Cellulose (2021) 28:11503–11517

https://doi.org/10.1007/s10570-021-04188-9 (0123456789().,-volV)
( 01234567
89().,-volV)

ORIGINAL RESEARCH

Deep eutectic solvent recycling to prepare high purity

dissolving pulp
Yuanhang Chen . Kuizhong Shen . Zhibin He . Ting Wu . Chen Huang .
Long Liang . Guigan Fang

Received: 5 May 2021 / Accepted: 12 September 2021 / Published online: 17 October 2021
Ó The Author(s), under exclusive licence to Springer Nature B.V. 2021

Abstract In this work, a novel acid-based deep
eutectic solvent (DES) recycling process was devel-
oped for the pretreatment of eucalyptus wood chips in
the preparation of high purity dissolving pulp. It was
found that a large amount of lignin and xylan in the
eucalyptus wood material were extracted in the
recycling treatment with lactic acid/choline chloride.
The dissolved lignin and xylan in DES were recovered
by precipitation in water, and the DES solvent was
regenerated by removing the water through evapora-
tion. The residual lignin and xylan in the DES pre-
treated wood chips were more readily removed in the
subsequent kraft cooking and bleaching processes,
yielding a high-purity dissolving pulp (DES-KP).
Compared with the conventional prehydrolysis kraft
based dissolving pulp (PHK), the DES-KP had a
higher a-cellulose (94.02% vs. 89.81%) and a
markedly higher Fock reactivity (98.94% vs.
60.97%). In addition, the DES pretreatment was also
combined with oxygen-alkali cooking and ECF
bleaching process, to prepare a dissolving pulp
(DES-BP) with a high pulping yield (44.40%). The
DES-KP pulping process was advantageous compared
with the conventional PHK process in terms of the

Y. Chen  K. Shen  T. Wu  C. Huang  G. Fang

L. Liang  G. Fang e-mail: fangguigan@icifp.cn
Key Lab of Biomass Engineering and Material Jiangsu
Province, National Engineering Lab. for Biomass Z. He
Chemical Utilization, Key and Open Lab of Forest University of New Brunswick, Fredericton,
Chemical Engineering, Institute of Chemical Industry of NB E3B 5A3, Canada
Forest Products, Chinese Academy of Forestry, SAF,
Nanjing 210042, People’s Republic of China

K. Shen  C. Huang  G. Fang

Nanjing Forestry University, Nanjing 210037, People’s
Republic of China

K. Shen  G. Fang
Shandong Huatai Paper Co., Ltd, Dongying,
Shandong Province 257335, People’s Republic of China

K. Shen (&)  G. Fang (&)

Institute of Chemical Industry of Forestry Products, CAF,
Nanjing 210042, People’s Republic of China
e-mail: kuizhong@icifp.cn

123
11504
biorefinery concept, as the total utilization rate of the
biomass of the eucalyptus wood chips reached
69.74%. Therefore, the DES pretreatment is a promis-
ing process for efficient biomass utilization in dis-
solving pulp production.
Keywords Lactic acid/choline chloride 
Eucalyptus  Dissolving pulp
Introduction
With the increase of public awareness of environment
protection and economy sustainability, research on
sustainable utilization of wood biomass is increasing.
Efficient separation of the cellulose, hemicellulose and
lignin components of lignocellulosic biomass is the
key to success. Ionic liquid known as ‘‘green solvent’’
is an attractive solvent for separating lignocellulosic
biomass, with non-flammability, thermal stability and
chemical stability. But ionic liquids are complex and
expensive to prepare.
In recent years, deep eutectic solvents (DESs) have
been sought after by a large number of researchers
because of their physical and chemical properties
similar to ionic liquids but being cheaper, more
environmentally friendly, and simpler to prepare.
DES is a mixture of hydrogen bond donors and
hydrogen bond acceptors, which could be liquidized at
lower temperature (Francisco et al. 2012; Tan et al.
2020). Different types of DESs have great differences
in the solubility of lignin. Zhang et al. pretreated corn
cobs with different DESs and found that lactic
acid/choline chloride (15:1) and oxalic acid/choline
chloride (1:1) had the best lignin extraction effect
(93.1 and 98.5% respectively), and lactic acid/choline
chloride(10:1) had the second best effect (86.1%)
(Zhang et al. 2016). In addition, Francisco found that
lactic acid/choline chloride (10:1) had a good delig-
nification selectivity because lignin has a high solu-
bility in it (11.82 wt%), while cellulose is almost
completely insoluble in the solvent (Francisco et al.
2012). DES has high power to dissolve lignin, and its
dissolving capacity is strongly dependent on the
process conditions. In addition to the type and molar
ratio of DESs mentioned above, contact time and
temperature are also key factors affecting the solubil-
ity of lignin in DESs. Gou et al. used three kinds of
123
Cellulose (2021) 28:11503–11517
DESs (ChCl/methanol, ChCl/1,4-butanediol and
betaine/lactic acid) to treat xylose residues at different
temperatures (80, 100 and 120 °C), and the results
showed that the removal rate of lignin increased at
higher temperature (the removal rate changes were
from 54.0 to 63.5%, 30.7–54.0% and 64.5–81.6%,
respectively) (Guo et al. 2018). And according to the
research of Pan et al., the lignin content of the solid
residues treated at 130 °C were lower than that at
110 °C; at 130 °C, with the increase of DES pretreat-
ment time, the residual amount of lignin in the solid
residue reduced, but at 110 °C, the effect of time
change on the removal of lignin was insignificant (Pan
et al. 2017). When Procentese et al. used choline
chloride/glycerol and choline chloride/ethylene glycol
to treat agricultural food waste, it was concluded that
the biomass recovery and lignin content of the sample
decreased with the decrease of temperature. It was
worth noting that solid-liquid ratio had an effect on the
dissolution of lignin and polysaccharides (Procentese
et al. 2018). DES is a good organic solvent for lignin
dissolution, and when acid-based DES is used to treat
lignocellulose, it can not only effectively dissolve
lignin, but also remove hemicellulose. Tan et al.
utilized DESs, including lactic acid, maleic acid, citric
acid and other acids as HBD and choline chloride as
HBA to treat the empty fruit bunch of oil palm, and
found that all acidic DESs pretreatments removed
large portions of xylan (Tan et al. 2019). Another
advantage of DESs is that it can be recycled and reused
(Kumar et al. 2016; Zhang et al. 2012). After five
cycles of reuse of lactic acid/choline chloride (3:1), Li
et al. found that the recovery rate of DES reached 88%,
but the delignification ability had decreased (from 64.8
to 46.4%) (Li et al. 2018). Although the cost of DES is
low, the process is more expensive than conventional
treatment methods (such as acid treatment), which
limits its commercialization. The DES recycling
process still needs to be improved.
DES treatments have been applied in many fields,
such as enzymatic hydrolysis and nanocellulose
preparation. Shen et al. utilized lactic acid/choline
chloride to pre-treat eucalyptus, followed by enzy-
matic hydrolysis, and the enzymatically hydrolyzed
glucose yield reached 94.3%, which was a significant
increase of 9.8 times compared with the process
without DES pretreatment (Shen et al. 2019). Liu et al.
used chloride and lactic acid as pretreatment agents to
delignify moso bamboo for nanocellulose preparation
Cellulose (2021) 28:11503–11517
(Liu et al. 2019a). The results showed that lignin was
largely removed (94.39%). In addition, nanofibers
with a width of 20–80 nm were prepared through a
simple mechanical process, and the obtained nanofi-
bers produced a strong film with high tensile strength
(strength range of 163–213 MPa).
Dissolving pulp is a type of chemical pulp of high
alpha cellulose content, which is obtained by dissolv-
ing both lignin and hemicellulose from the wood raw
materials in the pulping process. Acid-based DES can
serve as a solvent to dissolve lignin and at the same
time as catalyst for hemicellulose hydrolysis. In
addition, DES can be recycled and reused for the
process to reduce chemical cost and environmental
impact, and therefore is a green pulping agent.
In present study we investigated the effect of
process conditions of lactic acid/choline chloride
(10:1) in treating eucalyptus wood materials to prepare
high-purity cellulose pulp. First, we optimized the
solid-liquid ratios in terms of lignin and hemicellulose
removal rates. Next, through recycling of DES, we
established an efficient process for lignocellulosic
biomass fractionation. Finally, the DES treatment
process was combined with pulping and bleaching
processes to produce high-quality dissolving pulp.
Materials and methods
Wood raw materials and DES preparation
Eucalyptus chips were obtained from the Hunan
Province Yunong Paper Co, Ltd. (Chenzhou, China).
For the optimization experiments of the DES pre-
treatment, the eucalyptus chips were soaked in water
overnight, and processed into a pulp slurry of 350 mL
CSF freeness with a disc refiner (BX-200-2, Beijing
Chunhui Xinji paper mechanical factory). The pulp
slurry was dried at 40 °C to obtain an air-dry
mechanical pulp (DMP) for subsequent treatments
with DES. The contents of glucan, xylan, acid-
insoluble lignin, and acid-soluble lignin of the DMP
sample were 39.01%, 14.3%, 25.95%, and 3.29%,
respectively. For dissolving pulp preparation experi-
ments, the eucalyptus wood chips were used directly
without refining treatment for the DES pretreatment.
DES was prepared by mixing lactic acid/choline
chloride with a molar ratio of 10:1 at 60 °C and 200
r/min for 2 h the constant temperature shaking
11505
incubator (ZWYR-2402, Zhicheng Analytical Instru-
ment Co., Ltd, Shanhai, China).
DES pre-treatment of the DMP wood material
In order to optimize the process conditions of DES
extraction including temperature, time and solid–
liquid ratio, the following experiments were carried
out. In the first set of experiments, 40 g DES was
mixed with to 2 g DMP and heated at 80, 110, 140 or
160 °C for 1, 2, 4, or 6 h. In another set of
experiments, 10, 20, 30 or 40 g DES was mixed with
2 g DMP, and heated at 110 °C for 6 h. By the end of
the reaction, the mixture was centrifuged at
10,000 rpm for 10 min and filtered with a G2 glass
filter. The filtrate was saved for analyses, and the solid
residue retained on the filer was washed with 50 mL of
ethanol, and then washed with hot water until the
filtrate was colorless. The solid residue was then air
dried in an oven at 40 °C.
In the 3rd set of experiments, 300 g of DES was
added to 30 g of DMP and heated at 110 °C for a total
reaction time of 6 h, with or without solvent recycling.
In one experiment, the reaction time was 6 h in one
stage, without solvent recycling. The second experi-
ment was carried out in two 3-h stages. After the first
3-h stage, the mixture was centrifuged at 10,000 r/min
for 10 min, and the supernatant was treated to recover
the dissolved lignin and to regenerate the DES solvent
according to the procedure described in the subsection
of Lignin recovery and DES solvent regeneration. The
regenerated DES solvent was then used for the second
3-h treatment of the DMP (the solid precipitate from
the centrifugation). In the third experiment, the DES
extraction treatment was split into three 2-h stages,
with DES solvent recycling for the 2nd and 3rd stages
as described above. In completion of the total 6 h of
DES treatment, the mixture was centrifuged at 10,000
r/min for 10 min, and the sediment was washed with
300 mL of ethanol and then with hot water until the
water was colorless. The spent ethanol was recovered
by rotary evaporation, and the residue liquid was
combined with the supernatant of the centrifugation to
give a final spend liquor for lignin recovery and DES
solvent regeneration.
123
11506
Lignin recovery and DES solvent regeneration
Ten volumes of water were mixed with the final spend
liquor from the DES extraction process, refrigerated
overnight and filtered out the precipitated lignin with a
G3 glass filter. The filtrate was treated with a vacuum
rotary evaporator at 60 °C to regenerate DES solvent
by boiling out the water. The regenerated DES was
labeld as CRL. The lignin retained on the filter was
washed with deionized water to neutrality, dried in an
oven at 60 °C (Alvarez-Vasco et al. 2016; Kim et al.
2018), and labeled as RL.
Lignin and carbohydrate analyses
The composition of the CRL were determined accord-
ing to the method provided by NREL (Sluiter et al.
2006). The liquid sample was hydrolyzed by acid, and
then the components in the liquid were analyzed by
HPLC. The amount of DES recovered was calculated
from the lactic acid content in CRL, and 1 g lactic acid
was equal to 1.155 g DES. The DES recovery rate was
calculated by the following equation.
DES Rec: Rate ¼ 1:155
Lactic acid content  Vol: of CRL
 :
Amount of DES added
The total amount of acid soluble lignin (ASL) and
acid insoluble lignin (AIL) in the sold samples (DMP
Average dissolution rate
mass of component in DMP  mass of component after DES treatment
¼
mass of DES  reation time
or RL) were determined in accordance with NREL
(Sluiter et al. 2008). The AIL was the solid residue
after acid hydrolysis that was measured by gravimetric
method, and the ASL in the acid-hydrolyzed filtrate
was measured using UV–vis spectrophotometer
((UV–Vis, T6 series, Beijing spectrum analysis Gen-
eral Instrument Co., Ltd., Beijing, China) at 205 nm.
The calculation of the lignin purity and recovery rate
was as follows.
123
Cellulose (2021) 28:11503–11517
AIL þ ASL
Ligninpurity ¼  100%
RL
Lignin Purityð%Þ  RLðgÞ
Lig: Rec: Rate ¼  100%
Lignin in woodðgÞ  Lignin in residueðgÞ
The pH of the black liquor from the pre-hydrolysis
kraft cooking process was adjusted to 2 by adding
dilute sulfuric acid (Tian et al. 2013), and the resulting
precipitate was recovered as kraft lignin (KL).
The molecular mass distribution of lignin was
obtained by GPC (Waters 1515). Tetrahydrofuran
THF) was used as a column eluent (1 mL/min flow
rate). The column temperature was maintained at
45 °C. The sample was injected through 0.45 lm
filter, and the lignin in the eluent was detected by 245
nm variable wavelength detector.
The cellulose hemicellulose contents in the solid
samples were determined in accordance with the
National Renewable Energy Laboratory Standard
Procedure (NREL) (Sluiter et al. 2008). The solid
sample underwent a two-step acid hydrolysis to
convert carbohydrates into monosaccharides, which
were determined by HPLC (Agilent1260 series, Agi-
lent Technologies, USA; Bio-Rad Aminex HPX-87 H
Column, Refractive index detector). The average
dissolution rate of each component was calculated
by the following formula:
FT-IR spectroscopy and X-ray diffraction (XRD)
For FT-IR analysis, the solid samples (DMP and RL)
were first freeze-dried and ground into powder. The
powder sample (0.01 g) was mixed with KBr (1 g),
ground and pressed at 10 MPa into thin disc. The thin
disc was then scanned with a Fourier Transform
infrared spectroscopy instrument (FT-IR, Nicolet
iS10, Thermo Fisher Scientific, Waltham, MA,
USA) in the frequency range of 4000–400 cm- 1 with
a resolution of 4 cm- 1.
Cellulose (2021) 28:11503–11517
For XRD analysis, the dried powder samples were
analyzed by a X-ray diffractometer (XRD) (D8-FOUS,
Bruker, Kalsruhe, Germany). The scattering angle (2h)
ranged from 10° to 50° with 0.02 increments. The
crystallinity index (CrI) was calculated as follow:
Crl ¼ ðI002  Iam Þ=I002  100%
where I002 was the intensity of the crystalline peak at
about 2h = 22°–23°, attributed to crystalline regions,
and Iam was the intensity at about 2h = 18°–19°,
attributed to amorphous regions.
Preparation of dissolving pulps by 3 different
processes
For comparison, 3 dissolving pulps were prepared by 3
different processes from the eucalyptus wood chips.
The first one was pre-hydrolyzed kraft pulp (PHK),
which was prepared by hydrothermal pretreatment of
eucalyptus wood chips at 160 °C for 2 h with a liquid-
to-wood ratio of 7:1, followed by kraft cooking with
21% alkali and 17% sulfidity, at 130 °C for 0.5 h and
at 160 °C for 2 h. The resultant pulp was bleached with
a DEP sequence (D: 0.9% ClO2, T = 70 °C, t = 2 h;
E: 0.3% NaOH, T = 70 °C, t = 1 h°°P: 0.3% H2O2,
0.05% MgSO4, 3% Na2SiO3, T = 70 °C, t = 1 h)
(Chen et al. 2020).
The second dissolving pulp was DES-KP. It was
prepared by DES pretreatment of eucalyptus wood
chips with lactic acid/choline chloride (10/1) DES
solvent, liquid–wood ratio of 10, total reaction time of
6 h at 110 °C in two 3-h stages with lignin removed
between the two stages. After the DES solvent
recycling pre-treatment, the same kraft cooking and
bleaching processes were carried out as described
above for the PHK dissolving pulp preparation.
The third dissolving pulp was labeled as DES-BP,
which was prepared by the same pretreatment as in
DES-KP preparation, but instead of kraft cooking the
pretreated wood chips were delignified by two stages
of oxygen delignification under the same conditions
(O–O: 0.5 MPa O2, 3% NaOH, 0.05% MgSO4,
T = 100 °C, t = 1 h). The resultant pulp was bleached
by a DEpD sequence (D: 0.9% ClO2, T = 70 °C,
t = 2 h; Ep: 2% H2O2, 4% NaOH, 0.5% DTPA, 2%
Na2SiO3; D: 0.5% ClO2, T = 70 °C, t = 2 h).
11507
Characterization of the dissolving pulps
The a-cellulose content was determined as the insoluble
fraction of cellulose pulp in consecutive treatments with
17.5 and 9.45% NaOH solutions according to TAPPI
T203. The Fock reactivity test was conducted according
to the method reported by Tian et al. (Tian et al. 2013).
Brightness was tested according to TAPPI T452 om-08,
and the whiteness test was conducted with a whiteness
meter (WS-SD, WENZHOU INSTRUMENTS &
METERS CO., LTD, Wenzhou, China). Viscosity was
measured using a capillary viscometer with CED as the
solvent by following TAPPI T230-94.
Results and discussion
Effect of process condition on the dissolution
of lignin, cellulose and hemicellulose in DES pre-
treatment
We first studied the effect of process conditions on the
dissolution of lignin, cellulose and hemicellulose in
DES solvent pretreatment, in order to maximize the
pre-treatment effect of DES on eucalyptus wood
materials. The eucalyptus DMP was treated with DES
at different temperatures (80, 110, 140 or 160 °C) and
for various periods of time (1, 2, 4 and 6 h), and the
composition changes of the samples after the DES
pretreatment were compared. The results were sum-
marized in Fig. 1. When the reaction time was short (1
or 2 h), higher temperature promoted lignin and
hemicellulose dissolution, but when the temperature
was too high (e.g. 160 °C) the loss of glucan was
significant. As reported by Song et al., cellulose was
severely degraded at a temperature above 140 °C
(Song et al. 2016). When the temperature was fixed,
increasing reaction time increased the dissolution of
lignin and the removal of xylan. It was interesting to
note that when the temperature was lower (B110 °C),
increasing the reaction time had negligible effect on
the loss of glucan. However, when the temperature
was too low (e.g. 80 °C), DES pretreatment had little
effect on the major components (lignin, hemicellulose
and cellulose) of the DMP eucalyptus wood material.
This may be attributed to the fact that the breakage of
the guaiacylglycerol-b-guaiacy ether bonds (b–O–4
model) in DES treatment occurred only at higher
temperature (Zheng et al. 2020). It was worth noting
123
11508
Fig. 1 Effect of the DES treatment process conditions on the lignin, xylan and glucan contents of the DMP wood material. The content
of each component was based on DMP mass
that when the reaction was longer than 4 h in the DES
treatment at a high temperature (160 °C), lignin
dissolution actually started to decrease, while the loss
of glucan was more pronounced. This observation may
be explained by the dehydration condensation of
polysaccharide at high temperature forming lignin-
like substances (Donohoe et al. 2008).
Effect of liquid-to-wood ratio
We also investigated the effect of the ratio of DES
solvent to DMP wood material (or liquid to wood
ratio) on the dissolution of lignin, xylan and glucan
from the lignocellulosic biomass. In Fig. 2, the
eucalyptus DMP wood material were treated with
the DES solvent at various liquid-to-wood ratios (5,
10, 15 and 20) for various periods of time (2, 3 and
6 h). As it can be seen in Fig. 2(1), the dissolution of
lignin and xylan increased with the increase of
reaction time, and the loss of glucan also increased.
When the reaction time was fixed, the dissolution of
lignin and xylan also increased, which was consistent
with the results reported by Alessandra Procentese
(Procentese et al. 2018). We also used the average
dissolution rates to evaluate the efficiency of DES
123
Cellulose (2021) 28:11503–11517
solvent in different cases, as shown in Fig. 2(2), (3),
(4). The average dissolution rate was calculated as the
average mass of a wood component (lignin, xylan or
glucan) dissolved per gram of DES and per hour of
reaction time. For example, at the same liquid-to-
wood ratio and reaction time, a higher average means
more mass of a wood component was dissolved in the
process. At the same liquid-to-wood ratio of 10, the
largest average dissolution rate was obtained with 3 h
of reaction time, in which 0.1145 g acid-insoluble
lignin was dissolved. When the liquid-to-wood ratio
was increased further to 15, the average dissolution
rate of acid-insoluble lignin decreased, and it
remained almost the same for 3 different reaction
times. This suggests that with a total pretreatment
reaction time of 6 h, the dissolved mass of acid-
insoluble lignin were similar in the 1-stage, 2-stage
and 3-stage processes. This also suggests that the
dissolution reaction of acid-insoluble lignin at a
liquid-to-wood ratio of 15 was a first-order reaction.
The results in Fig. 2 also show that a higher average
dissolution ratio was obtained at a lower liquid-to-
wood ratio. However, at a lower DES to wood ratio
(e.g. 5), the amounts of lignin and xylan dissolved in
the pre-treatment process were significantly lower.
Cellulose (2021) 28:11503–11517 11509

Fig. 2 Effect of DES

treatment time and liquid–
wood ratio on the contents of
lignin, xylan and glucan.
The content of each
component was based on the
original DMP mass

Effect of DES solvent recycling
Results in Fig. 2 show that the DES was more fully
utilized at a lower liquid-to-wood ratio in terms of
average dissolution rates. However, when the pre-
treatment with DES was carried out with a liquid-to-
wood ratio of 5 for 3 h, for example, only 0.0967 g of
acid-insoluble lignin was dissolved (the acid-insoluble
lignin content in the wood materials decreased from
0.2595 to 0.1628 g Fig. 2(2) and Fig. 1.), which was
compared to 0.1216 g with a liquid-to-wood ratio of

123
11510
Table 1 Solid and liquid components after multiple times DES processing
Process Solid Residuei
Glucan Xylan Lignin Lignin recovery
(%) (%) (%) rate (%)
1-stage 37.21 1.71 8.53 86.11
2-stage 34.99 1.46 7.34 72.42
3-stage 34.69 1.36 8.99 78.47
i. The solid residue was obtained by centrifugation after the last stage of DES pre-treatment, and the component contents were based
on DMP; ii. ‘–‘ Not detected.
20. Although DES was more efficiency with a smaller
liquid-to-wood ratio, the total mass of lignin dissolved
in the process was less. Therefore, the liquid-to-wood
ratio needs to be balanced between pre-treatment
effectiveness and solvent cost. DES solvent recycling
can substantially improve the cost-effectiveness.
Table 1 compares 3 different DES pre-treatment
processes (1-stage, 2-stage and 3-stage) in treating the
DMP wood material. The total treatment reaction time
was fixed at 6 h in all cases. The 2-stage process (3 h
for each stage) dissolved the most lignin (lowest lignin
content in the solid residue), followed by the 3-stage
process. The dissolution of glucan and xylan in the
3-stage process was slightly higher than the 2-stage
process. These results also indicated that the recycled
DES solvent was as active as the fresh DES. It was
worth noting that the 3-stage pre-treatment process
dissolved the most lignin, but the lignin recovery rate
was the lowest. Similar observation was reported by
Ao-Lin Li et al.: increased severity of DES pretreat-
ment could lead to increased loss of lignin (Li et al.
2018). This phenomenon may be attributed to the
additive effect of the severity of each stage of DES
treatment in the 2-stage process. The lignin was
determined as the acid insoluble substances that
cannot be dissolved in 72 and 4% sulfuric acid
solutions, and the substances dissolved in the acid
solutions that have absorption at 205 nm UV wave-
length. Pseudo-lignin formed through the condensa-
tion of oligosaccharides also had strong absorption at
205 nm. After prolonged acid treatment, polysaccha-
rides could condense to form pseudo-lignin, which
could not be distinguished from true lignin (Sanni-
grahi et al. 2011). In the process of acidic pretreatment
of biomass, cellulose was mainly degraded to glucose,
glucose was converted to HMF, and HFM could be
123
Cellulose (2021) 28:11503–11517
CRL
Xylose Glucose Furfural HMF Levulinic DES recovery
(mg) (mg) (mg) (mg) acid (mg) rate (%)
1771.54 –ii 0.195 9.373 28.13 77.36
ii
2261.73 – 2.402 9.232 18.46 92.86
ii
2750.99 – 0.268 12.855 231.39 86.34
degraded to levulinic acid; long chains of xylan in
hemicellulose was degraded into xylose, xylose was
converted into furfural, and branched chains in
hemicellulose could be degraded into mannose,
galactose, glucose and arabinose, etc. And among
them, hexoses were converted into HMF and levulinic
acid like glucose, and pentoses were converted into
furfural like xylose (Palmqvist and Hahn-Hägerdal
2000). It can be seen in Table 1 that the saccharides
and their degradation products (furfural, HMF and
levulinic acid) in the spent liquor from the 1-stage
process were significantly lower than those from the
2-stage and 3-stage processes, indicating that the
lignin recovered from the second and third stages
could contain more pseudo-lignin produced by the
condensation and degradation of polysaccharides.
X-ray diffraction (XRD) and FT-IR analyses
XRD was used to analyze the changes in the
crystallinity of the eucalyptus wood material after
DES treatment.
As shown in Fig. 3, after the DES treatment, the
peak position did not change, indicating that DES did
not cause any change to the cellulose crystal structure
(Liu et al. 2019b). The calculated cellulose crys-
tallinity index of the solid residue and the raw material
was shown in Fig. 3. After the DES treatment, the CrI
index of cellulose increased from 49.4 to 60.6%, 60.8
and 62.2%, respectively. The increase of crystallinity
index indicated the decrease of non-cellulose compo-
nents (hemicellulose and lignin) (Xu et al. 2016).
The FT-IR spectra of the eucalyptus DMP wood
material and the solid residue after DES treatment
were shown in Fig. 4. The presence of a strong
absorption peak at 1732 cm- 1 that was attributed to
Cellulose (2021) 28:11503–11517 11511

Fig. 3 XRD spetra of the wood material before and after DES pre-treatment. The 1 time, 2 times and 3 times referred to the 1-stage,
2-stage and 3-stage DES treatment processes, respectively. The total reaction time was 6 h in all cases

Fig. 4 The FT-IR spectra of

the wood material before
and after the DES pre-
treatment. The 1 time, 2
times and 3 times referred to
the 1-stage, 2-stage and
3-stage DES treatment
processes, respectively. The
total reaction time was 6 h
in all cases

123
11512 Cellulose (2021) 28:11503–11517

Table 2 Fiber morphology analysis of the eucalyptus wood and dissolving pulps prepared by three different processes
Average length (mm) Fines content (%) Average width (lm)
Ln Lw Lww Number average Weight average

Eucalyptus 0.710 0.789 0.856 9.600 1.880 19.9

PHK 0.582 0.663 0.733 15.612 3.949 15.3
DES-KP 0.525 0.623 0.718 27.378 8.460 15.3
DES-BP 0.516 0.614 0.704 28.459 9.148 15.3
Compared with the fibers of the original eucalyptus wood raw material, dissolving pulp fibers exhibited various degrees of fiber
deformation including kink, twist and swelling, as shown in Fig. 5. The microscopic images show that the PHK fibers seemed to be
shorter than the original eucalyptus wood fibers, but longer than the DES-BP and DES-KP fibers. The fiber morphology data in
Table 2 confirmed this observation: the weight average length (Lw) of the eucalyptus wood fibers was 0.789 mm, which was
compared to 0.663 mm, 0.623 mm and 0.614 mm for the PHK, DES-BP and DES-KP fibers, respectively. The micrographs in Fig. 5
also show that many fiber fragments were ‘‘stripped out’’ from the DES-KP and the DES-BP fibers, which contributed to the higher
fines content of the two dissolving pulps, as shown in Table 2. The fiber morphology changes of the DES-KP and DES-BP fibers
were somewhat similar to the effect of mechanical refining, which increased the specific surface area and accessibility of the fibers
(Mou et al. 2013; Pönni et al. 2013). These changes in fiber morphology were probably the cause of the improved Fock reactivity of
the dissolving pulps that were prepared with DES solvent recycling pretreatment, as shown in Table 3. The Fock reactivity of the
DES-KP and DES-BP dissolving pulps were 98.9% and 96.8% respectively, which were much higher than that of the PHK (60.97%)

Table 3 Characteristics of the dissolving pulps prepared by 3 different processes from Eucalyptus wood chips
Xylan Glucan Lignin Yield a-cellulose content Fock reactivity Brightness (% Viscosity
(%) (%) (%) (%) (%) (%) ISO) (mL/g)

PHK 3.31 83.7 0.583 38.59 89.81 60.97 87.99 436.2

DES- 1.70 88.0 0.560 37.01 94.02 98.94 88.98 396.3
KP
DES- 2.11 84.6 3.424 44.40 88.31 96.82 88.48 461.1
BP

Fig. 5 Micrographs of the fibers of the euclaytus wood and the dissolving pulps prepared by three different processes

123
Cellulose (2021) 28:11503–11517
Fig. 6 The FT-IR spectra of
kraft lignin (KL) and
recovered lignin (RL)
recovered from DES
pretreatment spent liquor
Table.4 Molecular weight of kraft lignin (KL) and recovered lignin (RL) from the spent liquor of DES pre-treatment
Lig. Rec. Rate (%) Mn (g/mol)
KL 67.0 1234
RL 72.4 1757
the overlapping bands of the C=O groups of the ester
and the protonated carboxylic acid, indicated that a
large amount of hemicellulose acetyl groups was
retained after the lactic acid/ChCl treatment (Liu et al.
2019b; Sirviö and Visanko 2017; Tian et al. 2020).
The intensity of peak 1235 cm- 1 (the vibration peak
of ether bond between lignin and hemicellulose)
weakened, indicating that lignin was removed in the
process (Pan et al. 2017). The characteristic peaks of
lignin at 1600 cm- 1, 1505 cm- 1 and 1462 cm- 1
were significantly weakened after the DES treatment,
which also indicated that DES treatment removed
lignin from the eucalyptus DMP wood material (Guo
et al. 2018). The peaks at 2900 cm- 1 and 898 cm- 1
were attributed to the C–H vibration of cellulose and
the C–O–C stretching vibration of the b-1,4 glycosidic
bond (Lim et al. 2019). Both peaks were significantly
11513
Mw (g/mol) Mz (g/mol) Mw/Mn
2648 5450 2.1450
4484 14,804 2.5521
enhanced after the DES treatments due to the removal
of lignin and xylan and the relative increase in glucan
content.
Dissolving pulp performance
To evaluate the effect of the 2-stage DES pretreatment
process on dissolving pulp production, 3 dissolving
pulps were prepared by 3 different processes from the
eucalyptus wood chips. The first one was conventional
pre-hydrolyzed kraft pulp (PHK), which was prepared
by hydrothermal pretreatment of eucalyptus wood
chips followed by kraft cooking and DEP bleaching.
The second dissolving pulp was DES-KP that was
prepared by the 2-stage DES pretreatment of the
eucalyptus wood chips followed by kraft cooking and
DEP bleaching. The DES pretreatment was carried out
123
11514
Fig. 7 Biorefinery strategies in the preparation of dissolving pulp by the DES-KP pulping process. (The purified lignin included acid-
soluble lignin and acid-insoluble lignin.)
with lactic acid/choline chloride (10/1) DES solvent at
a liquid-to-wood ratio of 10 in two 3-h stages with
lignin removed between the two stages, at 110 °C for a
total reaction time of 6 h. After the DES solvent
recycling pre-treatment, the same kraft cooking and
bleaching processes were carried out as described
above for the PHK dissolving pulp preparation. The
third dissolving pulp was labeled as DES-BP, which
was prepared by the same pretreatment as in DES-KP
preparation, except that instead of kraft cooking the
pretreated wood chips were delignified with two stages
of oxygen delignification and bleached by a DEpD
sequence.
Table 3 Also shows that the DES-KP that was
prepared with DES pre-treatment had higher glucan
content (88.0% vs. 83.7%), much better Fock reactiv-
ity (98.94% vs. 60.97%) substantial higher a- cellu-
lose content (94.0% vs. 89.8%) and significantly lower
xylan contents (1.7% vs. 3.31%), in comparison with
the PHK dissolving pulp. Compared with the PHK
process, the DES-KP process had much lower loss of
cellulose thanks to the DES solvent recycling pre-
treatment, which removed a large amount of hemicel-
lulose and lignin. The advantages of the DES-BP
process included a higher pulping yield (44.4%) and
lower environmental impact thanks to its greener
oxygen-alkali cooking and DEpD bleaching pro-
cesses. However, the DES-BP pulp had higher lignin
123
Cellulose (2021) 28:11503–11517
content (3.424%) and lower a-cellulose content
(88%), which were unsuitable for the production of
viscose fibers.
Recovery of lignin
Figure 6 shows the FT-IR spectra of the kraft lignin
(KL) and DES recovered lignin (RL). The broad band
at 3400 cm- 1 was attributed to aliphatic and phenolic
hydroxyl groups. 1600 cm- 1, 1513 cm- 1 and
1457 cm- 1 were characteristic peaks of lignin (Li
et al. 2017). The peaks at 1324 cm- 1 and 1212 cm- 1
represented syringyl lignin and guaiacyl lignin,
respectively. These two peaks showed presence of
the benzene ring structure of the lignin. The RL lignin
had a strong peak at 1324 cm- 1, indicating that the
DES treatment dissolved more syringyl lignin. The
1732 cm- 1 peak (ester C=O tensile vibration peak)
was intensified markedly in the infrared spectra of RL,
suggesting that the carboxyl and hydroxyl groups of
lignin were esterified in the acid DES pre-treatment
(Constant et al. 2016). In summary, most of the
characteristic absorption peaks of lignin remained
unchanged in the spectra of RL, indicating the lignin
structures were largely preserved (Tian et al. 2020).
Table 4 compares the mean molecular weight of the
KL and RL lignin. The RL lignin (Mw = 4484 g/mol)
had a higher mean molecular weight than the KL
Cellulose (2021) 28:11503–11517
lignin (Mw = 2648 g/mol), and the recovery rate of
the RL lignin was also higher (72.4% vs. 67.0%). The
phase transition from soluble to precipitated lignin
was found to be tightly linked to molecular weight of
lignin. It has been shown that high molecular weight
lignin precipitates more easily, while stable low
molecular weight lignin precipitates by attaching to
the surface of high molecular weight lignin, thus
making high molecular weight lignin easier to recover
(Norgren et al. 2001). Moreover, the RL lignin was
easier to be recovered than KL lignin, as it was
precipitated out from the spent liquor simply by
adding water without the need of a complex desulfu-
rization process. Molecular weight of lignin can also
provide information on the fragmentation and re-
condensation reactions of lignin in the pulping
processes. Lignin varied in molecular weight due to
the difference in the dissolving processes and the
difference in the extraction methods. In the sulphate
cooking process, lignin reacted not only with alkali,
but also with sodium sulfide. Lignin monosulfide
produced by the reaction with sodium sulfide could
also inhibit the secondary condensation of lignin
structural units, leading to smaller mean molecular
weight (Mw = 2648 g/mol) (Van Heiningen 2007).
Lactic acid/choline chloride DES solvent mainly
attacked the b–O–4 bond of lignin, breaking down
lignin macromolecules into small molecules, as illus-
trated in the Fig. 7. The RL lignin also had a wider
molecular weight distribution compared with the KL
lignin, as indicated by the Mw/Mn ratio (2.55 vs. 2.14)
(Shen et al. 2020). The DES pretreatment not only
contributed to the breakage of the C–O and C–C bonds
of the lignin macromolecules, but also caused the
dehydration and acylation of the OH groups on the
side chains of lignin and partial condensation of lignin
(Shen et al. 2020), which led to the larger molecular
weight of the RL lignin.
Biorefinery strategies in the preparation
of dissolving pulp by the DES-KP pulping process
According to the biorefinery concept, the mass balance
of the components of the eucalyptus wood material
were evaluated for the processes in the preparation of
dissolving pulp by the DES-KP pulping process. The
process flow chart and biomass streams were illus-
trated in Fig. 7. Starting with 100 g of dry eucalyptus
wood chips, 37.01 g of DES-KP dissolving pulp was
11515
produced as the key product, and 7.54 g of xylan,
0.77 g of furfural and 1.05 g of acetic acid were
recovered from the spent liquor as byproducts. About
803.76 g of lactic acid in the spent liquor (CRL) were
recovered and reused in the DES pre-treatment. In
addition, 23.38 g crude lignin (RL) was obtained from
the spent liquor by precipitation in water, 17.21 g of
which of was recovered as purified lignin. The overall
biomass recovery rate reached 69.74% based on the
mass of the feedstock (eucalyptus wood chips), which
was significantly higher than the number of 66.04%
obtained by Chen et al. who applied an acetic acid
pretreatment process for the production of kraft-based
eucalyptus dissolving pulp (Chen et al. 2020).
The scaling up of the DES-KP process for com-
mercial production of dissolving largely depends on
the cost of the DES solvent. Lactic acid and choline
chloride are commercially available, but their costs are
still high compared to conventional PHK process.
Recycling of DES in the process is a key to make the
process more cost-effectively. If the oligosaccharides
dissolved in the process can be converted into lactic
acid through a fermentation process (Jojima et al.
2010; Dien et al. 2002), the cost of DES solvent will
decrease significantly in the future.
Conclusions
A novel acid-based deep eutectic solvent (DES)
recycling process was developed for the pretreatment
of eucalyptus wood chips in the preparation of high
purity dissolving pulp. It was found that a large
amount of lignin and xylan in the eucalyptus wood
material were dissolved in the lactic acid/choline
chloride DES solvent recycling treatment at 110 °C
for 3 h. Furthermore, the residual lignin and xylan in
the pre-treated wood materials was more readily
removed in the subsequent kraft cooking and bleach-
ing processes, yielding a high-purity dissolving pulp
(DES-KP). Compared with the conventional prehy-
drolysis kraft based dissolving pulp (PHK), the DES-
KP dissolving pulp had a higher a-cellulose (94.02%
vs. 89.81%) and markedly higher Fock reactivity
(98.94% vs. 60.97%), which was a breakthrough
improvement of the reactivity of kraft-based dissolv-
ing pulp. In addition, the DES pretreatment could be
combined with green oxygen-alkali cooking and ECF
bleaching process to produce a DES-BP dissolving
123
11516
pulp with a higher pulping yield (44.40%). The DES-
KP pulping process provided a good strategy to utilize
the biomass of the eucalyptus wood chips more
efficiently in dissolving pulp production. The overall
biomass utilization rate of DES-KP process reached
69.74%, which was significantly higher than that
reported in the literature.
Acknowledgments This work was financially supported by
the Fundamental Research Funds of CAF
(CAFYBB2018GD002) and Taishan Industrial Experts
Programm (tscy20200213).
Declarations
Conflict of interest No potential conflict of interest was
reported by the authors.
Human and animal rights The study was approved by the
Ethics Committee of Institute of Chemical Industry of Forestry
Products. All procedures performed in studies involving human
were in accordance with the ethical standards of the institution
or practice at which the studies were conducted.
Informed consent Informed consent was obtained from all
individual participants included in the study.
References
Alvarez-Vasco C et al (2016) Unique low-molecular-weight
lignin with high purity extracted from wood by deep
eutectic solvents (DES): a source of lignin for valorization.
Green Chem 18:5133–5141. https://doi.org/10.1039/
C6GC01007E
Chen Y et al (2020) Comparative evaluation of organic acid
pretreatment of eucalyptus for Kraft dissolving. Pulp Prod
Mater 13:361
Constant S et al (2016) New insights into the structure and
composition of technical lignins: a comparative charac-
terisation study. Green Chem 18:2651–2665. https://doi.
org/10.1039/C5GC03043A
Dien BS, Nichols NN, Bothast RJ (2002) Fermentation of sugar
mixtures using Escherichia coli catabolite repression
mutants engineered for production of L-lactic acid. J Ind
Microbiol Biotechnol 29:221–227. https://doi.org/10.
1038/sj.jim.7000299
Donohoe BS, Decker SR, Tucker MP, Himmel ME, Vinzant TB
(2008) Visualizing lignin coalescence and migration
through maize cell walls following thermochemical pre-
treatment. Biotechnol Bioeng 101:913–925. https://doi.
org/10.1002/bit.21959
Francisco M, Bruinhorst AVD, Kroon MC (2012) New natural
and renewable low transition temperature mixtures
(LTTMs): screening as solvents for lignocellulosic bio-
mass processing. Green Chem 14:2153–2157. https://doi.
org/10.1039/C2GC35660K
123
Cellulose (2021) 28:11503–11517
Guo Z, Ling Z, Wang C, Zhang X, Xu F (2018) Integration of
facile deep eutectic solvents pretreatment for enhanced
enzymatic hydrolysis and lignin valorization from indus-
trial xylose residue. Bioresour Technol 265:334–339.
https://doi.org/10.1016/j.biortech.2018.06.027
Jojima T, Omumasaba CA, Inui M, Yukawa H (2010) Sugar
transporters in efficient utilization of mixed sugar sub-
strates: current knowledge and outlook. Appl Microbiol
Biot 85:471–480. https://doi.org/10.1007/s00253-009-
2292-1
Kim KH, Dutta T, Sun J, Simmons B, Singh S (2018) Biomass
pretreatment using deep eutectic solvents from lignin
derived phenols. Green Chem 20:809–815. https://doi.org/
10.1039/C7GC03029K
Kumar AK, Parikh BS, Pravakar M (2016) Natural deep eutectic
solvent mediated pretreatment of rice straw: bioanalytical
characterization of lignin extract and enzymatic hydrolysis
of pretreated biomass residue. Environ Sci Pollut R
23:9265–9275. https://doi.org/10.1007/s11356-015-4780-
4
Li T et al (2017) Deep eutectic solvents (DESs) for the isolation
of willow lignin (Salix matsudana cv. zhuliu). Int J Mol Sci
18:2266. https://doi.org/10.3390/ijms18112266
Li A, Hou X, Lin K, Zhang X, Fu M (2018) Rice straw pre-
treatment using deep eutectic solvents with different con-
stituents molar ratios: biomass fractionation,
polysaccharides enzymatic digestion and solvent reuse.
J Biosci Bioeng 126:346–354. https://doi.org/10.1016/j.
jbiosc.2018.03.011
Lim W, Gunny AAN, Kasim FH, AlNashef IM, Arbain D (2019)
Alkaline deep eutectic solvent: a novel green solvent
for lignocellulose pulping. Cellulose 26:4085–4098.
https://doi.org/10.1007/s10570-019-02346-8
Liu Q, Yuan T, Fu Q, Bai Y, Peng F, Yao C (2019) Choline
chloride-lactic acid deep eutectic solvent for delignifica-
tion and nanocellulose production of moso bamboo. Cel-
lulose 26:9447–9462. https://doi.org/10.1007/s10570-019-
02726-0
Liu Q, Yuan T, Fu Q, Bai Y, Peng F, Yao C (2019) Choline
chloride-lactic acid deep eutectic solvent for delignifica-
tion and nanocellulose production of moso bamboo. Cel-
lulose 26:9447–9462. https://doi.org/10.1007/s10570-019-
02726-0
Mou H, Iamazaki E, Zhan H, Orblin E, Fardim P (2013)
Advanced Studies on the Topochemistry of Softwood
Fibres in Low-Consistency Refining as Analyzed by FE-
SEM XPS and ToF-SIMS. BioResources. https://doi.org/
10.15376/biores.8.2.2325-2336
Norgren M, Edlund H, Wågberg L, Lindström B, Annergren G
(2001) Aggregation of kraft lignin derivatives under con-
ditions relevant to the process, part I: phase behaviour.
Colloids Surf A 194:85–96. https://doi.org/10.1016/
S0927-7757(01)00753-1
Palmqvist E, Hahn-Hägerdal B (2000) Fermentation of ligno-
cellulosic hydrolysates. II: inhibitors mechanisms of inhi-
bition. Bioresour Technol 74:25–33. https://doi.org/10.
1016/S0960-8524(99)00161-3
Pan M, Zhao G, Ding C, Wu B, Lian Z, Lian H (2017)
Physicochemical transformation of rice straw after pre-
treatment with a deep eutectic solvent of choline chloride/
Cellulose (2021) 28:11503–11517
urea. Carbohyd Polym 176:307–314. https://doi.org/10.
1016/j.carbpol.2017.08.088
Pönni R, Kontturi E, Vuorinen T (2013) Accessibility of cel-
lulose: structural changes and their reversibility in aqueous
media. Carbohyd Polym 93:424–429. https://doi.org/10.
1016/j.carbpol.2012.12.025
Procentese A, Raganati F, Olivieri G, Russo ME, Rehmann L,
Marzocchella A (2018) Deep eutectic solvents pretreat-
ment of agro-industrial food waste. Biotechnol Biofuels
11:37. https://doi.org/10.1186/s13068-018-1034-y
Sannigrahi P, Kim DH, Jung S, Ragauskas A (2011) Pseudo-
lignin and pretreatment chemistry. Energy Environ Sci
4:1306–1310. https://doi.org/10.1039/C0EE00378F
Shen X, Wen J, Mei Q, Chen X, Sun D, Yuan T, Sun R (2019)
Facile fractionation of lignocelluloses by biomass-derived
deep eutectic solvent (DES) pretreatment for cellulose
enzymatic hydrolysis and lignin valorization. Green Chem
21:275–283. https://doi.org/10.1039/C8GC03064B
Shen X et al (2020) Structural and morphological transforma-
tions of lignin macromolecules during bio-based deep
eutectic solvent (DES) pretreatment. ACS Sustain Chem
Eng 8:2130–2137. https://doi.org/10.1021/acssuschemeng.
9b05106
Sirviö JA, Visanko M (2017) Anionic wood nanofibers pro-
duced from unbleached mechanical pulp by highly efficient
chemical modification. J Mater Chem A 5:21828–21835.
https://doi.org/10.1039/C7TA05668K
Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D
(2006) Determination of sugars, byproducts, and degra-
dation products in liquid fraction process samples Golden.
National Renewable Energy Laboratory
Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D,
Crocker D (2008) Determination of structural carbohy-
drates and lignin in biomass. Lab Anal Proced 1617:1–16
Song W, Deng Y, Zhu H (2016) Research on Wheat Straw
Pulping with Ionic Liquid 1-Ethyl-3-Methylimidazole
Bromide. Kem Ind https://doi.org/10.15255/KUI.2016.032
Tan YT, Ngoh GC, Chua ASM (2019) Effect of functional
groups in acid constituent of deep eutectic solvent for
extraction of reactive lignin. Bioresour Technol
281:359–366. https://doi.org/10.1016/j.biortech.2019.02.
010
11517
Tan YT, Chua ASM, Ngoh GC (2020) Deep eutectic solvent for
lignocellulosic biomass fractionation and the subsequent
conversion to bio-based products–a review. Bioresour
Technol 297:122522. https://doi.org/10.1016/j.biortech.
2019.122522
Tian C, Zheng L, Miao Q, Nash C, Cao C, Ni Y (2013)
Improvement in the Fock test for determining the reactivity
of dissolving pulp. Tappi J 12:21–26
Tian D et al (2020) Acidic deep eutectic solvents pretreatment
for selective lignocellulosic biomass fractionation with
enhanced cellulose reactivity. Int J Biol Macromol
142:288–297. https://doi.org/10.1016/j.ijbiomac.2019.09.
100
Van Heiningen A (2007) Converting a Kraft Pulp Mill into an
Integrated Forest Products Biorefinery. Technical associ-
ation of the pulp and paper industry of Southern Africa
Journal. May
Xu G, Ding J, Han R, Dong J, Ni Y (2016) Enhancing cellulose
accessibility of corn stover by deep eutectic solvent pre-
treatment for butanol fermentation. Bioresour Technol
203:364–369. https://doi.org/10.1016/j.biortech.2015.11.
002
Zhang Q, Benoit M, De Oliveira Vigier K, Barrault J, Jérôme F
(2012) Green and inexpensive choline-derived solvents for
cellulose decrystallization Chemistry–A. Eur J
18:1043–1046. https://doi.org/10.1002/chem.201103271
Zhang C, Xia S, Ma P (2016) Facile pretreatment of lignocel-
lulosic biomass using deep eutectic solvents. Bioresour
Technol 219:1–5. https://doi.org/10.1016/j.biortech.2016.
07.026
Zheng Z, Eglin D, Alini M, Richards GR, Qin L, Lai Y (2020)
Visible light-induced 3D bioprinting technologies and
corresponding bioink materials for tissue engineering: a
review. Engineering-Prc. https://doi.org/10.1016/j.eng.
2020.05.021
Publisher’s Note Springer Nature remains neutral with
regard to jurisdictional claims in published maps and
institutional affiliations.
123