Technetium-99m recovery process in the presence of stannous Sn (II) with the aim to determine it is
necessary and sufficient quantity, that would provide a complete "recovery" of 99mTc in the reaction mixture
has been investigated. The studies were identified conditions for receiving radioactive nanopreparations
based on a modified molecule DTPA. The authors has been shown that the direct interaction of
99mTc eluate solution with a solution of a mixture of SnCl2 DTPAmod and radiochemical impurity content
unreduced 99mTc (VII) in the preparation labeled with nanocolloids is 2,7% and the yield nanocolloids
with the size of 100 nm is about 80%. The medical-biological testing of the agents 99mTc (IV) - DTPAmod
on test animals for the determination of the functional suitability for the scintigraphic imaging of lymph
nodes has been carried out.
When you are citing the document, use the following link http://essuir.sumdu.edu.ua/handle/123456789/3540

Nesterov, E.A.

Rogov, A.S.

Sadkin, V.L.

Scuridin, V.S.

Stasyuk, E.S.

Varlamova, N.V.

Electronic Sumy State University Institutional Repository

PROCEEDINGS OF THE INTERNATIONAL CONFERENCE  NANOMATERIALS: APPLICATIONS AND PROPERTIES Vol. 2 No 4, 04NABM02(4pp) (2013)   2304-1862/2013/2(4)04NABM02(4) 04NABM02-1  2013 Sumy State University Preparation and Medico-Biological Tests Nanocolloids Radiopharmaceuticals on the Based of Modified DTPA Labeled Technetium-99m  V. S. Scuridin, E.S. Stasyuk, E. A. Nesterov, A.S. Rogov, V.L. Sadkin, N. V. Varlamova  National  Research Tomsk Polytechnic University, 30 Lenin Ave, 634050 Tomsk, Russia  (Received 16 May 2013; published online 30 August 2013)  Technetium-99m recovery process in the presence of stannous Sn (II) with the aim to determine it is necessary and sufficient quantity, that would provide a complete "recovery"  of 99mTc in the reaction mix-ture has been investigated. The studies were identified conditions for receiving radioactive nanoprepara-tions based on a modified molecule DTPA. The authors has been shown that the direct interaction of 99mTc eluate solution with a solution of a mixture of SnCl2 DTPAmod and radiochemical impurity content  unreduced 99mTc (VII) in the preparation labeled with nanocolloids is 2,7% and the yield nanocolloids with the size of 100 nm is about 80%. The medical-biological testing of the agents 99mTc (IV) - DTPAmod on test animals for the determination of the functional suitability for the scintigraphic  imaging of lymph nodes has been carried out.  Keywords: Technetium-99m, Modified molecule DTPA, Molybdenum, Nanocolloid labeled technetium-99m.   PACS numbers: 47.57.J –, 62.23. –c   1. INTRODUCTION  In the world practice nanocolloid preparations la-beled with a short-lived radionuclide technetium-99m (99mTc) are widely used for diagnostic tests in oncology, cardiology, for the detection of inflammatory diseases of the musculoskeletal system, cirrhosis, hepatitis and other diseases. The use of radioactive nanocolloids in oncology is based on the possibility of the rapid and effective identifi-cation of "sentinel" lymph nodes (SLN), which are the first lymph nodes where the lymph  from a malignant tumor flows. These nodes, by filtering the afferent lymph, be-come a  "trap" for malignant cells, so their biopsy is an objective diagnostic criterion for distributing a malignant process. The optimal effective method of identifying areas of localization of SLN is scintigraphy or radiometry with using technetium-99m-labeled nanocolloids [1]. As a rule, nanocolloid preparations are made on the basis of compounds forming stable hydrosols. The deci-sive factor for the success is not so much  their chemical composition as the size of the nanoparticles. It is known that the optimum particle size for lymphoscintigraphy is 20-100 nm. Such particles are derived from tissues at a rate which does not allow them to penetrate into the bloodstream. On the other hand, particles with sizes less than 20 nm can easily pass into the bloodstream, which prevents the imaging of lymph nodes [2]. Most of the known nanoсolloids radiopharmaceuti-cals is a simple inorganic complexes with 99mTc rheni-um sulfide and antimony produced by fairly complex technology. Presupposition for the chelator DTPA radi-olabelling is the ability of the molecule itself as DTPA or derivatives thereof, to form sufficiently stable com-plexes with various metals, including compounds with technetium-99m. This determined the goal of our work.  2. METHODS OF SAMPLE PRODUCTION AND ANALYSIS  The determination of the radiochemical purity of obtained nanocolloidal preparations were carried out by the thin - layer chromatography. The scanning of the chromatograms was carried out on the facili-ty"Gamma-Scan-01A." On the computer screen was received the information about the location of  activity peaks of the labeled and free (unreacted) 99mTc com-pounds.  Determining the amount of technetium-99m-labeled nanocolloidal particles was carried out by the methods based on measuring the activity of the sus-pension before and after filtration through filters with desired pore sizes: 100 nm. For this purpose, three samples were chosen volume of 5 ml of the original solutions and filtrates for  the subsequent measure-ment of their activity, as well as samples - for chroma-tograms for evaluating the content of impurities in the filtrates of unreacted 99mTc (VII) in the tested product. Calculations of the yield of products with different particle sizes were determined using the following formulas:   100100исхисхА АСА   where Aис - the activity of the original suspension be-fore the filtration, A100 - the activity measured after the filtration through the filter of 100 nm. For the original of the production of the prepara-tion 99mTc (eluate) was used the chromatographic generator "99mTc-GT-TOM" produced by the Institute of Physics and Technologies in Tomsk Polytechnic University.  3. RESULTS AND DISCUSSION  Obtained in the first stage of research Nanocolloids based on chemically modified DTPA molecules exhibit a certain similarity with a protein molecule - the pres-ence of an amide (peptide) links, the existence in the zwitter-ionic form. Simple chemical transformation  V. S. SCURIDIN, E.S. STASYUK, E. A. NESTEROV ET AL. PROC. NAP 2, 04NABM02 (2013)   04NABM02-2 yielded a number of derivatives of DTPA-modified hy-drophobic groups. Molecule itself  DTPA, as well as its metal complexes, hydrophilic and are therefore not able to form colloidal particles. Adding hydrophobic frag-ments yielded water-insoluble complexes of DTPA-modified. At its core are amphiphilic compounds exist as zwitterions - ions are both peers once the cation-and anion-active surfactants. On the molecule's solubility and its ability to form colloidal particles of a defined size primarily affect its geometry features. Furthermore, the stability of colloi-dal systems affected by the ratio between the amount of hydrophilic and hydrophobic groups (hydrophilic-lipophilic balance, HLB). Computer simulations carried out with these data allowed to calculate the size of the molecules and their resulting ionic radius in order to give them the desired properties. It was found that the addition of a molecule of DTPA two long straight-chain aliphatic chains with an aromatic group that performed the role of a "bridge" between the molecule DTPA and aliphatic chains (C21 - C22), has given the ability of the modified DTPA form colloidal solutions at physiological pH values. Compounds with shorter chains have shown a great deal poorer. To create a desired solution nanocolloid particle size (100 nm), it was necessary to select an organic solvent or inorganic, acidic or alkaline. The solvents were test-ed following solutions: saline - solution 0.9% NaCl, 5% solution of NaHCO3, HClconts., dehydrated ethyl alco-hol. To test the solubility of the modified selected sub-stance in a individual bottle were added 5 mg of sample DTPAmod, and then each of the vials prepared adminis-tered solvents. The solubility of the substance was determined visually by using a dedicated set represent-ing a light booth with an optical device. The results are shown in Table 1. Table 1 − DTPAmod dissolution in different solvents  Solvent Process of dissolution 0,9 % solution NaCl It does not dissolve even under heating to 800С HClconcent. It does not dissolve even under heating to  800С 10 % solution Na2CO3 Dissolves when heated, to form opalescent solution Dehydrated alcohol Soluble without heating, to form a clear solution  The studies are shown, that the acid solutions DTPAmod not dissolve in anhydrous alcohol DTPAmod completely dissolved and does not form colloidal solu-tions. In alkaline solutions DTPAmod forms hazy opales-cent colloidal type. On this basis, as the solvent for preparing solution nanocolloid DTPAmod was chosen 5% solution of NaHCO3. The next step study was the determination of the av-erage particle size of the resulting colloidal solution. The essence of the method consists in determining the particle size by the optical density of the colloidal systems in dif-ferent parts of the visible spectrum (λ = 400 - 800 nm) using a spectrophotometer Unico 2802 (S). In the original colloidal solution was added 5% NaHCO3 solution and the solutions were prepared with different concentrations by DTPAmod: 0.333 mg/ml (solution number 1), 0.167 mg/ml (solution number 2), 0.083 mg /ml (solution number 3). Prepared in four 10x10 mm cuvettes were injected 3 ml of solution number 1, 2 and 3, and in one (reference solution) - 3 ml of a 5% solution of NaHCO3. Optical transmission study DTPAmod gels were measured at wavelengths λ from 400 to 800 nm. The error in determining the optical densi-ty D% sample solutions was < 1%. The resulting optical density values are shown in Table 2.  Table 2 − The values of optical density of the solutions with different concentrations nanocolloids  Длина волны, λ lg λ D lg D D lg D D lg D Раствор № 1 Раствор № 2 Раствор № 3 400 2,602 0,786 -0,105 0,265 -0,577 0,099 -1,004 450 2,653 0,611 -0,214 0,2 -0,699 0,074 -1,131 500 2,699 0,495 -0,305 0,159 -0,799 0,059 -1,229 550 2,740 0,411 -0,386 0,131 -0,883 0,048 -1,319 600 2,778 0,349 -0,457 0,11 -0,959 0,041 -1,387 650 2,813 0,301 -0,521 0,093 -1,032 0,035 -1,456 700 2,845 0,263 -0,580 0,081 -1,092 0,031 -1,509 750 2,875 0,233 -0,633 0,072 -1,143 0,027 -1,569 800 2,903 0,208 -0,682 0,064 -1,194 0,025 -1,602  Based on the data we plotted lgD = f (lg λ). Product according possible to determine the slope of X, which, accordingly, was 1.91, 2.03 and 1.98. According to the methodology used, the range of values for x = 0.2 - 3.8 average r (nm) is calculated using the formula:   R  -32,662Х+157,92 (1)  As a result, it was found that the radius of the col-loidal particles in the test solution number 1, 2 and 3 are 96, 91 and 93 nm. Hence the average particle radi-us was 93 nm, and the diameter of -187 nm. From this it follows that under the chosen conditions nanocolloid solution preparation, the concentration of active ingre-dient (DTPAmod) and the degree of dilution does not affect the average particle size. In order to reduce the particle size has been tried nanocolloids combination of two methods: heating and sonication. For this purpose, in two bottle introduced on 5 ml nanocolloids DTPAmod a concentration 0.222 mg/ml. The bottle were placed in an ultrasonic bath filled with hot water (t ≈ 70 ° C): the first bottle 10 min. And a second bottle for 40 min. Then spent deter-mining the optical density of the solutions for the above described method. Based on the data obtained and built according lgD = f (lg λ). The slope of the dependencies were respectively 2.56 and 2.88. Subsequent calcula- PREPARATION AND MEDICO-BIOLOGICAL TESTS NANOCOLLOIDS… PROC. NAP 2, 04NABM02 (2013)   04NABM02-3 tions on the formula (1) showed that the radius of the particles in solution of the first and second vials respec-tively decreased to 74 and 60 nm, and diameters - 148 and 120 nm. Since the technique used to evaluate only allowed the mean particle radius, to verify that the solution particles with a diameter less than 100 nm, the solu-tion of the first bottle was further passed through a filter with a pore size of 100 nm, followed by measuring the optical density of the filtrate, and determining the radius of the particles. The calculated value of the mean radius was about 44 nm. From that it was con-cluded that for obtaining solutions with predetermined nanocolloids value particle size (less than 100 nm) to carry out their filtration through a filter with a pore size of 100 nm.For radiolabeling of 99mTc in nanocolloid substance has been proposed a method of direct mixing of solutions containing DTPAmod with a solution of so-dium pertehnetatat obtained from chromatographic generator «99mTc-GT-TOM." But as shown by our study, in the absence of a reducing agent to a modified inter-action 99mTc DTPA is not happening. Regardless of the choice of a mobile phase a single peak is observed in the chromatograms of Rf = 0,9, which corresponds to the ions of 99m Tc-pertechnetate. As is known 99mTc present in the original eluate to with the highest degree of oxidation of (VII), does not have a high reactivity, however, in a low oxidation is sufficiently reactive. For restoring 99mTc (VII), present in the original eluate 99mTc, tin chloride dihydrate (II) (SnCl2 ∙ 2H2O) was used.Therefore, preliminary studies were carried out to establish the necessary and enough amount of Sn (II), which provides the complete recovery of 99mTc (VII) in the eluate from the 99Mo/99mTc-generator The results are shown in Table 3.  Table 3 − Changing the content of 99mTc (VII) in the eluate from the 99Mo/99mTс-generator depending on the concentration of tin (II)  Concentration (Sn(II)), mg/ml Tc(VII),% 0.1400 0.0 0.1050 0.0 0.0700 0.7 0.0350 3.0 0.0175 7.0 0.00875 10.0  The table shows that the optimum amount of Sn (II) in the reaction mixture provides the content 99mTc (VII) of less than 10% corresponds to a value in the range of from 0.00875 to 0.0175 mg / ml. Subsequent radiolabeling 99mTc nanokolloidnuyu substance carried in two ways: by direct mixing solu-tions containing DTPAmod, SnCl2 and 99mTc, and by dissolving the dry residue obtained after evaporation of solutions with DTPAmod and SnCl2, 99m Tc in the eluate. The mixture was then heated in a water bath (70-80 ° C) for 30 min. After cooling to room temperature in an ultrasonic bath, filtration of the products obtained through filters with a pore diameter of 100 nm. Then, samples were taken for chromatography. The results of these studies are shown in Table 4. From the presented table shows that the most in-teresting result is the direct interaction of 99mTc eluate solution with a solution of a mixture of sodium chloride and DTPAmod tin containing a certain amount of Sn. The output labeled nanocolloids based on the modified molecules DTPAmod with particle size less than 100 nm was 80% with a total content of radiochemical impuri-ties  unreduced 99mTc (VII) in the resulting formulation 2.7% (Fig. 1). This is a fairly good indicator, closely matching the regulatory requirements for these drugs. When interacting with 99mTc eluate dry residues de-rived from alcohol mixtures containing DTPAmod and SnCl2, radiochemical impurity contents  unreduced 99mTc (VII) obtained in preparation nanocolloid was about 6% and the yield nanocolloids labeled with a size less than 100 nm was 74%.  Table 4 − The results studies  Ingredients of mixture Activity, imp Contents 99mTc(VII), % Yield % [DTPAmod +Sn(II)] solution +99mTc 550 965 3,5  After heating 484 552 3,0  Filtrate 100 387 642 2,7 80 [DTPAmod +Sn(II)]dry.+99mTc 545 128 7,0  After heating 476 179 5,8  Filtrate 100 352 372 4,8 74    Fig. 1 − Radiochromatogramma 99mTc elution with a solution of a mixture of DTPAmod and SnCl2  Medico-biological tests of the colloidal drug on the basis of modified molecules DTPAmod labeled with 99mTc were carried out at the Research Institute of Oncology, SB RAMS Tomsk city, on the white line of male rats "Wistar" with the mass of 300-350. The ani-mal's body scintigram obtained at various time inter-vals are shown in Figure 2. On the scintigrams of 60 and 120 minutes is a clearly visible sentinel lymph node, located between the bladder and the place of injection. The level of accumulation of the drug in the lymph node was 3.67% of the total intro-duced activity, which is enough for its safe visualization. This result exceeds to the standard requirements for such drugs (0.5 - 1.7%) and proves the functional suita-bility of our synthesized labeled with technetium-99m, nanocolloid on the basis of modified molecules DTPAmod.   V. S. SCURIDIN, E.S. STASYUK, E. A. NESTEROV ET AL. PROC. NAP 2, 04NABM02 (2013)   04NABM02-4   Fig. 2 − The drug  distribution in the body of rat  at the introduction of a nanocolloid [DTPAmod  и SnCl2]: A) Immediately after injection, B) 30 minutes after injection, C) 60 minutes after injection, D ) 120 minutes after injection. The numbers  is indicated: 1 - lymph node, and 2 - Location of the drug  4. CONCLUSION  1. The research process of restoration of 99mTc in the presence of stannous Sn (II) have been studied. As a result,  it has been found that the optimum concentration of the reducing agent, Sn (II) in the radiopharmaceutical must be in the range from 0.00875 to 0.0175 mg / ml. 2. The conditions for obtaining radioactive nano-preparations on the basis of the modified molecule DTPA. It is shown that the direct interaction of 99mTc eluate solution with a solution of a mixture of SnCl2 DTPAmod and radiochemical impurity content unre-duced 99mTc (VII) in the preparation labeled with Nanocolloids is 2.7% and the yield nanocolloids with the size of 100 nm is about 80%. 3. Medico-biological tests of  labeled 99mTc on the molecules DTPAmod  have been carried out on rats in order to study their distribution in the organism of the experimental animals and determine the func-tional suitability for the scintigraphic imaging of lymph nodes. The level of accumulation of the drug in the lymph node is 3.67% of the total introduced activ-ity, which is enough for its safe imaging. This work has been carried out in the frame of state program «Sciense» № 3.1192.2011.   RERERENCES  1. P. Reimann, Phys. Rep. 361, 57 (2002).  2. S. Maza et al., QJ  Nuc.Med 30/5, 651 (2003). 3. C.B. Sampson, Textbook of Radiopharmacy theory and Practice (London, United Kingdom: Gordon and Breach; 1994).  А B C D 1 2 

Preparation and Medico-Biological Tests Nanocolloids Radiopharmaceuticals on the Based of Modified DTPA Labeled Technetium-99m

SocioEconomic Challenges Journal (Sumy State University)

PROCEEDINGS OF THE INTERNATIONAL CONFERENCE  
NANOMATERIALS: APPLICATIONS AND PROPERTIES 
Vol. 2 No 4, 04NABM02(4pp) (2013) 
 
 
2304-1862/2013/2(4)04NABM02(4) 04NABM02-1  2013 Sumy State University 
Preparation and Medico-Biological Tests Nanocolloids Radiopharmaceuticals on the Based 
of Modified DTPA Labeled Technetium-99m 
 
V. S. Scuridin, E.S. Stasyuk, E. A. Nesterov, A.S. Rogov, V.L. Sadkin, N. V. Varlamova 
 
National  Research Tomsk Polytechnic University, 30 Lenin Ave, 634050 Tomsk, Russia 
 
(Received 16 May 2013; published online 30 August 2013) 
 
Technetium-99m recovery process in the presence of stannous Sn (II) with the aim to determine it is 
necessary and sufficient quantity, that would provide a complete "recovery"  of 99mTc in the reaction mix-
ture has been investigated. The studies were identified conditions for receiving radioactive nanoprepara-
tions based on a modified molecule DTPA. The authors has been shown that the direct interaction of 
99mTc eluate solution with a solution of a mixture of SnCl2 DTPAmod and radiochemical impurity content  
unreduced 99mTc (VII) in the preparation labeled with nanocolloids is 2,7% and the yield nanocolloids 
with the size of 100 nm is about 80%. The medical-biological testing of the agents 99mTc (IV) - DTPAmod 
on test animals for the determination of the functional suitability for the scintigraphic  imaging of lymph 
nodes has been carried out. 
 
Keywords: Technetium-99m, Modified molecule DTPA, Molybdenum, Nanocolloid labeled technetium-
99m. 
 
 PACS numbers: 47.57.J –, 62.23. –c 
 
 
1. INTRODUCTION 
 
In the world practice nanocolloid preparations la-
beled with a short-lived radionuclide technetium-99m 
(99mTc) are widely used for diagnostic tests in oncology, 
cardiology, for the detection of inflammatory diseases of 
the musculoskeletal system, cirrhosis, hepatitis and 
other diseases. 
The use of radioactive nanocolloids in oncology is 
based on the possibility of the rapid and effective identifi-
cation of "sentinel" lymph nodes (SLN), which are the first 
lymph nodes where the lymph  from a malignant tumor 
flows. These nodes, by filtering the afferent lymph, be-
come a  "trap" for malignant cells, so their biopsy is an 
objective diagnostic criterion for distributing a malignant 
process. The optimal effective method of identifying areas 
of localization of SLN is scintigraphy or radiometry with 
using technetium-99m-labeled nanocolloids [1]. 
As a rule, nanocolloid preparations are made on the 
basis of compounds forming stable hydrosols. The deci-
sive factor for the success is not so much  their chemical 
composition as the size of the nanoparticles. It is known 
that the optimum particle size for lymphoscintigraphy is 
20-100 nm. Such particles are derived from tissues at a 
rate which does not allow them to penetrate into the 
bloodstream. On the other hand, particles with sizes 
less than 20 nm can easily pass into the bloodstream, 
which prevents the imaging of lymph nodes [2]. 
Most of the known nanoсolloids radiopharmaceuti-
cals is a simple inorganic complexes with 99mTc rheni-
um sulfide and antimony produced by fairly complex 
technology. Presupposition for the chelator DTPA radi-
olabelling is the ability of the molecule itself as DTPA 
or derivatives thereof, to form sufficiently stable com-
plexes with various metals, including compounds with 
technetium-99m. This determined the goal of our work. 
 
2. METHODS OF SAMPLE PRODUCTION AND 
ANALYSIS 
 
The determination of the radiochemical purity of 
obtained nanocolloidal preparations were carried out 
by the thin - layer chromatography. The scanning of 
the chromatograms was carried out on the facili-
ty"Gamma-Scan-01A." On the computer screen was 
received the information about the location of  activity 
peaks of the labeled and free (unreacted) 99mTc com-
pounds.  
Determining the amount of technetium-99m-
labeled nanocolloidal particles was carried out by the 
methods based on measuring the activity of the sus-
pension before and after filtration through filters with 
desired pore sizes: 100 nm. For this purpose, three 
samples were chosen volume of 5 ml of the original 
solutions and filtrates for  the subsequent measure-
ment of their activity, as well as samples - for chroma-
tograms for evaluating the content of impurities in 
the filtrates of unreacted 99mTc (VII) in the tested 
product. Calculations of the yield of products with 
different particle sizes were determined using the 
following formulas: 
 
 100100
исх
исх
А А
С
А
  
 
where Aис - the activity of the original suspension be-
fore the filtration, A100 - the activity measured after the 
filtration through the filter of 100 nm. 
For the original of the production of the prepara-
tion 99mTc (eluate) was used the chromatographic 
generator "99mTc-GT-TOM" produced by the Institute 
of Physics and Technologies in Tomsk Polytechnic 
University. 
 
3. RESULTS AND DISCUSSION 
 
Obtained in the first stage of research Nanocolloids 
based on chemically modified DTPA molecules exhibit a 
certain similarity with a protein molecule - the pres-
ence of an amide (peptide) links, the existence in the 
zwitter-ionic form. Simple chemical transformation 
 V. S. SCURIDIN, E.S. STASYUK, E. A. NESTEROV ET AL. PROC. NAP 2, 04NABM02 (2013) 
 
 
04NABM02-2 
yielded a number of derivatives of DTPA-modified hy-
drophobic groups. Molecule itself  DTPA, as well as its 
metal complexes, hydrophilic and are therefore not able 
to form colloidal particles. Adding hydrophobic frag-
ments yielded water-insoluble complexes of DTPA-
modified. At its core are amphiphilic compounds exist 
as zwitterions - ions are both peers once the cation-and 
anion-active surfactants. 
On the molecule's solubility and its ability to form 
colloidal particles of a defined size primarily affect its 
geometry features. Furthermore, the stability of colloi-
dal systems affected by the ratio between the amount of 
hydrophilic and hydrophobic groups (hydrophilic-
lipophilic balance, HLB). Computer simulations carried 
out with these data allowed to calculate the size of the 
molecules and their resulting ionic radius in order to 
give them the desired properties. It was found that the 
addition of a molecule of DTPA two long straight-chain 
aliphatic chains with an aromatic group that performed 
the role of a "bridge" between the molecule DTPA and 
aliphatic chains (C21 - C22), has given the ability of the 
modified DTPA form colloidal solutions at physiological 
pH values. Compounds with shorter chains have shown 
a great deal poorer. 
To create a desired solution nanocolloid particle size 
(100 nm), it was necessary to select an organic solvent 
or inorganic, acidic or alkaline. The solvents were test-
ed following solutions: saline - solution 0.9% NaCl, 5% 
solution of NaHCO3, HClconts., dehydrated ethyl alco-
hol. To test the solubility of the modified selected sub-
stance in a individual bottle were added 5 mg of sample 
DTPAmod, and then each of the vials prepared adminis-
tered solvents. The solubility of the substance was 
determined visually by using a dedicated set represent-
ing a light booth with an optical device. The results are 
shown in Table 1. 
Table 1 − DTPAmod dissolution in different solvents 
 
Solvent Process of dissolution 
0,9 % solution NaCl 
It does not dissolve even 
under heating to 800С 
HClconcent. 
It does not dissolve even 
under heating to  800С 
10 % solution Na2CO3 
Dissolves when heated, to 
form opalescent solution 
Dehydrated alcohol 
Soluble without heating, to 
form a clear solution 
 
The studies are shown, that the acid solutions 
DTPAmod not dissolve in anhydrous alcohol DTPAmod 
completely dissolved and does not form colloidal solu-
tions. In alkaline solutions DTPAmod forms hazy opales-
cent colloidal type. On this basis, as the solvent for 
preparing solution nanocolloid DTPAmod was chosen 5% 
solution of NaHCO3. 
The next step study was the determination of the av-
erage particle size of the resulting colloidal solution. The 
essence of the method consists in determining the particle 
size by the optical density of the colloidal systems in dif-
ferent parts of the visible spectrum (λ = 400 - 800 nm) 
using a spectrophotometer Unico 2802 (S). In the original 
colloidal solution was added 5% NaHCO3 solution and the 
solutions were prepared with different concentrations by 
DTPAmod: 0.333 mg/ml (solution number 1), 0.167 mg/ml 
(solution number 2), 0.083 mg /ml (solution number 3). 
Prepared in four 10x10 mm cuvettes were injected 3 ml of 
solution number 1, 2 and 3, and in one (reference solution) 
- 3 ml of a 5% solution of NaHCO3. Optical transmission 
study DTPAmod gels were measured at wavelengths λ from 
400 to 800 nm. The error in determining the optical densi-
ty D% sample solutions was < 1%. The resulting optical 
density values are shown in Table 2. 
 
Table 2 − The values of optical density of the solutions with different concentrations nanocolloids 
 
Длина 
волны, λ 
lg λ 
D lg D D lg D D lg D 
Раствор № 1 Раствор № 2 Раствор № 3 
400 2,602 0,786 -0,105 0,265 -0,577 0,099 -1,004 
450 2,653 0,611 -0,214 0,2 -0,699 0,074 -1,131 
500 2,699 0,495 -0,305 0,159 -0,799 0,059 -1,229 
550 2,740 0,411 -0,386 0,131 -0,883 0,048 -1,319 
600 2,778 0,349 -0,457 0,11 -0,959 0,041 -1,387 
650 2,813 0,301 -0,521 0,093 -1,032 0,035 -1,456 
700 2,845 0,263 -0,580 0,081 -1,092 0,031 -1,509 
750 2,875 0,233 -0,633 0,072 -1,143 0,027 -1,569 
800 2,903 0,208 -0,682 0,064 -1,194 0,025 -1,602 
 
Based on the data we plotted lgD = f (lg λ). Product 
according possible to determine the slope of X, which, 
accordingly, was 1.91, 2.03 and 1.98. According to the 
methodology used, the range of values for x = 0.2 - 3.8 
average r (nm) is calculated using the formula: 
 
 R  -32,662Х+157,92 (1) 
 
As a result, it was found that the radius of the col-
loidal particles in the test solution number 1, 2 and 3 
are 96, 91 and 93 nm. Hence the average particle radi-
us was 93 nm, and the diameter of -187 nm. From this 
it follows that under the chosen conditions nanocolloid 
solution preparation, the concentration of active ingre-
dient (DTPAmod) and the degree of dilution does not 
affect the average particle size. 
In order to reduce the particle size has been tried 
nanocolloids combination of two methods: heating and 
sonication. For this purpose, in two bottle introduced 
on 5 ml nanocolloids DTPAmod a concentration 
0.222 mg/ml. The bottle were placed in an ultrasonic 
bath filled with hot water (t ≈ 70 ° C): the first bottle 10 
min. And a second bottle for 40 min. Then spent deter-
mining the optical density of the solutions for the above 
described method. Based on the data obtained and built 
according lgD = f (lg λ). The slope of the dependencies 
were respectively 2.56 and 2.88. Subsequent calcula-
 PREPARATION AND MEDICO-BIOLOGICAL TESTS NANOCOLLOIDS… PROC. NAP 2, 04NABM02 (2013) 
 
 
04NABM02-3 
tions on the formula (1) showed that the radius of the 
particles in solution of the first and second vials respec-
tively decreased to 74 and 60 nm, and diameters - 148 
and 120 nm. 
Since the technique used to evaluate only allowed 
the mean particle radius, to verify that the solution 
particles with a diameter less than 100 nm, the solu-
tion of the first bottle was further passed through a 
filter with a pore size of 100 nm, followed by measuring 
the optical density of the filtrate, and determining the 
radius of the particles. The calculated value of the 
mean radius was about 44 nm. From that it was con-
cluded that for obtaining solutions with predetermined 
nanocolloids value particle size (less than 100 nm) to 
carry out their filtration through a filter with a pore 
size of 100 nm.For radiolabeling of 99mTc in nanocolloid 
substance has been proposed a method of direct mixing 
of solutions containing DTPAmod with a solution of so-
dium pertehnetatat obtained from chromatographic 
generator «99mTc-GT-TOM." But as shown by our study, 
in the absence of a reducing agent to a modified inter-
action 99mTc DTPA is not happening. Regardless of the 
choice of a mobile phase a single peak is observed in 
the chromatograms of Rf = 0,9, which corresponds to 
the ions of 99m Tc-pertechnetate. 
As is known 99mTc present in the original eluate to 
with the highest degree of oxidation of (VII), does not 
have a high reactivity, however, in a low oxidation is 
sufficiently reactive. For restoring 99mTc (VII), present 
in the original eluate 99mTc, tin chloride dihydrate (II) 
(SnCl2 ∙ 2H2O) was used.Therefore, preliminary studies 
were carried out to establish the necessary and enough 
amount of Sn (II), which provides the complete recovery 
of 99mTc (VII) in the eluate from the 99Mo/99mTc-
generator The results are shown in Table 3. 
 
Table 3 − Changing the content of 99mTc (VII) in the eluate 
from the 99Mo/99mTс-generator depending on the concentration 
of tin (II) 
 
Concentration (Sn(II)), 
mg/ml 
Tc(VII),% 
0.1400 0.0 
0.1050 0.0 
0.0700 0.7 
0.0350 3.0 
0.0175 7.0 
0.00875 10.0 
 
The table shows that the optimum amount of Sn (II) 
in the reaction mixture provides the content 99mTc (VII) 
of less than 10% corresponds to a value in the range of 
from 0.00875 to 0.0175 mg / ml. 
Subsequent radiolabeling 99mTc nanokolloidnuyu 
substance carried in two ways: by direct mixing solu-
tions containing DTPAmod, SnCl2 and 99mTc, and by 
dissolving the dry residue obtained after evaporation of 
solutions with DTPAmod and SnCl2, 99m Tc in the eluate. 
The mixture was then heated in a water bath (70-80 ° 
C) for 30 min. After cooling to room temperature in an 
ultrasonic bath, filtration of the products obtained 
through filters with a pore diameter of 100 nm. Then, 
samples were taken for chromatography. The results of 
these studies are shown in Table 4. 
From the presented table shows that the most in-
teresting result is the direct interaction of 99mTc eluate 
solution with a solution of a mixture of sodium chloride 
and DTPAmod tin containing a certain amount of Sn. 
The output labeled nanocolloids based on the modified 
molecules DTPAmod with particle size less than 100 nm 
was 80% with a total content of radiochemical impuri-
ties  unreduced 99mTc (VII) in the resulting formulation 
2.7% (Fig. 1). This is a fairly good indicator, closely 
matching the regulatory requirements for these drugs. 
When interacting with 99mTc eluate dry residues de-
rived from alcohol mixtures containing DTPAmod and 
SnCl2, radiochemical impurity contents  unreduced 
99mTc (VII) obtained in preparation nanocolloid was 
about 6% and the yield nanocolloids labeled with a size 
less than 100 nm was 74%. 
 
Table 4 − The results studies 
 
Ingredients of 
mixture 
Activity, 
imp 
Contents 
99mTc(VII), % 
Yield 
% 
[DTPAmod +Sn(II)] 
solution +99mTc 
550 965 3,5  
After heating 484 552 3,0  
Filtrate 100 387 642 2,7 80 
[DTPAmod 
+Sn(II)]dry.+99mTc 
545 128 7,0  
After heating 476 179 5,8  
Filtrate 100 352 372 4,8 74 
 
 
 
Fig. 1 − Radiochromatogramma 99mTc elution with a solution 
of a mixture of DTPAmod and SnCl2 
 
Medico-biological tests of the colloidal drug on the 
basis of modified molecules DTPAmod labeled with 
99mTc were carried out at the Research Institute of 
Oncology, SB RAMS Tomsk city, on the white line of 
male rats "Wistar" with the mass of 300-350. The ani-
mal's body scintigram obtained at various time inter-
vals are shown in Figure 2. 
On the scintigrams of 60 and 120 minutes is a clearly 
visible sentinel lymph node, located between the bladder 
and the place of injection. The level of accumulation of 
the drug in the lymph node was 3.67% of the total intro-
duced activity, which is enough for its safe visualization. 
This result exceeds to the standard requirements for 
such drugs (0.5 - 1.7%) and proves the functional suita-
bility of our synthesized labeled with technetium-99m, 
nanocolloid on the basis of modified molecules DTPAmod. 
 
 V. S. SCURIDIN, E.S. STASYUK, E. A. NESTEROV ET AL. PROC. NAP 2, 04NABM02 (2013) 
 
 
04NABM02-4 
 
 
Fig. 2 − The drug  distribution in the body of rat  at the introduction of a nanocolloid [DTPAmod  и SnCl2]: A) Immediately after 
injection, B) 30 minutes after injection, C) 60 minutes after injection, D ) 120 minutes after injection. The numbers  is indicated: 1 
- lymph node, and 2 - Location of the drug 
 
4. CONCLUSION 
 
1. The research process of restoration of 99mTc in 
the presence of stannous Sn (II) have been studied. 
As a result,  it has been found that the optimum 
concentration of the reducing agent, Sn (II) in the 
radiopharmaceutical must be in the range from 
0.00875 to 0.0175 mg / ml. 
2. The conditions for obtaining radioactive nano-
preparations on the basis of the modified molecule 
DTPA. It is shown that the direct interaction of 99mTc 
eluate solution with a solution of a mixture of SnCl2 
DTPAmod and radiochemical impurity content unre-
duced 99mTc (VII) in the preparation labeled with 
Nanocolloids is 2.7% and the yield nanocolloids with 
the size of 100 nm is about 80%. 
3. Medico-biological tests of  labeled 99mTc on the 
molecules DTPAmod  have been carried out on rats in 
order to study their distribution in the organism of 
the experimental animals and determine the func-
tional suitability for the scintigraphic imaging of 
lymph nodes. The level of accumulation of the drug in 
the lymph node is 3.67% of the total introduced activ-
ity, which is enough for its safe imaging. 
This work has been carried out in the frame of 
state program «Sciense» № 3.1192.2011. 
 
 
RERERENCES 
 
1. P. Reimann, Phys. Rep. 361, 57 (2002).  
2. S. Maza et al., QJ  Nuc.Med 30/5, 651 (2003). 
3. C.B. Sampson, Textbook of Radiopharmacy theory and 
Practice (London, United Kingdom: Gordon and Breach; 
1994). 
 
А 
B C D 
1 
2 


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Preparation and Medico-Biological Tests Nanocolloids Radiopharmaceuticals on the Based of Modified DTPA Labeled Technetium-99m

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