Quality assurance of 61Cu using ICP mass spectroscopy and metal complexation

Introduction 61Cu (T1/2 = 3.33 hr, Eβ= 1.22 MeV, 61.4 %) is an attractive isotope for positron emission tomography (PET) radiopharmaceutical agents such as ATSM and PTSM. Various separation processes have been reported for the production of 61Cu on a medium cyclotron using 13–22 MeV protons on natural and enriched 64Zn target materials [1,2]. This work, investigates production of 61Cu using both natural and enriched 64Zn targets and its separation. Three types of resins were used to assess for their efficiency and speed to separate the desired 61Cu from the 66,67,68Ga and 64Zn and for the recycling of 64Zn target material. The effective specific activity of purified 61Cu, was determined by ICP-MS and its titration with various polyaza and polycarboxylate complexing ligands. Material and Methods 1. Production and Separation Targets were irradiated by proton beam of IBA cyclotron 18/18MeV via the 64Zn(p,α) 61Cu and natZn(p,x) 61Cu reactions using an enriched 64Zn foil(15×15×0.05mm, ~50 mg) and natural foil (diameter 25 mm, 0.05 mm,~ 60 mg). Thirty minute irradiations were conducted with incident proton energies between 11.7–12.0 MeV and beam currents of 20 and 40 µA. Irradiated Zn targets were dissolved in 8M HCl at 150 oC then evaporated to dryness. Trace water to the resultant residue (twice) and resultant solutions evaporated to dryness. The residue was re-dissolved in 2ml of 0.01M HCl before loading onto a Cu-resin column (FIG. 1) Zn and Ga isotopes were collectively eluted using 30 ml of 0.01M HCl. The Cu was then removed using 1.5 ml of 8M HCl and passed directly onto a cation exchange followed by an anion exchange column. An additional 3 ml of 8M HCl was used to rinse the cation exchange column and ensure quantitatively removal of Cu (II) ions. The Cu was finally eluted from the anion exchange column using 3 ml of 2M HCl. The Cu solution was heated up at 150 oC until evaporated to dryness and 61Cu final product dissolved in 400–800 μL of 0.01M HCl. 2. Specific activity of 61Cu The specific activity (GBq/µmol) of the purified 61Cu was determined by ICP-MS and compared with that determined using dota, nota and di-amsar complexing ligands. For each 61Cu production run aliquot of final solution (100 µL) was left to decay before dilut-ing to 10 mL with 10% HNO3. Decayed samples were sent to ChemCentre (Curtin University) for ICP-MS analysis. Each sample was analysed for Cu, Al, Ca, Co, Fe, Ga, Ni, Si, and Zn, which are known to compete with Cu2+ for ligand complexation. Effective specific activity of the 61Cu was deter-mined by titrating various known concentration of ligands with 61Cu solution. The method is detailed in the literature [3]. Briefly, varying concentrations of each ligand was prepared in 0.1M sodium acetate buffer pH 6.5 to a total volume 20 µL. Fixed concentration of diluted 61Cu (0.01M HCl) in 10 µL was added to each ligand solution. The mixtures were vortexed then left to incubate at the room temperature for 30 mins. Two uL aliquots were withdrawn (in triplicate) from each reaction mixture and spot-ted on ITLC –SA. [Mobile phase: 0.1M NaCl: 0.1M EDTA (9:1) for Cu2+ and diamsar mixtures: Rf <0.2 Cu-diamsar; Rf > 0.8 free Cu2+ and 0.1M sodium acetate pH 4.5: H2O: MeOH: ammonium hydroxide (20:18:2:1 v/v) for Cu2+ dota and nota mixtures: Rf >0.8 Cu-dota and Cu-nota Rf < 0.2 free Cu2+]. Complexation of the 61Cu with each ligand was complete within 30 mins at room temperature. Concentration of Cu2+ was deter-mined from the 50% labelling efficiency. Results and Conclusion 1. Production and Separation The radioisotopes production from natZn target must be minimized by the optimum proton energy to reduce a radiation dose in the final product. The excitation functions of 66,67,68Ga ,65Zn and 61Cu are shown in FIG. 2. Proton beam energy of 11.7 MeV was used for both Zn targets to minimise the production of Ga isotopes and prevent formation of 65Zn. For the enriched 64Zn target (99.30%) higher proton energy could be used for the production of 61Cu allowing for increased yields and reduce radio contaminants. Previously, we used anion and cation exchange resin as described in the literature to separate the 61Cu [1]. Unfortunately the literature method was too long (up to 3 hours) and requiring high concentration of HCl and long evaporation times compromising achievable yields [4]. Thieme S. et al., 2013 [2] reported the successful use of Cu-resin for the separation of Cu radioisotopes and it was of interest to the current work to test this material for the separation of 61Cu in our hands. A cation, anion exchange and Cu-resin were combined into closed system to separate the 61Cu within 30 mins (FIG. 1). The system is designed to contain the transfer of solutions be-tween each column using simple plunger to force solution through and between each column. This system afforded an easy, reliable and fast separation of 61Cu that could be completed within 30 min. 2. Specific activity The specific activity of 61Cu was determined using ICP-MS and by titration with three ligands is summarized in TABLE 1. The ICP-MS data show values ranging from 9.2 to 32.4 GBq/μmol for 8 production runs. Specific activity determine using nota and dota were in all cases lower than the ICP MS data indicating some interference from the other metal ion contaminates such as Fe(ii/Iii), Ni (II), Ca (II), Zn (II), Ga (III). The specific activity determine using diamsar, which is known to be highly selective for Cu(II) (and Zn(II) and Fe(III)) in the presence of alkali and alkaline earth ions gave values significantly higher effective specific activity than that obtained using ICP MS. Variations in values can be explained by presence of contaminating metal ions