Instrumental Analysis I

Chem.3021

November 2022 1
 Chapter One
Introduction to Separation Science
· What is separation science?
 Separation is an operation in which a mixture is divided into at least two components with
different compositions, or two molecules with the same composition but different
stereochemical structure.

 A collection of techniques for separating components of complex mixtures of analytes

 Most separations are not an analytical technique in their own right, until combined with an
analytical detector (often a type of spectrometer)

 The separation process usually entails physical means

 What is Separation?
·Separations are key aspects of many modern analytical methods.
·Real world samples contain many analytes, most analytical methods do not offer
sufficient selectivity to be able to speciate all the analytes that might be present.

·Most separation methods involve separation of the analytes into distinct

chemical species, followed by detection.

·(a +b +c +d +…) (a) + (b) + (c) + (d) complete separation

·(a +b +c +d +…) (a) + (b) + ((c) + (d)) partial separation

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Classifying Separation Techniques

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 General Theory of Separation Efficiency

• The goal of an analytical separation is to remove either the analyte or the

interferent from the sample matrix.

• To achieve a separation there must be at least one significant difference in

chemical or physical properties between the analyte and interferent.

• Here selectivity is also very important

• Efficiency of separation is influenced both by the failure to recover all the

analyte and the failure to remove all the interferent.

• We define the recovery of analyte as RA 5

• where CA is the concentration of analyte remaining after separation,

and (CA)o is the initial concentration of analyte.

• A recovery of 1.00 means that none of the analyte is lost during the
separation.

• The recovery of the interferent, RI, is defined in the same manner

where CI is the concentration of interferent remaining after separation, and

• (CI)o is the initial concentration of interferent.
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 The degree of separation is given by a separation factor, SI,A, which is
the change in the ratio of interferent to analyte caused by the separation

 In an ideal separation RA = 1, RI = 0, and SI,A = 0.

 In general, the separation factor should be approximately 10–7 for the

quantitative analysis of a trace analyte in the presence of a macro
interferent,

 and 10–3 when the analyte and interferent are present in approximately

equal amounts.

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 The degree of separation is given by a separation factor, SI,A, which is the
change in the ratio of interferent to analyte caused by the separation

 In an ideal separation RA = 1,RI = 0, and SI,A = 0.

 In general, 7 ≤ SI,A≤ 10 for quantitative analysis of a trace analyte in the

presence of a macro interferent,

 and 3≤ SI,A≤ 10 when the analyte and interferent are present in approximately
equal amounts.

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Example

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 Separating Mixtures
• Substances in a mixture are combined physically, so processes that are based
on differences in physical properties are used to separate components.

• Numerous techniques have been developed to separate mixtures to study

components

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Some simple examples of separation
• Filtration
• Chromatography
• Centrifuging
• Evaporating
• Crystallization
• Dissolving
• Decantation
• Sieving
• Flotation
• Physical Means
• Others
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 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 on the filter paper

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Filtration can be used to
separate an insoluble
substance from a soluble
substance

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 Distillation
• Used to separate homogeneous
mixtures
• Based on differences in boiling
points of substances involved

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Evaporation can be used to
separate a solute from the
solvent in a solution

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 Centrifugation
• Spins sample very rapidly: denser materials after
go to bottom (outside) Before
• Separate blood into serum and plasma
• Serum (clear)
• Plasma (contains red blood cells Serum
‘RBCs’)
• Check for anemia (lack of iron) Blood

RBC’s

A B C
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 A magnet
• Can be used to separate a magnetic substance
from a non-magnetic substance

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 Chromatography

• This will be discussed later

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 Decantation
• Decanting is done to separate
particulates from a liquid by allowing
the solids to settle down to the bottom
of the mixture and pouring off the
particle-free part of the liquid.
Another method is to allow two
immiscible liquids to separate and the
lighter liquid is poured off.

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1) Mechanical separations: separations based on size and/or
density differences of different components in a mixture, for
separation of solid from liquid (e.g. filtration and centrifugation).
2) Diffusional separations (mass transfer operations):
separations based on molecular movement toward a favourable
phase, for separation of dissolved components (e.g. distillation,
absorption, extraction). (Note: Mass transfer is the transfer of
solute molecules from one point to anther or from one phase to
another.)
3) Membrane separations: use of a semipermeable membrane to
separate molecules with difference in size or some other
properties.
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