In Vitro

1
In Vitro Metabolic Mapping by NMR Spectroscopy
Part 1 of the Four-Dimensional Project
D. Duong Hau, Jiangoux Xia, Dr. David Wishart
Introduction
The tremendous amount of research produced within
the fields of genomics, proteomics, and
metabolomics over recent years have provided a
wealth of data concerning the biology of a
variety of organisms. However, our understanding
of the data its implications within disease
mechanisms, the relationships between data sets,
or even the significance of the data remains
highly in need. The ultimate goal of the 4D
project is to alleviate this need by developing
software that enables one to visually display
experimental data on a model organism, showing
changes over time and space. Currently, the
project is focused on generating in vitro
metabolomic maps of plasma and specific organ
tissues in the rat. Specifically, the metabolite
composition and concentration of the samples will
be determined and confirmed by NMR spectroscopy
and mass spectrometry. Data acquired for the in
vitro maps will be used to refine and test the
visualization models.
Visualizing the Pharmacokinetics
Of Acetylsalicylic Acid
Metabolic Studies of Hypoxic-Ischemic Damaged Rats
In addition to the metabolic study of rats, we
are also cooperating with Dr. Andrei Turinsky
(University of Calgary) to produce a program that
allows one to visualize the movement of
metabolites through the body over time. To date,
we have compiled the kinetic parameters involved
in the absorption, metabolism, and excretion of
acetylsalicylic acid and its derivatives. We
have used these parameters to produce theoretical
4D data that Dr. Turinsky has used to test his
human model software.
In collaboration with Dr. Jerome Yager
(University of Alberta), we have begun research
delving into the metabolic effects of
hypoxic-ischemic brain injury in rats.
Currently, we are developing a protocol for
sample preparation and analysis. Samples include
blood plasma, cerebral spinal fluid, and brain
tissue. A 1D proton NMR spectrum is collected for
each sample. The NMR data is processed and
analyzed using the program Chenomx NMR Suite 4.5,
which contains a library metabolites and their
respective NMR information. This allows one to
identify metabolites and their concentrations for
each sample. We will compare samples from normal
and brain-injured rats. Moreover, we will
analyze rats at different time points after
injury to map the metabolic changes that occur
over time. This research should provide a solid
basis for further four-dimensional studies.
Figure 2. Above are four screenshots from Dr.
Turinskys software, which will be able to
visually display distribution data of metabolites
or drugs over time.
Figure 1. A screenshot from the Chenomx NMR
Suite software. A 1D proton NMR spectrum rat
plasma is presented.
Figure 3. A graph depicting the theoretical
abundance of aspirin and its derivatives within
different compartments of the body over the
course of 4 hours.