A ready-to-use kit for the preparation of99mTc-phytate for liver scanning

Kits were developed-for the sterile labelling of phytate with^99mTc. The effect of molar ratio of phytate to stannous chloride, pH, dilution of the Sn-phytate with^99mTc-generator eluate, time of incubation, the shelf life of^99mTc-phytate and the storage time of Sn-phytate on the labelling yield of phytate with^99mTc was investigated using paper chromatography and gel chromatography column scanning method (GCS). Organ distribution was performed in rabbits and mice. Excellent human liver scans were obtained.     

Radiochemical quality control of99mTc-labelled radiopharmaceuticals

Two methods for the determination of radiochemical purity of preparations labeled with^99mTc are described. Paper chromatography was used for separation of reduced^99mTc and free pertechnetate from the labeled radiopharmaceutical. Whatman 3MM paper was used first with a acetone and then with 1N NaCl, as the mobile phase. Instant thin layer chromatography was performed for comparison. The electrophoretic method was also applied, using 0.05 M Na-acetate and 0.017 M NaCl. Biodistribution of^99mTc-DMSA in the organs of experimental animals was followed in order to verify chemical control methods.     

New99mTc-diiodine substituted IDA derivative (DIIODIDA) for hepatobiliary imaging

A new diiodine substituted IDA derivative, 2,4-diiodine-6-methyl IDA (DIIODIDA) was synthesized and labeled with^99mTc. It was established that^99mTc-DIIODIDA had high radiochemical purity. Biodistribution and influence of bilirubin on^99mTc-DIIODIDA biokinetics has been studied in rats and compared to the corresponding results for^99mTc-SOLCOIODIDA. Related to^99mTc-SOLCOIODIDA,^99mTc-DIIODIDA has much better biliary exretion (55.18 versus 43.63%). No change of^99mTc-DIIODIDA biokinetics, under influence of bilirubin was noticed. Biliary excretion of^99mTc-SOLCOIODIDA has been reduced for about 60%. The protein binding of^99mTc-DIIODIDA and^99mTc-SOLCOIODIDA were also determined, using in vitro method of precipitation. These results showed that^99mTc-DIIODIDA hepatobiliary imaging agent is superior to the presently used^99mTc-monoiodine IDA derivatives.     

Statistical analysis of the results of99mTc-MDP quality control

The methods used for control of radiochemical purity of^99mTc-MDP are presented. TLC method on silica gel, developed with methanol and acetone (11 v/v), was convenient for determination of^99mTcO ₄ ^– with the content of 2.6±1.2%. The reliable results on detection of^99mTc hydrolyzate (2.2±1.3%) and for another^99mTc-MDP complex (13.2±2.8%) were obtained by application of ITLC (SA), developed with Sn-MDP. By Sephadex G-25 column chromatography (1.5 cm×5 cm) the separation of^99mTcO ₄ ^– was not achieved. The range of normal^99mTc-MDP biodistribution values in the organs of experimental animals have been determined. The mean value of bone distribution was 8.4±1.13%/g, in muscles 0.071±0.033%/g, while uptake in liver and kidneys was below 5%. Chi-square test and P show that the results on biodistribution of^99mTc-MDP in liver, bones and muscles are arranged around their mean values, which is statistically allowed.     

Studies of the chemical and biological properties of the bone and acute myocardial imaging agent99mTc-PYP

Correlation between the in vivo distribution and the chemical formulation of^99mTc-PYP complex has been studied. We chose mice to evaluate in vivo biodistribution and gel chromatography column scanning technique for radiochemical analysis. The influence of the pH, Sn(II), pyrophosphate concentration and molar ratios of Sn: PYP on the labelling of pyp with^99mTc has been investigated in vitro and in vivo. The reaction between^99mTc and Sn-PYP was complete within a few minutes. The stability studies were evaluated against dilution. Induced myocardial infarction was evaluated in rats. The clinical evaluation showed excellent definition of sternum and ribs with little blood background activity with normal subjects. Discrete localization of abnormally high activity was shown in the site of recent infarction of the left ventricular myocardium.     

Effect of chemical condition of technetium-99m sodium pertechnetate on the active kit components and radiolabeling yield of Tru-Scint® ADTM monoclonal antibody kit

The chemical condition of^99mTc eluate obtained from a⁹⁹Mo-^99mTc generator is a function of the source, time elapsed after elution and age of the eluate. The radiochemical purity and stability of^99mTc labeled MAb-170 (Tru-Scint^®AD^TM, photoactivated monoclonal antibody kit) preparations was evaluated comparing pertechnetate source of known age and elution history. The effect of H₂O₂, a radiolytic impurity in^99mTc eluates, on the active kit components stannous ion and photoactivated MAb and radiolabeling, yield has been investigated. The lyophilized Tru-Scint^® AD^TM kit has been labeled with 20 to 80 mCi in 0.5 to 4.0 ml of Sodium Pertechnetate^99mTc Injection, USP. The eluates were obtained from three brands of generators and used up to six hours after elution. The kits were reconstituted either with Sodium Pertechnetate^99mTc Injection, USP or Sodium Chloride Injection, USP, 0.9% containing known amounts of H₂O₂. The reconstituted kits were analyzed for radiolabeling yield and radiochemical impurities, stannous ion and protein sulfhydryl group. The results indicated that the radiolabeling yield is a function of both the chemical condition of^99mTc eluate, generator brand and the radiolabeling parameters like reconstitution volume and activity. The observed radiolabeling yield differences did not depend on the amount of chemical technetium in the eluate. The major radiochemical impurities at 15-minute post labeling have been identified as the^99mTc-buffer complex and column adsorbed reduced^99mTc (^99mTc-Ad) species and not the unreduced^99mTcO ₄ ^– .     

Comparative study of different gels for quality control of99mTc-radiopharmaceutical kits

The available six types of Sephadex and one type of Sepharose have been applied in the separation of technetium fractions in^99mTc-labelled radiopharmaceuticals produced in our laboratory using the GCS technique. By this technique the chemical state and the percentage of^99mTc-fractions have been determined. The resolution efficiency of some gel types were found to be significantly influenced by the pH of the eluent. The results obtained from the experiments indicated that Sephadex G-25 Fine was the best and can be routinely used in the radiochemical analysis of the following kits:^99mTc-HSA,^99mTc-DTPA and^99mTc-HIDA and G-100 for^99mTc-PYP. With^99mTc-HSA and^99mTc-PYP kits, 0.9% NaCl eluents at pH 3.2 and pH 2 to 2.5, respectively, were found to be necessary for the separation of^99mTc-fractions. G-50 Fine was found to be the best gel between the others in the separation of^99mTc-fractions in testing of the weak radiopharmaceuticals,^99mTc-GH and^99mTc-MDP. The development of^99mTc-MDP with the eluent at the same pH as the preparation gives negligible interaction effect.     

Technetium-99m dextran kit: formulation and biological behaviour

A new formulation of stannous-dextran (Sn-Dx) freeze dried kit, containing 60 mg dextran (Dx-70) and 0.08 mg SnCl₂·2H₂O, to be labelled with^99mTc, has been developed. At pH 6.5–7.0. the labelling efficiency was greater than 95%. Gel chromatography column scanning technique was applied for radiochemical purity determination of^99mTc-Dx preparation and the degree of in vivo plasma protein binding. Not less than 70% of the administered activity was bound to plasma and remained constant over a 1h period. The biological behaviour after intravenous injection of^99mTc-Dx kit was characterized by high and efficient yield of the radiopharmaceutical. The preliminarly clinical results on normal subjects showed that the radiopharmaceutical could be a useful agent for scintigraphy of leg lymph vesel, pelvic and inguinal lymph nodes. The activity uptake in liver and kidney (60 min) was relatively very low, whereas the urinal bladder activity (30 min) represents the drainage of the activity entering the blood stream after interdigital injection of^99mTc-Dx.     

Standardization of quality control methods for99mTc radiopharmaceuticals

Physico-chemical characterization of^99mTc-radiopharmaceuticals is presented. Limiting pH values, iso-osmotic pressure and the apparent coefficient values between two immiscible phases are determined too. A selection of radiochromatographic methods /stationary or mobile phase/ for routine quality control of^99mTc radiopharmaceuticals for radiochemical purity was made. The methods chosen are simple, accurate, sufficiently sensitive and fast in operation. The mean values were determined for^99mTc radiopharmaceutical distribution per organs, characteristic for the tested preparates and for radiochemical purity, as well as the time interval from injection to sacrifice of the animals.     

Quality control and statistical analysis of biodistribution of commercial99mTc-IDA derivatives

The results obtained for radiochemical purity of ITLC (SA) and (SG) using different solvent systems and low voltage electrophoresis are presented in the paper. Radiochemical purities obtained for^99mTc-dimethyl IDA (^99mTc-HIDA) and^99mTc-diethyl IDA (^99mTc-EHIDA) are 98.1±0.6% and 98.7±0.5%, respectively. Variable^99mTc hydrolyzate contents, depending on the ionic strength of the eluents and on the time interval between labelling and analysis, have been obtained by Sephadex chromatography. The eluent containing Sn-EHIDA inhibits dissociation of^99mTc-EHIDA due to dilution of the preparation during elution of the column and yielding only a small percent of Sephadex bound fraction, as compared to other investigated eluents. The range of normal^99mTc-IDA biodistribution values in the organs of experimental animals and statistical significance of the difference between these two preparations have also been determined. The results obtained for^99mTc-HIDA and^99mTc-EHIDA in the liver are 33.9±5% and 25.7±4.7%, respectively p<0.01.     

Comparative quality control of some bone-seeking99mTc radiopharmaceuticals by gel chromatography

The radiochemical purity of the three osteopatic ligands:^99mTc(Sn)-PyP,^99mTc(Sn)-DPD and^99mTc(Sn)-MDP has been determined by gel chromatography on Sephadex. The results of the analyses strongly depend on the composition of the eluent. The dilution effect of pure saline as eluent was observed in all the preparations examined. The most sensitive was found to be^99mTc(Sn)-PyP. The retention of^99mTc activity bound to the gel matrix (^99mTc-hydrolyte) was over 30%. The diphosphonates were found to be more stable (retention 10–15%). The retention is substantially lower, i.e. a high recovery of the labeled complexes is obtained when the eluent contains the ligand. The best results are obtained when the eluent contains the same concentrations of ligand and reductant as in the labeled complex. There was no significant difference in the behavior of the given radiopharmaceuticals prepared as a fresh solution and in the freeze-dried kit.     

Effect of cuprous ion on labeling of three skeletal imaging agents with99mTc

The effect of increased content of copper on the radiochemical composition of three skeletal imaging agents:^99mTc(Sn)-methylene diphosphonate (MDP),^99mTc(Sn)-pyrophosphate (PYP) and^99mTc(Sn)-2,3-dicarboxypropane-1, 1-diphosphonate (DPD) was observed only in the case of^99mTc(Sn)-MDP. It was found that the radiochemical purity of this radiopharmaceutical falls to about 50% when the copper content reaches about 10^–5 mol dm^–3. According to the results of radiochemical and biological analyses, it could be concluded that with the increase of copper content, the content of free pertechnetate rises, too. The two other radiopharmaceuticals,^99mTc(Sn)-PYP and^99mTc(Sn)-DPD, were found to be stable under the given experimental conditions.     

Kinetic analysis of99mTc-d, 1-HMPAO decomposition and the method of stabilization

The kinetics of^99mTc-d, 1-HMPAO decomposition is studied using home kits. The results showed that^99mTc-d, 1-HMPAO decomposition is a first-order reaction. The decomposition constant k is found to be 0.017±0.007h^–1 under the experimental conditions of 20°C, 185MBq/ml, pH 7.0. The stability of^99mTc-d, 1-HMPAO is affected not only by pH and radioactive concentration, but also by temperature. Using Immol/l gentisic acid as a stabilizer, 740MBq/ml of^99mTc-d, 1-HMPAO can be stabilized for 3h with the radiochemical purity above 80%.     

A novel technique for the separation of99mTc from99Mo

A new technique for the separation of^99mTc from low specific activity⁹⁹Mo is reported. A separation based on the principle of precipitation of⁹⁹Mo as calcium molybdate has been investigated. On precipitating⁹⁹MoO ₄ ^2– from alkaline solution as calcium molybdate under controlled conditions, the^99mTcO ₄ ^– is found to remain quantitatively in the supernatant solution with little carry-over of⁹⁹Mo. This calcium molybdate (⁹⁹Mo) could be redissolved and reprecipitated at regular intervals, yielding^99mTc quantitatively in aqueous neutral solutions. Calcium molybdate precipitates containing up to 1.5 GBq of⁹⁹Mo and 130–180 mg of molybdenum were prepared and evaluated. The performance in terms of repeated^99mTc separation gave yields of 75–93% with acceptable readionuclidic and radiochemical purity.     

99mTc-dimercaptosuccinic acid: Analytical procedures for its radiochemical quality control

A procedure for the radiochemical purity control of^99mTc-(2,3-dimercaptosuccinic acid) (DMSA), used as a renal scintigraphic agent is described. The proposed chromatographic system entails the use of two successive solvents, first MEK and second aqueous solution of 5% glycine, on the same supporting medium Gelman ITLC-SG. The procedure is fast and leads to separation and estimation of free pertechnetate, hydrolyzed form of^99mTc and^99mTc — DMSA. This system is superior to the others reported in the literature as the spots of the different species are more distinguished and more concentrated. Its reliability has been studied using dimercaptosuccinic acid kits of different manufacturers and the results have been checked biologically.     

The quality of99mTc eluates

The possible effects of several protecting procedures on the quality of^99mTc eluates were investigated. The content of⁹⁹Mo in the eluates (⁹⁹Mo breakthrough) was expressed in (%) with respect to the total adsorbed⁹⁹Mo radioactivity and in () i.e. as the ratio of⁹⁹Mo and^99mTc radioactivities in each particular eluate. The radiochemical purity was expressed in (%) of^99mTc(VII) in the eluates. The content of Al³⁺ and Cu²⁺ as chemical impurities was also determined.     

Labeling of ursodeoxycholic acid with technetium-99m for hepatobiliary imaging

An adopted method for the preparation of high radiochemical purity ^99mTc-ursodeoxycholic acid (UDCA) was conducted with a high radiochemical yield up to 97.5 %. The reaction proceeds well using 2 mg UDCA, 50 μg tin chloride in solution of pH 8 at room temperature for 30 min. The radiochemical yield was up to 97.5 % as pure as ^99mTc-UDCA. Different chromatographic techniques (paper chromatography and electrophoresis) were used to evaluate the radiochemical yield and purity of the labeled product. Biodistribution studies were carried out in Albino Swiss mice at different time intervals after administration of ^99mTc-UDCA. The uptake of ^99mTc-UDCA in the liver gave the chance to diagnose it. The results indicate that the labeled compound cleared from the systematic circulation within 2 h after administration and majority of organs showed significant decrease in uptake of ^99mTc-UDCA. Finally, the liver uptake was high and the results indicate the possibility of using ^99mTc-UDCA for hepatobiliary imaging.     

Preparation of Sn-β-alanine: A new freeze-dried kit to be labelled with99mTc for liver scintigraphy

Sn-alanine kits were prepared in lyophilized form containing 7.02·10^–2 M -alanine and 5.5·10^–4M stannous chloride dihydrate. The optimal pH value of the preparation was found to be equal to 4.3–5.1. The radiochemical purity and the stability of^99mTc-alanine were assessed by gel filtration column scanning techniques (GCS) and thin layer chromatography, and the labelling yield of the complex was higher than 95%. The organ distribution data in mice showed that more than 90% of the injected dose had been accumulated in the liver. However, a negligible amount of radioactivity was detected in the non-target organs. The stability of^99mTc-alanine was followed for 5 hours and the Sn-alanine kit was stable for at least 3 months.     

Comparative radiochemical and kinetic study of tumorotropic and renal99mTc-DMSA

A procedure for obtaining a stable^99mTc(V)-DMSA kit and methods for its radiochemical and biological control are described. The effect of pH on radiopharmaceutical stability of the complex was studied. The kinetic parameters of^99mTc(V)-DMSA were determined on rats and compared to the corresponding values for renal^99mTC-DMSA. Clinical tests showed that^99mTc(V)-DMSA is suitable agent for detecting the primary medullar carcinoma of thyroid, as well as for detection of thyroid metastasis.     

Radio TLC in quality control and development of radiopharmaceuticals

Radio TLC has been used for determining the radiochemical purity and composition of two^99mTc-radiopharmaceuticals, available as kits and commonly used for diagnostic imaging. Moreover, the same technique has been applied to detect impurities which decrease the specific activity of¹³¹I-derivatized dermatan sulfate, a new potential radiopharmaceutical, and for establishing the best labeling conditions.