ORIGINAL PAPER
AKD-Modification of bacterial cellulose aerogelsin supercritical CO2
Axel Russler • Marcel Wieland •
Markus Bacher • Ute Henniges • Peter Miethe •
Falk Liebner • Antje Potthast • Thomas Rosenau
Received: 8 March 2012 / Accepted: 15 May 2012 / Published online: 29 May 2012
� Springer Science+Business Media B.V. 2012
Abstract Different approaches towards hydropho-
bic modification of bacterial cellulose aerogels with
the alkyl ketene dimer (AKD) reagent are presented. If
AKD modification was performed in supercritical
CO2, an unexpectedly high degree of loading was
observed. About 15 % of the AKD was bound
covalently to the cellulose matrix, while the other part
consisted of re-extractable AKD-carbonate oligomers,
which are novel chemical structures described for the
first time. These oligomers contain up to six AKD and
CO2 moieties linked by enolcarbonate structures. The
humidity uptake from environments with different
relative humidity by samples equipped with up to
30 % AKD is strongly reduced, as expected due to the
hydrophobization effect. Samples above 30 % AKD,
and especially at very high loading between 100 and
250 %, showed the peculiar effect of increased
humidity uptake which even exceeded the value of
unmodified bacterial cellulose aerogels.
Keywords Cellulose aerogel � Bacterial cellulose �Surface modification � Hydrophobization �Supercritical carbon dioxide � Alkyl ketene dimer
(AKD) � AKD-CO2-oligomers � Humidity uptake
Introduction
Aerogels from bacterial cellulose (BCAG) are fasci-
nating ultra-lightweight structures, exhibiting a hier-
archical, three-dimensional porous morphology made
of pure cellulose, combined with extraordinarily high
surface area and toughness (Liebner et al. 2010).
However, for some application fields, the character-
istics of pure bacterial cellulose do not translate into
the desired material properties: The natural hydrophi-
licity may be a drawback for some applications where
an interaction with hydrophobic structures is neces-
sary or hydrophilic substances have to be prevented
from interaction with the BCAG structure. Modifica-
tion of bacterial cellulose generally can follow the
rules of ordinary cellulose modification when the
morphology of the product is of no further interest.
However, modification of BCAG under simultaneous
preservation of the inherent structural features
requires special approaches. In principle, both deriv-
atization (covalent reagent bonding) and coating
(reagents are just physically attached) are suitable
strategies for BCAG modification. Because of the
fragile nature of BCAGs, techniques based on
A. Russler � M. Wieland � M. Bacher �U. Henniges � F. Liebner � A. Potthast � T. Rosenau (&)
Christian Doppler Laboratory for Advanced Cellulose
Chemistry and Analytics, University of Natural Resources
and Life Sciences Vienna, Muthgasse 18, 1190 Vienna,
Austria
e-mail: [email protected]
P. Miethe
FZMB GmbH Forschungszentrum fur Medizintechnik
und Biotechnologie, Bad Langensalza, Germany
123
Cellulose (2012) 19:1337–1349
DOI 10.1007/s10570-012-9728-y
supercritical carbon dioxide (scCO2) have to be
applied for appropriate drying to largely avoid
changes of the natural BC structure (Liebner et al.
2010; Haimer et al. 2008; Liebner et al. 2009, 2007,
2008; Gavillon and Budtova 2008). Due to its
intrinsically low viscosity and special dissolution
properties, scCO2 can be used as multi-functional
medium for processing of (bacterial) cellulose aero-
gels: drying, chemical reactions, and solvent for
impregnation or fixation of various substances within
the porous material without altering the natural
morphology (Liebner et al. 2010; Russler et al.
2011). Combining supercritical drying with derivati-
zation or surface coating in one step would represent a
highly efficient and advantageous approach.
Alkyl ketene dimer (AKD, 1) is a well known and
widely used sizing agent in paper production that is
used to hydrophobize the cellulose surface to increase
water repellency and thus to enable paper to be printed
and written on in a way that applied inks do not leach
into the paper material and the print remains sharp and
detailed (Roberts 1996; Ek et al. 2009). It is added in
the head box of the paper machine in rather small
amount of 0.1–0.5 %. It is prepared e. g. from natural
fatty acids and stearic acid. Conventional AKD is a
waxy material with a melting point of about 50 �C,
nevertheless also formulations exist which are liquid at
room temperature. Having alkyl chains of normally
16–18 C-atoms, AKD wax is not soluble in water and,
when added to the headbox in paper production, is used
in the form of an aqueous dispersion in cationically
modified starch. Application is possible within a pH
range of about 7–10, so AKD became quite important
for neutral and alkali paper formation systems, which
compare favorably to acidic systems with regard to
long-term stability of the produced paper.
AKD is proposed to react with the hydroxyl groups
of cellulose to form a b-keto ester moiety (2). The
reaction with ethanol to the corresponding ethyl ester
2a is of limited predictive power as a model for the
reaction with cellulose: the esterification is in fact
similar, but the accessibility of the cellulose surface is
of course largely different from that of the low-
molecular weight alcohol. AKD will slowly react with
water to a b-keto acid (3) that readily decarboxylates
to produce a ketone (4) (see Fig. 1). Why the reaction
with the OH-groups of cellulose is favored over the
reaction with excess water under normal application
conditions is not clear until now (Odermatt et al. 2003;
Seo et al. 2008a, b; Lindfors et al. 2007). Nevertheless
it is known that the sizing effect is not an instant one,
but needs some time to develop (Shen and Parker
2001; Shen et al. 2005; Zhang et al. 2007). It is
assumed that in this time the AKD is spreading over
the cellulosic surface.
AKD is a well-known, well-established and cheap
agent for hydrophobization of cellulose in papermak-
ing—we were interested in its applicability in mod-
ifying BCAG by means of a novel application system
with scCO2. Novel cellulosic aerogels with more
hydrophobic surfaces will be the result. These mate-
rials will be better compatible with synthetic-organic
matrices than non-modified cellulose aerogels, which
would open up new fields of applications. Different
processes and process step sequences are possible for
the modification of BCAG with AKD. They differ in
complexity, limitations in reagent (AKD) loading,
effectiveness and homogeneity of the modification.
Naturally, it should be the aim to reduce the technical
complexity and work input while maintaining the
BCAGs0 above-discussed structural features. The
different processes are given in Fig. 2 schematically.
All of these methods contain at least one process
step involving scCO2, which we have recently shown
to be a promising medium for production of aerogels
also from bacterial celluloses (Liebner et al. 2010). As
AKD is soluble in scCO2, it is possible to perform also
the reagent-loading itself in scCO2 as the solvent, not
only the drying step. Some modifications of cellulose
with AKD applying scCO2 are known in literature, but
the techniques applied and the cellulose types used
were different from our approach (Hutton and Parker
2009; Quan et al. 2009).
OO
R
R
OCell
O
R
O
OHR'
O
R
O O
R
OEt
R
R
R
O
R
O
Cell-OH
H2O
+ CO2
1
2
3 4
2a
Et-OH
Fig. 1 Reaction of AKD with cellulose (Cell-OH) or ethanol,
and the hydrolytic reaction with water, R and R0 being C14-alkyl
or C16-alkyl
1338 Cellulose (2012) 19:1337–1349
123
Experimental
General
All chemicals were available from commercial sup-
pliers. Ethanol (absolute, Merck) and n-hexane
(Merck) of reagent grade were used as received.
Distilled water was used throughout. TLC was
performed using Merck silica gel 60 F254 pre-coated
plates. Flash chromatography was performed on Baker
silica gel (40 lm particle size). All products were
purified to homogeneity (TLC/GC analysis). Bacterial
cellulose was obtained from FZMB GmbH, Bad
Langensalza, Germany (see Liebner et al. 2010;
Russler et al. 2011 for preparation and purification).
AKD starting material
Alkyl ketene dimer (AKD, technical grade, Herkules)
was further purified by flash chromatography using n-
hexane as the solvent. GCMS analysis confirmed the
remainder to be pure AKD of different chain lengths,
without any impurities of AKD ketone (4) or esters.
NMR and GCMS
1H NMR spectra were recorded at 400 MHz for 1H
and at 100 MHz for 13C NMR in CDCl3. Chemical
shifts, relative to TMS as internal standard, are given
in d values, coupling constants in Hz. 13C peaks were
assigned by means of APT, H,H-COSY, HMQC and
HMBC spectra. As AKD is a mixture of several
compounds (due to the different chain lengths of the
used fatty acids), resonances are sometimes superim-
posed, and multiplicities are only given when clearly
discernible. GCMS analysis was carried out on an
Agilent 6890 N/5975B in the ESI (70 eV) ionization
mode.
Compound characterization by NMR
AKD (1, starting material). 1H NMR: d 0.87 (t, 6H,
CH3), 1.22–1.40 (m, br, CH2–), 1.39 (m, 2H, CH2–
CH2–CH–COO), 1.77 (q, 2H, CH2–CH2–CH–COO),
2.12 (q, 2H, CH2–CH=C), 3.94 (t, 1H, CH2–CH–
COO), 4.68 (CH2–CH=C). 13C NMR: d 14.1 (CH3),
22.7 (CH3–CH2), 24.6 (CH2–CH2–CH–COO), 26.3
(CH2–CH2–CH–COO), 27.5 (CH2–CH=C), 29.1–29.7
(CH2), 31.9 (CH3–CH2–CH2), 53.7 (=C–CH–COO),
101.7 (CH2–CH = C), 145.6 (CH=C), 169.7 (COO).
AKD ethyl ester (2a). 1H NMR: d 0.87 (t, 6H, CH3
in alkyl), 1.20–1.36 (m, br, CH2–), 1.28 (3H, super-
imposed by m, O–CH2–CH3), 1.51–1.64 (m, 2H,
CH2–CH2–CO), 1.78–1.84 (q, 2H, CH2–CH2–CH–
COO), 2.38 (t, 2H, CH2–CH2–CO), 3.40 (t, 1H, CO–
CH–COO), 4.18 (q, 2H, O–CH2–CH3). 13C NMR: d14.1 (CH3 in alkyl), 14.2 (O–CH2–CH3), 22.7 (CH3–
CH2–CH2), 23.8 (CH2–CH2–CO), 28.2 (CH2–CH–
COO), 29.1–29.9 (CH2), 32.0 (CH3–CH2–CH2), 43.7
(CH2–CH2–CO), 59.3 (CO–CH–COO), 169.8 (COO),
205.7 (CO).
AKD-ketone (4). 1H NMR: d 0.88 (t, 6H, CH3 in
alkyl), 1.22-1.34 (m, br, CH2–), 1.55 (pent, 2H, CO–
CH2–CH2), 2.38 (t, 2H, CO–CH2–CH2). 13C NMR: d14.2 (CH3 in alkyl), 22.8 (CH3–CH2–CH2), 24.0
BCG BCAGscCO2
Drying mBCAG
AKD
ExtractionscCO2
DryingscCO2
Loading
BCG Loading Bath mBCAG
AKD
Solvent ExchangescCO2
Drying
BCG BCAGscCO2
Drying mBCAG
AKD
scCO2
DryingLoading Bath Solvent Exchange
1
3
2
Fig. 2 Overview of the
different process approaches
for AKD modification of
BCAGs (see the
experimental section for a
detailed description), BCG:
bacterial cellulose gel,
BCAG, bacterial cellulose
aerogel, mBCAG: AKD-
modified bacterial cellulose
aerogel)
Cellulose (2012) 19:1337–1349 1339
123
(CH2–CH2–CO), 29.4–29.8 (CH2), 32.0 (CH3–CH2–
CH2), 42.9 (CH2–CH2–CO), 211.9 (CO).
AKD-CO2 hexamer (5). ‘‘Terminal unit’’ denotes
the AKD unit with the ketone moiety, ‘‘proximal unit’’
the one with the free acid moiety. 1H NMR: d 0.87
(36H, CH3), 1.20–1.42 (CH2–), 1.39 (m, 10H, CH2–
CH2–CH–COO), 1.51–1.59 (2H, CH2–CH2–CO in
terminal unit), 1.78 (q, 10H, CH2–CH2–CH–COO),
1.78–1.82 (q, 2H, CH2–CH–COO in terminal unit),
2.12 (q, 10H, CH2–CH=C), 2.38 (t, 2H, CH2–CH2–CO
in terminal unit), 3.40 (t, 1H, CO–CH–COO in
terminal unit), 3.95 (t, 5H, CH2–CH–COO), 4.72 (t,
5H, CH2–CH=C). 13C NMR: d 14.1 (CH3), 22.7
(CH3–CH2), 23.8 (CH2–CH2–CO in terminal unit),
24.4–24.6 (CH2–CH2–CH–COO), 26.3–26.4 (CH2–
CH2–CH–COO), 27.5 (CH2–CH=C), 28.2 (CH2–CH–
COO in terminal unit), 28.9–30.4 (CH2), 31.9 (CH3–
CH2–CH2), 43.7 (CH2–CH2–CO in terminal unit),
53.7-53.8 (=C–CH–COO), 59.3 (CO–CH–COO in
terminal unit), 101.7–101.9 (CH2–CH=C), 145.5–
145.7 (CH=C), 169.3–169.6 (COO), 169.9 (COOH
in proximal unit), 205.7 (CO).
scCO2 systems
For the drying and AKD-modification of BCAG, a
high-pressure system was used. CO2 from a gas bottle
was pressurized in a high pressure pump and charged
into a 500 cm3 reactor. Temperature was set to 40 �C
and was controlled by a thermostat. The reactor was
equipped with a stirrer and a grid to store the aerogels
separated from the stirrer. The stirrer was not neces-
sary for the drying of the aerogels alone, but is
required for AKD-modification to insure uniform
distribution of the reagent. The system was equipped
with a two-stage separator unit and with the possibility
to recycle the gas.
Before supercritical drying and AKD modification,
bacterial cellulose aquogels have to undergo solvent
exchange. A solvent exchange against absolute etha-
nol was performed three times for at least 24 h. The
subsequent supercritical drying of the BCGs was
performed in the pressure vessel at 100 bar, 40 �C and
a CO2 flow of 3.5 kg/h with no stirring. Treatment
times were 1 h per centimeter of aerogel diameter. For
properties and morphology of the BC aerogels see:
Liebner et al. 2010; Russler et al. 2011.
In the case of AKD-modification, the same proce-
dure as for the supercritical drying was used, with the
following modifications: pressure and time were
varied (see text), no CO2 flow was applied, and
stirring was applied at 350 min-1.
The simple setup of the pressure system guaranteed
proper cleanability and good repeatability. Neverthe-
less, since precipitated AKD can block valves and
tubes, it is advisable to flush the apparatus with
suitable organic solvents (chloroform, petrol ether)
from time to time, and to release the pressure by a
valve directly, without separators (Odermatt et al.
2003; Seo et al. 2008; Hutton and Parker 2009; Quan
et al. 2009).
SEM
Scanning electron microscopy (SEM) was performed
on a Phillips XL 30 ESEM (Environmental Scanning
Electron Microscope, ESEM) at an acceleration
voltage of 10 kV with different magnifications.
FTIR
Fourier transform infrared spectroscopy (FTIR) was
performed on a Bruker Vertex 70 HTS-XT in the
Attenuated Total Reflectance (ATR) mode with 32
scans per measurement between 4000 and 400 cm-1.
Humidity uptake
The humidity uptake of the samples was measured
gravimetrically after storage in desiccators of con-
trolled humidity for at least 72 h. The desiccators were
filled with phosphorous pentoxide for 0 % relative
humidity, with saturated solutions of calcium chloride
for 30 %, of ammonium nitrate for 65 % and of
potassium sulfate for 98 % relative humidity, and
were stored at room temperature.
Modification approaches according to Fig. 2
In the case of direct supercritical loading, the AKD
wax was charged directly into the pressure vessel
together with BCAG. The BCAG samples were put
into the internal wire caskets to prevent them from free
floating in the supercritical fluid. The system was then
pressurized at different pressure levels for 60 min.
The stirring was started when supercritical conditions
were reached, and was stopped during pressure
release before dropping below supercritical conditions.
1340 Cellulose (2012) 19:1337–1349
123
The AKD-modified BCAG was immediately subject to
analytical characterization, extracted after some curing
time (1 or 2 weeks at room temperature) or further
stored at ambient conditions. The extraction of AKD
from loaded samples (to test for physical adsorption vs.
covalent binding) was performed batchwise with neat
n-hexane (20-fold sample volume) for 24 h in a shaker,
and this extraction was repeated twice. As indicated by
the dotted line in Fig. 2, a combination of BCG drying
with AKD loading is possible if the scCO2 apparatus is
equipped with separate pressure chambers connected
with valves, so that after drying scCO2-dissolved AKD
can be introduced.
According to the second approach of Fig. 2, the
BCG was pretreated by solvent exchange to ethanol
and then subject to a loading bath, containing different
amounts of AKD dissolved in n-hexane. The modified
BCG was directly dried supercritically in the high
pressure system. Alternatively, another solvent
exchange step to ethanol, a non-solvent for AKD,
was done beforehand. The conditioning in the loading
bath should last long enough, depending on the sample
dimensions, to ensure homogeneous distribution of the
loading medium throughout the sample, at least 1 h
per centimeter of aerogel.
The third way in Fig. 2 for the loading with AKD
was starting from already dried BCAG. Modification
of this aerogel with AKD was performed by AKD
solutions in n-hexane of set concentrations, so that a
defined amount of AKD was homogeneously distrib-
uted throughout the BC matrix. To obtain aerogels
from the solvogels, it was necessary to apply another
scCO2-drying step, either with or without preliminary
solvent exchange to ethanol.
Results and discussion
BCAG modification with AKD (‘‘AKD loading’’)
The supercritical loading of AKD onto BCAG (path 1
in Fig. 2) was found to be a very flexible approach.
The degrees of loading, i.e. the ratio between mass of
AKD to mass of original, non-modified BCAG, was
varied in a wide range from about 30 % to over 250 %.
When the pressure in the vessel is reduced below
supercritical conditions, the dissolved AKD would be
expected to precipitate homogeneously within the
whole pressurized volume. If, for example, the density
of BCAG is 10 mg/cm3 and the density of the agent
to be precipitated into the system is 2 mg/cm3, a
maximum degree of loading of about 20 % is
expected. Such a result coming close to the ‘‘theoret-
ical’’ value is obtained, for instance, for inert long-
chain aliphatics and triglycerides as the substances to
be deposited within the aerogel matrix. However, in
the case of AKD, we found in all cases a higher
loading of the BCAG than calculated from the mass of
AKD put into the pressure system, with values ranging
up to 250 %.
Three mechanisms can be summoned to account for
this peculiar behavior. The first is the reaction of AKD
with the cellulose. The contribution of this mechanism
to the enrichment effect, however, must be rather
small since 85 ± 3 % of the deposited AKD can be re-
extracted with n-hexane from the cellulosic bodies
(independent of the degree of loading), so that a
maximum of 15 ± 3 % of the initially deposited AKD
can be covalently bound to cellulose, thus being non-
extractable. The second mechanism of the enrichment
as described might be a preferred precipitation within
the nano-scale pore system of the BCAG due to
capillary effects. However, it is not clear why such
effects would be operative for AKD, but not for
paraffin waxes or triglycerides (which do not show the
enrichment effect). Hence, this mechanism is rather
unlikely. The third mechanism is an oligomerization
of AKD. By polymer–polymer (polymer-oligomer)
interactions, these bigger molecules would be prefer-
ably adsorbed on the cellulose surface, so that as the
net effect the aerogel is enriched with AKD moieties at
the same time depleting the supercritical solvent of
AKD. The initial trigger for assuming that such a
reaction could have occurred was the observation that
re-extracted AKD was no longer monomeric, but
oligomeric. This prompted us to look into the mech-
anism more closely.
Mass spectrometric analysis of the n-hexane
extracts of the deposited AKD, showed the presence
of larger AKD-derived molecules (5), which however
were not just AKD-oligomers, but appeared to contain
additional CO2-moieties, readily identifiable by a
mass difference of 44 and its multiples. Apparently,
the supercritical medium did not behave as inert
solvent for AKD, but participated in a reaction,
promoting an oligomerization. Such oligomerization
is not observed in aqueous suspension or in organic
solutions in common solvents, so that this effect must
Cellulose (2012) 19:1337–1349 1341
123
be a peculiarity of the scCO2 medium. NMR analysis
showed the absence of non-reacted AKD. However,
the expected a-alkyl-substituted b-keto acid, as the
usual product of AKD ring opening (see Fig. 1), was
not present. Instead, a b-enolate structure was found,
which is linked to the carboxylic acid motif via a
carbonate moiety. By two-dimensional experiments
(HSQC and HMBC), the structure of the oligomers as
enolcarbonate anhydrides of b-keto-carboxylic acids
were unambiguously confirmed (Fig. 3) (Rosenau and
Russler 2012). No direct linkages between the enolate
of one AKD molecule and the carboxylic acid of the
next one are contained—the two molecules are always
connected via one CO2 as ‘‘spacer’’, i.e. in the form of
an enolcarbonate. The substances can thus be per-
ceived as ‘‘co-oligomers of AKD and CO2’’ in a 1:1
ratio.
The formation mechanism can reasonably be
assumed to be a CO2-induced opening of the AKD0sfour-membered oxetan ring. A transient structure with
negatively charged enol carbonate oxygen is attacking
the next AKD molecule under ring opening to give a
dimer, of which the formed enolate motif is once more
attacking another CO2 molecule. This results in a new
enol carbonate that reacts with the next AKD, and so
on. Finally the observed oligomers are built up (see
Fig. 3). Whether ionic structures indeed occur as
intermediates or just as transition state cannot be
answered conclusively at present. The observation of
only minor amounts (\0.5 % of re-extracted material)
of an addition product of AKD and CO2, a cyclic
organic carbonate with a stable six-membered ring
structure (6), disfavors the presence of the zwitterionic
intermediate as independent species. If indeed exist-
ing—even as highly transient species—the hypothetic
zwitterion would preferably react by intramolecular
recombination to the cyclic carbonate (6), rather than
by attacking another AKD molecule to form linear
oligomers (5). The near-absence of this cyclic car-
bonate by contrast to the larger amount of oligomers
formed argue against formation of a free zwitterionic
intermediate.
The presence of a cellulosic surface with small
amount of water present appeared to be imperative for
OO
R
R
OO O
O
R
O
O
O
R
O
OO
O
R
O
O
O
R
OOH O
O
R
O
O
O
R
O
O
OO
R
R
OHCell
CO2
H2O
OO
O
R
OOH O
O
R
O
O
O
R
R RR RR R
RR RO
n
O O
R
O
R
O
O OH
R
O
R
O
O_
Cellulosesurface AKD, scCO2
oligomers detected for 0 < n < 4
1
5 (hexamer)
5 (general formula)
putative ionicintermediate
AKD, scCO2
Fig. 3 Formation of AKD-CO2 ‘‘co-oligomers’’ as found in the
n-hexane extracts of AKD-modified bacterial cellulose aerogels.
Middle row: Grey shades show the CO2-derived units, while the
dotted boxes denote ring-opened AKD motifs. The oligomers
contain an acid (left) and a ketone (right) terminus. Lower row:
general formula of the AKD-CO2-compounds emphasizing the
‘‘oligomeric’’ character by showing the repeating AKD-CO2
‘‘repeating unit’’
1342 Cellulose (2012) 19:1337–1349
123
the oligomer formation: no such products were
observed when working with cellulosic model com-
pounds that were dry and non-hydrated, whereas
addition of small amounts of water (one equivalent per
hydroxyl group) induced the oligomer formation once
more. By contrast, larger amounts of water, in turn,
prevent oligomer formation from the beginning.
Polyhydroxylic compounds, such as polyvinyl alcohol
and even silica gel appeared to favor the oligomeri-
zation, and always required small amounts of water for
this reaction to proceed. Non-hydroxylic surfaces did
not at all induce AKD-oligomerization, independent
of the presence of water traces. The exact formation
mechanism of the oligomeric AKD compounds and
the peculiar water and surface effects are currently
under study, and an account will be given as soon as a
conclusive mechanism is available.
The AKD loading values of 30–250 % did not
represent a possible minimum or maximum, respec-
tively. Values beyond these limits were just not tested.
These variations were performed by changing the
amount of AKD charged into the pressure vessel. Also
variation of the pressure (100 bar to 300 bar) had
some influence on the degree of loading, see Figs. 4, 5.
For AKD concentrations below 5 mg/cm3, the solu-
bility of AKD in the supercritical fluid increased with
increasing pressure until about 150 bar and then
moderately decreased. Above 5 mg/cm3 AKD con-
centration, the solubility increased over the whole
tested pressure range, although above 200 bar the
increase was less pronounced than at lower pressures.
Thus, the actual degree of loading of the gel is
determined by both the starting AKD concentration
and the working pressure.
The application of the two other techniques for the
loading of BCG/BCAG with AKD (path 2 and 3 in
Fig. 2) resulted in much lower degrees of loading.
This is especially due to the fact that for the production
of aerogels—either by loading via loading bath or by
filling the BCAG with an AKD-containing solution—
a subsequent supercritical drying step is required.
During this drying excessive AKD was dissolved in
the supercritical fluid and removed.
Homogeneity of modification according
to pressure vessel geometry and sample geometry
The degree of loading of BCAG under supercritical
conditions was influenced also by the geometry of the
setup, i.e. the vessel containing the supercritical
medium. BCAG located in the upper part of the
pressure reactor showed somewhat lower degrees of
loading than samples located near the bottom. This
might be due to a limited solubility of AKD in the
supercritical CO2 fluid: when the pressure was
increased to 250 bar, the solubility increased and the
inhomogeneity of deposition got smaller (see Figs. 4,
5). At conditions of limited solubility (lower pres-
sures) the samples nearer to the bottom face an
environment with a higher concentration of AKD due
Fig. 4 Dependency of
AKD loading on pressure
and sample location in the
pressure vessel, AKD
density in the supercritical
medium: 2 mg/cm3
Cellulose (2012) 19:1337–1349 1343
123
to the presence of a suspension of non-dissolved AKD
particles that adhere to the BCAG and remain attached
beyond pressure reduction to subcritical conditions.
At a pressure of 250 bar, no inhomogeneities of the
material within a batch are present any longer.
However, differences in loading among samples of
different batches—despite similar AKD concentra-
tions in the supercritical medium—are still noticeable
(see Figs. 4, 5). Altogether, the samples with higher
AKD concentration (e.g. 10 mg/cm3) showed a more
homogeneous and better reproducible modification
than samples loaded at lower pressures.
These findings show that the control of the degree
of loading is not as easy to control as was expected. In
an upright reaction vessel, the circulation of the
supercritical fluid was not sufficient, even when
stirring was applied. Tumbling of the supercritical
vessel solved the problem of inhomogeneities within a
batch, but the experimental setup and the procedure
became more complicated.
Another solution to the problem of batch inhomo-
geneity is the preparation according to the other two
methods proposed, i.e. paths 2 or 3 in Fig. 2. In these
cases, no differences were observed according to the
location of the samples within the pressure system.
Some differences can also be seen, if homogeneity
is not considered with regard to the sample position in
the pressure vessel, but with respect to the degree of
modification within or across an aerogel body. As
weight analyses of different regions of individual
samples are tedious to perform for larger sample
numbers, we used FTIR measurements for a semi-
quantitative analysis. Figure 6 shows the FTIR spectra
of pure BCAG (bottom) and pure AKD (top), as well
as of BCAG loaded with 15 % AKD and 70 % AKD
loading. The sample with a content of 15 % AKD was
produced by extraction of the higher loaded sample
with n-hexane after initial AKD loading. Some
characteristic IR bands of AKD are indicated by
arrows in Fig. 6, the most prominent ones are the
intense aliphatic stretching bands at 2,916 and
2,848 cm-1, which are also prominently found in the
AKD-modified BCAG. The bands at 1,848, 1,720 and
1,467 cm-1 originate from AKD0s oxetanone struc-
ture. Some of its structural motifs, such as the enol
ether structure and the C(=O)O motif, are retained
after covalent attachment to cellulose or formation of
the CO2-containing oligomers (see Fig. 3), while
others are canceled, such as the four-membered ring.
In the case of 70 % loading, both AKD oligomers (5)
and cellulose-bound AKD (2) contribute to the
spectrum, while only the latter adds to the spectrum
in the case of 15 % loading (the oligomers are
removed by extraction). The prominent bands at
2,916 and 2,848 cm-1 are most suited for quantifica-
tion of AKD on the BCAG matrix.
Figure 7 (left) shows the characteristic part of the
FTIR spectra of different regions (surface, outer part,
core) of a BCAG sample supercritically loaded with
70 % AKD. The high surface coverage with AKD
Fig. 5 Dependency of
AKD loading on pressure
and sample location in the
pressure vessel, AKD
density in the supercritical
medium: 10 mg/cm3
1344 Cellulose (2012) 19:1337–1349
123
(in the form of oligomers) is evident. After extraction
with n-hexane (Fig. 7, right), this surface coverage is
removed, but a loading of 15 % remained persistently.
This portion corresponds to AKD covalently bound to
the cellulose surface, while the major, extractable part
was just physically adsorbed. After extraction, the
loading differences between surface and core vanish,
proving that covalent modification was uniform over
the cellulose matrix, while deposition of the physi-
sorbed portion occurred only on the surface, but not in
the interior. As mentioned above, the amount of
covalently bound AKD was nearly constant at about
15 ± 3 %, independent of the respective conditions of
supercritical loading. By contrast, physically adsorbed
AKD and its CO2-containing oligomers, which are just
deposited on the surface in a re-extractable manner,
can be enriched up to 250 % (and likely beyond) in the
loaded BCAG sample.
In SEM pictures of the samples with 70 % loading
(see Fig. 8) AKD-oligomer agglomerates can be seen.
In the sample subsequently extracted with n-hexane,
which contained 15 % covalently bound AKD, no
such agglomerates were still visible, but a fibrillar
structure with smooth surface coating was observed
(Fig. 9).
Just from weight determination, it could evidently
not be clear whether the amount of covalently
bound AKD (approx. 15 %) consisted of ‘‘proper’’
monomeric AKD in the form of the expected b-
ketoester or whether it was made up of covalently
bound CO2-AKD oligomers. We therefore used per-
acetylation of the sample with subsequent 1H NMR
analysis in CDCl3, which showed a DSAKD of 0.07
(± 0.03). A mass percentage of 15 % monomeric
AKD (C16, M = 494 g mol-1) would mean a theo-
retical DSAKD of 0.05, while substitution with AKD-
hexamers (5) translates into a theoretical DS(AKD)6 of
0.008. The experimental value of 0.07 clearly indi-
cates that only monomeric AKD is bound to the
cellulose matrix—or at least that the amount of
covalently linked oligomeric AKD is negligibly small.
Thus, the loading of the cellulose matrix did occur
5001000150020002500300035004000
Wavenumber cm-1
Fig. 6 FTIR spectrum of AKD-loaded modifications of bacte-
rial cellulose aerogels with 70 % AKD and 15 % AKD. Spectra
of pure AKD (top) and pure bacterial cellulose (bottom) are
given for comparison. y-axis: relative absorbance
2700280029003000
Wavenumber cm-1
2700280029003000
Wavenumber cm-1
Fig. 7 Left: characteristic section of an FT-IR spectrum of
surface, outer part and core of a BCAG loaded with an average
of 70 % AKD, indicating different AKD contents in different
aerogel parts. Right: extraction of the aerogel with n-hexane
leaves only covalently bound AKD behind, which is uniformly
distributed throughout the material, eliminating the observed
loading variations between different aerogel parts. y-axes:
relative absorbencies
Cellulose (2012) 19:1337–1349 1345
123
with monomeric AKD, but not with AKD oligomers—
in contrast to the physically absorbed, extractable
residue which consists only of such oligomers.
It should be noted that hydrolytic cleavage of the
bound AKD molecules and subsequent analysis—
which at a first glance appears to be the method of
choice to characterize the covalently bound AKD
fraction—is not a feasible approach here: the oligo-
meric AKD—if present—will fragment under these
conditions into CO2 and the equivalent amount of
monomeric AKD residues, rendering monomeric and
oligomeric AKD forms indistinguishable by this
method. Under aqueous hydrolysis conditions, the
monomeric AKD-derived ketone (3) is formed from
CO2-AKD oligomers such as (5), under non-aqueous
conditions (e.g. gaseous HCl in ethanol) the corre-
sponding AKD-derived ethyl ester (2a) is produced.
Hydrolysis thus offers no way to distinguish between
bound monomers and bound oligomers.
All these findings led us to the conclusion that only
a relatively minor (about 15 %), non-extractable part
of AKD monomers was covalently bound to the
cellulose. The starting content of AKD, after loading
but before extraction, is itself variable in wide ranges,
but does not, however, influence the eventual amount
of covalently bound AKD, i.e. the degree of loading
after the extraction. If an AKD-loading in percentages
higher than 15 % was found, the reason was always a
physical deposition of AKD oligomers in addition to
the chemically bound part.
Fig. 8 ESEM pictures (left 5.000x, right 20.000x) of a BCAG sample scCO2-loaded with 70 % AKD, exhibiting agglomerates of
AKD-oligomers within the BCAG matrix, besides the coating of the BC fibrils
Fig. 9 ESEM pictures (left 5.000x, right 20.000x) of a BCAG sample scCO2-loaded with 15 % AKD (70 % initial loading and
subsequent extraction with n-hexane), exhibiting smooth fibrillar coating and no visible agglomerates
1346 Cellulose (2012) 19:1337–1349
123
Samples prepared according to the other methods
described in Fig. 2 (paths 2 and 3) showed only
negligible differences between different parts of the
samples, provided the curing time in case of method
No. 2 was long enough ([1 h) to allow a homogeneous
distribution of AKD in the solvogels by diffusion.
Humidity uptake
As AKD is providing hydrophobic properties if
applied to cellulosic material in the form of paper, it
was reasonable to assume that is has the same effect
also on cellulosic aerogels. We tested the humidity
uptake of different modified BCAG samples by
monitoring the weight gain during storage in con-
trolled atmospheres with different relative humidities
(rH). After storage of the prepared samples for some
time at ambient conditions, they were put into a
desiccator over phosphorous pentoxide to completely
dry the samples to weight constancy. This condition
was set as the starting point. The weight re-gain of the
samples when placed in different humidity environ-
ment (rH 30, 65, 98 %) is displayed for selected
specimens in Fig. 10. Pure AKD powder which was
placed in the humidity controlled desiccators as a
reference showed no weight gain at all. Samples
extracted with n-hexane having an effective AKD
loading of just 15 % showed results similar to the
sample with 29 % AKD loading. If dried again by
storing at 0 % relative humidity for several days, all
samples returned to their initial, constant weight.
At relative humidities of 30 %, none of the samples
tested showed significant humidity uptake. The high-
est weight gain observed was just 0.6 %, compared to
a weight gain of 2.5 % for non-modified BCAG
samples. Also at a rH of 65 %, all modified samples
stayed below 1 % weight gain; the non-modified
sample showed a mass increase of 3.4 % here. In an
atmosphere saturated with water vapor (98 % rH),
non-altered BCAG samples showed a weight gain of
20 %. Modified samples with loadings up to 70 %
stayed below a weight gain of 8 %. The weight gain of
a sample with 29 % AKD was 5 %, that of a specimen
with 15 % AKD only 2 %. All these samples showed
thus a pronouncedly decreased humidity uptake. So
far, the outcomes reflect the expectations. AKD-
modified BCAG samples with a loading higher than
70 % showed a rather unexpected behavior: the
humidity uptake increased quite drastically; samples
with very high loading (102 and 212 %) even signif-
icantly exceeded the level of unmodified BCAG. The
weight gain of all samples was completely reversible.
This ‘‘humidity enrichment’’ effect was readily repro-
ducible. It generally occurs only at near-saturated
relative humidities (rH [ 95 %), only at AKD-load-
ings above 70 %, and it increases further with
increasing AKD amounts in the modified BCAG.
The production of samples with such very high
AKD loadings is not possible according to the
Fig. 10 Humidity uptake
by BCAG samples with
different degrees of AKD
loading at room temperature
in atmospheres of different
relative humidities (0, 30, 65
and 98 % rH)
Cellulose (2012) 19:1337–1349 1347
123
alternative modification methods (paths 2 and 3 in
Fig. 2). Samples with the effect of increased humidity
uptake at very high AKD-loads could thus only be
obtained by the supercritical deposition method (path
1 in Fig. 2) just discussed.
The peculiar effect that the humidity uptake
increases despite (or better: due to) surface-deposited
AKD agglomerates can be assumed to be caused by
some special morphological features created upon
loading under supercritical conditions, which boost
a kind of water condensation already beneath the
saturation point. As shown above, the high AKD loads
can be explained with globular agglomerates of AKD-
oligomers which appear to cause the condensation
effect, whereas simply a thicker surface coverage does
not cause this effect. High AKD-percentages do not
necessarily translate into increased surface hydro-
phobization, but rather into increased concentrations
of strongly hydrophobic spots. We are currently
following routes to visualize water droplet formation
in the material and to account for the water enrichment
in cooperation with material physicists.
Conclusions
AKD, a well known sizing agent for paper, can also be
used for the modification of BCAG. There were three
main routes followed to perform the modification, all
are based on super-critical CO2 which is able to
preserve the delicate porous network structure of
cellulosic aerogels without morphological changes.
The direct loading of BCAG with AKD dissolved in
the supercritical fluid is the most elegant and most
flexible variant, which also allows especially high
degrees of loading.
About 15 % of the deposited AKD—this number
being largely independent of starting AKD concen-
tration and system pressure—was bound to the
cellulose surface in a covalent way—this part of the
AKD loading remains non-extractable. Covalently
bound AKD is predominantly monomeric as seen by
derivatization/NMR studies, and is homogeneously
distributed throughout the BCAG. With the scCO2
approach, degrees of loading up to 250 % can be
reached. The additional AKD beyond the limit of
15 % is bound by physisorption only and thus is
extractable. The main part of the extractable portion is
composed of an CO2-AKD oligomer (dimer to
hexamer), which forms spot-like agglomerates on
the BCAG surface, but does not penetrate into the bulk
material by contrast to the covalently fixed AKD part.
The BCAG samples loaded with up to 70 % AKD
exhibited hydrophobic behavior with reduced humid-
ity uptake at all relative humidity levels. The uptake
was reduced to one tenth up to one quarter of non-
modified BCAG. An interesting reverse effect, which
is currently being scrutinized, was observed for BCAG
with very high AKD loads of above 70 % up to 250 %.
The humidity uptake at nearly 100 % rH increased
significantly, by far surpassing that of genuine
BCAGs. This peculiarity might be due to superhy-
drophobicity effects of the AKD-oligomer deposits
inducing water condensation.
The AKD-modification of bacterial cellulose aero-
gels in scCO2 media is a convenient and flexible way
to alter the BCAGs0 properties towards increased
hydrophobicity and compatibility with hydrophobic
polymers. The finding of AKD-CO2-oligomers
formed under conditions of supercritical processing
is novel. At present, it cannot yet be decided whether
this behavior is positive or negative with regard to the
general processability and applicability of AKD in
scCO2. The chemical nature and the properties of the
AKD oligomers are topics of current studies in our
group.
Acknowledgments We would like to thank the Austrian
Christian-Doppler-Society, Vienna and the FZMB GmbH, Bad
Langensalza, Germany, for financial support within the
‘‘Christian Doppler Laboratory for Advanced Cellulose
Chemistry and Analytics’’ and Walter Klug for recording the
SEM pictures.
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