7.

Radiochemical methods of
Analysis

By Birhanu M.
Dec 2014

10/20/2023 Radiochemical Methods of Analysis 1

 Introduction
In nuclear medicine, drugs containing
radioactive metals, metal complexes, and
metal conjugates are used for diagnosis and
therapy of various diseases.

Radioactivematerials used as
pharmaceuticals include
◦ small organic and inorganic molecules and
◦ macromolecules such as monoclonal antibodies
and antibody fragments that are attached to
radioac-tive metals. 10/20/2023 Radiochemical Methods of Analysis 2
Introduction…
The successful use of radiochemicals
needs a basic understanding of
◦ Radiation
◦ Radioactivity and
◦ The nature and characteristics of
instruments to detect and quantitate radiation

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 Introduction…
ATOMIC STRUCTURE, NUCLEAR STABILITY,
AND RADIOACTIVITY
Bohr’s planetary model of the atom consists of a
dense positively charged nucleus surrounded by
negatively charged electrons (e) in orbits of well
defined energy states.

The nucleus consists of positively charged protons

and neutral particles called neutrons.

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Introduction…
The protons and neutrons are held together by very
a strong nuclear force of attraction, effective at
very close distances (approximately
).

The electroneutrality of the atom is maintained

by the orbital electrons, which are equal in
number to that of the protons.

This number is called the atomic number, Z.

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 Introduction…
Any configuration of protons and neutrons is
called a nuclide.

Thereare three nuclides of the element

hydrogen with atomic number one.

The three nuclides of hydrogen are called

isotopes of hydrogen.

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Introduction…
Tritium with an N/Z ratio of two is unstable.

When the N/Z ratio becomes higher, the

nucleus become unstable and results in the
disintegration of the nucleus so as to achieve a
stable N/Z ratio and therefore a stable nucleus.

This process is called radioactive decay.

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Introduction…

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 Introduction…
 This radioactive process can be spontaneous in some
naturally occurring nuclides; then these elements are
said to be naturally radioactive.

 When such instability is brought about by bombarding

stable nuclides with high-energy particles, it is called
artificial radioactivity.

 In general, when the N/Z ratio is greater than 1.6, the

radioactive nuclide readjusts to a stable ratio of N/Z
with the release of energy and particles of matter.
10/20/2023 Radiochemical Methods of Analysis 9
 Introduction…
Radioactive Decay
Different radioactive species undergo
disintegration at different rates.

The rate of this decay or activity is

◦ characteristic of the individual nuclide and
◦ proportional to the number of radioactive
nuclides

10/20/2023 Radiochemical Methods of Analysis 10

Introduction…
The proportionality constant is called the
decay constant and is denoted by

The decay constant is a measure of the

probability that a certain radioactive nucleus
will disintegrate within a specified time
interval.

10/20/2023 Radiochemical Methods of Analysis 11

 Introduction…
These disintegrations are characteristic of the
nuclide and are unaffected by
◦ pressure
◦ temperature
◦ concentration and
◦ other physical or chemical properties of the
radio-nuclide.

This rate constant is conveniently denoted in terms

of t1/2, or halflife.
10/20/2023 Radiochemical Methods of Analysis 12
Introduction…
The halflife of a radionuclide is the time
required for the sample activity to decrease to
half its initial value

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Introduction…

Other commonly used terms in nuclear

medicine and pharmacy are
 average (mean) halflife,
 biological halflife, and
 effective halflife

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Introduction…
Average half-life
 is the mean lifetime of a nuclide, and it
is equal to 1.44 x t1/2

Biologic halflife, tb,

 is the time required for the body to
eliminate half the administered dose by
normal biological process of elimination.

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Introduction…
Effective halflife (teff)
is a measure of how fast the body eliminates the
radioactive material by the combination of
biological elimination and radioactive decay:

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 Decay of Radionuclide
 The radioactive decay process involves the emission of
radiation
 This radiation is dependent on the mode of decay of the
particular radionuclide.

 Radiation resulting from any decay process can be

classified as
◦ Alpha (α),
◦ Beta minus (β-),
◦ Positron (β+) and
◦ Gamma (γ) radiation.
10/20/2023 Radiochemical Methods of Analysis 17
Decay of Radionuclide
Alpha () decay
Alpha () decay is characterized by the emission
of an alpha particle from the nucleus.
  particle is a helium ion containing two protons and
two neutrons bound together in the nucleus.
A4 2
A
ZX Z2 Y  He 4
2

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Decay of Radionuclide…
 The  particles are monoenergetic, and their range
in matter is very short (on the order of 10-6 cm)
due to their mass, thus leaving much of its energy
on a very small area (only a few cell diameters)
 Used only for therapeutic purposes
 Their clinical use is very limited, and they are
mainly used for research purposes

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Decay of Radionuclide…
Beta-Minus (Negatron) Decay
 Beta-minus (-) decay characteristically occurs with
radionuclides that have an excess number of neutrons
(‘‘neutron rich’’) compared with the number of protons

◦ i.e., has a higher N/Z ratio compared to the

stable nucleus
 In - decay, a neutron (n) decays into a proton (p)
and a - particle
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 Decay of Radionuclide…
 After - decay, the atomic number of the daughter nuclide
is one more than that of the parent nuclide; however, the
mass number remains the same for both nuclides

A
Z X A
Y  β 
Z 1
-

An antineutrino ( ) is an entity almost without mass and


charge and is primarily needed to conserve energy in the
decay

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Decay of Radionuclide…

10/20/2023 Radiochemical Methods of Analysis 22

 Decay of Radionuclide…
Beta emitters represent different energy levels,
and have different range in matter (40 – 100μm)
depending on their energy.

 Beta emitting radionuclides are used in

radiopharmaceuticals mainly for therapeutic
purposes
10/20/2023 Radiochemical Methods of Analysis 23
Decay of Radionuclide…
Positron or β+ Decay
 Nuclei that are ‘‘neutron deficient’’ or ‘‘proton
rich’’ (i.e., have an N/Z ratio less than that of the
stable nuclei) can decay by β+ -particle emission
accompanied by the emission of a neutrino (V),
which is an opposite entity of the antineutrino.

10/20/2023 Radiochemical Methods of Analysis 24

Decay of Radionuclide
 In β+ decay, a proton transforms into a neutron by
emitting a β+ -particle
The daughter nuclide has an atomic number that is
one less than that of the parent
 Since a β+ particle can be emitted with energy
between zero and decay energy, the neutrino
carries away the difference between decay energy
and β+ energy.
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Decay of Radionuclide…
A
Z X Y  β  
A
Z-1


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Decay of Radionuclide…
Gamma radiation
A nucleus can remain in several excited energy
states above the ground state
All these excited states are referred to as isomeric
states and decay to the ground state
The decay of an upper excited state to a lower
excited state is called the isomeric transition

10/20/2023 Radiochemical Methods of Analysis 27

 Decay of Radionuclide…
 In isomeric transition, the energy difference between
the energy states may appear as gamma (γ) rays.
Gamma radiation is characterized as
electromagnetic radiation
When used in diagnostic radiopharmaceuticals,
gamma rays are powerful enough to be detected
outside the body of the patient

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 ANALYSIS OF RADIOCHEMICALS

Radiochemical methods of analysis are more

sensitive than other chemical methods.

Most spectral methods can quantitate at the parts-

per-million (ppm) level

Atomic absorption and some HPLC methods with

UV, fluorescence, and electro-chemical methods can
quantitate at the parts-per billion (ppb) levels.

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ANALYSIS OF
RADIOCHEMICALS…
Itis possible to attain quantitation levels lower
than ppb levels of elements by radiochemical
analyses.

Radiochemical analysis, in most cases, can be

done without separation of the analyte

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ANALYSIS OF
RADIOCHEMICALS…
Radionuclides are identified based on the
characteristic decay and the energy of the
particles

Radiochemical methods of analysis include

◦ Tracer methods,
◦ Activation analysis and
◦ Radioimmunoassay techniques.

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ANALYSIS OF
RADIOCHEMICALS…
A. Tracers and Tracer Methods of Analysis

 Radiochemical tracers or radiotracers are compounds

labeled with radioisotopes.

 Fortracer methods, the compound to be measured or a

suitable reagent is radiolabeled.

 A measurement of the redistribution of tracer

within such a sample–reagent reaction system provides
the required quantitative analytical information.

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ANALYSIS OF
RADIOCHEMICALS…

Example: Chromate has been determined

by precipitating it with radioactive Ag +
(Ag-111) of a known activity. The
limited reactant is the analyte, here CrO 4-
2
. Determining the activity of the
precipitate of Ag2CrO4 allows for the
determination of the amount of
chromate.

10/20/2023 Radiochemical Methods of Analysis 33

ANALYSIS OF
RADIOCHEMICALS…
Major advantages of tracer methods are
 high sensitivity,
 simplicity, and
 speed.
Radiotracers are more commonly used for
◦ following mechanisms of biological and/or
chemical processes or
◦ if there is need to eliminate complicated
separation procedures, especially in biological
processes.
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ANALYSIS OF
RADIOCHEMICALS…
B. Activation analysis
isa process in which a target trace element in a
sample matrix is irradiated with particles in a
nuclear reactor.
As a result, an activated radionuclide is
formed.

The characteristic particles or γ -Rays emitted are

used for qualitative identification and, more
often, for quantitative measurement.
10/20/2023 Radiochemical Methods of Analysis 35
ANALYSIS OF
RADIOCHEMICALS…
The most common activation analysis is
neutron activation analysis (NAA).

In this technique, a sample containing the

element is irradiated with neutrons in a
reactor.
After irradiation, gamma-Emissions ensue
from the decaying radionuclide.
These are quantitated by using appropriate
semiconductor radiation detectors.
10/20/2023 Radiochemical Methods of Analysis 36
ANALYSIS OF
RADIOCHEMICALS…
Detecting gamma-Rays of a specific energy
identifies the radionuclide.

These particular energy values correspond to

unique energies characteristic of the decaying
radionuclides.

For example, when 24 Na decays to 24Mg, the

gamma-Rays released have unique energies of
1.268 and 2.754MeV.

10/20/2023 Radiochemical Methods of Analysis 37

ANALYSIS OF
RADIOCHEMICALS…
All methods of activation analysis are
◦ very accurate
◦ sensitive, and
◦ a precision of approximately 2%RSD is
easily attainable.
As many as 60 different elements that can form
radionuclide can be analyzed using NAA.

NAA finds wide application in a number of other

fields, and these are summarized in Table 6.
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10/20/2023 Radiochemical Methods of Analysis 39
ANALYSIS OF
RADIOCHEMICALS…
C. Radioimmunoassay
Radioimmunoassay (RIA) is a technique based
on the formation of antigen–antibody complex.

An antigen is typically a protein of molecular

weight greater than 10,000 that stimulates the
production of antibody in an animal body.

The antigen subsequently binds with the

antibody.
10/20/2023 Radiochemical Methods of Analysis 40
 ANALYSIS OF
RADIOCHEMICALS…
Antigen is usually measured in the patient’s
sample, and the antigen becomes the analyte.

To an antibody, a mixture of labeled and

unlabeled antigen is added in excess such that
the quantity of antibody needed to bind is allowed
to be insufficient.

As a result, both types of antigen compete with

the limited amount of antibody in the sample.
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ANALYSIS OF
RADIOCHEMICALS…
The reaction in an RIA mixture can be
described as follows.

10/20/2023 Radiochemical Methods of Analysis 42

ANALYSIS OF
RADIOCHEMICALS…
To a constant amount of labeled antigen and
antibody, increasing amounts of unlabeled antibody
are added.
The initial amount added is still in excess of the
antibody needed for binding.
As a result of competing reactions of the labeled
and unlabeled antigen, the greater the
concentration of the unlabeled antigen added, the
less is the amount of bound labeled complex
(Ag–Ab complex) and hence greater is the free
(unbound) antigen.
10/20/2023 Radiochemical Methods of Analysis 43
ANALYSIS OF
RADIOCHEMICALS…
 After incubation to equilibrium at a specified
temperature and time, unique to the system, separation
of the free labeled Ag the fraction of the bound labeled
antigen is determined by measuring the activity of the
radioactive nuclide.

 By plotting the percent of bound labeled antigen

versus the concentration of antigen added, the
concentration of the unknown antigen can be determined.

 Severalradionuclides such as 14C, 3H, 131I, 32P, 75Se,

59Fe, 99Mo, and 57Co have been used for RIA.

10/20/2023 Radiochemical Methods of Analysis 44

ANALYSIS OF
RADIOCHEMICALS…
 The RIA technique is applied in assays of hormones,
steroids, peptides, aminoglycosides such as tobramycin
and gentamycin, insulin, many immunoglobulins,
different types of viral heptitis, plasma catecholamines,
angiotension-converting enzymes, many vitamins
including vitamine B12, human growth hormones,
many folate derivatives, and others.

 Many commercially available RIA kits, unique to each kit,

contain series of standards with known concentrations of
unlabeled antigen, a vial of suitable labeled antigen, a vial
of antibody solution, and appropriate precipitants or other
analytical aides.
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Thank you

10/20/2023 Radiochemical Methods of Analysis 46