Isotope Dilution Analysis

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 1

 Isotope Dilution Analysis
Isotope Dilution Analysis

There are certain analytical methods that can be applied to all fields of tracer use.
Foremost among these isotope dilution analysis(IDA). The method of isotope dilution
comprises the addition of known amounts of isotopically-enriched substance to the
analyzed sample. Mixing of the isotopic standard with the sample effectively "dilutes" the
isotopic enrichment of the standard and this forms the basis for the isotope dilution
method.

A known quantity of a compound containing an unknown quantity of a particular element

is mixed with a spike (a known weight of a radioactive isotope of the element). The
specific activity (disintegrations per second per kilogram) of the spike is known precisely,
so the isotopic composition of the mixture can be used to calculate the amount of the
element in the sample. A small amount of the mixture is isolated from the sample,
weighed, and its specific activity measured. The concentration of the inactive element in
the sample may be estimated by the dilution of the radiotracer. Isotope dilution analysis
can be applied to all elements that have two or more naturally occurring isotopes (about
80% of all elements), provided that a spike enriched in one of the isotopes of that element
is available.

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 2

 Isotope Dilution Analysis
Direct IDA
In direct IDA, we are faced with the problem of determining the amount of some
inactive material A in a system. Let us define this unknown amount as x grams. To
the system containing x grams of inactive A, we add y grams of active material A*
Of known activity D. Thus, we know the specific activity of the added active material,
S1. That is,

After thoroughly mixing the active material A* with the inactive A in the system,
one isolates, not necessarily quantitatively, and purifies a sample of the mixture of A
and A* and measures its specific activity, S2. Clearly, conservation of material says that

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 3

 Isotope Dilution Analysis
This is the basic equation of direct isotope dilution analysis. The unknown amount x
of material A is given in terms of the amount y of added labeled material A* and the
two measured specific activities S1and S2.

Example Problem: Let us consider a practical problem to illustrate the use of

this technique. A protein hydrolysate is to be assayed for aspartic acid. Exactly
5.0 mg of aspartic acid, having a specific activity of 0.46 µCi/mg, is added to
he hydrolysate. From the hydrolysate, 0.21 mg of highly purified aspartic acid,
having a specific activity of 0.01 µCi/mg, can be isolated. How much aspartic
acid was present in the original hydrolysate?

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 4

 Isotope Dilution Analysis
Thus, by isolating a small fraction of the added aspartic acid and measuring the
diminution in its specific activity, the aspartic acid content of the original sample can
be determined. Note this example involved a large change in specific activity upon
dilution. Poor experimental design or other circumstances may lead to a small
change in specific activity upon dilution. In such cases, the results obtained from
IDA involve a small difference between two large numbers and are quite uncertain.

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 5

 Isotope Dilution Analysis
Inverse IDA
Inverse IDA is a simple variant on the basic direct IDA. In inverse IDA, we measure
the change in specific activity of an unknown radioactive material A* after diluting it
with inactive A. Specifically, let us assume that we have q milligrams (where q is
unknown) of a radioactive substance A* whose specific activity is known (i.e., Sq =D/q). (Sq
can be measured by isolating a small portion of A*, weighing it, and measuring its activity.)
Let us add r milligrams of inactive A to A* and thoroughly mix the A and A*. Suppose that
we then isolate and purify some of the mixture and measure its specific activity Sr. Note
that Sr =D/(q + r). And so, we have

The above equation is the basic equation of inverse isotope dilution analysis and
indicates that the unknown amount q of active material A* can be deduced by adding r
grams of inactive material A to A* and measuring the specific activities before and after
the addition Sq and Sr, respectively.
A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 6
 Isotope Dilution Analysis
Double IDA
Another variant on the basic isotope dilution technique is that of double isotope
dilution, first proposed by Block and Anker (1948). It is used in reverse IDA where the
specific activity of the original unknown radioactive
material A* cannot be measured for some reason. Hence a second dilution is
made to determine the specific activity of the original sample. Consider a system
containing an unknown amount q of some active substance A* whose specific activity A*
cannot be measured. Take two equal aliquots of this unknown substance A*. Add r
milligrams of inactive A to one aliquot and p milligrams of inactive A to the other aliquot.
Measure the specific activities of the two aliquots, Sr andSp, respectively. For the first
sample of specific activity Sr, we have
A major difficulty
with double isotope
dilution analysis is
that, because of the
double dilution, the
specific activities
involved become low
and therefore more
uncertain.

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 7

 Isotope Dilution Analysis
Common Notes
First, reagents and tracers of high purity are necessary. Two techniques are usually
preferred for the separation: precipitation and solvent
extraction. As a purification step, precipitation has the advantage that the precipitate
can easily be weighed at the time of separation, thereby allowing a quick determination of
the specific activity. Precipitation techniques are used for the isolation of inorganic
components.
Solvent extraction is a frequently employed technique in isotope dilution analysis. It gives
very clean separations, resulting in high-purity samples. It has the disadvantage of
requiring further chemical processing to determine the mass of material isolated and the
specific activity.
Second, is incomplete isotopic exchange, in which the active and inactive atoms do not
mix. This lack of exchange can be due to differing physical and chemical states of tracer
and inactive materials. Steps must be taken to ensure complete exchange.
One must also be sure that the labeled position in any
compound is relatively inert. If the atom in question is very labile, one can get a
reduction in specific activity without any dilution having taken place. To compare
specific activities, all samples must be counted under identical conditions with
proper corrections for self-absorption in samples of varying mass.

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 8

 Isotope Dilution Analysis
Advantages
Isotope dilution analysis is a highly sensitive, selective analytical method capable of high
precision. It offers the opportunity to determine the amount of material present in a
system without the need for a quantitative separation of the material from the system.
Partial loss of analyte during preparation is compensated for since physical and chemical
interferences are not an issue and will cancel out as they will affect each isotope
identically. Isotope dilution is classified as a method of internal standardisation, because
the standard (isotopically-enriched form of analyte) is added directly to the sample. In
addition, unlike traditional analytical methods which rely on signal intensity, isotope
dilution employs signal ratios. It is free of interference from other elements present and its
accuracy is governed by the calibration of the spike solution.

Disadvantages
– Generally only applicable to multiple-isotopic elements
– Need an enriched isotope spike for the analyte of interest - not always available or
sometimes at very high cost
– Need two interference free isotopes
– VERY time consuming

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 9

 Neutron Activation Analysis
ACTIVATION ANALYSIS
Activation analysis is an analytical technique that allows one to determine the
amount of a given element X contained in some material Y. The basic steps in
the activation technique are as follows:

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 10

 Isotope Dilution Analysis
activation analysis. [From Corliss
Schematic representation of

(1963).]

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 11

 Neutron Activation Analysis
Neutron activation analysis, discovered in 1936, stands at the forefront of techniques
used for quantitative multi-element analysis of major, minor, trace, and rare elements.
NAA allows the measurement of ~60 elements in small samples. The lower limit of
detection is of the order of parts per million to parts per billion depending on the analyzed
element and the activity of the bulk sample matrix.
The principle involved in neutron activation analysis consists of first irradiating a sample
with neutrons to produce specific radionuclides. After the irradiation, the characteristic
gamma rays emitted by the decaying radionuclides are quantitatively measured using
gamma spectroscopy, where the gamma rays detected at a particular energy are indicative
of a specific radionuclide's presence. Data analysis then yields the concentrations of
various elements in the samples being studied.
The following table provides a list of elements that may be quantitatively analyzed using
neutron activation analysis:
Sensitivity (ppb) Element
5 Dy, Eu
5-50 In, Lu, Mn
50-500 Au, Ho, Ir, Re, Sm, W
Ag, Ar, As, Br, Cl, Co, Cs, Cu, Er, Ga, Hf, I, La, Sb, Sc, Se, Ta, Tb, Th,
500-5000
Tm, U, V, Yb
Al, Ba, Cd, Ce, Cr, Hg, Kr, Gd, Ge, Mo, Na, Nd, Ni, Os, Pd, Rb, Rh, Ru,
5000-5x104
Sr, Te, Zn, Zr
5x104-5x105 Bi, Ca, K, Mg, P, Pt, Si, Sn, Ti, Tl, Xe, Y
5x105-5x106 F, Fe, Nb, Ne

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 12

 Neutron Activation Analysis

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 13

 Neutron Activation Analysis
The measured count rate (R) of the gamma rays from the decay of a specific
isotope in the irradiated sample can be related to the amount (n) of the original, stable
in the sample through the following equation

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 14

 Neutron Activation Analysis

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 15

 Neutron Activation Analysis
Applications of NAA
NAA can be used for a large variety of applications; the following represent some
examples for scientific uses in different disciplines:
• Archeology- Sourcing of clays and pottery
• Biology- Toxins in Fish and agricultural products,- Trace elements in oil and lipids,
• Che istry- Contaminants in salts, pure crystals and metals
• E gi eeri g- Composition and contaminants in metals, thin film deposits plastics
• Fore sics- Analysis of bullets, and other crime scene materials (paint, glass, metals)
• Geology- Sourcing and composition of igneous rocks, sediments and basalts
• Medici e- Toxins and trace elements in hair, skin and nail samples

A. K. M. Nur Alam Siddiki, Department of Chemistry, University of Dhaka.(www.nuralam.net/nuclear) 16