Identification and characterization of related substances in EVT-401 by hyphenated LC-MS techniques

  • J Pharm Anal. 2017 Aug;7(4):223-230. doi: 10.1016/j.jpha.2017.03.008.
Binan Zhu  1 ,  Yuting Lu  1 ,  Leilin Chen  2 ,  Binbin Yu  3 ,  Yuexin Liu  1 ,  Min Song  1 ,  Taijun Hang  1
Affiliations
  • 1. Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education; Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China.
  • 2. Department of Chemical Drug Control, Hubei Institute for Food and Drug Control, Wuhan 430074, China.
  • 3. Department of Pharmacy, Taizhou First People's Hospital, Taizhou 318200, China.
PMID: 29404042 DOI: 10.1016/j.jpha.2017.03.008
Abstract

A sensitive and selective method was developed for the separation and characterization of related substances (RSs) in EVT-401 by hyphenated LC-MS techniques. Complete separation of the RSs was achieved with an Inertsil ODS-SP column (250 mm×4.6 mm, 5 µm) by linear gradient elution using a mobile phase consisting of 0.2% formic acid solution, methanol and acetonitrile. EVT-401 was found to be susceptible to acid, alkaline and oxidative stresses, while relatively stable under photolytic and thermal dry stress conditions. Fourteen RSs including six process-related substances and eight degradation products were detected and identified in EVT-401 with positive ESI high-resolution TOF-MS analysis of their parent ions and the corresponding product mass spectra elucidation, and some of them were further verified by chemical synthesis and NMR spectroscopy. The specific LC-MS method developed for separation, identification and characterization of RSs is valuable for EVT-401 manufacturing process optimization and quality control.

Keywords
Degradation products; EVT-401; LC–MS; Related substances.
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