The research area is in the field of pharmaceutical analysis and in
particular it concerns the development and application of
chromatographic techniques (HPLC and GC-MS) and capillary
electrophoresis (CE) in analysis of bioactive compounds.
The specific subjects of research are:
1) Development and application of electrokinetic systems based on
complex pseudostationary phases (mixed micelle,
micelle-cyclodextrin, microemulsions, proteins).
2) Development and application of chromatographic methods in
analysis of bioactive compounds in complex samples.
3) Development and application of capillary isoelectric focusing for characterization of therapeutic proteins (in particular monoclonal antibodies).
1. Development and application of electrokinetic systems based
on complex pseudostationary phases.
Improved separation selectivity in electrokinetic systems can be
achieved by supplementing the electrophoretic running buffer with
additives able to establish reversible interactions with the
solutes to be separated. Among the additives, surfactants are very
successful; they are used at concentration higher the critical
micelle concentration (cmc). Under these condition a micellar
pseudostationary phase is obtained. In this field the research
activity has been addressed to the development of new micellar
systems based on mixed micelle (from different surfactants) such as
sodium dodecyl sulfate (SDS) and bile salts. Further, the role of
cyclodextrins as chiral additives in electrokinetic systems has
been considered. Finally, microemulsion based on oils such as
heptane and octane in the presence of surfactants and
co-surfactants, have been applied as separation pseudostationary
phases. The possible role of the components of the considered
pseudostationaty phases can be hypothesized by means of the
evaluation of characteristic chromatographic parameters such as
selectivity, resolution, capacity factor, and efficiency;
furthermore, also physico-chemical parameters such as zeta
potential, ionic radius of the micelle aggregates,
inclusion/complexation constants and kinetic constants can be
determinated in order to characterize the complex systems.
The use of proteins as additives to the running buffer belongs to
this research field; proteins can be considered as useful chiral
selectors.
In general the development of all of these described complex
systems can be advantageously carried out by means of chemometric
and Quality by Design approaches: this part of the research
activity is in collaboration with a team of the
University of Florence. The proposed CE systems allowed
to achieve interesting and useful methods for related substances
analysis in pharmaceuticals.
2. Development and application of chromatographic methods (HPLC,
GC-MS, CE and CE-MS) in analysis of drugs and biomoelcules
(including metabolites) in complex real samples.
The development, validation and application of chromatographic
methods in pharmaceutical analysis is a field of the present
research. Sensitive, reproducible and accurate determination
of the active components require for efficient analytical
techniques such as GC-MS, HPLC-MS and CE, also used in integrated
approaches and in combination with sample prep methods as solid
phase microextraction (SPME). In addition a new approach to
the hyphenation CE-MS is developing for the direct
insertion of the CE effluent to the electron impact
(EI) source.
3. Capillary isoelectricfocusing (CiEF)
CiEF is developed and applied to study and characterize the charge variants of therapeutic proteins and in particular of monoclonal antibodies, allowing the determination of the isoelectric points of the charge variants. This characterization is of fundamental importance in the quality control of biotechnological drugs
