•Pre-concentration of Pb-203 and Pb-212 via extraction chromatography resin was optimized for automated radiopharmaceutical production.•An HPLC isolation of Pb-203 and Pb-212 labeled peptides from unlabeled precursors produced specific activities near theoretical maximums.•The approach is robustly adaptable to automation and a cassette-based system was customized for clinical radiopharmaceutical production.A method for preparation of Pb-212 and Pb-203 labeled chelator-modified peptide-based radiopharmaceuticals for cancer imaging and radionuclide therapy has been developed and adapted for automated clinical production. Pre-concentration and isolation of radioactive Pb2+ from interfering metals in dilute hydrochloric acid was optimized using a commercially-available Pb-specific chromatography resin packed in disposable plastic columns. The pre-concentrated radioactive Pb2+ is eluted in NaOAc buffer directly to the reaction vessel containing chelator-modified peptides. Radiolabeling was found to proceed efficiently at 85 °C (45 min; pH 5.5). The specific activity of radiolabeled conjugates was optimized by separation of radiolabeled conjugates from unlabeled peptide via HPLC. Preservation of bioactivity was confirmed by in vivo biodistribution of Pb-203 and Pb-212 labeled peptides in melanoma-tumor-bearing mice. The approach has been found to be robustly adaptable to automation and a cassette-based fluid-handling system (Modular Lab Pharm Tracer) has been customized for clinical radiopharmaceutical production. Our findings demonstrate that the Pb-203/Pb-212 combination is a promising elementally-matched radionuclide pair for image-guided radionuclide therapy for melanoma, neuroendocrine tumors, and potentially other cancers.

Metastatic melanoma is the most aggressive and lethal form of skin cancer. Emerging evidence suggests that differences in melanoma metabolism relative to nonmalignant cells represent potential targets for improved therapy for melanoma. Specifically, melanoma cells exhibit increased mitochondrial electron transport chain (ETC) activity and concomitant hyperpolarized mitochondrial membrane potential relative to nonmalignant cells. We have synthesized several new fluorescent lipophilic vinylpyridinium cations built from tetraarylethylene scaffolds that target mitochondria via attraction to the hyperpolarized mitochondrial membrane potential. Mitochondria-specific accumulation in melanoma cells relative to normal human fibroblasts was demonstrated using confocal fluorescence microscopy and resulted in the disruption of oxidative metabolism leading to melanoma specific cell death in vitro. Thus, the pyridinium tetraarylethylene platform represents a promising new mitochondrial-targeted delivery vehicle with potential imaging and therapeutic properties.

Naturally-occurring radioactive materials (NORM) associated with unconventional drilling produced fluids from the Marcellus Shale have raised environmental concerns. However, few investigations into the fundamental chemistry of NORM in Marcellus Shale produced fluids have been performed. Thus, we performed radiochemical experiments with Marcellus Shale produced fluids to understand the partitioning behavior of major radioelements of environmental health concern (uranium (U), thorium (Th), radium (Ra), lead (Pb), and polonium (Po)). We applied a novel radiotracer, 203Pb, to understand the behavior of trace-levels of 210Pb in these fluids. Ultrafiltration experiments indicated U, Th, and Po are particle reactive in Marcellus Shale produced fluids and Ra and Pb are soluble. Sediment partitioning experiments revealed that >99% of Ra does not adsorb to sediments in the presence of Marcellus Shale produced fluids. Further experiments indicated that although Ra adsorption is related to ionic strength, the concentrations of heavier alkaline earth metals (Ba, Sr) are stronger predictors of Ra solubility.

We have developed a new chromatographic method to efficiently separate and isolate neptunium (Np) and protactinium (Pa), based on the selective extraction of protactinium by primary alcohols. The effectiveness of the new technology is demonstrated by efficient separation of 233Pa from parent radionuclide 237Np, using a hydrochloric acid mobile-phase medium. Our new approach reproducibly isolated 233Pa tracer with a yield of 99 ± 1 % (n = 3; radiochemical purity 100 %) and enabled chemical recovery of 237Np parent material of 92 ± 3 % (radiochemical >99 %) for future 233Pa tracer preparations. Compared to previous methods, the new approach reduces radioactive inorganic and organic waste; simplifies the separation process by eliminating cumbersome liquid–liquid extractions; and allows isolation of radiochemically-pure fractions in less than 1 h.

Analysis of stable gallium in nuclear materials has applications in nuclear fuel characterization and nuclear forensics. The use of positron-emitting gallium isotope 68Ga as a tracer for Ga recoveries for analyses in materials containing actinides was explored. A radiochemical method for the separation of Ga, Pu, U, Th, and Am using commercially-available extraction chromatography resins was developed and evaluated. The method effectively allows precise determination of Ga yield (97 ± 3 %) in the analysis of stable Ga (spike recovery 101 ± 1 %) and radioactive Pu (radiochemical yield, 82 ± 10 %; spike recovery, 96 ± 3 %), while also providing pure elemental fractions of other actinides relevant to materials encountered in the analysis Pu-containing materials.

The rapid proliferation of horizontal drilling and hydraulic fracturing for natural gas mining has raised concerns about the potential for adverse environmental impacts. One specific concern is the radioactivity content of associated “flowback” wastewater (FBW), which is enhanced with respect to naturally occurring radium (Ra) isotopes. Thus, development and validation of effective methods for analysis of Ra in FBW are critical to appropriate regulatory and safety decision making. Recent government documents have suggested the use of EPA method 903.0 for isotopic Ra determinations. This method has been used effectively to determine Ra levels in drinking water for decades. However, analysis of FBW by this method is questionable because of the remarkably high ionic strength and dissolved solid content observed, particularly in FBW from the Marcellus Shale region. These observations led us to investigate the utility of several common Ra analysis methods using a representative Marcellus Shale FBW sample. Methods examined included wet chemical approaches, such as EPA method 903.0, manganese dioxide (MnO2) preconcentration, and 3M Empore RAD radium disks, and direct measurement techniques such as radon (Rn) emanation and high-purity germanium (HPGe) gamma spectroscopy. Nondestructive HPGe and emanation techniques were effective in determining Ra levels, while wet chemical techniques recovered as little as 1% of 226Ra in the FBW sample studied. Our results question the reliability of wet chemical techniques for the determination of Ra content in Marcellus Shale FBW (because of the remarkably high ionic strength) and suggest that nondestructive approaches are most appropriate for these analyses. For FBW samples with a very high Ra content, large dilutions may allow the use of wet chemical techniques, but detection limit objectives must be considered.

Ribonucleic acid (RNA) aptamers with high affinity and specificity for cancer-specific cell-surface antigens are promising reagents for targeted molecular imaging of cancer using positron emission tomography (PET). For this application, aptamers must be conjugated to chelators capable of coordinating PET-radionuclides (e.g., copper-64, 64Cu) to enable radiolabeling for in vivo imaging of tumors. This study investigates the choice of chelator and radiolabeling parameters such as pH and temperature for the development of 64Cu-labeled RNA-based targeted agents for PET imaging. The characterization and optimization of labeling conditions are described for four chelator–aptamer complexes. Three commercially available bifunctional macrocyclic chelators (1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid mono N-hydroxysuccinimide [DOTA-NHS]; S-2-(4-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid [p-SCN-Bn-NOTA]; and p-SCN-Bn-3,6,9,15-tetraazabicyclo [9.3.1]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid [p-SCN-Bn-PCTA]), as well as the polyamino-macrocyclic diAmSar (3,6,10,13,16,19-hexaazabicyclo[6.6.6] icosane-1,8-diamine) were conjugated to A10-3.2, a RNA aptamer which has been shown to bind specifically to a prostate cancer-specific cell-surface antigen (PSMA). Although a commercial bifunctional version of diAmSar was not available, RNA conjugation with this chelator was achieved in a two-step reaction by the addition of a disuccinimidyl suberate linker. Radiolabeling parameters (e.g., pH, temperature, and time) for each chelator–RNA conjugate were assessed in order to optimize specific activity and RNA stability. Furthermore, the radiolabeled chelator-coupled RNA aptamers were evaluated for binding specificity to their target antigen. In summary, key parameters were established for optimal radiolabeling of RNA aptamers for eventual PET imaging with 64Cu.

A strain-induced copper-free click reaction mediated by a new and easily prepared cyclooctyne derivative was used to efficiently assemble a DOTA−biotin adduct capable of radionuclide (68Ga) uptake. This synthetic strategy offers a potentially general and convenient means of preparing targeted radiolabeling and radiotherapeutic agents.

Attachment of DOTA to a novel monofluoro-cyclooctyne facilitates bioconjugation to an azide-modified peptide via Cu-free click chemistry. The resulting conjugate was radiolabeled with 111In to afford a potential targeted molecular imaging agent with high specific activity and an excellent radiochemical purity.

•Lead-203 can be used as a radiochemical tracer for lead-210 analysis.•Elimination of stable lead carrier allows for smaller amounts of resin and reagents.•Lead-203 tracer allows for the analysis of complex matrices for lead-210.•Liquid scintillation counting may be used and possesses important benefits.Determination of Pb-210 (210Pb) in aqueous solution is a common radioanalytical challenge in environmental science. Widely used methods for undertaking these analyses (e.g., ASTM D7535) rely on the use of stable lead (Pb) as a yield tracer that takes into account losses of 210Pb that inevitably occur during elemental/radiochemical separations of the procedures. Although effective, these methods introduce technical challenges that can be difficult to track and potentially introduce uncertainty that can be difficult to quantify. Examples of these challenges include interference from endogenous stable Pb in complex sample matrices; contamination of stable Pb carrier with 210Pb; and high detection limits due to counting efficiency limitations. We hypothesized that many of these challenges could be avoided by the use of the electron-capture, gamma-emitting isotope, 203Pb as a chemical yield tracer in the analysis of 210Pb.A series of experiments were performed to evaluate the efficacy of 203Pb as a tracer. Four different matrices were analyzed, including a complex matrix (hydraulic-fracturing produced fluids); and samples comprising less complicated matrices (i.e., river water, deionized water, and tap water). Separation techniques and counting methodologies were also compared and optimized. Due to a relatively short-half life (52 h), 203Pb tracer is effectively massless for the purposes of chemical separations, allowing for reduced chromatography column resin bed volumes. Because 203Pb is a gamma emitter (279 keV; 81% intensity), recovery can be determined non-destructively in a variety of matrices, including liquid scintillation cocktail. The use of liquid scintillation as a counting methodology allowed for determination of 210Pb activities via 210Pb or 210Po; and recoveries of greater than 90% are routinely achievable using this approach. The improved method for the analysis of 210Pb in aqueous matrices allows for the analysis of complex matrices, at reduced cost, while providing greater counting flexibility in achieving acceptable detections limits.Download high-res image (289KB)Download full-size image

•210Po accumulates in lake bottom sediments downstream of water treatment facility.•210Po approximately 10-fold higher than parent 238U.•Increased levels of 210Po appear to be a natural phenomenon.•More studies are needed to understand the bioaccumulation pathways of 210Po.Coal is an integral part of global energy production; however, coal mining is associated with numerous environmental health impacts. It is well documented that coal-mine waste can contaminate the environment with naturally-occurring radionuclides from the uranium-238 (238U) decay series. However, the behavior of the final radionuclide in the 238U-series, i.e., polonium-210 (210Po) arising from coal-mine waste-water discharge is largely unexplored. Here, results of a year-long (2014–2015) field study, in which the concentrations of 210Po in sediments and surface water of a lake that receives coal-mine waste-water discharge in West Virginia are presented. Initial measurements identified levels of 210Po in the lake sediments that were in excess of that which could be attributed to ambient U-series parent radionuclides; and were indicative of discharge site contamination of the lake ecosystem. However, control sediment obtained from a similar lake system in Iowa (an area with no coal mining or unconventional drilling) suggests that the levels of 210Po in the lake are a natural phenomenon; and are likely unrelated to waste-water treatment discharges. Elevated levels of 210Po have been reported in lake bottom sediments previously, yet very little information is available on the radioecological implications of 210Po accumulation in lake bottom sediments. The findings of this study suggest that (Monthly Energy Review, 2016) the natural accumulation and retention of 210Po in lake sediments may be a greater than previously considered (Chadwick et al., 2013) careful selection of control sites is important to prevent the inappropriate attribution of elevated levels of NORM in lake bottom ecosystems to industrial sources; and (Van Hook, 1979) further investigation of the source-terms and potential impacts on elevated 210Po in lake-sediment ecosystems is warranted.

Naturally-occurring radioactive materials (NORM) associated with unconventional drilling produced fluids from the Marcellus Shale have raised environmental concerns. However, few investigations into the fundamental chemistry of NORM in Marcellus Shale produced fluids have been performed. Thus, we performed radiochemical experiments with Marcellus Shale produced fluids to understand the partitioning behavior of major radioelements of environmental health concern (uranium (U), thorium (Th), radium (Ra), lead (Pb), and polonium (Po)). We applied a novel radiotracer, 203Pb, to understand the behavior of trace-levels of 210Pb in these fluids. Ultrafiltration experiments indicated U, Th, and Po are particle reactive in Marcellus Shale produced fluids and Ra and Pb are soluble. Sediment partitioning experiments revealed that >99% of Ra does not adsorb to sediments in the presence of Marcellus Shale produced fluids. Further experiments indicated that although Ra adsorption is related to ionic strength, the concentrations of heavier alkaline earth metals (Ba, Sr) are stronger predictors of Ra solubility.