title: HPLC method development, validation, and impurity characterization of a potent antitumor indenoisoquinoline, LMP776 (NSC 725776).
abstract: An HPLC method for the assay of a DNA topoisomerase inhibitor, LMP776 (NSC 725776), has been developed and validated. The stress testing of LMP776 was carried out in accordance with International Conference on Harmonization (ICH) guidelines Q1A (R2) under acidic, alkaline, oxidative, thermolytic, and photolytic conditions. The separation of LMP776 from its impurities and degradation products was achieved within 40 min on a Supelco Discovery HS F5 column (150 mm × 4.6 mm i.d., 5 μm) with a gradient mobile phase comprising 38-80% acetonitrile in water, with 0.1% trifluoroacetic acid in both phases. LC/MS was used to obtain mass data for characterization of impurities and degradation products. One major impurity was isolated through chloroform extraction and identified by NMR. The proposed HPLC assay method was validated for specificity, linearity (concentration range 0.25-0.75 mg/mL, r = 0.9999), accuracy (recovery 98.6-100.4%), precision (RSD ≤ 1.4%), and sensitivity (LOD 0.13 μg/mL). The validated method was used in the stability study of the LMP776 drug substance in conformance with the ICH Q1A (R2) guideline.
authors.affiliation: SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA. Electronic address: jennie.wang@sri.com.
authors.forename: Jennie
authors.initials: J
authors.lastname: Wang
authors.affiliation: SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA.
authors.forename: Mingtao
authors.initials: M
authors.lastname: Liu
authors.affiliation: SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA.
authors.forename: Chun
authors.initials: C
authors.lastname: Yang
authors.affiliation: SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA.
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Wu
authors.affiliation: SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA.
authors.forename: Euphemia
authors.initials: E
authors.lastname: Wang
authors.affiliation: Pharmaceutical Resources Branch, DCTD, NCI, 6130 Executive Blvd., Bethesda, MD 20892, USA. Electronic address: liup@dtpepn.nci.nih.gov.
authors.forename: Paul
authors.initials: P
authors.lastname: Liu
mesh_terms: D000970:Antineoplastic Agents / Q000032:analysis*; D052117:Benzodioxoles / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007546:Isoquinolines / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D015203:Reproducibility of Results
chemical_list: D000970:Antineoplastic Agents; D052117:Benzodioxoles; D007546:Isoquinolines; C578795:indimitecan
keywords: Forced degradation; HPLC validation; Impurity and degradation product characterization; LMP776; NMR; NSC 725776