Регуляторные исследования и экспертиза лекарственных средств (May 2025)

Quantitative Determination of Major Active Components in Polymyxin B Medicinal Products by NMR Spectroscopy

  • N. E. Kuz’mina,
  • S. V. Moiseev,
  • I. Yu. Yakupov,
  • S. I. Kuleshova

DOI
https://doi.org/10.30895/1991-2919-2025-15-2-206-212
Journal volume & issue
Vol. 15, no. 2
pp. 206 – 212

Abstract

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INTRODUCTION. Polymyxin B is a natural multicomponent antibiotic used in its sulfate form for the treatment of infections caused by multidrug-resistant Gram-negative bacteria. Pharmacopoeial standards for polymyxin B medicinal products include limits for the sum of polymyxins B1, B2, B3, and B1-I and for individual polymyxins B3 and B1-I. Since polymyxins B1 and B2 differ in their nephrotoxicity, it is important to establish individual limits for their content in polymyxin B medicinal products. Establishing individual limits for polymyxins B1 and B2 requires determining the component composition of various polymyxin B samples. However, high-performance liquid chromatography (HPLC) may not always provide accurate results because of the lack of reference standards for individual polymyxin B components.AIM. This study aimed to investigate the possibility of quantifying polymyxins B1, B2, B3, and B1-I in polymyxin B medicinal products by nuclear magnetic resonance (NMR) spectroscopy.MATERIALS AND METHODS. The study focused on a reference standard for polymyxin B sulfate, as well as polymyxin B medicinal products from different manufacturers. NMR measurements were performed on an Agilent DD2 600 MHz NMR spectrometer. The study used high-performance liquid chromatography (HPLC) on an Agilent 1200 HPLC system as the reference method.RESULTS. The 13C NMR spectrum contained signals characteristic of polymyxins B1, B2, B3, and B1-I. The authors determined the mole fractions of polymyxins B1, B2, B3, and B1-I by the internal normalisation method using the integrated intensities of the characteristic NMR signals. The results of NMR and HPLC quantification of the specified components in polymyxin B samples practically coincided (with due regard to the confidence intervals).CONCLUSIONS. According to the results, the 13C NMR analytical procedure can identify and quantify the major active components in polymyxin B sulfate medicinal products without reference standards for individual polymyxins.

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