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Preparation of clinical-scale 99Mo/99mTc

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specific activity 99Mo and nanocr....

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DOI: 10.1016/j.nucmedbio.2012.03.010 · Source: PubMed

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Contents lists available at SciVerse ScienceDirect

Nuclear Medicine and Biology

j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / n u c m e d b i o

Preparation of clinical-scale 99Mo/ 99mTc column generator using neutron activated

low specific activity 99Mo and nanocrystalline γ-Al2O3 as column matrix
Rubel Chakravarty, Ramu Ram, Ashutosh Dash ⁎, M.R.A. Pillai
Radiopharmaceuticals Division, Bhabha Atomic Research Centre, Mumbai-400085, India

a r t i c l e i n f o a b s t r a c t

Article history: Introduction: Preparation of clinical-scale 99Mo/99mTc generator using (n,γ) activated low specific activity 99Mo
Received 13 February 2012 and nanocrystalline γ-Al2O3 as a high capacity sorbent matrix is attempted.
Received in reviesd form 19 March 2012 Methods: Nanocrystalline γ-Al2O3 was synthesized by ‘solid state mechanochemical’ reaction of aluminum
Accepted 26 March 2012 nitrate with ammonium bicarbonate. Experimental parameters were optimized to effectively separate 99mTc
from 99Mo using this sorbent as the column matrix. The performance features of a 13 GBq (350 mCi) 99Mo/99mTc
Keywords:
generator using this sorbent and 99Mo produced by (n,γ) route having specific activity 12.9–18.5 GBq/g were
γ-Al2O3
Nanocrystalline
evaluated for 10 days.
99
Mo/99mTc generator Results: The sorbent possessed the requisite selectivity for 99Mo and demonstrated a maximum sorption capacity
Low specific activity 99Mo (LSA 99
Mo) of 200 ± 5 mg Mo/g, which is ~10 times higher than that of ordinary acidic alumina. The overall yield of 99mTc
Elution was N 80%, with radionuclidic purity N 99.99% and radiochemical purity N99%. The yield of 99mTc varied from 7.8
to 2.1 GBq in the eluate for the six days of operation of the generator. The radioactive concentration of 99mTc
eluted was adequate for the formulation of radiopharmaceuticals. The performance of the generator remained
consistent over an extended period of 10 days. The eluted 99mTc was suitable for the formulation of 99mTc-DMSA
and 99mTc-EC resulting in high radiolabeling yields (N 98%).
Conclusion: The effectiveness of γ-Al2O3 as a new generation sorbent in the development of clinically useful 99Mo/
99m
Tc generator using low specific activity 99Mo and yielding 99mTc with adequate radioactive concentration and
high purity suitable for formulation of radiopharmaceuticals is demonstrated.
© 2012 Elsevier Inc. All rights reserved.

1. Introduction abundant availability of clinical grade 99mTc [5–10]. Hence, several

Considering the immense diagnostic values offered by the
different technetium-99m ( 99mTc) tracers for the last fifty years,
this radioisotope will continue to be the workhorse of diagnostic
nuclear medicine in the foreseeable future [1]. Hence, there is a need
to ensure the reliable supply of 99mTc to support the diagnostic
imaging that is being done the world over. The world wide application
of 99mTc in nuclear medicine was possible due to the availability of
highly efficient 99Mo/ 99mTc column generators [2,3]. These column
chromatographic generators used a small bed of acidic alumina to
reliably hold the 99MoO42 − ions and allowed 99mTcO4− to be eluted in a
pure form in high radioactive concentration (N0.74 GBq (20 mCi)/
mL). As 99Mo used in the preparation of the column generators was of
fission origin having a specific activity of ~370 TBq (10 4 Ci)/g, the low
capacity of alumina (2–20 mg Mo/g) [4] for taking up molybdate ions
posed no problem for the preparation of clinically useful generators.
The disruption of the availability of the fission 99Mo in the last few
years has posed serious challenge to the comfortable situation of
⁎ Corresponding author. Fax: + 91 22 2550 5151.
E-mail address: adash@barc.gov.in (A. Dash).
alternative approaches for production of 99Mo [11], 99mTc [12] or
replacement of 99mTc with PET tracers [9,13] are seriously being
explored. However, for very strange reasons the utilization of the
more easily producible (n,γ) activated 99Mo is overlooked [14]. A
pertinent point that is ignored is that irrespective of the specific
activity of 99Mo used in the generator, 99mTc produced is no-carrier-
added (NCA) yielding 99mTc formulation suitable for clinical use.
There are over 75 operating research reactors in the world capable of
making low specific activity 99Mo (LSA 99Mo) [15] and hence there is
an urgent need to devise means to enhance the use of 99Mo produced
by (n,γ) route.
Numerous routes for the development of 99Mo/ 99mTc generators
using (n,γ) 99Mo have been investigated over the years [16]. However,
there is an overwhelming preference for column generator among the
users as it is the most simple and elegant method to get clinically
useful 99mTcO4− with minimum manipulation. Hence, there is need to
have high sorption capacity sorbents compatible for making column
generators using LSA 99Mo. In this context, the polymeric zirconium
compound (PZC) reported by Tanase et al. having high sorption
capacity (200 mg g − 1) was an important milestone [17]. On a similar
theme, alumina functionalized with a sulfate moiety synthesized by
an intricate sol-gel synthesis route, having high 99Mo capacity

0969-8051/$ – see front matter © 2012 Elsevier Inc. All rights reserved.
doi:10.1016/j.nucmedbio.2012.03.010
 R. Chakravarty et al. / Nuclear Medicine and Biology 39 (2012) 916–922
(217 mg/g) has also been tried [18]. Clinically useful generators are
yet to be deployed using the above technologies. Hence, the
development of high capacity sorbents offering improved properties
for the preparation of 99mMo/ 99mTc column generators remains an
interesting goal.
Nanocrystalline metal oxides possess large surface areas, defect-
rich morphology, large porosities, and small crystallite sizes that
collectively result in extremely high reactivity and enhanced sorption
capacities [19,20]. Hence, the use of nanomaterial based sorbents as
column matrices for the preparation of chromatographic 99Mo/ 99mTc
generator is a promising approach [21,22]. In this paper, we report the
utilization of nanocrystalline γ-Al2O3 as a column material in the
preparation of 99Mo/ 99mTc generators. Synthesis of the nanomaterial,
assessment of its sorption properties and preparation of a 99Mo/ 99mTc
generator having loading up to 13 GBq (350 mCi) of 99Mo are
reported. Suitability of the eluted 99mTcO4− for radiolabeling was
assessed by formulating lyophilized kits of dimercaptosuccinic acid
(DMSA) and N,N-ethylene-L,L-dicysteine (EC) with eluted 99mTc.
2. Materials
Reagents such as hydrochloric acid, ammonium hydroxide,
ammonium carbonate etc. were of analytical grade and were procured
from S. D. Fine Chemicals, India. Aluminum nitrate (+ 99.9%) was
purchased from BDH Chemicals, Mumbai, India. HPLC grade water
was purchased from E. Merck, Germany. Paper chromatography (PC)
strips (3MM Chr, 20 mm width) were purchased from Whatman
International Limited, England. Lyophilized kits of DMSA and EC
were obtained from the Board of Radiation and Isotope Technology
(BRIT), India.
An HPGe detector (Canberra Eurisys, France) coupled to a multi-
channel analyzer was used for analysis of 99mTc in the presence
of 99Mo. The chemical analysis for the trace level of Al 3 + ions was
done using Inductively Coupled Plasma-Atomic Emission Spectrosco-
py (ICP-ES JY-238, Emission Horiba Group, France).
3. Experimental
3.1. Synthesis of γ-Al2O3
γ-Al2O3 was synthesized by solid state mechanochemical reaction
of aluminum nitrate and ammonium carbonate, as reported by us
earlier [20]. In brief, the procedure involved mixing of aluminum
nitrate and ammonium bicarbonate in an agate mortar and grinding
for 30 min at room temperature. The ground mixture was dried in
an oven at 100 °C for 5 h, followed by calcination at 700 °C for 2 h.
The γ-Al2O3 thus formed was washed with deionized water to remove
the traces of unreacted water soluble precursors and the granular
material was dried under an infrared lamp.
The material was synthesized in several (N10) batches and
the structural characterization of the nanomaterial synthesized in
each batch was carried out adopting the procedure reported by us
earlier [20].
3.2. Chemical stability of γ-Al2O3
The chemical stability of γ-Al2O3 was assessed in mineral acids
such as HCl, HNO3, H2SO4 and NaOH up to concentrations of 2 M. 1 g of
the dry sorbent material was placed in 50 mL solvent of interest and
kept for 24 h with continuous shaking at room temperature. The
nanomaterial was filtered and the level of Al metal ions released in the
solutions was determined by Inductively Coupled Plasma-Atomic
Emission Spectroscopy (ICP-AES). The chemical stability of γ-Al2O3
under continuous column-flow conditions was determined by passing
100 mL solutions of different mineral acids and bases through a
917
chromatographic column packed with 2 g of the nanomaterial. The
level of Al 3 + ions in the effluent was determined by ICP-AES analysis.
3.3. Production of 99
Mo
The target material, natural MoO3 (15 g) in the form of powder
was packed in an Al capsule, sealed by cold-welding method, checked
for leakage by quality control procedures prescribed by the Atomic
Energy Regulatory Board of India and neutron irradiated at a position
having neutron flux of ~ 9.6 × 10 13 n cm − 2 s − 1 in the Dhruva reactor
for 1 week. After irradiation, the target was dissolved in 50 mL of 2 M
NaOH solution at 80 °C. The resultant solution was evaporated to near
dryness and then reconstituted with 50 mL of 0.1 N NaOH solution.
The activity of the 99Mo solution was assayed in an ion-chamber.
3.4. Sorption properties of γ-Al2O3
The sorption characteristics of γ-Al2O3 were studied adopting
standard radiometric techniques as reported earlier [22]. The
distribution coefficients (Kd) of MoO42 − and TcO4− ions were
determined by batch process at different pH, using 99Mo and 99mTc
radiotracers and the results are reported as averaged values of
triplicate tests. In order to determine the time required to attain
sorption equilibrium, the Kd values of MoO42 − ions were studied as a
function of time. The static sorption capacity of γ-Al2O3 was
determined by batch equilibration experiments using MoO42 −
solutions of known concentration (10 mg Mo mL − 1 ) spiked
with 99Mo radiotracer. The dynamic sorption capacity and break-
through pattern were studied by passing sodium molybdate solution
(25 mg Mo mL − 1) at pH 3, spiked with 99Mo radiotracer through
borosilicate glass columns of dimension 6 cm × 1 cm (i.d.) with
sintered disc (G0) at the bottom containing 5 g of γ-Al2O3 at a flow
rate of 2 mL min − 1. From the feed 99Mo solution (25 mg Mo mL − 1),
2 mL was kept as reference and the eluted solution was collected in
2 mL aliquots. The radioactivity associated with the reference solution
(Ao), as well as each fraction (Ai) was measured in an HPGe detector
by counting the 181 keV peak of 99Mo. The sorption capacity of γ-
Al2O3 for Mo under dynamic conditions was estimated by the study of
the breakthrough profile determined using these data. In order to
determine the practical sorption capacity of γ-Al2O3, 50 mL of sodium
molybdate solution (25 mg Mo mL − 1) at pH 3, spiked with 99Mo
radiotracer was passed through the column containing 5 g of γ-Al2O3
at a flow rate of 2 mL min − 1. Subsequently, the column was washed
with 200 mL of 0.9% solution and the activities of the effluent and the
washing were measured. The sorption properties of γ-Al2O3 synthe-
sized in different batches were studied.
3.5. Development of 99
Mo/ 99mTc generator
The schematic diagram of the 99Mo/ 99mTc generator is shown in
Fig. 1. All the operations were carried out in the closed cycle system
using connecting tubes. The column matrix was pre-conditioned at pH
3 by passing 0.001 N HNO3 solution. A borosilicate glass column of
dimension 6 cm × 1 cm (i.d.) with a sintered disc (G2) at the bottom
was packed with 5.5 g of γ-Al2O3 and kept in a lead shielded
container. A small piece of glass wool was placed on top of the sorbent
bed to prevent disturbance of the bed during passage of eluents.
Input/output connections were made with standard teflon tubings of
1 mm inner diameter and connectors. The generator column,
connectors and connection tubings were integrated within a small
portable lead shielded unit throughout experimental use for radio-
protection purpose. Only the feed and output vials were accessible
externally. A disposable 0.22 μm membrane filter was attached to the
generator column output by teflon tubing. The column was then
loaded with 13 GBq (350 mCi) of 99Mo solution maintained at pH 3.
The loaded column was washed with 500 mL of 0.9% NaCl solution.
918 R. Chakravarty et al. / Nuclear Medicine and Biology 39 (2012) 916–922

for column operation. The mechanochemical process adopted for the

preparation of nanomaterial is facile, fast and can be conveniently
adapted for the preparation of adequate quantities of nanosorbents
needed for the preparation of column generators.
The structural characterization studies adopting the reported
procedure revealed that the material is nanocrystalline in γ-phase,
consisting of agglomerated particles (median agglomerate size
~ 23 μm) made of primary particles of size about 5–6 nm and
possessing surface area of 252 ± 10 m 2/g [20]. The synthesis yield
and the characteristics of the nanomaterial remained steady in all the
batches, thereby demonstrating the reproducibility of the synthetic
pathway in the generation of a consistent γ-Al2O3 matrix material.

4.2. Chemical stability of γ-Al2O3

The chemical stability studies by solubility tests showed that

γ-Al2O3 was insoluble in mineral acids and alkalis up to 2 M as only
negligible amount of Al ions (b0.1 ppm) were detected in the filtrate
when analyzed by ICP-AES. This is well below the permissible limit
(b10 ppm) of Al 3 + ions in 99mTc [23]. From the perspective of
radionuclide generator applications, this is significant for two reasons.
First, it indicates that the sorbent packed in a column could be used for
repeated elution for an extended period of time without chemical
degradation. Secondly, it ensures that the use of this material does
99
Fig. 1. Schematic diagram of the Mo/99mTc generator assembly.
not lead to addition of chemical impurities in the eluate.

99
4.3. Production of Mo
Subsequently, after allowing adequate time (24 h) for 99mTc build up,
the column was eluted with 0.9% NaCl solution at a flow rate of
Typically in one batch, ~ 177.6 GBq (4.8 Ci) of 99Mo was produced
2 mL min − 1. In order to examine the elution profile, the eluate was
when 15 g of natural MoO3 was irradiated at a flux of 9.6 × 10 13
collected as 2 mL aliquots and each fraction was counted for gamma
n cm − 2 s − 1. The specific activity of 99Mo produced was ~17.6 GBq
activity. The performance of the generator was evaluated for a period
(475 mCi)/g of Mo at the end of radiochemical processing.
10 days, by periodic elution at 24 h intervals.

3.6. Quality control of 99m

Tc eluate 4.4. Determination of the distribution ratio (Kd) values of MoO42 −
and TcO4− ions
The evaluation of the radionuclidic purity, radiochemical purity
and labeling efficacy of 99mTc was carried out adopting the procedures The feasibility of separation of 99mTc from 99Mo was ascertained by
as described earlier [22]. The presence of trace levels of Al 3 + ions as examination of the data on distribution ratios. Determination of
chemical impurities in 99mTc eluate was estimated by the analysis of distribution ratio of 99Mo and 99mTc ions as a function of pH was
the decayed 99mTc samples using Inductively Coupled Plasma-Atomic studied to understand the affinity of the sorbent for these ions and the
Emission Spectroscopy (ICP-AES). results are summarized in Table 1. The results indicate that γ-Al2O3
has high affinity for MoO42 − ions and negligible affinity for TcO4− ions
across all pH ranges studied. It is apparent that the Kd values for both
4. Results
the ions attained the maximum value at around pH 3 and hence this
pH is appropriate for sorption of 99Mo. Looking at the distribution
4.1. Synthesis of γ-Al2O3
ratio (Kd) in 0.9% NaCl, it is clear that 99mTc has negligible Kd value
(0.2 ± 0.1) whereas 99Mo has very high Kd value (12276 ± 521). The
A mechanochemical approach was used for the synthesis of γ-
observed Kd values ensures that 99mTc can effectively be eluted using
Al2O3. In this method the reactants i.e. aluminum nitrate and
0.9% NaCl solution with no contamination of 99Mo.
ammonium carbonate, were taken in stoichiometric ratio and were
thoroughly ground. During grinding, a self-initiated and self-
sustained reaction starts with the evolution of CO2 gas and forms
Table 1
a precursor, which was dried at 100 °C for 5 h and then subjected Distribution ratios (Kd) of 99
MoO42 − and 99m
TcO4− in γ-Al2O3.
to calcination at 700 °C for 2 h. The particle size in the reaction
Medium (pH) Kd
depends on the rates of nucleation and growth of the product.
Nanoparticles are formed when the growth of product clusters stops 99
MoO42 − 99m
TcO4−
or the nucleation rate is faster than the growth of product clusters; 1 12129 ± 578 3±2
otherwise, bulk materials will be produced. The present solution-free 2 22295 ± 718 14 ± 3
mechanochemical reaction by grinding is maintained at the nucle- 3 26611 ± 228 35 ± 5
4 25960 ± 778 29 ± 4
ation stage of the product because the reaction occurs only at the
5 218525 ± 594 2.7 ± 0.8
contacting surface of solid reactant particles and the amount of 6 11174 ± 818 0.3 ± 0.1
reactants involved in the reaction is far smaller than in traditional 7 10874 ± 628 0.1 ± 0.1
solution synthesis. The evolution of gases during calcination contrib- 8 1890 ± 199 0.6 ± 0.2
uted to increase in the porosity in the material. The material exhibited 0.9% NaCl 12276 ± 521 0.2 ± 0.1

good mechanical strength, granular properties and was amenable n = 3, ‘±’ indicates standard deviation.
 R. Chakravarty et al. / Nuclear Medicine and Biology 39 (2012) 916–922 919

30000 Amount of sorbent = 5 g

1.0 Breakthrough capacity
(Ai /Ao ~0.01) = 160 mg Mo/g
25000
0.8
20000

0.6

Ai /Ao
15000
Kd

0.4
10000

0.2
5000

0.0
0 0 10 20 30 40 50 60 70 80
0 5 10 15 20 25 30 Volume of Mo solution (25 mg Mo/mL) passed (mL)
Time (min)
Fig. 3. Breakthrough profile of Mo from the γ-Al2O3 column by passing MoO42 − solution
Fig. 2. Study of the sorption kinetics of 99
MoO42 − on γ-Al2O3. (pH 3, 25 mg Mo/mL) at a flow rate of 2 mL/min.

4.5. Determination of the time of equilibration
The time dependence of 99Mo sorption was studied by following
the distribution ratio (Kd) at different time intervals which were an
indication of the progress of the sorption process. The results of the Kd
values studied as a function of time for 99Mo given in Fig. 2 reveal that
the Kd value increased with increase in contact time, remained almost
constant (~ 25000) after 10 min. It indicates that sorption equilibrium
was reached within 10 min. The pores of the γ-Al2O3 matrix probably
allow the material to have rapid and complete access to the solution
containing 99Mo.
4.6. Determination of sorption capacity of γ-Al2O3
The sorption capacity of γ-Al2O3, indicative of the ability of the
sorbent to retain 99Mo, depends on the availability of sorption sites
within the matrix. The sorption capacities of γ-Al2O3 were deter-
mined both under static and dynamic conditions. The static sorption
capacity for γ-Al2O3 was estimated to be 205 ± 10 mg Mo per g of
sorbent (n = 10). This result clearly illustrates the superiority of this
material over bulk alumina with respect to sorption capacity of 2–
20 mg Mo per g of alumina [4].
In addition to the static sorption capacity, the dynamic sorption
capacity needs to be taken into account when designing a chromato-
graphic column. In order to determine the sorption capacity under
dynamic conditions, the breakthrough profile of 99Mo was studied at
pH ~ 3. A typical breakthrough curve is depicted in Fig. 3 in which a
chromatographic column containing 5 g of γ-Al2O3 was used and the
compared to the bulk alumina, which shows a maximum sorption
capacity of ~ 20 mg Mo/g [4]. This study serves as baseline data and
helped in designing chromatographic column generator.
99
4.7. Development of Mo/ 99mTc generator
In order to translate the laboratory findings into real conditions,
process demonstration runs were carried out using ~ 13 GBq
(350 mCi) of (n,γ) 99Mo. It is the physical, chemical, and radiolytic
stability that determines the suitability of the γ-Al2O3 for long
term application. The practical utility and separation capability
of 99mTc were monitored, by following the elution profile of the
generator which is depicted in Fig. 4. A reasonably sharp elution
profile of 99mTc was observed. The obtained results show the first
2 mL fraction containing negligible amount of 99mTc activity could be
discarded and N80% of 99mTcO4− was obtained within the next 6 mL
volume of normal saline. Thus, 99mTc could be eluted from the
generators with radioactive concentration sufficient for clinical use.
In order to assure the repeated use of the generator, the elution
of 99mTcO4− was carried out after every 24 h.
Table 2 summarizes the results of operation of the 99Mo/ 99mTc
generator in a typical process demonstration run. It was observed that
generator gives consistently high yields (N82% of the theoretical
50
99m
1. Yield of Tc (%)
100
2
99m
2. Cumulative yield of Tc (%)
2. Cumulative yield of 99mTc (%)

solution was passed at a flow rate of 2 mL/min. From the break- 40 80

through curve, it is possible to determine the capacity at various
1. Yield of 99mTc (%)

values of Ai/Ao. It was observed that the breakthrough point was

reached after 156±6 mg of Mo (n = 5) was quantitatively retained by 30 60
1 g of sorbent.
Prior to the preparation of the 99Mo/ 99mTc generator, it was 1
essential to determine the maximum achievable sorption capacity of 20 40
γ-Al2O3 under generator column conditions in order to avoid
breakthrough of 99Mo during elution of 99mTc. After loading the
10 20
γ-Al2O3 column with 99Mo under dynamic conditions, it was washed
with 200 mL of 0.9% NaCl solution, to remove the unadsorbed and
loosely held 99Mo. Examination of the practical sorption capacity of 0 0
γ-Al2O3 under the given set of experimental conditions was 0 2 4 6 8 10
determined to be 148±3 mg Mo/g of sorbent. It was observed that Volume of 0.9% NaCl solution (mL)
under the given set of experimental conditions, the columns with the
γ-Al2O3 sorbent have much higher dynamic sorption capacity as Fig. 4. Elution profile of 99m
TcO4− from 99
Mo/99mTc generator.
920 R. Chakravarty et al. / Nuclear Medicine and Biology 39 (2012) 916–922

Table 2
99
Elution performance of the 12.95 GBq (350 mCi) Mo/99mTc generator over a period of 1 week.

Elution No. Time of Theoretically expected Activity of 99mTc Elution yield (%) 99
Mo impurity
growth (h) activity of 99mTc GBq (mCi) obtained GBq (mCi) in 99mTc (%)

1 22 8.8 (239) 7.8 (212) 89 5 × 10− 3

2 24 6.8 (183) 6.0 (161) 87 2 × 10− 3
3 25 5.3 (142) 4.5 (122) 85 4 × 10− 3
4 24 4.1 (110) 3.6 (96) 87 3 × 10− 3
5 24 3.2 (86) 2.6 (71) 82 4 × 10− 3
6 23 2.5 (67) 2.1 (57) 85 3 × 10− 3

yields) of 99mTc over a period of 1 week. Adopting fractionated elution
approach over a period of 1 week, it was observed that the radioactive
concentration of 99mTc in the fraction containing highest radioactivity
of 99mTc was ~ 1.85 GBq/mL and it reduced to ~ 0.6 GBq/mL towards
the end of the week (Fig. 5). The radioactive concentration of 99mTc
eluted during the entire week was thus adequate for the formulation
of radiopharmaceuticals. It is worthwhile to note that during several
process demonstration runs, no operational problems such as
reduction of flow rate, packed bed compression and channeling in
packed column were observed. Appearance of fines or suspended
solids in the effluent from the packed bed due to potential
degradation of the sorbent was also not observed.
99m
4.8. Quality control of
In order to utilize 99mTc obtained from the generator for medical
applications, the presence of radionuclidic impurities of the parent
radioisotope ( 99Mo) must be within the prescribed limit of b10 − 2%
of 99mTc [23]. 99mTc eluted from the generator was checked in an HPGe
detector to see the peaks due to the γ-rays corresponding to 99Mo
gamma activity. The γ-spectrum did not show the presence of such
peaks, thereby confirming the absence of 99Mo in the eluted 99mTc. The
analysis of the decayed samples of 99mTc by γ-spectroscopic technique
revealed that the level of 99Mo present in the eluate was below 10 − 3%
in all the batches of 99mTc. The radiochemical purity of 99mTc
in pertechnetate form as determined by paper chromatography [22]
was N99% which was within the prescribed limits as per the
pharmacopoeias [23]. The level of Al 3 + ions present as chemical
impurity in the 99mTc eluate was determined to be b0.1 ppm which is
less than the prescribed limit [23]. The negligibly small level of
chemical impurity present in 99mTc would not interfere in the
subsequent radiolabeling procedures.
60
Radioactive concentration of 99mTc (mCi/mL)
In order to examine the suitability of 99mTcO4− for radiolabeling
studies, it was used for the preparation of 99mTc-DMSA and 99mTc-EC
using lyophilized kits and the complexation yields were estimated to
be N98% in both the cases (results not presented).
5. Discussion
The wide availability of alumina based 99Mo/ 99mTc column
generators prepared using fission produced 99Mo on an economical
scale has been the major factor responsible for the growth of SPECT
in the past. Over 95% of the world's supply of fission 99Mo is produced
in only six nuclear reactors in the world and repeated and unex-
pected shutdowns of the above ageing reactors since 2007 have
Tc eluate challenged the above scenario [5–9,14]. Moreover, most of these
reactors use highly enriched uranium (HEU) targets for the produc-
tion of fission 99Mo. HEU used as target for 99Mo production as
well as found in the waste after the isolation of 99Mo, poses serious
proliferation risks as it can be used for nuclear weapons and
other explosive devices. Although the use of low enriched uranium
(LEU) targets have been advocated as a near-term solution to reduce
the security concerns, there are technical, logistical and econo-
mic implications as both the target design and chemical processing
for recovering 99Mo are different when LEU is used. An alternative
to the fission based production is to use 99Mo produced by (n,γ)
route, which is technologically less demanding, inexpensive and the
least intricate for post irradiation processing with negligible
generation of radioactive waste [14,15]. However, the specific
activity of (n,γ) 99Mo (~ 11–37 GBq/g) produced by irradiation of
natural MoO3 target is much lower than that of fission 99Mo (~ 370
TBq/g) [24].
The use of low specific activity (n,γ) 99Mo (LSA 99Mo) for the
preparation of 99mTc is very important in the current scenario as
several more reactors which are currently not in the 99Mo produc-
tion circle can be effectively utilized to enhance the availability

Day 1
Volume of each fraction = 2 mL of 99Mo and 99mTc [14,15]. However, a paradigm shift in the current
Day 2 technologies used for availing 99mTc is essential in order to keep the
50
Day 3 continued supply of clinical grade 99mTc from LSA 99Mo. Owing to
Day 4
the limited sorption capacity of bulk alumina (2-20 mg Mo/g) [4], the
40 conventional column chromatographic approach using a bed
Day 5 of acidic alumina cannot be used for the preparation of clinical-
30 Day 6 scale 99Mo/ 99mTc generators using LSA 99Mo. In order to overcome
the constraint on the specific activity of 99Mo for the preparation
of 99Mo/ 99mTc generators, numerous alternative technologies based
20 on solvent extraction [25], sublimation [26], electrochemical method
[27] and post-elution concentration [28] have been proposed.
10 However, none of these techniques can match the simplicity of a
single column generator operation and they have their inherent
limitations which may often restrict their widespread applicability
0
for clinical use.
0 1 2 3 4 5 6
Though the zirconium molybdate- 99Mo ‘gel-generator’ using
Fraction Number
LSA 99Mo has been advocated as one of the alternatives to the alumina
Fig. 5. Radioactive concentration of 99m
Tc in fractionated elution over a period of based chromatographic 99Mo/99mTc generator using fission 99Mo [29],
1 week. the preparation of the ‘gel’ requires an elaborate complex procedure
 R. Chakravarty et al. / Nuclear Medicine and Biology 39 (2012) 916–922
involving precipitation, filtration, drying, gel fragmentation in hostile
radiation environment necessitating a dedicated facility with sophisti-
cated well-equipped hot cells and trained manpower. Further the need
to obtain 99mTc activity from the generator with adequate radioactive
concentration puts a limit on the minimum specific activity of 99Mo that
can be useful. The developments over the last two decades have shown
that it has not been easy to take full advantage of the early promises and
as a result, this approach has not yet been embraced by major generator
manufacturers. Hence, development of chromatographic generators
using high capacity sorbents to utilize LSA 99Mo is an important option.
In our earlier studies, we demonstrated that nanomaterial based
sorbents due to higher surface area are capable of exhibiting high
sorption capacity coupled with ability to achieve sorption equilibrium
rapidly and hence are suitable matrices for the preparation of column
generators [20–22,30,31]. As acidic alumina is already in use for the
preparation of 99Mo/99mTc generators for more than three decades, we
explored the use of nanocrystalline γ-Al2O3 for the same purpose as it
will be more readily acceptable to the regulators.
The mechanochemical synthesis developed is an effective, facile,
robust, inexpensive and green process that can be easily adapted for
large-scale production of the column matrix. The most striking
feature of nanocrystalline γ-Al2O3 is its high capacity for Mo as
compared with that of bulk alumina. This higher capacity sorption is
pivotal in realizing the use of LSA 99Mo produced in medium flux
research reactors where this technology can find application. Other
inherent benefits are that the rigid, open pore structure of nano γ-
Al2O3 facilitates rapid attainment of sorption equilibrium, imparts
chemical stability and enhances the longevity of the sorbent while
in use.
In devising a radionuclidic generator for medical use, an obvious
choice for eluting reagent is 0.9% NaCl due to its isotonicity with blood.
The decay of 99Mo to 99mTc is not accompanied by any serious
disruption of the chemical bonds. As the molybdate ions start
transforming into pertechnetate ion ( 99mTcO4−), which has only −1
charge, the binding would get weaker and an easy displacement
of 99mTcO4− is expected and it gets eluted easily with normal 0.9%
saline. From the perspective of the end-user, the reported procedure
retains the simple-to-operate qualities of the chromatographic
generator. The 99mTc availed is similar in quality and applicability
from that produced by the traditional alumina based generator
containing fission 99Mo. The radioactive concentration and purity
of 99mTc availed from this generator are adequate for labeling the
lyophilized radiopharmaceuticals kits.
The present procedure described could be a viable option for
accessing 99mTc and have high potential for centers having capability
for production of (n,γ) 99Mo. The capacity of the generator produced
will be governed by the specific activity of 99Mo. Although the specific
activity of 99Mo available with us is ~ 16.7 GBq (450 mCi)/g of Mo due
to the flux position available in our reactor, there are reports of regular
availability of 37–74 GBq (1–2) Ci 99Mo/g of Mo from many other
research reactor centers having higher operational neutron flux for
radioisotope production. In such cases, 99Mo/ 99mTc generator having
capacity N37 GBq (1 Ci) can be easily prepared and these generators
are adequate for most clinical use even assuming decay of part of
the 99Mo in the generator while it is transported from the
manufacturer to the user end.
6. Conclusion
The potential utility of nanocrystalline γ-Al2O3 as a viable sorbent
with significantly high 99Mo retention capacity for the preparation of
clinically useful 99Mo/ 99mTc generator is demonstrated. Present
investigations prove that the solid state mechanochemical approach
is suitable for the preparation of nanocrystalline γ-Al2O3. The rapid
attainment of 99Mo sorption equilibrium, selectivity towards 99Mo
and high capacity are powerful attributes of nano γ-Al2O3 sorbent that
921
are exploited for the preparation of 99Mo/ 99mTc generators of
moderate activity (up to 350 mCi) using low specific activity
(~16.7 GBq (450) mCi/g) 99Mo. Scaling up of the generator capacity
is feasible by using 99Mo produced in higher flux research reactors
or using enriched 98Mo for irradiation.
Acknowledgments
The authors are grateful to Mr. P. V. Joshi and his group for
providing 99Mo. Thanks are due to Analytical Chemistry Division,
BARC for ICP-AES analysis of the solutions. The help received from Mr.
Pritam Bansode in the preparation of the diagrams used in this
manuscript is acknowledged.
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