Title:
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Probing reactions at the liposome/H2O interface with bi-functional spin labels
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In this thesis we will present the results obtained in a research project aimed at using EPR
spectroscopy to determine the kinetic properties of reactions takfug place at the interface
between H20 and an amphiphilic supramolecular ag~egates. The choice of EPR
spectroscopy is related mostly to its high sensitivity (the absolute EPR lower limit is around
10-12 M of spins). The possibility to measure the diffusion parameters and flexibility of the
molecules making up the amphiphilic aggregate was also anothe[ aspect that appe.aled to us: .
it was possible, in a single experiment, to determine a series ofdata that would have required
at least two separate spectroscopic methods to access.
Chapter I reviews all the relevant literature in the EPR field as well as of phospholipid
membranes and their characteristics with a particular attention on the reactions taking place
at the H20/amphiphilic aggregate interphase.
The design and preparation of the molecules involved in this investigation is discussed in
Chapter IT along with the characterization of the model molecules used for the optimization
of the synthetics passages involved in the prepar~tion of the spin probes employed in the
subsequent analyses.
In Chapter III the problem of the characterization of the liposomes is addressed. Dynamic
Light Scattering and EPR line shape simulation results are presented along with cw-EPR
biradical distance measurement procedures for the determinati~n of the distance between the
two nitroxides embedded in the hydrophobic phase.
The kinetic analyses -for the thiol-disulphide exchange reaction are discussed in Chapter
IV. Preliminary studies for the optimization of the analytical protocol are presented for the
reaction taking place in homogenous phase in MeOH and in the presence of phospholipid
amphiphilic aggregates. Along with the data collected for egg yolk phosphatidylcholine with
a 0-1 molar ratio ofcholesterol, systems employing micelles and synthetic phospholipids are
presented in various conditions of pH. All kineti~s were recbrded under pseudo 1st order
conditions using different concentrations of2-mercaptoethanol.
Chapter V describes the results obtained using spin labels in the characterization of
organogelators. Monoradical and biradic~l systems were employed in order to compare the
different kinds ofinformation from these two different nitroxide systems.
In Chapter VI all the experimental procedures are reported along with the corresponding
characterization for the target molecules prepared.
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