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Annual Report of China Institute of Atomic Energy 2006
Isotopes
Development of 124Xe Gas Target System
for 123I Production by Cyclone-30
LUO Zhi-fu, YIN Wei, ZHANG Bao-qi, LIU Yu-ping, DENG Xue-song,
LIU She-yang, LIANG Cheng-hu, LIU Geng-shou, ZHAO Gui-zhi
Same as the main preparation method for high purity of 123I in other countries, we would use also
99.9% of 124Xe gas via 124Xe(p, x) reaction and 123Xe decay to get the objective product 123I.
Because the cyclotron facility in CIAE only has two beam outlets for solid and liquid targets, we
have to construct a new one for gas target. Therefore, a new 30 MeV beam line is extracted from the mid
way to the solid target with 75° bending by a bending magnet and focused to the gas target window.
The general purpose of our project is to construct a 124Xe gas target system, 123I washing out and
concentration system with a batch yield of no less than 3.7×1010 Bq (1 Ci) 123I. The chemical form of the
product is Na123I in the solution of 0.02 mol/L NaOH, radionuclide purity≥99.8%, radiochemical purity
－
≥95% for ion I .
The core of the 124Xe gas target technology is the safe transfer of the 124Xe gas and the stable
running of the components at the high vacuum and high pressure.
The whole system is composed by the following parts.
1) Target body. It is machined with metal aluminum having a chamber of 20 mm×500 mm and a
cooling water jacket. 124Xe gas loading pressure is 3 bars and irradiating pressure about 7 bars. The
target is equipped with the pressure and temperature sensors for the on-line detection of the bombarding
situation. The target chamber is connected with cyclotron vacuum line by two molybdenum foils which
are cooled by He flow.
2) 124Xe storage and transfer system. This system is composed mainly a storage bottle, two trapping
bottles and corresponding LN2 Dewar. 124Xe loading and recovering is realized by the way of cryogenic
transfer.
3) Helium cooling loop. This system includes helium pump, heat exchanger and the pressure and
temperature sensors, etc. which offers helium flow for target windows cooling.
4) 124Xe containment. It is a vacuumed 350 L stainless steel tank for collecting the 124Xe from
cyclotron vacuum chamber just in case of target window broken. The collected 124Xe can be partly
recovered cryogenically.
5) 123I washing out and concentration system. It is assembled in a small hot cell. After the
bombardment and 4-6 h decay, formed 123I is absorbed on the inner wall of the target chamber. As soon
as the 124Xe has been received in the storage bottle, the 123I would be washed out with de-oxygen sterile
water and be purified and concentrated. And Na123I in 0.02 mol/L NaOH solution would be the final
product.
6) Control system. It is a set of PLC hard and soft ware system for the automatic and manual
control of the whole facility.
FUNDAMENTAL AND APPLIED FUNDAMENTAL RESEARCH·Isotopes
183
Progress on Preparation of 103Pd Seed Source
DENG Xue-song, LI Guang, WANG Wei, YUE Wen-qiang,
CHEN Da-ming, CHEN Yu-qing
103
Pb has very good nuclide properties, such as half life of 16.9 d and X-ray energy of 21-23 keV, it
is an ideal radioisotope for the use of cancer local treatment (brachytherapy). 103Pd is produced by using
the pulse electroplated Rh target on cyclone-30 cyclotron. 103Pd is deposited on the surface of metal bar
with the dimensions of the seed source of 0.8 mm×4.5 mm by the way of electroplating, and the typical
activity of individual 103Pd seed source is in the range of 18.5-74 MBq.
HYNIC-Anx13 Labelled With 99Tcm for Imaging of Apoptosis
LI Hong-yu, HU Ji, LIANG Ji-xin, CHEN Bao-jun, LU Jia, LUO Lian-zhe
Apoptosis, also known as programmed cell death, is an indispensable component of normal
human growth and development, immunoregulation and homeostasis. Apoptosis is nature’s primary
opponent of cell proliferation and growth. In the course of apoptosis, phosphatidylserine (PS) is rapidly
exposed on the cell’s outer surface. Annexin V, an endogenous human protein, has a high affinity for
membrane- bound PS. Annexin V, therefore, is a sensitive marker of the early to intermediate phases of
apoptosis. Recently, radiolabelled Annexin V has received increasing interesting for detecting apoptosis
in vivo. Particularly, 99Tcm labelled Annexin V has been developed and successfully utilized in clinical
trials.
HYNIC-Anx13 is an Annexin V fragment which has been modified with HYNIC. The sequence of
this peptide is as following:
HYNIC-Ala-Gln-Val-Leu-Arg-Gly-Thr-Val-Thr-Asp-Phe-Pro-Gly(OH)
In this report, the 99Tcm labelling of Annexin V fragment modified with HYNIC (HYNIC-Anx13)
using Tricine, EDDA or EDDA/Tricine as coligands is described. The effects of the pH, the amount of
reducing agent (SnCl2·H2O), the reaction time and temperature on 99Tcm labelling were investigated. The
methods of HPLC and ITLC used for the analysis of radiochemical purity of labelled conjugates were
developed. Biodistribution studies in normal mice and gamma imaging of apoptosis in rats induced by
cyclophosphamide were performed. The labelled conjugates are stable in aqueous solution and serum
solution in vitro, but they are not stable when challenged with cysteine and in vivo.
99Tcm
labelled
HYNIC-Anx13 shows rapid blood clearance and renal excretion. The uptakes in target organs in model
rats are significantly higher than those in control rats (p<0.05), but the T/NT ratios are relatively low. .
184
Annual Report of China Institute of Atomic Energy 2006
Synthesis and Biodistribution of Carborane-Containing Porphyrins
as Tumour Targeting Agents for BNCT
YANG Ting, LUO Zhi-fu, SHEN Lang-tao, LI Hong-yu, LIANG Ji-xin
Boron neutron capture therapy (BNCT) is based on the nuclear reaction between boron-10 and
low-energy thermal neutrons to yield high linear energy transfer  particles and recoiling lithium-7
nuclei. Clinical interest in BNCT has focused primarily on the treatment of high- grade gliomas.
Two boron drugs have been used clinically, sodium mercaptoundecahydro-closo-dodecaborate and
L-p-boronophenylalanine. The major challenge in the development of boron delivery agents has been
the searching of selective tumor targeting agents.
In this report，the synthesis of a novel boronated porphyrin where four closocarborane cages were
appended to the porphyrin macrocycle through four ester bonds between four OHs of bisglycol of
deuteroporphyrin and four carboxylic acids chlorides was described. This compound with many boron
atoms is expected to be a potential BNCT agent.
The intermediate and final products were characterized by IR, 1HNMR, MS and elemental analysis.
In this report，the tritium labeling of BOPP with 10% Pd/C was described. The radiochemical purity
of the labeled compound is higher than 95% by methods of TLC. The biodistribution of 3H-BOPP in
mice model bearing NHC-1 shows that the tumor uptake of 3H-BOPP is high, but the blood clearance is
slow.
Development of Immunoradiometric Assay Kit for Insulin
DONG Hong-guang, JIA Juan-juan, LIU Yi-bing, XU Wen-ge
The detection of insulin level in blood is important to the assessment of β-cells’ function, the
therapy plan of diabetes mellitus and the diagnosis of insulinoma. Insulin is the first human hormone to
be determined by radioimmunoassay (Yallow and Berdon, 1959). Despite the enormous improvements
and developments in immunoassay in the subsequent years, RIA still plays an important role in China.
Due to interfere of breakdown products of insulin, the immunoradiometric assay using two monoclonal
antibodies has a higher specificity than RIA.
We use two monoclonal antibodies that bind to two different epitopes on the insulin molecule. The
immunoassay is specific. The detection limit is 2.8 mIU/L. The working assay range is 5-160 mIU/L; the
CV of intra- assay and inter-assay is below 5% and 10%, respectively. The recovery is 92.3%-112.6%;
the correlation coefficient between measured and expected values is 0.996 after serial dilutions of the
sample with high concentration of insulin. The value for normal samples (n=39) is 4.7-14.3 mIU/L and
the whole incubating time is 3 h at 37 ℃.
FUNDAMENTAL AND APPLIED FUNDAMENTAL RESEARCH·Isotopes
185
Establishment of Enzyme-Linked Immunosorbent Assay
for Sulfaquinoxaline
LIU Chao, LIU Yi-bing, HAN Shi-quan, XU Wen-ge, Zhang Li-ling
An indirect competitive enzyme-linked immunoassay (ELISA) was developed using polyclonal
antibody to measure the residues of SQ in food. The range of standard curve is 0.3-100 g/L. The
sensitivity of the assay is 0.2 g/L. The intra-assay and inter-assay CVs are 7.72%-8.43%, and 9.92%10.79%, respectively. The recovery of SQ in milk, honey, chicken and eggs are 93.1%-106.7%, 69.0%103.5%, 40.0%-73.6%, 50.0%-84.0%, respectively. The correlation coefficient between measured and
expected value is 0.997 after serial dilutions of the samples with high concentration of SQ.
Synthesis of Novel Chelating Agents for Actinide Ions
SHEN Lang-tao, DENG Xin-rong
The synthesis of novel chelating agents for actinide ions is very important to the therapy of internal
pollution of uranium and plutonium. This report describes the reactions between 1, 3, 5-benzenetricarbonyl trichloride and desferrioxamine B (DFO) in aqueous solution. Firstly, DFO was protected as
FeDFO, then, 1, 3, 5-benzenetricarbonyl trichloride was mixed with FeDFO in the ratio of 1︰3. Based
on the experiments, most of 1, 3, 5-benzenetricarbonyl trichloride reacted with one or two equivalents of
FeDFO and produced compound 1 and 2 (Fig .1), only a few of 1, 3, 5-benzenetricarbonyl trichloride
reacted with three equivalents of FeDFO and produced compound 3. The IR and MS of these products
were identified. Compound 1-3 are novel chelating agents for actinide ions. It is worth further studying
the various chemical properties of these compounds and their decorporating efficacy for actinide ions.
Fig. 1 Structures of compounds 1-3
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Annual Report of China Institute of Atomic Energy 2006
Preparation and Biodistribution in Mice
of 99Tcm-Labelled Fatty Acids
LIANG Ji-xin, HU Ji, CHEN Bao-jun, LUO Lian-zhe,
LI Hong-yu, SHEN Lang-tao, LUO Zhi-fu
Long free chain fatty acids are the major energy source of the normal myocardium. Approximately
60%-80% of adenosine triphosphate (ATP) are produced in aerobic myocardium derived from fatty acid
β oxidation.
The measurement of regional difference in the uptakes and retention of radiolabelled fatty acids
using nuclear medicine techniques could provide significant information on myocardium energy metabolism in vivo, and thus could be a valuable approach in diagnoses of several heart diseases. The aim of
the present work is to develop the radiolabelling fatty acids based on 99Tcm carbonyl chemistry for heart
imaging.
Undecanoic acids functionalised with iminodiacetatic acid and cysteine were radiolabelled with
99
[ Tcm (CO)3(H2O)3]+ intermediates, and their radiolabelling condition was studied. Radiolabelling of
CYST FAC11(S-cysteine undecanoic acid) and IDA FAC11 (N, N-di(carboxymethyl) amino undecanoic
acid) could be carried out in high yields using the [99Tcm(CO)3(H2O)3]+ intermediate under optimized
conditions. Biodistribution of 99Tcm (CO)3-CYST FAC11 and 99Tcm (CO)3-IDA FAC11 were performed
in normal mice.
The result shows that two 99Tcm-labelled compounds has similar profile in terms of high initial
radioactivity uptake and rapid washout of radiotracers in the heart. 99Tcm (CO)3-IDA FAC11 is mainly
excreted via hepatobiliary system in contrast to 99Tcm (CO)3-CYST FAC11, which excreted from urinary
system. It may be in part prone to the more lipophilicity of 99Tcm (CO)3-IDA FAC11than that of
99Tcm (CO) -CYST FAC11.
3
Preparation and Imaging Studies
of 99Tcm-EDDA/HYNIC-Lys0-TOCA
LUO Lian-zhe, HU Ji, CHEN Bao-jun, LI Hong-yu, LIANG Ji-xin, CHEN Yang
The new somatostatin analogue
99Tcm-EDDA/HYNIC-Lys0-TOCA
was designed and synthesized.
The biological studies were carried out in normal and tumor-bearing nude mice in order to evaluate its
potential application as an imaging agent for somatostatin receptor positive tumor.
The labelling efficiency of
99Tcm-EDDA/HYNIC-Lys0-TOCA
is approximately 96% under the
optimized conditions, and more than 99% could be reached after Sep-Pak column purification. The
radiolabelled complex shows satisfactory stability in vitro. 99Tcm-EDDA/HYNIC-Lys0-TOCA displays a
rapid blood clearance (about 20 min of half-life) and the predominant excretion via the kidney and
urinary system in normal mice.
Meanwhile, the substantial uptake of radioactivity in stomach, lung and adrenals was observed. The
FUNDAMENTAL AND APPLIED FUNDAMENTAL RESEARCH·Isotopes
biodistribution in tumor-bearing nude mice of
187
99
Tcm-EDDA/HYNIC-Lys0-TOCA suggests that there is
high radioactivity uptake in tumor and rapid blood clearance, and thus high T/NT ratios of tumor to
heart, tumor to blood and tumor to muscle are obtained at 1 h post injection. The radio-tracer remains
almost stable in urine as assayed by HPLC. Planar gamma imaging allows contrasting visualisation of
tumours at 1 h and 2 h post injection.
All these results indicate that 99Tcm-EDDA/HYNIC-Lys0-TOCA is a novel promising candidate for
somatostatin receptor -positive tumor imaging.
Chemiluminescence Immunoassay
for Total Prostate-Specific Antigen
ZHANG Xue-feng, LIU Yi-bing, HAN Shi-quan
Prostate-specific antigen (PSA) is a 30 kDa serine protease produced by epithelial cells of the
prostate. It is a reliable marker for monitoring treatment of prostate cancer (PCa), and it is also widely
used for early diagnosis and screening. The enzyme-linked immunosorbent assay (ELISA) for PSA is in
widespread use, and the final readout of the enzyme product is commonly made spectrophotometrcally
using a chromogenic substrate in 96-well microtitre plates. There is now a general movement towards the
development of chemiluminescence because of its high sensitivity, specificity, precision and accuracy.
Therefore we established a PSA detection system based on chemiluminescence immunoassay, using HRP
as the label.
The chemiluminescence reaction of luminol with hydrogen peroxide is used in this process. The
limit of detection is established as 0.12 g/L (n=10, mean of zero standard + 2S.D.) and the analytical
recovery of PSA is 83.8%-118.7%. The intra-assay and inter-assay coefficient of variation (CV) varies
from 4.4%-5.0% and 6.2%-11.7%, respectively. The experimental correlation coefficient of dilution is
0.999. Compared with immunoradiometric assay (IRMA) kits, the correlative equation is y = 1.07x +
0.68, and R=0.97. The standard range for the method is 1.5-80 mg/L, and it presents good linearity.
Determination of Seventeen Trace Impurities in Tellurium Oxide
by Atomic Emission Spectrometry
ZHAO Xiu-yan, JING Lie, LU Rong, YE Zhao-yun, JIANG Hua, ZHAO Yan
The determination of trace Pb, Mn, Ba, Sn, Fe, Al, etc. in tellurium oxide was carried out by atomic
emission spectrometry (AES). The sample was directly loaded into a cup electrode with fine neck.
The experimental result shows the method is simple, rapid and accurate. The range of the recovery
is 80%-120%, and relative standard deviation (RSD) is smaller than 8%.
The method has been applied to the determination of the sample with satisfactory results.
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Annual Report of China Institute of Atomic Energy 2006
Establishment of Chemiluminescence Immunoassay for Insulin
JIA Juan-juan, LIU Yi-bing, HAN Shi-quan
Insulin is the principle hormone responsible for the control of glucose metabolism. It is synthesized
in the β-cells of the islets. The mature insulin molecule comprises two polypeptide chains, the A chain
and B chain (21 and 30 amino acids, respectively). The detection of insulin in serum is significant in
diagnosis and pathological research of diabetes mellitus. Chemiluminescence immunoassay is a highly
sensitive method that enables non-isotopic detection in immunoassay. In many hospitals chemiluminescense immunoassay kits has become popular due to its high sensitivity, specificity, wider detection limit
and automatic operation.
We develop a solid phase two-site chemiluminescence immunoassay. It is based on the direct
sandwich technique in which two monoclonal antibodies are directed against separate antigenic determinates on the insulin molecule. The sensitivity of the assay is 1.96 mIU/L, and the linear calibrator is in
the range of 5-160 mIU/L. The CVs of intra- assay and inter-assay are both below 6%. The correlation
coefficient between measured and expected values is well after serial dilutions of the samples with high
concentration of insulin. The value for normal samples (n=39) is 2.84-9.08 mIU/L and the whole
incubating time is 1 h at 37 ℃.
Establishment of Chemiluminescence Immunoassay
of Human Ovarian Cancer Antigen 125
FENG Shu-yuan, HAN Shi-quan, LIU Yi-bing
Measurement of Cancer Antigen 125 (CA125) assay values has useful index as an aid in monitoring the response to therapy for patient with epithelial ovarian cancer.
This abstract describes a new chemiluminescence immunoassay method to detect the concentration
of CA125 in the blood sample. One monoclonal antibody against CA125 was immobilized on 96 wells
plate. Another HRP labeled antibody was added along with standards and samples. After 3 h incubation,
plate was washed with PB three times. A new type of chemiluminescent substrate was added into the
well before measuring the result.
The lowest detection limit is 1.78 U/mL, the CVs of intra assay and inter assay are both below 10%.
The recovery is from 100.9% to 106.7%; the correlation coefficient between measured and expected
values is 0.999 after serial dilutions of the sample with FBS. The value for normal samples (n=33) is
9.49 U/mL.
FUNDAMENTAL AND APPLIED FUNDAMENTAL RESEARCH·Isotopes
189
Establishment of Competition Inhibition ELISA
for Sulfamethoxazole
FENG Ting-ting, LI Zi-ying, LIU Yi-bing, XU Wen-ge, GUAN Guo-ying, HAN Shi-quan
Sulfonamides are chemotherapeutical reagents widely used in human as well as in veterinary
medicine for the treatment of bacterial infection. They are also used as growth-promoting feed additives.
Adverse effects include allergenic effects, carcinogenicity, and antibiotic resistance of microorganisms.
Therefore, to protect the consumer’s health, the use of sulfonamides has been regulated in China, and a
maximum residue limit of 100 ng/g in edible tissue for all sulfonamides combined has been established.
ELISA is a simple, rapid and quantifiable screening method for sulfonamides. A sulfamethoxazolespecific ELISA was developed to analyze sulfonamide sulfamethoxazole (SMX) residues in animalbased food products. The principle for the measurement of SMX is SMX in the sample and
enzyme-labeled SMX bind to antibody in solid phase competitively. And signal falls with increasing
SMX concentration. The sensitivity of the assay is 0.067 μg/L. The intra- assay and intro-assay CVs of 3
samples are lower than 10% and 20%, respectively. The analytical recoveries of milk, meat, egg and
honey are in the range of 94.9%-116.0%, 82.0%-107.6%, 73.0%-90.0% and 61.9%-94.0%, respectively.