Industrial Crops & Products 115 (2018) 315–322

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Industrial Crops & Products

journal homepage: www.elsevier.com/locate/indcrop

The reactivity of lignin carbohydrate complex (LCC) during manufacture of T

dissolving sulfite pulp from softwood
Raghu Deshpandea,c,1, Nicola Giummarellab,1, Gunnar Henrikssona,b, Ulf Germgårda,
⁎
Lars Sundvallc, Hans Grundbergd, Martin Lawokob,
a
Karlstad University, SE-65188 Karlstad, Sweden
b
Wallenberg Wood Science Center, School of Chemical Science and Engineering, Royal Institute of Technology, KTH, 100 44 Stockholm, Sweden
c
MoRe Research, SE-89122 Örnsköldsvik, Sweden
d
Domsjö Fabriker, SE-89186 Örnsköldsvik, Sweden

A R T I C L E I N F O A B S T R A C T

Keywords: The presence of covalent bonds between lignin and polysaccharides was investigated in dissolving pulps made
Lignin carbohydrate complexes with one-stage and two-stage acidic sulfite pulping for 100% pine heartwood raw material. The covalent bonds
Sulfite pulping between lignin and pulp polysaccharides occurred mainly to xylan and glucomannan and were of the phenyl
Pine heartwood glycosides and γ–esters types. The α-ethers that are common in wood were missing in the studied pulp samples.
Universal fractionation method
Based on these findings and known lignin reactions during sulfite pulping, a mechanism explaining the absence
Size exclusion chromatography
of the α-ethers is discussed. It is suggested that the lignin carbohydrate bonds may play a vital role in lignin
recalcitrance.

1. Introduction hemicellulose than softwoods (conifers), where the hemicellulose xylan

In order to build a sustainable society, it is of central importance to
increase the use of renewable resources and decrease that of non-re-
newable raw materials, such as petroleum. Furthermore, it is important
that the materials based on the renewable resources are biodegradable,
or at least biocompatible. This is a problem with many synthetic
polymers, which today are accumulating in sea and other environments
with unclear consequences (Wright et al., 2013). In this context, ma-
terials based on wood cellulose, such as regenerated cellulose, cellulose
derivatives and nanocellulose, can therefore be foreseen to get an in-
creased importance in the future (Kamm and Kamm, 2004).
There are three main technologies for producing such pulps, that
normally are called dissolving pulps: prehydrolysis kraft pulping, one-
stage acidic sulfite pulping and two-stage acidic sulfite pulping, all of
which give different pulp properties (Woodings, 2001). The sulfite
pulps are often regarded to be higher qualities than the prehydrolysis
kraft pulps for certain applications; for instance, sulfite pulps have
higher reactivity in the viscose process (Bajpai, 2012). Another im-
portant parameter is the choice of raw material for the pulping. Hard-
woods (woody eudicotyledons) have a different composition of
dominates in the former, and the hemicellulose glucomannan dom-
inates the later (Sjöström, 1993). Glucomannan are to a great extent
degraded at higher temperature under alkali conditions, such as kraft
pulping or hot alkali extraction, whereas xylan is more stable (Sjöström,
1993). Thus, softwood can compete as raw materials for making dis-
solving pulps. However, for the Scandinavian industry, also the choice
of the species for softwood raw material is important, since the heart-
wood of pine contains phenolic extractives, such as pinosylvin, which is
a highly conjugated aromatic structure and might act as a nucleophile
in acidic conditions; thereby preventing efficient delignification and
lignin condensation reactions (Fengel and Wegener, 1984; Sixta, 2006;
Sjöström, 1993). Thus, pure spruce wood is preferred for one-stage
acidic sulfite pulping. Although pine wood is easily available and is
cheaper than spruce, there is limited knowledge on studies of pine
wood and especially pine heartwood during acid sulfite pulping.
However, for the two-stage acid sulfite pulping where the first pulping
stage is performed at higher pH (around pH 5), pine can also be used as
a raw materials for dissolving pulp production. At these pH conditions
condensation reactions are minimized and lignin become extensively
sulfonated. This also lowers the risk for condensation reactions in a

Abbreviations: DMSO, dimethyl sulfoxide; L/W, liquor to wood ratio; LCC, lignin carbohydrate complexes; LC, lignin carbohydrate; LiBr, lithium bromide; NMR, nuclear magnetic
resonance; RI, refractive index; SEC, size exclusion chromatography; UV, ultraviolet; GE, gamma ester; PG, phenyl glycoside; BE, benzyl Ether
⁎
Corresponding author.
E-mail addresses: raghu.deshpande@more.se (R. Deshpande), nicolag@kth.se (N. Giummarella), ghenrik@kth.se (G. Henriksson), ulf.germgard@kau.se (U. Germgård),
lars.sundvall@more.se (L. Sundvall), hans.grundberg@domsjo.adityabirla.com (H. Grundberg), lawoko@kth.se (M. Lawoko).
1
Contributed equally to this work.

https://doi.org/10.1016/j.indcrop.2018.02.038
Received 17 October 2017; Received in revised form 28 December 2017; Accepted 11 February 2018
Available online 22 February 2018
0926-6690/ © 2018 Elsevier B.V. All rights reserved.
R. Deshpande et al. Industrial Crops & Products 115 (2018) 315–322

later more acidic stage. Furthermore, initial pulping at higher pH also

removes a large fraction of the pinosylvin.
However, it has been demonstrated that covalent bonds between
lignin and polysaccharide, also called lignin carbohydrate complexes
(LCC’s) most likely acts as an obstacle for the complete delignification
both in kraft pulping and in subsequent oxygen delignification
(Karlsson and Westermark, 1996; Karlsson et al., 2001; Lawoko et al.,
2004); since virtually all lignin still in the pulps were covalently bound
to the polysaccharides. Furthermore studies (Lawoko et al., 2006) have
demonstrated the presence of lignin polysaccharide networks in paper
grade sulfite pulp from softwood. Interestingly, this study suggested
that the reactivity of LCC in sulfite pulping differs from that of kraft
pulping, since the nature of the covalent networks appeared to be dif-
ferent, with more bonds to xylan in the former case. Studies of LCC in
dissolving pulps are rare, but their presence has been demonstrated
(Gübitz et al., 1998). Fig. 1. Cooking temperature profile for two-stage sulfite (dotted line) and acid sulfite
In this study, we investigate the presence and development of lignin (continuous line) cooking.
carbohydrate complexes during one-stage sulfite and two-stage sulfite
pulping of pine heartwood to produce dissolving pulp.
cook the autoclaves were cooled rapidly by submerging them into cold
water of about 10 °C to stop further reactions. Pulp and liquor samples
2. Material and methods were collected after each cook. The pulp samples obtained after cooking
were washed overnight with distilled water and dried at 45 °C in an
2.1. Wood sample preparation oven to air dry equilibrium conditions. A Wiley Mill was then used to
grind the dried samples of cooked chips into a wood powder that passed
For preparing 100% pine (Pinus) heartwood chips, a pine tree with a through slots of 0.4 mm (40 mesh).
diameter of around 24 cm was collected from the Örnsköldsvik forest
area in Sweden. The outer sapwood was separated from heartwood in
2.4. Pulp and liquor analysis
the sawmill and the heartwood was chipped in MoRe Research chipper.
All the chips obtained were classified in a chip classifier which had a
After cooking experiments, all the pulp samples were analyzed for
series of trays (Ø 45 mm,//8 mm, Ø 13 mm, Ø 7 mm, Ø 3 mm and < Ø
different chemical composition using standard procedure as shown in
3 mm) and chips retained on the 13 mm tray were used for the ex-
Table 2.
perimental trials.
The carbohydrate monomer values obtained were recalculated to
the initial carbohydrate polymers using the correlations of Meier
2.2. Cooking liquor preparation (Meier, 1958). Using the softwood data, the carbohydrate content of the
pulp was calculated as the percentage of wood based on the monomer
Lab prepared bisulfite and acid sulfite cooking acids were used in all results of the carbohydrate analysis according to Eqs. (1)–(3) below,
experimental trials, the composition of which is given in Table 1. The
starting pH for the bisulfite cooking liquor was 4.5 and for acid sulfite Cellulose = Glucose − (1/3.5) Mannose (1)
cooking liquor it was 1.5 pH, when measured at room temperature. The
Glucomannan = Mannose (1 + 1/3.5) + Galactose (2)
base (cation) charge was 5% measured as Na2O on wood basis for bi-
sulfite cook and 2.5% for acid sulfite cook. Xylan = Xylose + Arabinose (3)

2.3. Cooking procedure

2.5. Wood mill preparation
The cooking experiments were carried out in a lab digester using
Pine heartwood and sulfite pulp samples of particle size (below
autoclaves and circulation type digester. The heating for autoclaves
40 mesh) were converted to fine powder of approximately 30 μm by ball
were carried out in a glycol bath and circulation digester used steam as
milling technique after removal of extractives according to earlier
heating media. The startup temperature was 100 °C and the L/W-ratio
studies done Giummarella et al., (2016).
was always 4.6 in the impregnation stage. Temperature profile for
single acid sulfite cooking at 142 °C and two- stage sulfite cook is shown
in Fig. 1. The dotted line for two-stage sulfite cook represents the first 2.6. LCC’s fractionation
impregnation stage with bisulfite cook (pH-4.5, 154 °C, L/W ratio-4.6)
up to two-hour cooking time; which is then continued with acid sulfite LCC fractionation was performed as recently described
cook as second stage (pH-1.5, 142 °C, L/W ratio-2.6). At the end of each (Giummarella et al., 2016) with slight variation at first stage. Precisely,
after extraction (80 °C, 4 h) of the ball milled sample with deionized
Table 1
The composition of the two cooking acids used in the experiments. Table 2
Testing methods used for the wood and pulp samples.
Lab prepared bisulfite Lab prepared acid sulfite
cooking acid cooking acid Tests Methods

Total SO2, % 2.7 3.8 Acetone extract ISO 14453

Combined SO2, % 1.5 0.6 Klason lignin Tappi T-UM 250
Free SO2, % 1.2 3.2 Lignin (UV) Tappi T 222
pH 4.5 1.5 Arabinose, Galactose, Glucose, Mannose and Xylose SCAN-CM 71: 09*
Base charge as Na2O, % (% 5 2.5 Total-S SCAN-CM 57
on wood basis)
* Calculated as anhydrous sugar.

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water (mwater:msample = 10:1), the sample was centrifuged and the su- pinosylvin in heartwood is well known and it leads to lignin con-
pernatant, Extract 1, was directly freeze dried. The pellet was treated as densation through reactions with lignin moieties in acid sulfite cooking
previously described in order to obtain Precipitate 1 by addition of 20% conditions (Sjöström, 1993; Sixta, 2006). So, it was interesting to in-
in volume of water after complete dissolution in the same amount in vestigate the LCC behavior when using 100% pine heartwood with
weight of[Amim]Cl-DMSO (1[Amim]Cl: 1DMSO = w:w) system. Finally, respect to one-stage and two-stage acid sulfite pulping. Simultaneously,
Precipitate 2 was obtained similarly by precipitation in ethanol of the we obtain information of relevance to understanding the role of lignin
solute dissolved in DMSO-water solution (vdmso:vwater = 1:1) while structure in recalcitrance delignification.
Precipitate 3 by addition of three times the volume solution of water.
3.1. Chemical compositions and mass balance
2.7. Enzymatic hydrolysis
The chemical composition of the pine heartwood and the pulp ob-
An amount of 200 mg of Extract 1 obtained from two-stage sulfite tained after different pulping conditions were analyzed for carbohy-
pulp was incubated in presence of 50 mM phosphate buffer (pH = 7) for drate, lignin and extractives contents as shown in Table 3. Pinosylvin
48 h at room temperature under continuous stirring. The enzyme do- was also present in relatively high amounts with respect to wood ex-
sage was of 100 μl and the enzymes, all from Novozyme, were respec- tractives.
tively Pulpzyme® HC (a product rich in xylanase activity), Gamanase The pine heartwood and the pulp samples obtained after different
(1000 VHCU/g, a product rich in mannanase activity) and Fibercare® (a sulfite pulping conditions were fractionated according to the procedure
product rich in cellulase activity). Enzyme treated and untreated sam- for LCC fractionation described by Giummarella and Lawoko (2016).
ples were analyzed by size exclusion chromatography. Mass balance analysis for all pulp samples show recovery yields all over
above 90% (Table 4).
2.8. SEC-DMSO/0.5% LiBr Chemical wood analysis including sugar composition, Klason lignin
and extractive contents were performed on all of the fractions (Table 4).
Size-exclusion chromatography (SEC) of LCC’s fractions were per- It is evident for the pine heartwood that the hemicelluloses were en-
formed on a SEC-curity 1260 system (Polymer Standards Services, riched in the E1 fraction, cellulose in the P1 fraction and Lignin in the
Mainz, Germany) coupled with a dual system detector (UV and RI) P2 fraction. However, most of the lignin was present in the P1 fraction,
using the same columns, standards and procedure as Duval (Duval which was the largest by mass. This result was similar to that reported
et al., 2015). for spruce wood (Giummarella et al., 2016). A similar fractionation
applied to the pulps however shows a different tendency with respect to
yield and composition of the fractions. The P1 fraction progressively
2.9. 2D HSQC-NMR analysis
diminishes from one stage to two stage pulping, while the E1 increases.
The compositional changes of the growing E1 fraction indicate dom-
For 2D-NMR, roughly 100 mg of freeze dried fractions were dis-
inance of cellulose. This tendency can be explained by that the cellulose
solved in 700 μl of deuterated DMSO-d6 or DCCl3 in the case of acety-
is depolymerized during the pulping and becomes more water-soluble
lated samples, derivatized at same condition reported by Lu and Ralph
during LCC fractionation. In this context, the two-stage pulping is more
(Lu and Ralph, 2003). NMR spectra were recorded and processed as
severe on cellulose degradation. Further support for this hypothesis is
described elsewhere (Giummarella et al., 2016). DMSO (δC/δH = 39.5/
derived from the SEC data (Fig. 2).
2.5 ppm) and chloroform (δC/δH = 77.3/7.2) ppm) were used as in-
Looking at lignin content, it can be seen that the temperature had an
ternal reference.
influence on lignin sulfonation and lignin removal from the wood.
During sulfite pulping the lignin is sulfonated and it is gradually re-
3. Results and discussion
moved from the pulp with cooking time. The hot water solubility ex-
periment (E1 fraction) can be used as a tool to determine the amount of
When performing chemical pulping of softwood, it is known that
lignosulfonate formed during different sulfite cooking conditions which
delignification becomes slow at about 90% of the lignin removal during
can be extracted with hot water (UV results*).
one-stage acid sulfite pulping (Sjöström, 1993; Rydholm, 1965) and
around 96–97% during two-stage pulping. The reasons for this could be
3.2. Molar mass distributions of LCC fractions
the presence of non-reactive lignin structures (Rydholm, 1965), the
evolution of lignin condensation reactions leading to repolymerization
The E1 fraction and the P2 fraction were completely soluble in the
(Sixta, 2006), or the existence of stable covalent bonds between lignin
eluent solvent system (0.5% LiBr/DMSO) but the P1 was only partially
and carbohydrates (Lawoko et al., 2006).
soluble. The insoluble fraction was probably restricted by its large
In this work, we investigate the role of lignin carbohydrate bonds
molar mass.
using pine heartwood as starting wood raw material. This is because the
For the pine heartwood (Fig. 2), it is generally observed that some
acid sulfite pulping is sensitive to certain wood raw material like pine
lignin (UV detector signal) co-eluted with most of the material (RI de-
because of phenolic extractives which is present in high concentration
tector signal). Yet the lignin content in the fractions was in the range of
in heartwood of pine (see Table 3) (Sjöström, 1993). The presence of
9–40% (Table 4, columns 1–3). This is a strong indicator of covalent
linkages between lignin and carbohydrates. It should however be noted
Table 3
Wood and different sulfite pulps composition.
that in the case of the P1 which was only partly soluble, the dissolved
fraction consists mainly of hemicelluloses and lignin, the cellulose
Pine heartwood 142 °C, Acid Two stage cooking fraction constituting the insoluble material. Thus, the deduction of
(Wood) sulfite (Pulp) (Pulp) lignin carbohydrate bonds from the SEC chromatograms is specifically
Yield, % 100 65.5 61.5
related to lignin-hemicellulose bonds. This is further substantiated
Lignin, % 26.6 16.3 11.2 when one follows the cellulose degradation during pulping by SEC (P1,
Cellulose, % 35.0 ± 1.4 34.5 ± 0.5 33.0 ± 1.5 Fig. 3). It is clear that the high intensity RI peak (representative of
Glucomannan, % 12.6 ± 0.2 3.0 ± 0.1 3.95 ± 0.1 cellulose) is not associated with a UV peak. However, the involvement
Xylan, % 6.9 ± 0.02 2.0 ± 0.02 2.51 ± 0.12
of non-cellulosic glucans in LC linkages cannot be disregarded. It is
Extractives, % 11.8 4.7 3.6
Pinosylvin, mg/kg 2411 127 12.3 evident from the molar mass parameters of pine heartwood fractions
reported in Table 5, that the P2 fraction represents the highest number

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Table 4
Mass balance on wood or pulp basis, relative carbohydrate and lignin content of different fractions.

Pine heartwood 142 °C, Acid sulfite Two stage cooking

E1 P1 P2 P3 E1 P1 P2 E1 P2

Mass balance (pulp/wood %) 18 52 21 2 28 5 40 70 12

Arabinose, % 5.9 1.0 1.7 n.a 0.1 0.0 0.0 0.0 0.0
Galactose, % 10.2 1.4 3.2 n.a 0.1 0.0 0.0 0.1 0.1
Glucose, % 13.7 62.6 38.3 n.a 49.8 57.9 75.6 66.8 80.8
Xylose, % 17.3 3.7 7.0 n.a 8.4 0.4 1.1 5.8 1.9
Mannose, % 43.2 4.0 10.3 n.a 10.0 0.9 1.9 7.0 2.4
Lignin, % 9.7 (11) 27.3 39.5 n.a 31.6 (35) 40.7 21.4 20.3 (25) 14.9

(Obtained by UV); n.a = not analyzed.

average molar mass (Mn) and the E1 the lowest. The P1 is intermediate
but it should be noted that solubility was poor for this fraction and we
attribute this to high molar mass insoluble fraction.
The SEC profiles of the fractions obtained from the pulps (Fig. 3)
were somewhat different from those of the original pine heartwood.
Firstly, most of the carbohydrate fractions seemed to be free of lignin
since the UV and RI were not superimposed. In general, it was difficult
to deduce the existence of lignin carbohydrate bonds from the elution
profiles.
To further investigate the type of the carbohydrate linked to lignin,
different carbohydrate degrading enzymes, with known specificities
were applied on E1 fraction from two-stage process, followed by SEC
analysis. The working hypothesis here is that the enzymatic treatment
should not affect the elution profile of lignin (observed by UV) unless
the lignin was covalently attached to the partially depolymerized car-
bohydrate. The enzymes used were cellulase (Fibercare), mannanase
(Gamanase) and xylanase (Pulpzyme). As shown in Fig. 4, the enzy-
matic depolymerization of the different carbohydrates surely did affect
the lignin SEC profiling as well as shifting of UV signal, yet the enzymes
do not degrade the lignin. This indicated that covalent bonds exist to at
least xylan, glucomannan and possibly also to glucans. Furthermore,
both the effect of enzyme contamination as well as the enrichment of
fractions recalcitrant to the enzymatic treatments could explain the
wider distribution of the curves towards higher molecular weight.
3.3. Analysis of lignin structure and lignin carbohydrate bonds
To get some insight on lignin structure and lignin carbohydrate
bonds, the fractions were analyzed by 2D HSQC NMR. The assignments
for the pine heartwood were made according to the literature
(Balakshin et al., 2011; Balakshin et al., 2007; Toikka et al., 1998; Del
Río et al., 2016). For the pulp fractions, lignin inter-unit linkages were
assigned according to Marques et al. (2009) and Khokarale et al.
(2016).
Semi-quantitative analysis was achieved by using the integrals of
the C2 aromatic signal as internal standard for the native fraction, and
the C2 and C2-Sulf + C5-Sulf as internal standards for the sulfite pulps.
The Eqs. (4) and (5) below were used for the calculations:
I. Spine = G2 (4)
G2 (αsulf ) + G5 (αsulf ) ⎤
I . Spulps = G2 + ⎡
⎢
⎣ 2 ⎥
⎦ (5)

Fig. 2. SEC elution patterns for pine heartwood fractions.

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Fig. 3. SEC results for 142 °C fractions (left) and for 60 mins fractions (right).

Where: (Giummarella et al., 2016). In E1 fraction, weak signals from γ − esters

– I.Spine = Internal standard adopted for the semi-quantitative 2D
HSCQ analysis of pine samples (Fig. 5),
– I.Spulps = Internal standard adopted for the semi-quantitative 2D
HSCQ analysis of pulp samples (Fig. 6),
– G2 = C2/H2 cross correlation signal in Guaiacyl units,
– G5(αsulf) = C5/H5 cross correlation signal in Guaiacyl units with
sulfonated group in α position,
– G2(αsulf) = C2/H2 cross correlation signal in Guaiacyl units with
sulfonated group in α position.
Eq. (1) Internal standards calculation for semi-quantitative analysis
of 2D HSQC NMR spectra of pine (up) and sulfite pulps (down).
From the Pine heartwood analysis, CH correlations from the 3 LCC
types were observed similarly of previous studies carried on Spruce
were detected while phenyl glycosides were detected in all fractions
except P2. Benzyl ethers were detected in P1 and P3 fractions. The P2
fraction seemed to consist mainly of the bulk of hemicellulose not
bound to lignin. Alternatively, if any LC bonds were present in this
fraction, then the signals were below detection limit. In the pulps
(Fig. 6), the β-O-4 sub-units with a hydroxyl group on the α − carbon
was identified. The sulfonated β-O-4, which if present should have the β
CH correlation at 4.76/82 ppm and the α CH correlation at 4.39/
66.4 ppm according to a recent publication (Miles-Barrett et al., 2017)
was not detectable. This was expected from the chemistry of lignin
dissolution of sulfite pulps, since sulphonated lignins are water soluble
and would end up in the liquor.
The lignin in the E1 fraction of the acid sulfite pulps had a β-O-4
content of 12% and that of the E1 from the 2-stage pulping was 8%. The
lower content of aryl ether linkages in the latter was likely due to a

Table 5
Polydispersity (D), Mn and Mw values for LCCs fraction of pine, 142 °C –Acid sulfite and Two-stage cooking pulps.

Pine heartwood 142 °C, Acid sulfite Two stage cooking

E1 P1 P2 P3 E1 P1 P2 E1 P2

Mn, g/mol 4800 6100 13200 3700 1800 4600 3200 1700 2600
Mw, g/mol 13000 30900 93000 8700 7700 15100 12700 6200 7100
D 2.72 5.04 7.02 2.33 4.22 3.26 4.03 3.77 2.72

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Fig. 4. SEC elution pattern of E1 two-stage sulfite pulp (black

line) and after Fibercare® (cellulase), Gamanase (Mannanase), and
Pulpzyme® HC (xylanase) enzyme treatment looking at UV (left)
and RI (right) detectors. The shift towards lower molecular weight
after the treatment in both detectors indicates the presence of
covalent bonds between lignin and polysaccharides.

more favorable hydrolysis of β-O-4 at the higher temperature. The ex-
tent of hydrolysis can be realized since the β-O-4 content of the bulk of
lignin, which is present in the P1 fraction of native pine heartwood
fractions was about 52%. That lignin was more depolymerized in two-
stage pulping was also supported by the SEC data, where it is observed
that the 2-stage fraction had a lower molar mass.
The lignin-carbohydrate (LC) bond types identified were the phenyl
glycosides (PG) and gamma esters (GE). These were only observed in
the E1 fractions. The P2 fractions had lower contents of lignin and if LC
bonds were present in them, they were below the detection limits. The
benzyl ethers, which were present in the native wood had been cleaved.
A likely explanation for this is that ethers are broken during the sul-
fonation of lignin according to the mechanism in Fig. 7. Xylan and
pectin carry carboxylic acid groups that maybe responsible for ester
linkage formation to lignin. However, since only xylan was detected in
the pulps, it plays a more important role in lignin carbohydrate net-
works in sulfite pulps. This substantiated previous observations
(Lawoko et al., 2006).
According to the proposed mechanisms of LC bond formation in the
wood cell, the benzyl ethers and benzyl esters could form through nu-
cleophilic addition to the quinone methide intermediate of lignin
polymerization. However, of the two LC bond types formed by the
proposed mechanisms, only benzyl ethers have been detected by 2D
HSQC analysis. The absence of benzyl esters has been attributed to the
migration of the uronic acid esters from the alpha to the gamma posi-
tion (Evtuguin et al., 2005) (Fig. 8). This may explain why gamma
esters (GE) were detected in the pulps. Besides, sulfonation of the pri-
mary alcohol located at the gamma carbon is not as common as that of

Fig. 5. 2D HSQC of LCC’s precipitated fractions from pine heartwood in d6-DMSO (original) and CDCl3 (acetylated). Lignin inter-unit linkages and lignin-carbohydrate (PG, γ − ester,
BE) bonds are assigned. In the anomeric region of carbohyrdates, Xyl=Xylan, Man=Mannose, Gal=Galactose, 4OMGA=4-O-Methyl glucuronic acid. The subscript “t” stands for the
carbohydrate in the terminal reducing end and non-reducing end. Regarding lignin assignment G=guaiacyl, −OCH3=Methoxy, DBO=Dibenzodioxicin SD=Spirodienone. β-O-4 signal
are colored in red. Ac=Acetylated and the number in subscript indicates the carbon number either in the aromatic or sugar ring. Similarly C2-4 Carbs stand for the signaĺs region of C2, C3
and C4 in carbohydrates.

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Fig. 6. 2D HSQC spectra of sulfite pulps in d6-DMSO where β-O-4 (in red) and lignin-carbohydrate (PG, γ − ester) bonds are assigned. In the right side it has been expanded the aromatic
region guaiacyl unit. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

the secondary alcohol at the benzylic carbon (Gellerstedt and Gierer,
1971).
Although the cleavage of alkyl-aryl ether linkages has been pro-
posed (Gellerstedt and Gierer, 1971), the cleavage of benzyl ether (BE)
by sulfonation reaction has not been discussed and would have a
technical significance in the context of dissolving pulp production and
future bio refineries targeting hemicellulose or lignin extractions. As
such a more detailed investigation is required.
The results obtained in this work indicate that lignin-carbohydrate
bonds of the ester and glycoside type may in part explain why lignin is
retained in pulps. In fact, we observe that residual lignin carbohydrate
complexes are significantly sulfonated. Hence, sulfonation alone is not
enough for lignin dissolution. The cleavage of stable LC bonds is equally
important to release sulfonated lignin into the liquor. This applies for
both the one- and two-stage pulping.
4. Conclusions
• Covalent bonds between lignin and polysaccharides exist both in the
pine heartwood and in the sulfite dissolving pulps.
• The covalent bonds are mainly between lignin and hemicelluloses.
• The covalent bonds from lignin are both to xylan and glucomannan.
• In the original pine heartwood the LCC were phenyl glucosides, γ-
esters and α-ethers but in the pulps the α-ethers have mostly dis-
appeared.
• Covalent bonds between lignin and wood polysaccharides might be
a factor that limits delignification in sulfite pulping.
5. Technical implications
The present data indicate that covalent bonds between lignin and
hemicellulose present in wood survive the dissolving type sulfite
pulping. However, it appears as the α-ethers are broken in the sulfite
pulping. This reaction is probably of large importance in the removal of
the lignin during sulfite pulping. In an earlier study, it was noted that
the bonds between the lignin and xylan was dominating in the sulfite
pulps (Lawoko et al., 2006); the data in the present work, where γ-
esters play an important role, might also give an explanation since

Fig. 7. Suggested mechanism where LCC of ether type are broken by acidic sulfonation.

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R. Deshpande et al.
Fig. 8. Hypothetical mechanism of uronosyl migration from benzylic to gamma carbon (Evtuguin et al., 2005). In this mechanism the originally formed α ester (iii) is formed by the
reaction between a carbohydrate-bound carboxylic acid and the quinomethide structure (ii) formed by the coupling of radicals (i). The hydroxyl on the γ-carbon performs a nucleophilic
attack on carboxyl carbon and forms a cyclic intermediate (iv), before an ester to the γ-carbon eventually is formed. That structure (v) is more favorable than structure (iii) might be due
to that the gamma ester is more energetically favorable.
xylan carries a major part of the carboxylic acids in wood in the form of
4-O methyl glucuronic acid. Dissolving pulp production involves re-
taining only α-cellulose and hence this study will help pulp mill in
understanding the LCC that prevails sulfite cooking conditions and
optimize their process conditions to produce high purity cellulose
pulps.
Acknowledgements
This study was performed within the Industrial Graduate School
VIPP (Values Created in Fibre-Based Processes and Products) with fi-
nancial support from the Knowledge Foundation and Knut and Alice
Wallenberg research foundation, Sweden. Thanks are due to the fi-
nancial contribution from Domsjö Fabriker and MoRe Research in
Örnsköldsvik, Sweden. The authors are also grateful for the support of
the project from the Kempe Foundations in Örnsköldsvik, Sweden.
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