Identification and quantitative analysis of genotoxic impurities in rifampicin: Development and validation of a targeted LC-MS/MS method for 1-amino-4-methylpiperazine

J Pharm Biomed Anal. 2025 Jan 1:252:116459. doi: 10.1016/j.jpba.2024.116459. Epub 2024 Sep 6.

Abstract

Rifampicin, essential for long-term tuberculosis treatment, requires rigorous control of non-therapeutic impurities due to their potential adverse, including mutagenic effects. Reports on control strategies for genotoxic impurities in rifampicin have been limited. This study introduced an analytical method to identify potential genotoxic impurities from the synthesis of raw materials. The structure of the 25-deacetyl-23-acetyl-rifampicin genotoxic impurity was confirmed using nuclear magnetic resonance, high-resolution mass spectrometry (HRMS), and high-performance liquid chromatography (HPLC). An HPLC-HRMS method was established and validated for detecting another genotoxic impurity, 1-amino-4-methylpiperazine, adhering to the International Council on Harmonization guidelines, which include specificity, linearity, detection and quantification limits, accuracy, precision, and robustness. These developments improve the quality control strategy for genotoxic impurities in rifampicin, ensuring product safety.

Keywords: 25-deacetyl-23-acetyl rifampicin; Genotoxic impurities; High-resolution mass spectrometer; Nuclear magnetic resonance; Rifampicin.

Publication types

  • Validation Study

MeSH terms

  • Antibiotics, Antitubercular / analysis
  • Antibiotics, Antitubercular / chemistry
  • Antibiotics, Antitubercular / toxicity
  • Chromatography, High Pressure Liquid / methods
  • Drug Contamination* / prevention & control
  • Limit of Detection
  • Liquid Chromatography-Mass Spectrometry
  • Magnetic Resonance Spectroscopy / methods
  • Mutagens* / analysis
  • Piperazines / analysis
  • Piperazines / chemistry
  • Quality Control
  • Reproducibility of Results
  • Rifampin* / analysis
  • Rifampin* / chemistry
  • Tandem Mass Spectrometry* / methods

Substances

  • Rifampin
  • Mutagens
  • Piperazines
  • Antibiotics, Antitubercular