title: HPLC method development, validation, and impurity characterization of a potent antitumor nucleoside, T-dCyd (NSC 764276).
abstract: An HPLC method for the assay of an anticancer nucleoside, 4'-thio-2'-deoxycytidine (T-dCyd, NSC 764276), has been developed and validated. The stress testing of T-dCyd was carried out in accordance with ICH guidelines Q1A (R2) under acidic, alkaline, oxidative, thermolytic, and photolytic conditions. The separation of T-dCyd from its impurities and degradation products was achieved in 40min on a Luna®Phenyl-Hexyl column (150mm×4.6mm i.d., 3μm) with a gradient elution using ammonium phosphate buffer (pH 3.85) and methanol as the mobile phase. The gradient starts from 2% and ends at 80% of methanol. Detection is by UV at 282nm. LC-QTOF/MS was used to obtain mass data for characterization of impurities and degradation products. The proposed HPLC assay method was validated for specificity, linearity (concentration range 0.25-0.75mg/mL, r≥0.9998), accuracy (recovery 98.1-102.0%), precision (RSD≤1.5%), and sensitivity (LOD 0.1μg/mL). The developed method was suitable for the quality control and stability monitoring of the T-dCyd drug substance.
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. Electronic address: jennie.wang@sri.com.
authors.forename: Jennie
authors.initials: J
authors.lastname: Wang
authors.affiliation: Pharmaceutical Resources Branch, DCTD, NCI, 9609 Medical Center Drive, 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*; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis; D004340:Drug Contamination; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards; D013872:Thionucleosides / Q000032:analysis*
chemical_list: D000970:Antineoplastic Agents; D013872:Thionucleosides; D003841:Deoxycytidine; C070453:2'-deoxy-4'-thiocytidine
keywords: 4′-thio-2′-deoxycytidine; Forced degradation; HPLC validation; Impurity and degradation product characterization; NSC 764276; T-dCyd