Banca de DEFESA: ANA ROBERTA PEREIRA JOHNSON DOS ANJOS

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : ANA ROBERTA PEREIRA JOHNSON DOS ANJOS
DATE: 26/03/2026
TIME: 09:00
LOCAL: VIDEOCONFERÊNCIA
TITLE:

Quality-by-Design Approach in the Development of a Green Method by UHPLC-ESI-TOF-MS for the Simultaneous Analysis of the Synthetic Peptides Stigmurin, Stiga8, TsAP-2, and TISTH

KEY WORDS:

analytical quality by design; method development; peptides; Tityus stigmurus; UHPLC-MS-ESI-TOF.

PAGES: 165
BIG AREA: Ciências da Saúde
AREA: Farmácia
SUMMARY:

The growing therapeutic interest in peptides derived from the venom of the Tityus stigmurus scorpion and its analogs demands the development of robust, selective, and reproducible analytical methods for their simultaneous characterization. In this context, this thesis aimed to develop and optimize a high-performance chromatographic method for the simultaneous separation of four synthetic peptides (Stigmurin, two analogs, and TistH) using the Analytical Quality by Design (AQbD) approach. Initially, a risk assessment was conducted to identify the critical quality attributes and operational parameters impacting analytical performance. Subsequently, Design of Experiments (DoE) was applied in sequential screening and optimization stages, considering variables such as column type, mobile phase composition, temperature, and flow rate. Analyses were performed using ultra-high-performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry (UHPLC-ESI-ToF-MS). Experimental data were processed via analysis of variance (ANOVA) and statistical modeling, with the construction of response surfaces to determine the Method Operable Design Region (MODR). The results demonstrated that columns featuring core-shell technology exhibited superior performance, providing higher chromatographic efficiency, adequate peak symmetry, and resolution greater than 2 for all critical pairs. The organic mobile phase composition was identified as the factor with the greatest influence on chromatographic resolution and retention time, with an optimal range established between 40% and 43% (v/v). Additionally, robust operational conditions were defined, involving temperatures between 20 and 40 °C and flow rates between 0.25 and 0.40 mL min⁻¹, enabling analytical run times between 6 and 20 minutes. The statistical models showed significance, high coefficients of determination, and adequate predictive capacity. The assessment of analytical sustainability indicated that the developed method meets the principles of green chemistry. It is concluded that the application of AQbD enabled the development of a flexible, robust, and high-performance analytical method, applicable to peptide analysis and potentially extensible to other peptide mixtures, contributing to the assurance of quality, reliability, and reproducibility in bioanalytical assays.

COMMITTEE MEMBERS:
Presidente - 1492900 - CICERO FLAVIO SOARES ARAGAO
Externa ao Programa - ***.920.424-** - DAYANNE LOPES PORTO - UFRN
Interno - 1544647 - MATHEUS DE FREITAS FERNANDES PEDROSA
Externo à Instituição - RUI OLIVEIRA MACEDO - UFPB