An alkaline kit formulation to obtain [99mTc]MAG3 in high radiochemical yields

An alkaline kit formulation (pH 9) to obtain [^99mTc]MAG₃ with radiochemical puritives over 98% has been developed, avoiding the addition of filtered air to the vial, the use of large amounts of^99mTc activity (i.e., 3.7 GBq) or the reconstitution of large volumes. The use of this radiopharmaceutical in mice showed a minimal accumulation in the hepatobiliary system (0.37±0.3% I.D., 1 h postinjection). However, in rabbits we always obtained good image quality.     

Technetium-99m-diethyl-IDA instant kit: Labelling kinetics and biological properties

A formulation of stannous-diethyl-IDA freeze-dried kit, containing 50 mg diethyl-IDA and 0.4 mg hydrated stannous chloride, to be labelled with technetium has been developed for hepatobiliary scintigraphy. Gel chromatography column scanning technique has been applied for determination of technetium fractions in the preapration. The optimal pH value of the preparation with a high hepatobiliary specificity was found between 5.5 to 6.0. Effect of ligand to metal ratio on the stability of the prepared Sn-diethyl-IDA solution prior mixing with technetium has been investigated. The reaction conditions (initial pH) between the ligand and stannous ion affects the percent hydrolysis of the labelled compund, i.e. the formation rate of the complex. The organ distribution data of^99mTc-diethyl-IDA in mice for 60 minutes post injection were satisfactory. The radiopharmaceutical exhibits rapid blood clearance, great hepatic clearance and very short hepatocyte transit time. Uptake of the radiopharmaceutical in various organs of mouse at 5 minute post injection showed that the greater part of the injected radioactivity has distributed between liver and intestine. However, about 12% and 60% of the injected dose has been found in liver and intestine respectively. The renal uptake of the HB agent in mice is relatively low and decreases with time to become about 0.3% at 60 minutes post injection. Blood clearance data of the radiopharmaceutical kit in rabbits showed that the HB agent is-rapidly cleared, since the initial decrease was very fast with a half-time of 1.5 minutes.     

Preparation and evaluation of99mTc-thiodiglycolic acid (TDGA) for renal function studies

Methods for the preparation and analysis of a new renal radiopharmaceutical,^99mTc-thiodiglycolic acid (TDGA), are reported. The kit for Tc-TDGA contains a lypophilized acidic (pH 2.5) mixture of 5 mg TDGA and 0.05 mg SnCl₂·2H₂O. Acetate buffer has been found to be a suitable solvent for paper chromatography of^99mTc-TDGA. The results of the quantitative organ distribution studies in rats and rabbits indicated the characteristics of^99mTc-TDGA to be intermediate to the renal tubular agent¹³¹I-Hippuran and the GFR agent^99mTc-DTPA.Parts of this work have been presented at the Indo-US Seminar on Radiopharmaceuticals-cum-17th Ann. Conf. of Soc. Nucl. Med. (India) held at Bangalore in Jan. 1986 and at the 18th Ann. Conf. of Soc. Nucl. Med. (India held at Madras in Dec. 1986. Also a part of this work has been included in the dissertation of N. Ramamoorthy submitted to the University of Bombay for the Ph. D. degree in chemistry.     

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.     

A kit of human polyclonal IgG for the diagnosis of infectious processes

The aim of this work was to obtain a freeze-dried kit for direct^99mTc-labeling of human polyclonal IgG. The labeling procedure was carried out by Schwarz's method. The best yields of^99mTc-IgG were obtained by using sodium pyrophosphate decahydrate as a weak chelating agent. Performed tests showed the stability of the radiopharmaceutical up to 24 hours. Plasma clearance in rats was fitted to a biexponential curve withT _1/2α=(0.1 ±0.9) h andT _1/2β=(10±3) h. The organs with higher uptake of radiopharmaceutical were lung, kidneys and blood. In a rabbit model the abscess target/background ratio was 3–6 according to time of the scintigraphic images. Thirty patients with musculoskeletal infection were studied. Twenty-one lesions were detected and confirmed by culture/biopsy.     

Preparation,molecular modeling and biodistribution of 99mTc-phytochlorin complex

Phytochlorin [21H, 23H-Porphine-7-propanoicacid, 3-carboxy-5-(carboxymethyl)13-ethenyl-18-ethyl-7,8-dihydro-2,8,12,17-tetramethyl-,(7S,8S)] was labeled with ^99mTc and the factors affecting the labeling yield of ^99mTc-phytochlorin complex were studied in details. At pH 10, ^99mTc-phytochlorin complex was obtained with a high radiochemical yield of 98.4 ± 0.6 % by adding ^99mTc to 100 mg phytochlorin in the presence of 75 μg SnCl₂·2H₂O after 30 min reaction time. The molecular modeling study showed that the structure of ^99mTc-phytochlorin complex presents nearly linear HO–Tc–OH unit with an angle of 179.27° and a coplanar Tc(N1N2N3N4) unit. Biodistribution of ^99mTc-phytochlorin complex in tumor bearing mice showed high T/NT ratio (T/NT = 3.65 at 90 min post injection). This preclinical study showed that ^99mTc-phytochlorin complex is a potential selective radiotracer for solid tumor imaging and afford it as a new radiopharmaceutical suitable to proceed through the clinical trials for tumor imaging.     

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 ^– .     

Effect of experimental conditions on the in vitro stability of99mTc (Sn)-pyrophosphate

The in vitro stability of^99mTc (Sn)-PyP as a function of experimental conditions of the preparation of the kit and time elapsed after labeling has been tested. The preparation was protected by using nitrogen-purged reactant solutions and kit vials and by ascorbic acid. The samples under nitrogen are stable for 6 h when the content of^99mTc-pertechnetate raises to 5%. The best stability was achieved by addition of 5 g of ascorbic acid per ml of the kit (content of^99mTc-pertechnetate about 0.5%). To accelerate the decomposition, exogenous hydrogen peroxide was used. In this case it was found that the presence of 10 g of ascorbic acid inhibits the effect both of oxygen and peroxide (6 g H₂O₂/ml of the kit). Radiochemical purity of^99mTc (Sn)-PyP remains practically unchanged for 6 h (content of^99mTc-pertechnetate about 0.5%).     

99mTc-DMSA complex preparation: the effect of pH and tin(II) chloride amount on reaction

Labelling of meso-2,3-dimercaptosuccinic acid (DMSA) with technetium-99m was reinvestigated. Dependence of the ^99mTc-DMSA complex formation on the molar ratio of DMSA:reducing agent (SnCl₂·2H₂O) and pH was studied. Five different types of ^99mTc-DMSA complexes were determined. Especially three different complexes were established in the clinically used and prepared DMSA kit labelled with ^99mTc under alkaline condition. This radiopharmaceutical is used as imaging agent of the primary medullary carcinoma in the thyroid gland and different metastasis types. The existence of all complexes was observed by paper chromatography, paper electrophoresis and high performance liquid chromatography.     

Comparative evaluation of four different99mTc-sulfur colloid kits used for liver scanning

The distribution of four different commercially available A, B, C, D kits (^99mTc-sulfur colloid) for hepatoimaging was compared in mice by organ radioassay and in rabbits for blood clearance. The distribution of kits A and C (single step kits) was assessed in the human by blood clearance, external liver, spleen measurement, and scintillation camera imaging. Kit A reaches a high concentration in liver within 15–20 minutes with relatively high surrounding tissue background, and superior spleen scintiphotos. However, when kit C was used, a high activity concentration in the liver was reached within 10–15 minutes with low tissue background and faint visualization of the radiotracer in the spleen. Blood clearance of the four^99mTc-sulfur colloids was determined in rabbits. The data obtained indicated that the four hepatoagents exhibit rapid blood clearance but the initial decrease of blood activity curve of kit D was relatively faster than the other three hepatic agents. The biodistribution is similar for the four^99mTc-S-colloids but the blood retained higher activity residue using kit A compared with others. The formation of^99mTc-sulfur colloid using kits B, D (multistep kits) involves many steps after the addition of^99mTcO ₄ ^– to the reagent. These procedures are time consuming, required facilities at the medical institutions and give rise to the radiation exposure. While single step kits A and C have the same diagnostic value, the use of kit C allows a reduction of absorbed radiation, which may be useful in the liver exploration in children.     

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.     

The effect of selected preparation variables on the radiochemical purity of <Superscript>99m</Superscript>Tc-EDDA-HYNIC-TOC

[^99mTc-EDDA-HYNIC-D-Phe¹, Tyr³]-Ocreotide (^99mTc-EDDA-HYNIC-TOC) increasingly emerges to be an alternative tool for somatostatin receptor (SSTR) scintigraphy of neuroendocrine tumours. The high quality of this radiopharmaceutical and its uniformity are very important facts for application of this preparation in clinical practice. Various factors may influence the radiochemical purity (RCP) of certain reagent kits. Some of these include the amount of activity added to the reagent kit, heating time and the age of the formulated kit. The effect of these factors on RCP of ^99mTc-EDDA-HYNIC-TOC has been investigated using high performance liquid chromatography (HPLC) and instant thin layer chromatography (ITLC).     

The stability of99mTc phosphorus compounds in plasma both in vivo and in vitro

The in vivo and in vitro stability of^99mTc hydroxyethlylidene diphosphonate, 99^mTc methylenediphosphonate and^99mTc pyrophosphate in plasma has been studied using paper chromatographic technique as the analytical tool. The results indicate that the amounts of^99mTc activity found both at the origin and R_f range of^99mTcO₄ ^? for in vivo experiments are slightly greater than those for either in vitro or control experiments. However, this amount of^99mTc activity represents about 0.16–0.4% of the injected dose. Therefore, it is suggested that^99mTc phosphorus radiopharmaceuticals are stable in vivo and neither oxidation nor hydrolysis of these bone imaging agents occurs in the blood.     

Labelling and quality control of cysteine with99mTc and biological distribution

An instant kit of cysteine (amino acid) to be labelled with^99mTc was prepared. Optimal conditions were found, and a procedure to prepare the kit ready to use in liophilized form to gain the highest labelling yield. More than 95% labelling yield was obtained when^99mTc (TcO ₄ ^– ) eluted from^99mTc-generator was added to the contents of the kit. Each kit contains 0.66 mg of SnCl₂·2H₂O as stannite and 66 mg cysteine in lyophilized form. The formulation of cysteine tin (kit) was stable for nearly three months giving labelling yield more than 95%. Using GCS technique, different species of technetium and labelled cysteine were identified when Sephadex (G-50, G-25) was applied. Biodistribution of the labelled preparation revealed that kidney was the target organ. The ratio of accumulated dose in kidneys/liver was greater than 2.     

HPLC investigation of the radiochemical purity of <Superscript>99m</Superscript>Tc-MIBI

Technetium-99m-sestamibi (^99mTc-MIBI) is a small, lipophilic and cationic compound used for myocardial perfusion imaging. In addition, ^99mTc-MIBI has been shown to be useful in identifying several types of tumors, such as breast, lung and thyroid cancers. The high quality of this radiopharmaceutical and its uniformity are very important facts for application of this preparation in clinical practice. The monograph for ^99mTc-MIBI recommends at least 90% radiochemical purity (RCP) for clinical use. Various factors may influence the RCP of certain reagent kits. Some of these include the amount of activity added to the reagent kit, heating time and the age of the formulated kit. The effect of these factors on RCP of ^99mTc-MIBI has been investigated using high performance liquid chromatography (HPLC) and instant thin layer chromatography.     

Biodistribution of A14C-and99mTc-labeled N-substituted nitrosourea (CCNU) in an animal tumor model

One formulation of¹⁴C labeled and another of^99mTc labeled 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) were administered i.v. to tumor (glioma) bearing rats. The radiopharmacokinetics of¹⁴C-CCNU were followed up to 24 hours post injection. On a per organ basis the blood, liver, small bowel, kidney cortex and muscle contained most of the activity. Optimum tumor to organ ratios occurred at 4–12 hours. The^99mTc-CCNU biodistribution was determined at 4 hours and compared to^99mTc-NaTcO₄. Tumor capsule to brain (29.5) and to muscle (10.59) ratios suggest^99mTc-CCNU to be a potential tumor seeking agent. Funded in part by USPHS Grant RR-05486-12.     

Preparation of 99mTc-cefoperazone complex, a novel agent for detecting sites of infection

Prompt localization of infection sites is essential for initiating appropriate therapeutic measures. There have been major advances in the management of patients suffering from infective and/or inflammatory disorders as a result of introduction of newer drugs with high sensitivity and specificity. Since the last decade, ^99mTc-ciprofloxacin was used as a biologically active radiopharmaceutical to diagnose inflammation but it has some problems related to radiochemical purity and stability. The aim of this study is to develop simple and easy formulation of cefoperazone (other broad spectrum antimicrobial agent) with ^99mTc a ready to use labeling kit for infection imaging. The optimum condition that gives high labeling yield of ^99mTc-cefoperazone complex, 97.9%, was achieved by using 3 mg cefoperazone, 100 μg Sn(II), at pH 8 and 10-minute reaction time. For in vivo binding of ^99mTc-cefoperazone pharmacokinetic studies were carried in experimentally induced infection, in the left thigh, using Staphylococcus aureus in rats. Both thighs of the rats were dissected and counted and the ratio of bacterial infected thigh/contralateral thigh was then evaluated. The time for maximum accumulation of ^99mTc-cefoperazone at the site of infection (T/NT = 4.5) was 45-minute post intravenous injection, followed by gradual decline. So, ^99mTc-cefoperazone complex is simple and stable preparation for infection imaging after 45-minute post injection.     

Comparison of some physicochemical and pharmacokinetical parameters of 99mTc-PAH, 99mTc-MAG3 and 131I-OIH

The purpose of this study was to compare some physicochemical characteristics as well as pharmacokinetic behavior of ^99mTc-PAH, as a novel renal agent, with ^99mTc-MAG₃ and ¹³¹I-OIH. ^99mTc-PAH was prepared from lyophilized kit by adding ^99mTcO₄ ^–. Labeled complex was stabile and high radiochemical purity radiopharmaceutical, with a low percentage of protein bound to human albumin and hydrophilic character. In spite of its smaller renal uptake, ^99mTc-PAH gave satisfactory renal images. ^99mTc-PAH showed faster urinary elimination than ^99mTc-MAG₃ and similar to those one for ¹³¹I-OIH. The comparison of pharmacokinetic parameters of ^99mTc-PAH, ^99mTc-MAG₃ and ¹³¹I-OIH indicated the favorable characteristics of ^99mTc-PAH.     

Labeling of famotidine with 99mTc and biodistribution studies on rats

Famotidine, a gastrointestinal drug was labeled with ^99mTc and its radiopharmaceutical potential was observed on male Albino Wistar rats. Labeling yield was over 95%. Average specific activity and n-octanol/water partition coefficient (lipophilicity) were approximately 74 MBq/mg-0.66 GBq/mg and 3.4, respectively. Biodistribution studies were performed on Albino Wistar rats. The activity per gram tissue was calculated and time-activity curves were generated. The majority of the activity was observed in stomach, small intestines and kidneys. Liver and kidneys showed lower uptake compared to other H₂-receptor rich tissues. Results show that ^99mTc-famotidine is H₂receptor specific. This revised version was published online in July 2006 with corrections to the Cover Date.     

Exchange labeling of proteins:99mTc labeled transferrin

^99mTc-labeled transferrin was prepared by reduction of^99mTcO ₄ ⁻ ; with stannous DTPA or stannous citrate followed by equilibration of the technetium chelate with human transferrin. The rate of transfer of^99mTc to transferrin in the presence of 0.015M citrate buffer was dependent on pH in the order pH 2.1> pH 7.2> pH 4.1> pH 5.9. The incorporation rate was inversely proportional to the concentration of DTPA and citrate buffer. The replacement of citrate buffer by acetate buffer or oxalate buffer reduced drastically the formation of^99mTc-labeled transferrin at pH 4.1. The formation of^99mTc-labeled transferrin prepared from the reduction of^99mTcO ₄ ⁻ with stannous citrate was faster than that from the reduction with stannous DTPA in the presence of 0.015M citrate buffer and pH 2.5. Equilibration of transferrin with^99mTc-labeled pyrophosphate did not produce^99mTc-labeled transferrin at pH 4.5. The ligand exchange labeling of^99mTc to transferrin in 0.015M citrate did not cause appreciable denaturation of the protein at all pH values. This method also enabled labeling of the protein in a low concentration (2.6·10⁻⁴ M) via tin reduction. Sequential external imaging of the^99mTc-labeled transferrin in Sprague-Dawley rats bearing Walker-256 carcinosarcoma showed optimal tumor localization occurred at 3 hr after injection. In spite of this,^99mTc-labeled transferrin does not appear to be a suitable imaging agent because of the low tumor to blood ratio of^99mTc (0.50±0.17) at 3 hr post injection. This is similar to that of^{6 7}Ga-citrate (0.43±0.15%).