Optimized analysis for related substances in spiramycin based on high performance liquid chromatography with hybrid particle column and characterization of its impurities by single heartcut two-dimensional liquid chromatography coupled with quadrupole time-of-flight mass spectrometer

• Published in: Journal of pharmaceutical and biomedical analysis 2025-01, Vol.252, p.116486, Article 116486
• Main Authors: Yao, Yuan, Wang, Qian, Lu, Yue, Zhang, Jinlin, Yao, Weifeng, Yuan, Yaozuo
• Format: Article
• Language: English
• Subjects: Acetonitriles - chemistry; Chromatography, High Pressure Liquid - methods; Drug Contamination; Impurity profiling; Related substances; Reproducibility of Results; Sensitivity and Specificity; Spiramycin; Spiramycin - analysis; Spiramycin - chemistry; Tandem Mass Spectrometry - methods; Two-dimensional liquid chromatography coupled with quadrupole time-of-flight mass spectrometer
• ISSN: 0731-7085; 1873-264X; 1873-264X
• DOI: 10.1016/j.jpba.2024.116486

This article described the development and validation of a method for spiramycin related substances based on hybrid particle column. The chromatographic conditions were as follows: water - 0.2 mol/L dipotassium hydrogen phosphate (the pH value adjusted to 9.5 using a 1 mol/L KOH solution) - acetonitrile - methanol (10: 60: 28.5: 1.5, v/v/v/v) as mobile phase A, water - 0.2 mol/L dipotassium hydrogen phosphate (pH 9.5) - acetonitrile - methanol (10: 30: 57: 3, v/v/v/v) as mobile phase B and gradient elution was performed. Compared with previous analytical methods, this method has strong specificity, excellent sensitivity and stability, which could be used for the daily testing of related substances of spiramycin. Furthermore, impurities above 0.1 % were characterized using two-dimensional liquid chromatography coupled with quadrupole time-of-flight mass spectrometer (2D LC-QTOF-MS/MS) and there were 6 impurities reported for the first time. •Optimized HPLC-UV method was developed for impurity analysis of spiramycin samples.•2D LC-QTOF-MS/MS method was developed for the characterization of impurities in spiramycin.•6 new impurities were reported for the first time.