Radiochemical Methods

7.
Radiochemical methods of
Analysis
By Sisay K.
07/09/2024 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.
 Radioactive materials used as pharmaceuticals include
◦ small organic and inorganic molecules and
◦ macromolecules such as monoclonal antibodies and

antibody fragments that are attached to radio-active
metals. 07/09/2024 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

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.
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.

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

nuclide
 There are three nuclides of the element hydrogen with

atomic number one
 The three nuclides of hydrogen are called isotopes of

hydrogen
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

Introduction…

Introduction…
 This radioactive process can be spontaneous --said to be
naturally radioactive elements.
 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
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

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.

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.
 Thisrate constant is conveniently denoted in terms of t1/2, or

halflife.

Introduction…
 The
halflife of a radionuclide is the time required for the
sample activity to decrease to half its initial value

Introduction…
A sample of material contains 1 mg of iodine- 131.This
isotope is used in small dose in thyroid cancers therapies, it
is also a dangerous product of nuclear fission if released in to
the environment. Iodine- 131 have half-life of 8.02 days.
1. The number of atoms of iodine-131 initially present.
2. Number of iodine-131 atoms present after 50 days.

Introduction…
 Other commonly used terms in nuclear medicine and
pharmacy are
 Average (mean) halflife,
 Biological halflife, and
 Effective halflife

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.

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:

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.07/09/2024 Radiochemical Methods of Analysis 18

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
Z X  AZ42Y  42 He2

 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

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
 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
Tc = Technetium, Mo = Molybdenum

 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

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.

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

A
Z X Y  β  
A
Z-1


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

 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

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.
Analysis of Radiochemicals…
 It is 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

 Radionuclidesare identified based on
◦ The characteristic decay and
◦ The energy of the particles
 Radiochemical methods of analysis include

◦ Tracer methods
◦ Activation analysis and
◦ Radioimmunoassay techniques.

A. Tracers and Tracer Methods of Analysis
 Radiochemical tracers or radiotracers are compounds

labeled with radioisotopes.
 For tracer 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

 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

B. Activation analysis
 is a 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

Rnalysis 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.
 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.

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 the Table
below.
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

 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
 The reaction in an RIA mixture can be described as
follows

 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
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.
ANALYSIS OF RADIOCHEMICALS…
 Severalradionuclides such as 14C, 3H, 131I, 32P, 75Se,
59Fe, 99Mo, and 57Co have been used for RIA.
The RIA technique is applied in assays of

◦ hormones
◦ 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,07/09/2024
and others.
Radiochemical Methods of Analysis 46
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7.

07/09/2024 Radiochemical Methods of Analysis 1

 In nuclear medicine, drugs containing radioactive metals,

 Radioactive materials used as pharmaceuticals include

◦ small organic and inorganic molecules and

◦ macromolecules such as monoclonal antibodies and

 The successful use of radiochemicals needs a basic

◦ The nature and characteristics of instruments to

07/09/2024 Radiochemical Methods of Analysis 3

 Bohr’s planetary model of the atom consists of a dense

 The nucleus consists of positively charged protons and

 The electroneutrality of the atom is maintained by the

 This number is called the atomic number, Z.

 Any conﬁguration of protons and neutrons is called a

 There are three nuclides of the element hydrogen with

 The three nuclides of hydrogen are called isotopes of

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

 When the N/Z ratio becomes higher, the nucleus become

07/09/2024 Radiochemical Methods of Analysis 7

07/09/2024 Radiochemical Methods of Analysis 8

 When such instability is brought about by bombarding stable

 The rate of this decay or activity is

07/09/2024 Radiochemical Methods of Analysis 10

 The decay constant is a measure of the probability that a

07/09/2024 Radiochemical Methods of Analysis 11

 Thisrate constant is conveniently denoted in terms of t1/2, or

07/09/2024 Radiochemical Methods of Analysis 12

07/09/2024 Radiochemical Methods of Analysis 13

isotope is used in small dose in thyroid cancers therapies, it

is also a dangerous product of nuclear fission if released in to

the environment. Iodine- 131 have half-life of 8.02 days.

1. The number of atoms of iodine-131 initially present.

2. Number of iodine-131 atoms present after 50 days.

07/09/2024 Radiochemical Methods of Analysis 14

07/09/2024 Radiochemical Methods of Analysis 15

Biologic halﬂife, (tb)

07/09/2024 Radiochemical Methods of Analysis 16

Effective halﬂife (teff)

07/09/2024 Radiochemical Methods of Analysis 17

 Radiation resulting from any decay process can be

◦ Beta minus (β-),

◦ Positron (β+) and

◦ Gamma (γ) radiation.07/09/2024 Radiochemical Methods of Analysis 18

07/09/2024 Radiochemical Methods of Analysis 19

 The  particles are monoenergetic, and their range in

07/09/2024 Radiochemical Methods of Analysis 20

Beta-Minus (Negatron) Decay

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

 After - decay, the atomic number of the daughter

07/09/2024 Radiochemical Methods of Analysis 23

 Beta emitters represent different energy levels, and have

 Beta emitting radionuclides are used in

07/09/2024 Radiochemical Methods of Analysis 24

07/09/2024 Radiochemical Methods of Analysis 25

07/09/2024 Radiochemical Methods of Analysis 26

07/09/2024 Radiochemical Methods of Analysis 27

07/09/2024 Radiochemical Methods of Analysis 28

07/09/2024 Radiochemical Methods of Analysis 29

 Most spectral methods can quantitate at the parts-per-million