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In-Depth Information
urinary samples,
3
e
6
conjugation of reduced
thiols with various maleimide analogs was
employed for increasing the ionization ef
ANALYSIS OF METABOLITE
PROFILES USING CAPILLARY
ELECTROPHORESISeMASS
SPECTROMETRY
ciency
in conjunction with online sample preconcentra-
tion, allowing for sensitive and comprehensive
determination of thiol redox status in plasma.
7
Furthermore, various puri
Capillary Zone Electrophoresise
Electrospray IonizationeMass
Spectrometry
Capillary zone electrophoresis (CZE) resolves
analytes based on their differences in electropho-
retic mobility, which is a function of the charge-
to-size ratio. CZE equipped with a two-spectral
channel laser-induced
cation approaches,
including methanol deproteinization, ultra
ltra-
tion, and solid phase extraction, were used prior
to the analysis of metabolite pro
les in human
HT29 colon cancer cells.
8
Important differences
were observed in the metabolomic pro
les
obtained from solid phase extraction and meth-
anol deproteinization samples, indicating poten-
tial bias as the result of different puri
uorescence detector has
been employed for the simultaneous studies of
two glycosphingolipid metabolic pathways in
single primary neurons with unparallel detec-
tion sensitivities and at least six orders of magni-
tude of dynamic ranges.
1
Due to its high
throughput and excellent resolving power, the
coupling of CZE with electrospray ioniza-
tion
e
mass
cation
strategies.
A mouse multiple-tissue metabolome data-
base, including the analyses of cerebra, cerebella,
thymus, spleen, lung, liver, kidney, heart,
pancreas, testis, and plasma from a single
mouse, was developed using CZE-ESI-MS.
9
Matrix-assisted
(ESI-MS)
e
enabled
spectrometry
laser
desorption/ionization
metabolomic pro
ling of single cells and subcel-
lular structures such as single R2 neuron and
metacerebral cell from Aplysia californica.
2
CZE is particularly applicable for the separa-
tion of highly charged and hydrophilic metabo-
lites that may not be retained in reversed-phase
liquid chromatography (RPLC). The identi
time-of-
ight imaging mass spectrometry
(MALDI-TOF-IMS) was combined with CZE-
ESI-MS to determine contents of individual
metabolites in serial tissue sections obtained
from livers of super immune de
cient mice.
10
The combination of MALDI-TOF-IMS with
CZE-ESI-MS was further employed for the visu-
alization of spatiotemporal energy dynamics of
hippocampal neurons by the analysis of
energy-related metabolites during a kinetic-
induced seizure.
11
CZE-ESI-MS was demonstrated for perform-
ing quantitative metabolome pro
ed
metabolic features in human urine generally
exhibit an m/z value in the range of 50 to 150
using CZE-ESI-MS. In contrast, 95% of the attri-
butes detected by LC-MS have an m/z value
above 150. CZE-ESI-MS therefore appears to be
highly complementary to LC-MS, in providing
the characterization of different
ling of colon
and stomach tumor tissues.
12
Quanti
classes of
cation of
94 metabolites in colon and 95 metabolites in
stomach involved in glycolysis, the pentose phos-
phate pathway, the tricarboxylic acid (TCA) and
urea cycles, and amino acid and nucleotide
metabolisms resulted in the identi
metabolites.
3
CZE-ESI-MS was demonstrated for the search
of metabolic indicators from urine in chronic
patients with complex regional pain syndrome,
4
the comprehensive pro
ling of free estrogens
and their glucuronide/sulfate conjuagtes,
5
and
the discovery of urinary biomarkers for hepato-
toxicity induced by drug therapy or exposure
to toxicants.
6
cation of
several cancer-speci
c metabolic traits. For the
quanti
ed
standards, a multivariate strategy was intro-
duced to derive a quantitative relationship
cation of metabolites without puri
In addition to the analysis of
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