J Sci Food Agric 1998, 77, 38È44

Hydration Properties of Xylan-Type Structures:

an FTIR Study of Xylooligosaccharides
Marta Kac— ura kova ,1* Peter S Belton,2 Reginald H Wilson,2
Ja n Hirsch1 and Anna Ebringerova 2
1 Institute of Chemistry, Slovak Academy of Sciences, 842 38 Bratislava, Slovakia
2 Institute of Food Research, Norwich Laboratory, Norwich Research Park, Colney, Norwich, NR4 7UA, UK
(Received 14 October 1996 ; revised version received 13 June 1997 ; accepted 4 September 1997)

Abstract : An FTIR study of the e†ect of hydration as a function of relative

humidity on the structurally di†erent xylooligosaccharide models is presented.
The IR spectral pattern changes are strongly a†ected by the type and position of
the glycosidic linkages and reÑects the hydration properties of the models.
( 1998 SCI.

J Sci Food Agric 77, 38È44 (1998)

Key words : xylooligosaccharides ; FTIR spectroscopy ; hydration ; hemicelluloses

INTRODUCTION FTIR spectroscopy has been shown to be a powerful

Of all the polysaccharide components of plant cell walls
perhaps the least attention is paid to the molecular
structure of xylans, and very little is known about the
role of the non-cellulosic matrix polysaccharides
(Mackie 1985). Xylans largely occur as complex poly-
saccharides, the structure of which vary with the nature
and degree of branching of the b-(1 ] 4)-D-xylopyranan
main chain (Aspinall et al 1983). Xylans character-
istically carry side chains of 4-O-methyl-D-glucuronic
acid and L-arabinose. Polysaccharides usually have a
strong affinity for water, and in the solid state these
macromolecules may have disordered structures which
can easily be hydrated. The interaction of water mol-
ecules with amorphous zones increases the mobility of
macromolecular chains and hence modiÐes the swelling
and glass transition properties (Fringant et al 1995).
Water sorption studies of polysaccharide-based
materials have concentrated mainly on starch and
related compounds (Fringant et al 1995).
* To whom correspondence should be addressed.
Contract/grant sponsor : Slovak Grant Agency.
Contract/grant number : 2/4148.
Contract/grant sponsor : The British Council, Slovakia.
Contract/grant sponsor : BBSRC.
( 1998 SCI. J Sci Food Agric 0022È5142/98/$17.50.
tool for the study of the physicochemical and conforma-
tional properties of carbohydrates (Mathlouthi and
Koenig 1986 ; Wilson et al 1988 ; Gilli et al 1994 ; Kac—u-
rakova et al 1994 ; Kac—urakova and Mathlouthi 1996).
Oligosaccharides have been shown to be suitable
models for understanding the conformation in the solid
state and in aqueous solution, as well as intermolecular
interactions. b-D-Xylobiosides and xylooligosaccharides
which consist of a (1 ] 4)-linked-b-D-xylopyranoside
substituted at O-2 or O-3 with a terminal b-D-xylo-
pyranosyl or a-L-arabinofuranosyl residue were used in
a previous study (Kac—urakova et al 1994). It was shown
that the presence of a (1 ] 3)-linkage in xylan molecules
and in the corresponding oligosaccharide models had
an e†ect on the IR spectral pattern of compounds in the
freeze-dried state. These changes, occurring mainly in
the 1200È900 cm~1 region, have been shown to be
useful for xylan type recognition, and even for composi-
tional analysis. However, the origin of the e†ect has not
been elucidated. Based on this study we hypothesise
that the glycosidic linkage type, and position, could
play an important role also in the swelling and gelling
mechanism of xylans. To date, water sorption e†ects on
methylglucuronoxylan as a Ðlm has been studied using
IR spectroscopy (Kalutskaya 1988). In this system the
water sorption has been shown to inÑuence molecular
38
Printed in Great Britain
An FT IR study of xylooligosaccharides 39

orientation resulting in a more homogeneous system of
rotamers of OH side groups as reÑected by the intense
bands in the 1110È1030 cm~1 region.
In order to gain information on the interactions of
various xylan-type structures with water we have con-
tinued the investigation of xylooligosaccharides which
represent structural units of xylans particularly with
regard to the e†ect of the glycosidic linkage type and
position. The present study was undertaken with the
objective of attempting to interpret the FTIR spectra of
crystalline and amorphous solids, as well as aqueous
solution of xylooligosaccharides in order to obtain
information on their structure, especially in relation to
the glycosidic bond and the e†ect of hydration, as well
as on xylooligosaccharideÈwater interactions.
EXPERIMENTAL
Materials
Xylooligosaccharide models 1È6 containing b-D-xylo-
pyranosyl and a-L-arabinofuranosyl units were synthe-
sised as reported earlier 1, 2 and 3 (Kovac— 1980), 4
(Hirsch and Kovac— 1979), 5 (Kovac— et al 1979) and 6
(Hirsch et al 1984). Table 1 lists the compounds investi-
gated.
Methods
Amorphous Ðlms on silver chloride plate
The absorption spectra of the Ðlms equilibrated in a
controlled humidity atmosphere for 24 h were collected
on a Nicolet Magna 750 FTIR spectrometer as an
average of 256 scans and at a resolution of 4 cm~1. The
solutions of the model compounds in doubly distilled
water were dropped on the surface of a silver chloride
plate and dried over phosphorus pentoxide for 72 h.
The thin amorphous Ðlms of model compounds formed
on the surface of the plate were not completely dry in
all cases even after a prolonged drying time (up to 30
days). No time dependence of the spectra of selected
samples was found (ie shown on Fig 1) which reÑects no
inÑuence of the laboratory conditions on hydrated
samples during spectral recording.
The corresponding salt solutions and relative humidi-
ties (RH) used for equilibration were : NaBr, 22% ;
Ca(NO )2É4H O, 56% ; NaCl, 76% ; KHSO , 84% and
3 2 4
K Cr O , 98%.
2 2 7

TABLE 1
List of methyl-b-glycoside oligosaccharide models

Compound Structure

1 Xylp-a-(1 ] 4)-Xylp-(1-Me)
2 Xylp-b-(1 ] 4)-Xylp-(1-Me)
3 Xylp-b-(1 ] 2)-Xylp-(1-Me)
4 Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 2)]-Xylp-(1-Me)
5 Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 3)]-Xylp-b-(1 ] 4)-Xylp-(1-Me)
6 Xylp-b-(1 ] 4)-[Araf-a-(1 ] 3)]-Xylp-(1-Me)

Fig 1. Time dependence of spectra quality control ; sample no 3 at 76% RH.

40 M Kac— ura kova et al

RESULTS AND DISCUSSION neous system of rotamers of OH side groups, occurs in

The observed FTIR frequencies of studied models and
the tentative assignment of the absorption bands are
listed in Table 2. The spectral changes during the
hydration of the (1 ] 4)-a-D-xylobioside (1) showed fre-
quencies of the absorption bands as seen on Fig 2. The
swollen samples gave broader OH bands of lower inten-
sity but a higher intensity at 1640 cm~1 for the HOH
vibrational mode compared to dry state. With respect
to the results of Kalutskaya (1988) it can be shown that
molecular deorientation, resulting in a less homoge-
this model in two steps at 22 and 98% RH.
The characteristic FTIR spectrum of amorphous
dried (1 ] 4)-b-D-xylobioside (2) is replaced by a well-
resolved spectrum with sharp bands as the RH changes
from 22 to 84% (Fig 3). In addition to the band multi-
plets in the whole 1500È500 cm~1 range, sharp bands
also appear in the 3600È3400 cm~1 together with a
doublet at 1697 and 1635 cm~1. In agreement with the
xylan X-ray study of Atkins et al (1992) the higher
hydration results in a crystalline structure, however, at
98% RH the spectrum becomes less well resolved and

TABLE 2
Observed FTIR frequencies and band assignment of the hydration treated xylooligosaccharide
modelsa

Frequency (cm~1) No of compound Assignment Comments

3617È3461 2, 3, 6 l(O-H) Water of crystallinity

Free OH, (C4-O4)b
3378È3384 1, 2, 4, 5 l(O-H) Amorphous water, liquid
3355 3, 6 l(O-H) Water of hydration
3293È3278 l(O-H) Water of hydration
3002 CH combination tone
2988È2862 l(C-H)
1697 2 d(HOH) ?
1651 4 d(HOH) Water of crystallinity
1645 1È6 d(HOH) Adsorbed water
1635 2 d(HOH) Water of hydration
1615 3 d(HOH) ?
1477 3 d(CH) d(CH )
2
1459È1447 d(CH) d (CH )
as 3
1428È1407 d(CH)
1390È1364 d(CH) d (CH )
s 3
1336È1202 d(CH),d(OH)
1174 4 l(CC), l(CO), l(COC)
1165È1156 2, 3, 4, 6 l(CC), l(CO), l(COC)
1149 1 l(CC), l(CO), l(COC) a-g l
1125È1093 asym in-phase ring
1078È1070 3, 4, 6 l(CC), l(CO)
1049È1045 1, 2, 3, 5 l(CC), l(CO)
1039È1021 4, 6 l(CC), l(CO), d(COH)
1002 6 Araf
994È988 1È4 d(CH ), d(COH)
3
979È968 3, 4, 6 d(CH ), d(COH)
3
939 1 type 1c a-g l
906 6 ring Araf
899È895 1È5 type 2, d(C-1-H)c
882 3 (1 ] 2) g l
865, 813 6 a-g l
770 1 type 3c a-g l
677, 668 3, 4 l(CCH), d(CH)w (1 ] 2) g l
660È600 1, 2, 5, 6 l(CCH), d(CH)w
595È405 d(COC), d(OCO)

a Key : g l, glycosidic link ; l, stretching ; d, bending ;

b Mathlouthi et al (1994).
c Mathlouthi and Koenig (1986).
An FT IR study of xylooligosaccharides 41

Fig 2. FTIR spectrum of Xylp-a-(1 ] 4)-Xylp-(1-Me) (1) Ðlm on AgCl plate. Ordinate in arbitrary units.

the bands at around 3384È3300 cm~1 and 1640 cm~1
increase. At this RH the sample became a viscous liquid
and we assume that the structure collapses.
The (1 ] 2) linked b-xylobioside (3) exhibits similar
spectra to those of 2. The di†erence is that, in this case,
hydration causes the crystalline structure to change in
two discrete steps. The Ðrst occurs at 22È56% and
second between 76È84% RH (Fig 4). The dry and
hydrated crystalline forms of 4-O-methyl-D-glucuron-
oxylan have been described by Marchessault and Liang

Fig 3. FTIR spectrum of Xylp-b-(1 ] 4)-Xylp-(1-Me) (2) Ðlm on AgCl plate. Ordinate in arbitrary units.

Fig 4. FTIR spectrum of Xylp-b-(1 ] 2)-Xylp-(1-Me) (3) Ðlm on AgCl plate. Ordinate in arbitrary units.
42
(1962). The intensity of the anomeric region band at
900 cm~1 was described (Marchessault and Liang 1962)
as being of low intensity in the case of the dry crystal
and of high intensity for the highly crystallinic hydrate.
Our IR results for model 3 are thus in good agreement
with their Ðndings. Another band which might be
related to the crystalline state is that at 668 cm~1. The
OH stretching and the deformation band at 1615 cm~1
are sharp from 22 to 84% RH. Similarly to 2, at 98%
RH the compound becomes amorphous, as clearly
demonstrated in both l(OH) and d(OH) regions. In
both cases (2 and 3) water is Ðxed into the crystalline
lattice probably due to strong water mediated hydrogen
bonds.
The trimer Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 2)]-Xylp-(1-
Me) (4) has quite sharp Maxima in the 1500È500 cm~1
range, however, none of the peaks seen for 2 and 3 in
the l(OH) and d(HOH) regions are present. In addition,
an absorption band at 677 cm~1 can be seen in the well
resolved spectra of 4 mainly at 76 and 84% RH. At the
highest RH the bands become less resolved and it is
Fig 5. FTIR spectrum of Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 2)]-Xylp-(1-Me) (4) Ðlm on AgCl plate. Ordinate in arbitrary units.
Fig 6. FTIR spectrum of Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 3)]-Xylp-b-(1 ] 4)-Xylp-(1-Me) (5) Ðlm on AgCl plate. Ordinate in arbitrary
units.
M Kac— ura kova et al
assumed that the crystallinity is lost (Fig 5). No signiÐ-
cant band frequency or intensity change as a function of
hydration was observed.
The tetramer Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 3)]-Xylp-b-
(1 ] 4)-Xylp-(1-Me) (5) has one branching b-D-xylo-
pyranosyl unit in position 3. The sample has a higher
degree of polymerisation in comparison to the afore-
mentioned models. It gives spectra (Fig 6) which are
typical for amorphous compounds with fewer IR bands
when compared to those of crystalline compounds (ie
models 2 and 3). The water vapour treated samples were
viscous liquids. The swelling caused an increase of the
d(HOH) vibrational mode at 22 and 56% RH and
broadening of bands in the l(OH) regions up to 98%
RH. Beyond 56% RH the spectra are poorly resolved.
The substitution in position 3 for model 5 in compari-
son to the substitution in position 2 in model 4, shows
that a di†erent hydration process is occurring. The
absence of an inter-residual OH3 . . . O@5 hydrogen bond
in model 5 which is supposed to exist in d(1 ] 4)-D-
xylans (Nieduszynski and Marchessault 1972 ; Atkins
An FT IR study of xylooligosaccharides
1992) may contribute to the peculiar behaviour of O-3
substituted xylooligomer.
The model Xylp-b-(1 ] 4)-[Araf-a-(1 ] 3)]-Xylp-(1-
Me) (6) represents the structural unit of a-L-arabino-(b-
D-xylans). In contrast to 5 the position 3 in the xylooli-
gomer is substituted by an a-L-arabinofuranosyl unit.
The spectral changes seen in 6 as a function of RH
reÑects two discrete crystalline forms (Fig 7). In contrast
to the disaccharide model 3 at 22 and 56% RH water
adsorption occurs within an amorphous structure as
indicated by the increase of l(OH) and d(HOH) bands.
Crystallisation only starts at 76% RH and at 84% RH
is already completed. At 98% RH the structure is again
amorphous but the poorly resolved spectrum shows
higher disorder in the structure than in the dry amorp-
hous structure up to 56% RH. In this model there is no
band at 1150 cm~1 and only a weak band in the l(OH)
region. These spectral features may indicate the di†erent
hydration of the arabino-xylo-oligosaccharide structure
in comparison with the xylo-oligosaccharide models.
We may conclude that the models of the a-linked
Xylp-a-(1 ] 4)-Xylp-(1-Me) (1), the higher oligosac-
charide Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 3)]-Xylp-b-(1 ] 4)-
Xylp-(1-Me) (5), and the Xylp-b-(1 ] 4)-[Araf-b-(1 ] 3)]
-Xylp-(1-Me) (6) up to 56% RH remain in the amorp-
hous state during the hydration treatment. The Xylp-b-
(1 ] 4)-Xylp-(1-Me) (2), Xylp-b-(1 ] 2)-Xylp-(1-Me) (3),
Xylp-b-(1 ] 4)-[Xylp-b-(1 ] 2)]-Xylp-(1-Me) (4) and
partly the 6 create crystalline structures represented by
sharp bands in the 1500È500, 3600È3200 and 1700È
1600 cm~1 ranges, when hydration occurs, up to 84%
RH. However, no models remain in a crystalline state at
98% RH.
Since the model compounds 2, 3 and 6 were studied
in the previous paper (Kac—urakova et al 1994) we may
compare both results. In this previous paper the spec-
tral character of xylo-oligosaccharides and xylans at
around 1150 and 1000 cm~1 was described. Our new
FTIR experiments showed that in the IR spectra of
models 2, 3, 4 (22È84% RH) and 6 (76 and 84% RH) the
Fig 7. FTIR spectrum of Xylp-b-(1 ] 4)-[Araf-a-(1 ] 3)]-Xylp-(1-Me) (6) Ðlm on AgCl plate. Ordinate in arbitrary units.
43
bands in both regions in question are well resolved and
sharp. However, in the case of 1 and 5 (whole RH
range), and 6 (up to 56% RH) the bands disappear or
are shifted to lower frequencies in the main multiplet.
Based on our knowledge we suggest that the existence
or non-existence of the 1150 and 1000 cm~1 bands in
the IR spectra of the xylo-oligosaccharides and prob-
ably also xylan-type polysaccharides are related to
hydration. In order to support our hypothesis further
FTIR experimental work is planned on polysaccharide
models. These facts also indicate that the hydration
depends on the structural features and support the
special role of the O3 branching point discussed earlier
(Kac—urakova et al 1994).
The spectral shapes of the dried amorphous Ðlm and
the amorphous Ðlm obtained in most cases (as men-
tioned in discussion) at 98% RH are not identical. In
the former case the compounds show a higher degree of
order with a more homogenous system of rotamers of
OH side groups reÑected by the intense bands in the
1110È1030 cm~1 region.
The H O deformation mode centered at 1640 cm~1
2
occurs mainly in the amorphous state and crystalline
spectra measured in KBr and belongs to the adsorbed
water. However, new bands can be seen on both low
and high frequency side of this band in the 1700È
1600 cm~1 range in the case of the crystallised Ðlm.
They can be assigned as vibrational modes of the water
of crystallinity and water of hydration, respectively,
which also creates the sharp bands in the OH stretching
region. We assume that the sharp OH bands in the
stretching region at 3617 to 3455 cm~1 (see Figs 3, 4
and 7 and Table 2) may represent the non-hydrogen
bonded OH groups (or more) occurring in the di- and
tri-mer models 2, 3 and 6 similarly to the spectrum of
sucrose where the band observed at 3550 cm~1 has
been assigned as an O4-H4 stretching vibration
(Mathlouthi et al 1994). The splitting of the OH band
would also suggest that water is important in the crystal
structure and is highly ordered (models 2, 3 and 6 at
44
higher RH). In the case of 6 only one sharp band at
higher RH can be seen at 3510 cm~1 while no sign of a
sharp water band exists. In this case the non-hydrogen
bonded OH may be internal, within the molecule.
Another band which might be related to the crys-
talline state is at 677 and 668 cm~1 in the case of
models 3 and 4, respectively, and is probably due to
(1 ] 2) link contribution.
CONCLUSION
The water-vapour treatment on the xylo-
oligosaccharide models results in IR spectral pattern
changes which are strongly a†ected by the type and
position of the interglycosidic linkages and reÑects the
hydration properties of the models at the discrete rela-
tive humidity steps. The studied xylo-oligosaccharide
models can be divided into two categories : those which
readily crystallize into highly ordered hydrated struc-
tures (having prevalence of 1 ] 4 and/or 1 ] 2, b glyco-
sidic links), and those which do not crystallise (1 ] 3
and a-link contributions).
ACKNOWLEDGEMENTS
Authors MK, JH and AE are grateful for Ðnancial
support from the Slovak Grant Agency (grant no
2/4148) and The British Council, Slovakia (MK). PSB
and RHW thank the BBSRC for funding.
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