QUANTITATIVE DETERMINATION OF METHYL-4-CHLOROBUTYRATE, A POTENTIAL GENOTOXIC IMPURITY, CONTENT IN MOXIFLOXACIN HCL BY GC-EI-MS

K. APARNA; K. VIJAYA RACHEL; K. M. V. NARAYANA RAO

doi:10.22159/ijap.2024v16i5.51551

Authors

K. APARNA Department of Biochemistry/Bioinformatics, GIS, VSP, GITAM University, Visakhapatnam-530045, India https://orcid.org/0000-0002-2239-8641
K. VIJAYA RACHEL Department of Biochemistry/Bioinformatics, GIS, VSP, GITAM University, Visakhapatnam-530045, India https://orcid.org/0000-0001-7099-3915
K. M. V. NARAYANA RAO Analytical Research and development, Aragen Life Sciences Pvt Ltd, Hyderabad-5000076, India https://orcid.org/0000-0002-8339-1750

DOI:

https://doi.org/10.22159/ijap.2024v16i5.51551

Keywords:

Fluoroquinolone, Moxifloxacin, Methyl-4-Chlorobutyrate, Genotoxic impurity, Quality control, GC-EI-MS

Abstract

Objective: Methyl-4-Chlorobutyrate (M4CB), a genotoxic impurity, was identified in the active pharmacological components of the fourth-generation fluoroquinolone, moxifloxacin (MXFN). There has not yet been a report on the analysis of the M4CB impurity content in the MXFN molecule. Consequently, a Gas Chromatography-Electron Ionization-Mass Spectrometry (GC-EI-MS) method was established that has the ability to identify and measure M4CB impurity content at ppm level.

Methods: The column exploited in M4CB impurity assay was a Dura Bond 624 (DB-624) type stationary column. Temperatures of 220 °C and 280 °C were consistently maintained at the injection and detection sites, respectively. The helium, as carrier gas, in split mode with ratio of 1:7 was used. The column's flow rate remained steady around 2.0 ml/min. The mass spectrometer was operated in Single Ion Monitoring (SIM) mode at m/z = 74.

Results: The impurity M4CB is generated during the manufacturing process of cyclopropanamine, which is an intermediary molecule in the manufacturing process of MXFN. This new GC-EI-MS approach can measure the M4CB at 0.9452 ppm in MXFN samples with a 500 mg/ml concentration following International Council for Harmonisation (ICH) standards. Very low quantification limits (0.9452 ppm), high linearity (range=0.945 ppm to 5.625 ppm; regression coefficient= 0.9999), and a reasonable recovery range (94.60-94.63%) were all provided by this new validated GC-EI-MS approach. Three batches were analysed for M4CB content by new GC-EI-MS approach and found that none of the batches contained M4CB impurity.

Conclusion: The GC-EI-MS approach has excellent applicability in the quality assurance testing of MXFN for M4CB content since it was adequate in terms of linearity, precision, sensitivity, accuracy, specificity, and robustness.

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