Avaliação da robustez de um método analítico por cromatografia líquida de ultra eficiência desenvolvido para quantificação de stigmurina utilizando Box-Hehnken design

Abstract

Stigmurine is an antimicrobial peptide of pharmaceutical interest that has been gaining prominence for the production of new medicines. However, its a recently elucidated molecule and little has been reported in the literature about analytical methodologies aimed at its quality control. Thus, the aim of this paper is perform a study of the robustness of a method by Ultra High Performance Liquid Chromatography (UHPLC) developed for the quantification of Stigmurine using box-behnken design (BBD) as design of experiments (DoE) and response surface analysis and desirability function as statistical tool for data analysis. The method proved to be robust for the quantification of stigmurin with Relative Standard Deviation (%DPR) < 2% for the response chromatographic peak area, however it was possible to observe that the %FMB factor influences the alteration of the system's suitability parameters (time of retention, resolution and tailing factor) more significantly than the other factors, but has little influence on the chromatographic peak area. The method under analysis had a predominantly desirable function (D) > 0.5 for the tested regions, and the regions that promote greater desirability are close to the highest value for %FMB and lowest values for temperature and flow, reaching up to a value of D > 0.7.