
The Brodbelt research program involves the development of quadrupole
ion trap mass spectrometry for a variety of interdisciplinary
applications related to bioanalytical, organic, and inorganic
areas. Research efforts are currently focused in three
areas: A) evaluation of aspects of molecular recognition by
development of new mass spectrometric methods, B) development
of
photodissociation methods for the structural characterization of
peptides, proteins, and DNA, and C) the application of
extraction, separation and mass spectrometric strategies for the
identification and quantification of phytochemicals in food, urine and
plasma for studies of biotransformation and bioavailability.
A)
Molecular recognition
The phenomenon of molecular recognition
is important
in many biological and chemical systems, such as those responsible for
drug actions, enzyme catalysis, and ion transport. Advances
in
the field of molecular recognition have stimulated a growing need for
new analytical methods for characterization of the structures of
host-guest complexes and determination of binding
selectivities.
One method which has shown great promise for the analysis of
supramolecular assemblies and more complex quantitative applications is
electrospray ionization-mass spectrometry (ESI-MS). In the
ESI
method, complexes formed in solution are sprayed into the gas phase,
and the types of complexes and distribution of complexes are
analyzed. This method has been used to study the binding
properties of lariat ethers, calixarenes, molecular clefts, crown
ethers, and a variety of other novel macrocycles. In recent
years, we have been examining DNA interactive agents to identify the
binding sites and stoichiometries of drug/DNA complexes and to measure
the sequence and drug selectivity.
B)
Infrared Multiphoton Dissociation
for Analysis of Biological Molecules
Infrared
multiphoton dissociation
is being developed to characterize the structures of biological
molecules, with special emphasis on peptides, identification of peptide
modifications, and sequencing of DNA. Infrared photodissociation has
proven to be a universal way to energize ions so that they dissociate
via structurally-informative pathways, and ways to make this method
more energy-tunable are under development.
C) Analysis of
Phytochemicals in
Food, Urine and Plasma
Phytochemicals, such as flavonoids,
limonoids, and
anthocyanins, are chemopreventive compounds found naturally in plants
that exert positive health effects as dietary antioxidants. Recent
epidemiological and clinical studies have provided strong evidence that
phytochemicals offer protection against cancer, heart disease, and
aging-related problems. Citrus fruit, kale, onions, berries,
tea
and soy are some of the best natural sources of flavonoids. Our goal is
the development of electrospray ionization/quadrupole ion trap mass
spectrometry for the sensitive detection of phytochemicals in foods
like citrus and kale, and in plasma and urine after consumption, thus
allowing the metabolism, uptake, distribution, and bioavailability of
these compounds to be mapped. Electrospray ionization, in conjunction
with tandem ion trap mass spectrometry and photodissociation methods,
are being developed to characterize the structures of complex
molecules, to detect trace quantities and metabolites in biological
fluids, and to distinguish isomers. The use of metal
complexation
is being explored as an alternative to conventional protonation as a
means to efficiently generate ions.