Wo t o In

Mass Spectrometry
rodelphini
prev io us and
reports flavanols,' also reflecting the large number of molecules eluting in that
concernizi
ESI class, in agreement with 'dins
anthocyanins, p
zone. The matrix effect can either enhance or suppress ion
reported to ionize peaks due to the presence of molecules . that facilitate or inhibit
[14,301
highlypolarmetabolitesandotherpolyphenols. Furthermore,ESIionization.
is These include salts, acids or bases at high
1291 did
concentrations,
stilbenes,
tes ettictently,[2, I 71
andas peptides, which as
may
well as carbohydrates, lipids, amines
interfere with droplet size reduction
not yield any data for non-polar metabolites i
during ionization.
c either
' However, our results t .rt dicated that
146 471

well as non-polar molecules.1311 However, our analysis

method because the extraction protocol favored the re
of strongly, moderately and weakly polar metabolites. previously reportedAPCI is less susceptible to matrix
Moreover, recent experiments have shown that APCI is
suitable for the analysis of strongly polar metabolites,
including endogenous fructose and sorbitol in human nerve effectssugtahrasninEhSuIn, laans
r321
tissues, the glucose-d2/glucose ratio in human serum and e limited matrix
plasma, [331 and the presence of monosaccharides and for the quantification of ,_
effects associated with APCI
disaccharides in matrices such as orange juice, a well-
characterized sugar solution, and a solution derived
makes this an attr serum.132-331
from the enzymatic hydrolysis of sunflower seeds. 1-141 An The greater linear range and rnor
APCI source has also been used to investigate the for the simultaneous analysis of strongly
differentiation of rmderivatized monosaccharides as an and moderately
alternative to ESI-NIS.1351
APCI is less efficient for the ionization of molecules with a by a high content of sugars active method
high molecular weight, probably due to the preferential transit polar metabolites in fruit extraacntsd,
of small molecules in a vapor and the strong fragmentation in w
strongly hi
I
MS , which also makes each product ion more difficult to chareptc).thaarraacitiepiilizaeticci organic acids, but a
identify. Although APCI is considered as a soft ionization lower content of moderately polar
method, [2/361 the intrinsic ionization mechanism and the metabolites. The more limited matrix effects
nature of the target molecule can increase the degree of
associated with AP CI m ad e it possibl e to
fragmentation.137] ESI is also considered as a soft ionization
method due to the production of molecular ions without
observe a trend in sugar accumulation
fragmentation in MS 1. Indeed, we observed a strong and tartaric acid depletion in the berries during
correlation between the ESI class and molecules with a high ripening, as well as vintage-specific effects,
rniz ratio, confirming that ESI allows the detection of which were difficult or impossible to discern
metabolites with higher molecular weights.r 21 ESI has been using the ESI method.
favored because metabolites are easier to identifyr 2'381 and the
method also covers a greater metabolic range, e.g. as
shown during investigations of the antioxidant capacity
or phenolic composition of grape berries, juices and
wines.E39121 However, moderately polar metabolites and even The tight correlation between ESI and
more for strongly polar metabolites are subject to extreme moderately and weakly polar molecules
matrix effects when an ESI source is used.r221 included a correlation with adducts. It is not
The chromatography conditions we applied were based yet clear how adducts form during ESI, and the
around the typical C18 stationary phase and a gradient of process may take place in the liquid droplets or,
water and methanol, resulting in the elution of strongly polar less likely, in the vapor state.E 481 An intriguing
molecules such as sucrose and tartaric acid at the beginning of hypothesis is that adducts form immediately
the run. These molecules were therefore ideal to compare the before the droplets are disrupted, because the
performance of APCI and ESI in terms of ionizing polar rapidly declining droplet volume facilitates
metabolites. ESI showed a greater sensitivity for these twointeractions between solute molecules. In our
standard compounds, with LODs and LOQs lower than thecase, chloride and formic acid molecules in
APCI process, suggesting that ESI would be better for the the chromatography solvents could interact
detection of sucrose and tartaric acid traces in plant matrices.with berry metabolites during droplet
However, APCI achieved a broader lin' ear range for both
evaporation, leading to the formation of
sucrose and tartaric acid, suggesting this method may be
adducts during negative ionization.
better to assess the levels of these two metabolites in different
matrices, thus allowing a comparison among samples. Similar
considerations
epicatechin and
moderately polar metabolites, such
might be referred for the if'antatvrrix.eicstiegfaafcetiecddt:
 Unfortunately, there is no validated approach to predict the amount of a specific adduct that
will be formed, either based on the molecular structure of the analyte or the ionization
conditions.1491 Moreover, the abundance of adducts varies substantially amona experimental
replicates so the resulting data are unreliableP1
APO produced fewer adducts than ESI in our experiments a phenomenon that has been
reported previously with APCI in positive ion mode.i 481 Two hypotheses have been proposed to
explain this phenomenon, based on the observation that protonation (and probably also
deprotonation) occurs in the vapor phase during APCI. The first hypothesis is that sodium
ions form inefficiently in the vapor state because sodium is difficult to vaporize, and the
second hypothesis is that sodiuin adducts are formed by weak internctions that are destroyed by
the heated APCI interface. 1481 We can neither confirm nor exclude either of these hypotheses
because the positive and negative modes of APO involve different processes. 1511
However, several studies have shown
produces adducts in ne that APCI also
gative ion mode/134'51-531 so further investigations are required to
determine the mechanism of adduction. However, we observed a reproducibility of adduct
formation among the different sam les and even b tNA
two technical
The emp replicates of a sing le sample.
Tee: the
loyment of a standard RP C18 column leads to poor retention and separation of
highly
last pastpolar
decade,compounds.
issue and In the
dettc:sle,
including amides orr different strategies have
t on o fbeen applied to the addition
all carbarnates, to the polar grow ps,'
owed a better sep a r a tions'oof t e classical
1 RP
columns
examples, pot( r embedded poiar com 0 , 1541
li
mbedded pha - p unds. For
wileyonlinelibr
ary.comijournalircm se or polar end -
caPPed phase
Rapid Corwin'''.

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