4/28/2024 Radiochemical Methods of Analysis 1

RADIOCHEMICAL METHODS

OF ANALYSIS
Introduction…
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• Radioactive materials used as pharmaceuticals include

Small organic and inorganic molecules and

Macromolecules such as monoclonal antibodies and antibody

fragments that are attached to radioactive metals.

• In nuclear medicine, drugs containing radioactive metals,

metal complexes, and metal conjugates are used for

diagnosis and therapy of various diseases.
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The successful use of radiochemicals needs a basic

understanding of
Radioactivity

Radioisotopes

Radiation

Radioactive decay

The nature and characteristics of instruments to detect and

quantitative radiation, and

Other related terms

Common Radioactive Emissions Penetrability
Alpha particles may be completely

stopped by a sheet of paper,

Beta particles by aluminum

shielding.

Gamma rays, however, can only

be reduced by much more

substantial obstacles, such as a
very thick piece of lead.
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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 (Z) and

neutral particles called neutrons (N).

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• 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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• 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.
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• 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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• This radioactive process can be spontaneous that 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 re-adjusts to a stable ratio of N/Z with the
release of energy and particles of matter.
All the elements heavier than Bismuth (at N=126 and Z=83)
Radioactivity
• Radioactive decay is the process in which an unstable atomic

nucleus loses energy by emitting radiation in the form of particles

or electromagnetic waves.

• There are numerous types of radioactive decay; The general idea:

• An unstable nucleus releases energy to become more stable

 Radioactive Decay
• A neutron is very unstable; Alone neutron will spontaneously decay into a
proton + an electron.
• If you have a lot of neutrons, within 11 minutes ½ of them will have
decayed
• Particles that decay are said to be radioactive
• Alone neutron is radioactive
• 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
Radioactive Decay………..
Detection of Radioactivity and Concept of Half-Life

• Geiger-Muller counter – instrument which measures radioactive

decay by registering the ions and electrons produced as a radioactive
particle passes through a gas-filled chamber
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Radiation Detection Instruments

There are three types of detectors that are most commonly used, depending on

the specific needs of the device.

These are:

 Gas-Filled Detectors,

 Scintillators, and

 Solid State detectors

Each has various strengths and weaknesses that recommend them to their own

specific roles.
 Radioactive Decay………..
• Scintillation counter
instrument which
measures the rate of
radioactive decay by
sensing flashes of light
that the radiation
produces in the detector
Half-life
 Time required for half of the
original sample of radioactive
nuclides to decay
 Half of the radioactive parent atoms decay after one half-life.
 Half of the remainder decay after another half-life and so on……..
Carbon Dating
 The radioactivity of once living things decreases at a predictable
rate
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• 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.

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• 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

half-life.
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• The half-life of a radionuclide is the time required for the

sample activity to decrease to half its initial value

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• Other commonly used terms in nuclear medicine and

pharmacy are

• Average (mean) half-life,

• Biological half-life, and

• Effective half-life
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A. Average half-life is the mean life-time of a nuclide, and it is

equal to 1.44 x t1/2

B. Biologic half-life ( tb) is the time required for the body to

eliminate half the administered dose by normal biological
process of elimination.
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C. Effective half-life (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

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Decay of Radionuclide…
I. 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
Z X Y  He
Z 2
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…

II. 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
Z 1Y  β  -

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

charge, and it is primarily needed to conserve energy in the
decay
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Decay of Radionuclide…
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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
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Decay of Radionuclide…

III. 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.
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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…
IV. 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

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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…..

• 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

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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.
• 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.
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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
• It 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.
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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 follow from the decaying
radionuclide.
• These are quantitated by using appropriate semiconductor
radiation detectors.
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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 24Na decays to 24Mg, the gamma-Rays

released have unique energies of 1.268 and 2.754MeV.

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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 the table below.
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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.

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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.
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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.
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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.
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Analysis Of Radiochemicals…..
Several radionuclides 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, and others.