title: Characterization, HPLC method development and impurity identification for 3,4,3-LI(1,2-HOPO), a potent actinide chelator for radionuclide decorporation.
abstract: 3,4,3-LI(1,2-HOPO), 1,5,10,14-tetra(1-hydroxy-2-pyridon-6-oyl)-1,5,10,14-tetraazatetradecane), is a potent octadentate chelator of actinides. It is being developed as a decorporation treatment for internal contamination with radionuclides. Conventional HPLC methods exhibited speciation peaks and bridging, likely attributable to the agent's complexation with residual metallic ions in the HPLC system. Derivatization of the target ligand in situ with Fe(III) chloride, however, provided a single homogeneous iron-complex that can readily be detected and analyzed by HPLC. The HPLC method used an Agilent Eclipse XDB-C18 column (150 mm × 4.6mm, 5 μm) at 25°C with UV detection at 280 nm. A gradient elution, with acetonitrile (11% to 100%)/buffer mobile phase, was developed for impurity profiling. The buffer consisted of 0.02% formic acid and 10mM ammonium formate at pH 4.6. An Agilent 1200 LC-6530 Q-TOF/MS system was employed to characterize the [Fe(III)-3,4,3-LI(1,2-HOPO)] derivative and impurities. The proposed HPLC method was validated for specificity, linearity (concentration range 0.13-0.35 mg/mL, r = 0.9999), accuracy (recovery 98.3-103.3%), precision (RSD ≤ 1.6%) and sensitivity (LOD 0.08 μg/mL). The LC/HRMS revealed that the derivative was a complex consisting of one 3,4,3-LI(1,2-HOPO) molecule, one hydroxide ligand, and two iron atoms. Impurities were also identified with LC/HRMS. The validated HPLC method was used in shelf-life evaluation studies which showed that the API remained unchanged for one year at 25°C/60% RH.
authors.affiliation: Pharmaceutical Development Department, Biosciences Division, SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, United States.
authors.forename: Mingtao
authors.initials: M
authors.lastname: Liu
authors.affiliation: Pharmaceutical Development Department, Biosciences Division, SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, United States. Electronic address: jennie.wang@sri.com.
authors.forename: Jennie
authors.initials: J
authors.lastname: Wang
authors.affiliation: Pharmaceutical Development Department, Biosciences Division, SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, United States.
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Wu
authors.affiliation: Pharmaceutical Development Department, Biosciences Division, SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, United States.
authors.forename: Euphemia
authors.initials: E
authors.lastname: Wang
authors.affiliation: Chemical Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States.
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Abergel
authors.affiliation: Chemical Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States.
authors.forename: David K
authors.initials: DK
authors.lastname: Shuh
authors.affiliation: Chemical Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States; Department of Chemistry, University of California, Berkeley, CA 94720-1460, United States.
authors.forename: Kenneth N
authors.initials: KN
authors.lastname: Raymond
authors.affiliation: Pharmaceutical Resources Branch, DCTD, National Cancer Institute, NIH, 9609 Medical Center Dr., Room 4W-206, Bethesda, MD 20892, United States.
authors.forename: Paul
authors.initials: P
authors.lastname: Liu
mesh_terms: D008671:Actinoid Series Elements; D002614:Chelating Agents / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004340:Drug Contamination; D006573:Heterocyclic Compounds, 1-Ring / Q000032:analysis* / Q000737:chemistry; D011728:Pyridones / Q000032:analysis* / Q000737:chemistry; D011868:Radioisotopes; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C000594124:1,5,10,14-tetra(1-hydroxy-2-pyridon-6-oyl)-1,5,10,14-tetraazatetradecane; D008671:Actinoid Series Elements; D002614:Chelating Agents; D006573:Heterocyclic Compounds, 1-Ring; D011728:Pyridones; D011868:Radioisotopes
keywords: 3,4,3-LI(1,2-HOPO); HPLC method development and validation; Impurity and degradation product characterization; Metal chelation; NSC 749716; Speciation