Quality by Design application to the development of analytical procedures in the pharmaceutical and nutraceutical field for the determination of drugs and bioactive molecules

This dissertation is nested in the field of analytical method development for molecules of pharmaceutical and nutraceutical interest, with the aim to devise analytical procedures chiefly through a chemometric approach set in the Quality by Design (QbD) framework. It explored extraction techniques alongside Liquid Chromatography-Mass Spectrometry (LC-MS) and Capillary Electrophoresis-Ultraviolet detection (CE-UV) methods, all directed either at the quality control/assurance or at the process development area. The work herein presented primarily focuses on exploiting multivariate techniques in designing capillary electrophoresis methods and extraction methodologies, notably Online-SPE-LC-MS. These approaches have been applied to tackle various challenges, ranging from Quality Control (QC) in pharmaceuticals to the optimization of processes. By incorporating multivariate methods, we were able to achieve comprehensive insights from the systems being studied. This not only led to more time and cost-effective solutions but also provided a deeper understanding of the interplay between the various factors being examined. The study undertook multiple projects that leaned on multivariate techniques to enhance CE methods for simultaneous detection of active components and their related substances in medicinal products. Two projects were aimed at the multivariate development of a CE method geared towards QC analysis of the respective Active Pharmaceutical Ingredients (APIs) in commercial drug product formulations. In one venture, the multivariate approach was pivotal in setting up a rapid CE procedure tailored for quantifying APIs in a range of drug experimental formulations. Another initiative involved a multivariate-based thorough comparison and optimization of several extraction methods on a matrix saturated with polyphenols. The concluding endeavor focused on establishing a novel automated On-line SPE system, incorporating heat to boost the desorption phase, paving the way for subsequent LC-MS evaluations. The three CE investigations focused on the analysis of Trimecaine, Omeprazole, and their respective related substances, as well as Nebivolol. The extraction study delved into an extensive assortment of antioxidants, anthocyanins, and polyphenols sourced from freeze-dried bilberry. The setup of the online SPE technique was geared towards examining the breakdown products of these phenolic substances in human bodily fluids, specifically urine and plasma, while also establishing a novel thermally-assisted extraction in the LC context. Following optimization, the pharmaceutical quality control methodologies were validated in compliance with the latest International Council for Harmonisation (ICH) guidelines1–3. This verification phase was crucial in confirming the accuracy and reliability of these methods for their intended purposes. For Trimecaine and Omeprazole, the methodologies conformed to the ICH criteria for impurity and API separation and quantification. The employed multivariate strategies, strongly endorsed by those same guidelines, enabled us to delineate robust, model-driven operational zones (or Model Operable Design Regions, MODR), ensuring these methodologies were both efficient and trustworthy. The Nebivolol procedure, also developed under the aegis of ICH principles, succeeded in assessing the API across a spectrum of experimental orally dissolving tablet (ODT) formulations. Furthermore, this procedure resulted effective in observing within those same formulations the API’s long-term stability, and to follow its dissolution in different biologically relevant media, like simulated saliva and gastric fluid. As for the extraction technique enhancement focused on bilberry polyphenols, subsequent LC-MS evaluations revealed distinct polyphenolic concentration patterns between Ukrainian (the sourced freeze-dried pooled sample for the study) and Italian bilberries. The final venture centered on the design of an innovative SPE-online approach. Notably, this was the first instance in which heat was employed in order to aid in the desorption of analytes within an LC framework. Subsequently, this approach was employed to isolate and measure phenolic by-products (stemming from bilberry extract metabolism) present in human plasma and urine.