner flask and
Buchner funnel.
Separating Mixtures
Distillation a way of obtaining a pure solvent from a solution
(boil, evaporation : steam, cooled : condenses,
solvent is collected in another vessel)

Distillation
Used to separate
homogeneous
mixtures
Based on
differences in
boiling points of
substances
involved
solution boiled in distilling flask and
vapour/steam cooled and condensed in a Liebig
condenser
condenser consists of jacket of cold water with
coldest water entering bottom of jacket and
circulating out through the top ensuring that
coldest part of condenser is just before the
vapour escapes and that the jacket is completly
full of water
condensed solvent distillate
all impurities left in distilling flask
Simple vs Fractional
Distillation
Simple distillation is used to separate solvent
from a non-volatile solute (i.e. solute with
high boiling point). Eg. Water from salt
solution.
Fractional distillation is used to separate
mixture of miscible liquids, eg. Mixture of
ethanol and water.

Fractional Distillation
How fractionating column
works
May be filled with glass beads, plates or a spiral.
These provide a large surface area for vapour to
condense on.
Liquid with lowest boiling point will distill over to
the condenser first
Vapours of liquids with higher boiling points
condense along fractionating column and re-
enters the round-bottomed flask

Fractional Distillation
Fractional Distillation
Industrial Applications
oil refineries
separating the various components of crude oil or
petroleum
Sublimation
process used to separate a solid that sublimes
from one that does not
Eg:
mixture of iodine + copper;
mixture of ammonium chloride + sodium chloride
When a mixture of a solid that sublimes and a
solid that does not is heated, the solid that
sublimes will turn into a vapour and separates
from the other solid which remains in the
container.
Sublimation
Evaporation can
be used to separate
a solute from the
solvent in a solution.

Only for solids that
will NOT
decompose on
heating.

Eg. sodium
chloride (common
salt) from salt
solution
A separating funnel can be
used to separate two
liquids which are
immiscible, For eg. Oil &
Water
Chromatography
Separates components of a mixture
based on ability of each component
to be drawn across the surface of
another material and based on their
solubility
Mixture is usually liquid and is usually
drawn across chromatography paper
Separation occurs because various
components travel at different rates
Components with strongest
attraction for paper travel the least
and components with the highest
solubility travel the most
Important Concepts in
Paper Chromatography
Capillary Action the movement of liquid within the spaces of a
porous material due to the forces of adhesion, cohesion, and surface
tension. The liquid is able to move up the filter paper because its
attraction to itself is stronger than the force of gravity.

Solubility the degree to which a material (solute) dissolves into a
solvent. Solutes dissolve into solvents that have similar properties.
(Like dissolves like) This allows different solutes to be separated by
different combinations of solvents.

Separation of components depends on both their solubility in the
mobile phase and their differential affinity to the mobile phase and the
stationary phase.
Simple Example of Paper Chromatography
using Marker Pens

Chromatography
Examples of Chromatography

Gas Chromatography
Used to determine the chemical
composition of unknown
substances, such as the different
compounds in gasoline shown by
each separate peak in the graph
below.
Paper Chromatography
Can be used to separate the
components of inks, dyes, plant
compounds (chlorophyll), make-
up, and many other substances
Liquid Chromatography
Used to identify unknown plant
pigments & other compounds.
Thin-Layer Chromatography
Uses thin plastic or glass trays to
identify the composition of pigments,
chemicals, and other unknown
substances.
What if the substances you are
interested in are colourless?
In some cases, it may be possible to make the
spots visible by reacting them with something
which produces a coloured product. A good
example of this is in chromatograms produced
from amino acid mixtures.
Suppose you had a mixture of amino acids and
wanted to find out which particular amino acids
the mixture contained. For simplicity we'll
assume that you know the mixture can only
possibly contain five of the common amino
acids.

What if the substances you are
interested in are colourless?
A small drop of a solution of the mixture is
placed on the base line of the paper, and similar
small spots of the known amino acids are placed
alongside it. The paper is then stood in a suitable
solvent and left to develop as before. In the
diagram, the mixture is M, and the known amino
acids are labelled 1 to 5.
The position of the solvent front is marked in
pencil and the chromatogram is allowed to dry
and is then sprayed with a solution of ninhydrin.
Ninhydrin reacts with amino acids to give
coloured compounds, mainly brown or purple.

What if the substances you are
interested in are colourless?
The left-hand diagram shows the paper after the
solvent front has almost reached the top. The
spots are still invisible. The second diagram
shows what it might look like after spraying with
ninhydrin.
What if the substances you are
interested in are colourless?
There is no need to measure the R
f
values
because you can easily compare the spots in the
mixture with those of the known amino acids -
both from their positions and their colours.
In this example, the mixture contains the amino
acids labelled as 1, 4 and 5.
And what if the mixture contained amino acids
other than the ones we have used for
comparison? There would be spots in the mixture
which didn't match those from the known amino
acids. You would have to re-run the experiment
using other amino acids for comparison.

Chromatography - Applications
analyzing ink dyes for forgery cases
analyzing food dyes to ensure that only permitted
colorings are used in foodstuffs
checking whether pesticides on vegetables exceed
safe levels
detecting trace levels of drugs in urine samples
separation of amino acids, proteins &
carbohydrates.
analysis of drugs, hormones, vitamins & brain
amines.
determination of molecular weight of proteins.
isolation of plant pigments from roots and leaves.


Uses for Chromatography
Chromatography is used by scientists to:

Analyze examine a mixture, its components, and their
relations to one another
Identify determine the identity of a mixture or components
based on known components
Purify separate components in order to isolate one of interest
for further study
Quantify determine the amount of the a mixture and/or the
components present in the sample

Uses for Chromatography
Real-life examples of uses for chromatography:
Pharmaceutical Company determine amount of each
chemical found in new product
Hospital detect blood or alcohol levels in a patients blood
stream
Law Enforcement to compare a sample found at a crime
scene to samples from suspects
Environmental Agency determine the level of pollutants in
the water supply
Manufacturing Plant to purify a chemical needed to make a
product
R
f
Factor

Retention Factor
It is a number that represents how far a
compound travels in a particular solvent.

It is measured by measuring the ratio of the
distance traveled by the compound and the
distance traveled by the solvent.

Rf ( Retention Factor )Value
distance traveled by component from application point
Rf = -----------------------------------------------------------------------------
distance traveled by solvent from application point

R = zero, - Solute remains in the stationary phase and thus it
is immobile.
R = 1 - Solute has no affinity for the stationary phase and
travels with the solvent front.

Separating Mixtures
Centrifuging solid particles from a suspension
(liquid spun very quickly in a centrifuge pushes particles to sides and liquid can poured or
filtered off)