Development and validation of a stability-indicating LC-UV and LC-MS/MS methods for quantitative analysis of anisomycin and identification of degradation products

Ljiljana Tolić,Svetlana Grujić,Mila Laušević

doi:10.1590/s2175-97902018000217491

Abstract

Multifunctional drug anisomycin was subjected to forced degradation in accordance with International Conference on Harmonisation (ICH) guidelines for the first time. The drug was exposed to the recommended stress conditions of hydrolysis (acidic, alkaline and neutral), oxidation, thermal stress and photolysis, in order to investigate its stability. Optimized LC–MS/MS method was validated as recommended by ICH Q2(R1) guideline with respect to the specificity, accuracy, precision, limits of detection and quantitation, linearity and robustness. Anisomycin exhibited high instability under alkaline and thermal (neutral hydrolysis) conditions. It showed moderate stability under acidic, neutral, oxidative, thermal (acidic hydrolysis) and photolytic conditions, with the lowest degradation level observed in the case of light and oxidation stress. Formation of the same degradation product, identified as deacetylanisomycin, was observed under all applied stress conditions.

Highlights

Anisomycin, known as flagecidin, is a pyrrolidine antibiotic isolated from two species of Streptomyces, identified as S. griseolus and S. roseochromogenes (Sobin, Tanner, 1954)
Stability of anisomycin was examined by performing forced degradation study that includes investigation of susceptibility of the drug to hydrolysis, oxidation, light and thermal stress, in accordance with International Conference on Harmonisation (ICH) Q1A(R2) and Q1B guidelines
The simple and rapid stability-indicating Liquid chromatography (LC)–UV and LC–MS/MS methods were used for investigation of the drug degradation and identification of the degradation product

Summary

Introduction

Anisomycin, known as flagecidin, is a pyrrolidine antibiotic isolated from two species of Streptomyces, identified as S. griseolus and S. roseochromogenes (Sobin, Tanner, 1954). Anisomycin has an empirical formula of C14H19NO4 and its IUPAC name is 2-p-methoxyphenylmethyl-3-acetoxy-4hydroxypyrrolidine. In the previous studies it was shown that anisomycin exhibits multifunctional properties, it is still not clinically used. Anisomycin is a well-known inhibitor of protein synthesis (Grollman, 1967; Barbacid, Vazquez, 1974). Its effects on memory and influence on behavior have been widely reported (Cohen et al, 2006; Pena et al, 2014; Lopez et al, 2015). Anisomycin is proposed as a potential psychiatric drug. In some studies on animals it was shown that it affects protein synthesis in amygdala, a part of brain that influences memory.

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