Biomass and Bioenergy 143 (2020) 105824

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Biomass and Bioenergy

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Methane potentials and organic matter characterization of wood fibres

from pulp and paper mills: The influence of raw material, pulping process
and bleaching technique
E.-M. Ekstrand a,c,*, M. Hedenstrom¨ b, B.H. Svensson a,c, S. Shakeri Yekta a,c, A. Bjorn ¨ a,c

a
Department of Thematic Studies – Environmental Change and Biogas Research Center, Link¨oping University, SE-58183 Linkoping,
¨ Sweden b Department of Chemistry, Umeå University, 901 87, Umeå, Sweden
c
Biogas Research Center, Linkoping ¨ University, SE-58183, Linkoping, ¨ Sweden
that an observed positive effect of bleaching on the methane potential of softwood
CTMP fibres was likely related to a higher degree of deacetylation and improved
accessibility of the hemicel lulose. In conclusion, fibres from kraft and sulphite
ARTICLE INFO pulping are promising substrates for methane production irrespective of raw
material or bleaching, as well as fibres from CTMP pulping of hardwood.
Keywords:
Fibers
Softwood
Hardwood
Pulping
Bleaching
Anaerobic digestion

1. Introduction
ABSTRACT
high moisture content [3]. Different production routes within the pulp
During the process of pulp- and papermaking, large volumes of fibre-rich primary and paper industry leading to accumulation of primary sludge is shown
sludge are generated. Anaerobic digestion of primary sludge offers a substantial Large amounts of organic material are released during the process
potential for methane production as an alternative approach to the inefficient of pulp- and papermaking, resulting in high wastewater treatment costs
energy recoveries by commonly used incineration techniques. However, a
for the industry [1]. The first step in wastewater treatment at pulp and
systematic study of the importance of upstream process techniques for the
methane potential of pulp fibres is lacking. Therefore, biochemical methane
paper mills is primary sedimentation/flotation, which generates large
potentials were determined at mesophilic conditions for 20 types of fibres volumes of fibre sludge, often referred to as primary sludge (80–90 000
processed by a variety of pulping and bleaching techniques and from different raw tonnes in Sweden in 2012 [2]). The primary sludge is, however,
materials. This included fibres from kraft, sulphite, semi-chemical, chemical commonly dewatered and incinerated at low energy recovery due to the
thermo-mechanical (CTMP) and thermo-mechanical pulping plants and milled raw in Fig. 1, most of which may affect the possibility to produce biogas
wood. The pulping technique was clearly important for the methane potential, with
from this waste fraction. We have previously shown that there is
the highest potential achieved for kraft and sulphite fibres (390–400 Nml CH4 g substantial potential for methane production via anaerobic digestion
VS− 1). For raw wood and CTMP, hardwood fibres gave substantially more (AD) of fibres in pulp and paper mill wastewater [4]. In addition to the
methane than the corresponding softwood fibres (240 compared to 50 Nml CH4 g production of a valuable energy carrier in the form of methane, AD of
VS− 1 and 300 compared to 160 Nml CH4 g VS− 1, respectively). Nuclear magnetic organic waste leads to a reduction of the waste volume and reduces
resonance characterization of the organic content demonstrated that the relative the amount of sludge to be disposed of [5].
lignin content of the fibres was an important factor for methane production, and

Abbreviations: AD, anaerobic digestion; BMP, biochemical methane potential; CPMAS, cross-polarization magic angle spinning; CTMP, chemical thermo
mechanical pulping; ECF, elemental chlorine-free; Hw, hardwood; HRT, hydraulic retention time; HSQC, heteronuclear single quantum coherence; NMR, nuclear
magnetic resonance; NSSC, neutral sulphite semi-chemical; Sw, softwood; TCF, total chlorine-free; TMP, thermo-mechanical pulping; TOC, total organic carbon; TS,
total solids; VS, volatile solids.
* Corresponding author. Department of Thematic Studies – Environmental Change and Biogas Research Center, Linkoping ¨ University, SE-58183 Linkoping, ¨
Sweden.
E-mail addresses: eva-maria.ekstrand@liu.se (E.-M. Ekstrand), mattias.hedenstrom@umu.se (M. Hedenstrom), ¨ bo.svensson@liu.se (B.H. Svensson), sepehr.
shakeri.yekta@liu.se (S. Shakeri Yekta), annika.bjorn@liu.se (A. Bjorn). ¨

https://doi.org/10.1016/j.biombioe.2020.105824
Received 11 June 2020; Received in revised form 2 October 2020; Accepted 7 October 2020
Available online 27 October 2020
0961-9534/© 2020 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/).
E.-M. Ekstrand et al. fibres from kraft or sulphite pulping (Table 1). However, a systematic
study on the influence of raw material and commonly used pulping and
bleaching
techniques on the methane potential of this type of fibre is lacking. The
aims of the present study were to (i) assess the methane potential of
pulped fibres produced using common pulping or bleaching tech niques
for different wood materials, and (ii) elucidate the underlying
mechanisms for any observed differences in methane potentials of the
fibres. To address these aims, a systematic study of the methane po
tentials of fibres from mechanical, semi-chemical, kraft, and sulphite
pulp and paper plants was undertaken. The structural characteristics of
the fibre samples were studied using solid-state 13C cross-polarization
magic angle spinning (CPMAS) nuclear magnetic resonance (NMR)
spectroscopy in relation to the methane potentials. This method is a
well-established technique to characterize lignocellulosic materials by
generating chemical fingerprints of major constituents of wood samples
without the need for solvent extraction [10]. In addition, 2D
solution-state 1H–13C heteronuclear single quantum coherence (HSQC)
NMR was also performed, as the superior resolution of this technique
allows identification of the molecular structures in complex organic
samples – not possible using 1D NMR techniques [11,12].
Pulp and paper production is often carried out in campaigns, shifting
the production between unbleached and bleached qualities of pulp
while
Biomass and Bioenergy 143 (2020) 105824
Table 1
2.1. Fibre sampling and biochemical methane potential (BMP) tests
Fig. 1. Overview of the process steps
involved in pulp- and papermaking that
potentially affect the methane potential of
A summary of the different types of fibres investigated is presented
the primary sludge. Mechanical pulping in Table 2. When possible, both softwood fibres and hardwood fibres
techniques include thermo-mechanical pulp were sampled from the same pulping process. Fibre samples were
ing (TMP) and chemical thermo-mechanical collected from the wastewater after pulping and bleaching for the
pulping (CTMP), followed by bleaching thermo
with peroxide or sodium dithionite. Chemi mechanical pulping (TMP) and the softwood chemical thermo
cal pulping techniques include kraft and mechanical (CTMP) processes, whereas pulp was sampled directly
sulphite pulping, followed by elemental from the product line for kraft, sulphite and neutral sulphite semi
chlorine-free bleaching (ECF) or total chemical pulping (NSSC) processes. In addition, untreated wood chips
chlorine-free bleaching (TCF). Neutral sul
from hardwood and softwood were sampled, dried for 1 h at 105 ◦C and
phite semi-chemical pulping (NSSC) com
milled to a particle size of <1 mm (raw fibres). The collection of all
bines a mild chemical pulping step with a
samples followed the sampling procedures implemented by mill
mechanical refining step. Dashed lines
denote the wastewater streams from the personnel for regular process monitoring.
different process steps that are passed The methane potential was determined using biochemical methane
through primary clarification where residual potential (BMP) tests as described by Ekstrand et al. [4]. In short, the
fibres are collected as primary sludge. tests were performed using 320 ml glass bottles sealed with rubber
stoppers and aluminium screw caps. 20 ml of inoculum (digestate from
a municipal wastewater treatment plant, Linkoping, ¨ Sweden) was
2. Material and methods added together with fibre sludge, nutrient salts and tap water up to a
final volume of 100 ml. The organic loads in the batch bottles varied
depending on the degradability of the substrates, ranging betwen
varying the raw materials used (Fig. 1). If such shifts lead to large var
iations in the methane potentials of the pulped fibres, the use of these 2.3–3.5 g volatile solids (VS) per litre for chemically pulped fibre sam
fibres in a biogas production facility would lead to unwanted fluctua ples, 4–5 g VS/L for CTMP and TMP-pulped fibres, and 6.7–8 g VS/L
tions in methane production, as well as variations in the in the compo for raw fibres. This resulted in inoculum to substrate ratios of 1.4–2.0
sition of the digestate due to the accumulation of undigested fibres in for chemically pulped fibres, 0.9–1.4 for CTMP and TMP fibres, and
the digesters. There are indications in the literature that the methane 0.4–0.6 for raw fibres (see Appendix A for TS, VS and inoculum to
po tential of pulped fibres may vary depending on the pulping process substrate ratios for all samples). The ratio for CTMP, TMP and raw
used, and in particular, demonstrating high methane potentials for fibres is lower

Methane potential of fibres from different pulping processes according to previous studies. TMP = thermomechanical pulp, SGW
= stoneground wood, BMP = biochemical methane potential, CSTR = continuously stirred tank reactors.

Process Raw material Method T ( ◦C) Methane production Ref

Bleached kraft Softwood + birch BMP 35 210 m t VS [6] Bleached kraft Softwood + birch BMP 55 230 m3 t VS− 1 [6] Bleached kraft
3 − 1

Softwood + birch CSTR 55 190–240 m3 t VS− 1 [6] Unbl. + bleached Softwood + hardwood CSTR 37 210 ml g VS− 1 [7] TMP N/A BMP N/A
45 m3 t VS− 1 [8] Bleached kraft Softwood BMP 37 363 ml g VS− 1 [9] Bleached sulphite Softwood BMP 37 363 ml g VS− 1 [9] Bleached
 TMP Softwood BMP 37 59 ml g VS − 1 [9] Bleached SGW Softwood BMP 37 81 ml g VS− 1 [9] Recycle fibre N/A BMP 37 192 ml g VS − 1 [9]

2
E.-M. Ekstrand et al. Biomass and Bioenergy 143 (2020) 105824

Table 2 optimx package [15,16] in R [17]. The model parameters were evalu
Characteristics of fibres from different pulping processes, bleaching techniques ated by plotting the model together with the data, as well as plotting the
and raw materials. CTMP = chemical thermo-mechanical pulping, NSSC = residuals and the residual distribution. For the CTMP, TMP and raw fi
neutral sulphite semi-chemical pulping, TMP = thermo-mechanical pulping, bres, the modified Gompertz model could not adequately capture the
Unbl = unbleached, P = peroxide bleaching, ECF = elemental chlorine-free initial dynamics of methane formation, in which case the methane
bleaching, TCF = total chlorine-free bleaching, Hw = hardwood, Sw = softwood.
production rate between each measuring point was calculated by the
Pulping process Unbl. P ECF TCF slope of a linear function and the maximum value was reported as kmax.
Kraft Hw/Sw Hw/Sw Hw/Sw Sulphite Hw/Sw Hw/Sw CTMP Hw/Sw Hw/Sw
NSSC Hw 2.3. Solid-state and solution-state NMR
TMP Sw Sw
Raw fibre Hw/Sw
Metal ions in heterogenous samples interfere with the NMR
analyses of organic matter [18] so fibre samples were treated with
EDTA to remove metals that might be present. After freeze-drying, 800
than for the chemically pulped fibres due to their lignocellulosic prop mg of each sample was suspended in 40 ml EDTA solution with the
erties, and is in line with the recommendations for lignocellulosic ma concen
terials outlined by Holliger et al. [13] All tests were performed in tration of 8 g L− 1 and placed on a platform shaker at 200 RPM for 1 h.
triplicates and incubated in the dark at 37 ◦C. The gas production was Next, the samples were centrifuged at 10 000×g for 10 min and the
determined by pressure measurements (Testo 312-2 Precision manom supernatant was removed. The pellets were re-suspended in 50 ml of
eter, Testo Inc, USA), and its methane content was determined by gas milliQ water to remove any remaining EDTA and were centrifuged once
chromatography with a flame ionizing detection (GC-FID). All measured more. Lastly, the pellets were freeze-dried and stored at room temper
gas volumes were normalized to 1 atm pressure and 273 K and are ature until further analyses.
13
expressed as Normalized mL (Nml). After the completion of the BMP C CPMAS analyses were performed on a Bruker Avance III 500
tests, methane potentials were evaluated as the amount of methane MHz spectrometer equipped with a 4 mm MAS probe and a Bruker
produced per gram VS of substrate added, minus control (Nml CH4 g SamplePro solid-state sample changer. A 1 ms contact-pulse with a

VS− 1). TS and VS were measured in duplicate according to the ramped 1H amplitude (50–100%) and a 1H decoupling strength of 80
Swedish Standard method (SS 028113). The BMP test for each kHz were used. 3500 scans were collected with a relaxation delay of
sample was ended when the cumulative gas production curve levelled 1s, resulting in an experimental time of approximately 1 h per sample.
off, generally after 40–60 days of incubation. However, for a few Spectra were normalized to a constant sum and placed in a data matrix
samples, the BMP test was ended before the maximum potential was for subsequent multivariate analysis using Matlab 2014b (Mathworks
achieved (CTMP softwood, TMP softwood and raw fibres), since the Inc.). Principal Component Analysis (PCA) was performed in SIMCA
digestion time was too long (>96 days) to be reasonable in full-scale 14.1 (Umetrics).
applications. Student’s t-tests (independent samples) were used to Sample preparation for HSQC NMR analysis was performed as
determine statistical significance (α < 0.5 and α < 0.01) between BMP described by Shakeri Yekta et al. [19]. 2D 1H–13C HSQC NMR spectra
samples. were recorded at 298 K on a Bruker Avance III HD 600 MHz spec
Klason lignin was determined by applying acid hydrolysis of the fibre trometer equipped with a BBO cryoprobe and a SampleJet sample
samples, followed by a gravimetrical determination of the insoluble changer. 16 scans were recorded for each of the 256 t1 increments and
lignin (TAPPI T222). The remaining acid-soluble lignin was determined spectra were calibrated using the residual DMSO peak (δH = 2.49 ppm
spectrophotometrically at 204 nm (TAPPI UM 250). The composition of and δC = 39.5 ppm). All spectra were phase-corrected manually and
soluble compounds of the bleaching wastewater from the TMP mill was processed with Topspin 3.2 (Bruker Biospin, Germany). As CP-MAS
determined by mill personnel after solid-phase extraction (internal and HSQC NMR spectroscopy are inherently qualitative, the NMR
method, W35:97), followed by analyses of fatty- and resin acids, results were used to qualitatively analyse and compare the relative
lignans, sterols, stearyl esters, and triglycerides. Carbohydrates distribution of different molecular structures of the organic matter
included arabi among the samples.
nose, xylose, mannose, galactose, and glucose and were determined
according to an internal standard (C29:91) adapted from the method 3. Results and discussion
SCAN-CM71. Organic acids included butyric acid, propanoic acid, for
mic acid and acetic acid. Not all details of the analyses were shared by 3.1. Effects of the pulping process
the mill, so these results are presented primarily as a support to the
discussion of this study. The BMP tests showed that fibre residues from different pulping
processes carried very different methane potentials (Fig. 2). Sulphite-,
kraft-, NSSC-, and CTMP-pulped fibres showed methane potentials of
2.2. Kinetic modelling 400 ± 17, 390 ± 6, 280 ± 2, and 160 ± 8 Nml CH4 g VS− 1, respectively.
The lowest methane potential of 50 ± 1 Nml CH4 g VS− 1 was obtained
To estimate the maximal rate of methane production (kmax) during from raw fibres. Fibres from TMP gave a methane potential lower than
the BMP tests of the fibre samples, a modified Gompertz equation [14] the inoculum control, which indicates an inhibition of the AD process.
was fitted to the experimental data of each sample. The model used The methane production from kraft and sulphite fibres was relatively
was
(
kmax × e g VS− 1⋅day− 1 (Table 3). There was production started, ranging 0.8–1.2
)} days for kraft and sulphite fibres. The
CH4(t) = CH4, max × exp CH4,max(λ − t) + 1 methane production rate for NSSC
{ fast, with kmax ranging 130–200 Nml
a short lag phase (λ) before methane
− exp
methane production rate (Nml g VS− 1⋅day− 1), λ is the lag phase
where CH4(t) is the cumulative methane production (Nml g VS− 1) as a (days), and e is Euler’s number. The parameter kmax was normalized to
function of time (t in days), CH4,max is the maximum methane produc the amount of VS added. The parameter estimation was performed by
tion (i.e. the methane potential of the sample), kmax the maximum non linear least squares minimization (Nelder-Mead method) using the
 was lower, (kmax = 70 Nml g VS− 1⋅day− 1), with a lag phase of 1.4 days features of the degradation dynamics for CTMP- and TMP-pulped
(Table 3), which together with the significantly lower (p < 0.05) fibres nor for raw softwood fibres. In particular, the degradation
appears to follow a two-phase pattern for CTMP-processed
methane potential of 280 ± 2 Nml CH4 g VS− 1, demonstrated that
unbleached hardwood, TMP
NSSC pulped fibres were more difficult to degrade than chemically
pulped fibres. The modified Gompertz model could not capture all the

3
E.-M. Ekstrand et al. Biomass and Bioenergy 143 (2020) 105824

Fig. 2. Accumulation of methane minus control in biochemical methane potential tests on fibres from different pulping processes. All fibre samples were from
unbleached softwood except for NSSC, for which only unbleached hardwood was available. See Table 2 for abbreviations.
(Appendix B). Still, some anaerobic bacteria have been identified that
processed peroxide-bleached softwood and raw softwood (Appendix are able to depolymerize lignin, and if the lignin is modified or treated, it
B), with a secondary lag phase, followed by resumption of methane becomes more easily degradable by anaerobic microorganism
pro duction after approximately 13–14 days. One potential explanation (reviewed by Khan & Ahring [24]), and could thereby be a contributing
could be that easily degradable sugars and/or short fibres are degraded factor to the higher methane potential observed for the CTMP fibres
first, followed by colonization and degradation of larger, more recalci compared to
trant fibres. In part, this could be coupled to a feedback inhibition in the TMP. The kraft and sulphite process, and to some extent the NSSC
production of cellulases for the cellulose degraders, caused by the process, utilizes chemicals at high temperatures and pressures to break
presence of easily degradable sugars such as glucose or cellobiose up and dissolve the lignin. This removes most of the lignin in kraft fibres
(reviewed by [20]). After the depletion of such sugars, cellulose de (3% remaining, Appendix B), and is likely the main explanation for the
graders in anaerobic systems generally colonize cellulose fibres by higher methane potentials obtained for these fibres. The removal of
adhering to the surface of the fibres via complexed cellulase systems lignin was also clearly shown in the NMR analysis (see section 3.1.1).
called cellulosomes [20]. The reasoning that such colonization of cel Furthermore, chemically modified lignin has been shown to have higher
lulose fibres could be the explanation to the two-phase pattern seen in degradability than naturally occurring lignin during AD [25], and might
this paper is supported by another study, where anaerobic microor also contribute to a higher methane potential for chemically treated fi
ganisms from an AD reactor were suspended in a buffer/mineral me bres. Another important factor is the accessibility of the cellulose. One
dium deficient in easily degradable sugars together with filter paper. In purpose of lignin in plants is to provide a protective barrier against
this investigation, an increase in cellulase activity and a concomitant microbial degradation, and its removal would not only increase the
increase in biogas production could not be observed until after three more easily biodegradable portion of the fibres, but also give enzymes
days [21]. That the CTMP and TMP fibres were more difficult to better access to the cellulose (reviewed by Monlau et al. [26]).
degrade was supported by the estimated kmax being much lower for However, even though CTMP pulping did not remove any considerable
amount of lignin compared to the raw fibres (28% and 29% remaining,
CTMP and TMP fibres (5–30 and 3–20 Nml g VS − 1⋅day− 1,
respec
respectively) compared to the range of chemically pulped fibres
tively; Appendix B), the pulping likely increased the accessibility of the
(130–200 Nml g VS− 1⋅day− 1; Table 3). hemicelluloses and celluloses of the fibres by breaking up the lignin
TMP and CTMP are mechanical pulping techniques that result in a structures. This explains the higher methane potentials for these fibres
high fraction of lignin remaining in the fibres after pulping [22]. Lignin in compared to raw fibres.
its native form has a very low degradability during anoxic conditions
[23], since anaerobic microorganisms generally lack the oxidative en 3.1.1. NMR spectroscopy reveals differences in chemical composition
zymes and pathways required for depolymerization of high CPMAS spectra of kraft, CTMP and TMP fibres from unbleached
molecular-weight lignin [24]. This means that the microorganisms softwood are shown in Fig. 3. Peaks from all major wood components
cannot use the lignin as a substrate to any accountable extent in biogas are identified and annotated accordingly. Lignin is identified by pres
processes and have low access to the lignin-enclosed hemicellulose ence of broad peaks in the aromatic spectral region (110–160 ppm) and
and cellulose. Thus, the lignin content most likely explains the low a more well-defined peak from the methoxy group at 65 ppm [27].
potentials observed for the TMP and CTMP fibres. This was supported Peaks from cellulose are observed at 105 ppm (C1), 89 ppm (C4 from
by the chemical analyses showing relatively high lignin contents of crys talline cellulose), 84 ppm (C4 from amorphous cellulose), 70–75
32% and 25–28% of dry weight for unbleached TMP and CTMP fibres, ppm (C2, C3 and C5) and 66–62 ppm (C6) and are often the most
respec prominent peaks in spectra of lignocellulosic materials [28]. Peaks from
tively, which is comparable with the lignin content of the raw fibres hemicel lulose are mostly obscured by the dominant cellulose peaks but
 one distinct spectral feature originating from hemicelluloses is the while essentially no lignin is left in kraft fibres. These observations
hump on the cellulose C1 peak observed at 101 ppm [29]. The peak at corroborate the results from Klason lignin analysis (Appendix B). In
a chemical shift of 21 ppm originates from acetyl groups found in addition, hemicellulose is efficiently removed by the kraft process, as
hemicelluloses [28]. Comparison of the NMR spectra demonstrates evident from the disap pearance of both the resonance adjacent to
that part of the lignin fraction is still present in CTMP and TMP fibres cellulose C1 peak at 101 ppm

4
E.-M. Ekstrand et al. a–c, and samples marked with an asterisk were fibres collected from the
wastewater. Visual evaluation of the model fits (plots of model, residuals, re
Table 3 sidual distribution) is commented on under Notes.
The estimated parameter values obtained after non-linear regression to Comparisons of the methane potential of fibre from hardwood and
Biomass and Bioenergy 143 (2020) 105824
softwood used in the same pulping process were specifically carried
out for the kraft, sulphite, and CTMP processes, as well as for raw
3.2. Effect of raw material
wood (aspen and spruce). This comparison was not performed on the
biochemical methane potential (BMP) experimental data of the fibre samples. TMP and NSSC processes as these pulp mills did not shift their
CH4,max is the maximum methane production (Nml g VS− 1) predicted by the production be
model, kmax the maximum methane production rate (Nml g VS− 1⋅day− 1), λ is tween softwood and hardwood pulp. There were no significant differ
the lag phase (days), and kmax (Δy/Δx) is the maximum methane production ences in methane potentials for hardwood and softwood fibres from the
rate estimated based on the gradient between two measuring points. For kraft or sulphite processes, which yield pulp with very low lignin content
abbrevia
tions, see Table 2. Kraft pulp was sampled from three different mills, denoted
(day) (Fig. 5). Kinetic hardwood compared to 130 Nml g VS− 1⋅day− 1,
Sample CH4,max (ml g kmax
(Nml g 2
R kmax (Δy/Δx)
modelling of the BMP softwood for kraft fibres, respectively, but a
VS− 1)
VS− 1⋅day− 1) Notes data indicated a faster with a kmax of 180 and slower degradation
λ degrada tion of
dif
Kraft-Sw-Unbl*a 380 130 1,2 0,98
Kraft-Hw-Unbl*a 350 180 1,2 0,98 ference in lignin composition between hardwood and softwood dis
Kraft-Sw-Unblb 390 170 0,9 1,00 cussed below.
Kraft-Sw-TCFb 390 170 0,8 1,00 of hardwood compared to softwood for sulphite fibres (150 compared to
Kraft-Sw-Unblc 380 200 1,1 1,00 170 Nml g VS− 1⋅day− 1, Table 3). Previous studies have indicated that
Kraft-Sw-ECFc 380 200 0,8 1,00
available surface area is an important factor for enzymatic degradation
Sulphite-Sw-Unbl 380 170 1,0 0,99
[31], and as hardwood fibres are shorter and thinner than softwood fi
Sulphite-Sw-TCF 390 180 0,9 0,98
Sulphite-Hw-Unbl 390 150 1,0 0,99 bres [22], a higher methane potential and/or a faster degradation might
Sulphite-Hw-TCF 390 190 0,8 0,98 be expected for hardwood fibres. However, there seem to be conflicting
NSSC-Hw 280 70 1,4 0,98 results in the literature regarding pulp fibre length. Mooney et al. [32]
CTMP-Hw-Unbl* 280 50 2,0 0,91 30 (1) CTMP-Hw-P* 280 30 3,0 0,95 20 demonstrated a faster and higher digestibility of refined, shorter fibres
CTMP-Sw-Unbl 130 6 7,8 0,97 5 (2) CTMP-Sw-P* 160 8 5,7 0,95 7 (2)
of a softwood kraft pulp compared to untreated, longer fibres, whereas
CTMP-Sw-P 160 10 6,6 0,94 8 (2) TMP-Sw-Unbl* 260 16 51,8 − 0,01 3 (3)
TMP-Sw-P* 120 20 2,6 0,80 20 (1) Raw fibre-Sw 40 1 6,0 0,83 3 (2) Raw Del Rio et al. [33] could not show any difference in enzymatic degra
fibre-Hw 210 15 3,2 0,90 8 (2) dation of pre-treated and size-fractionated softwood fibres. Within the
(1) Poor fit to data. scope of this study, no difference in methane production rates or po
(2) Poor fit to data, BMP not completed. tentials could be observed for chemically pulped fibres originating from
(3) Unable to fit to data, inhibition of AD process. hardwood or softwood. Thus, shifts in raw material during kraft or
sulphite pulp and paper production would presumably have limited ef
and the acetyl peak at 21 ppm. The 13C CPMAS spectra of bleached fect on a full-scale biogas process.
and unbleached TMP revealed several peaks in the aliphatic region AD of raw fibres and CTMP pulped hardwood fibres gave rise to
(20–40 ppm) that were not present in any of the other spectra, with the significantly (p < 0.01) more methane than the corresponding softwood
exemption of the acetyl peak. fibres, i.e. 240 ± 8 compared to 50 ± 1 Nml CH4 g VS− 1 for raw fibres
To get an overview of the chemical compositions of all fibre samples and 300 ± 10 compared to 160 ± 8 Nml CH4 g VS− 1 for CTMP fibres
used in this study and relate that to the BMP results, a PCA was per (Fig. 5). The kinetic models indicated faster methane production rates,
formed on the solid-state 13C CPMAS NMR spectra of all samples. A with an estimated kmax of 20 compared to 7 Nml g VS− 1⋅day− 1 for
clear separation of mechanically pulped fibres and chemically pulped CTMP (bleached) and 8 compared to 3 Nml g VS− 1⋅day− 1 for
fibres was observed along the first principal component (explaining untreated fibres (Table 3, Appendix B). One contributing factor to these
88% of the variation; see Fig. 4a), implying that the pulping technique differences is likely that softwood (spruce) used at the mill naturally has
was the most important factor explaining the difference in chemical a higher lignin content than hardwood (27%, Appendix C, compared to
composition among the fibre samples. The loading plot (Fig. 4b) shows 20–22% [26]). Furthermore, overlays of 13C CPMAS spectra of soft-
that the variation was largely related to the differences in aromatic and and hard wood fibres – either raw (Fig. 6a) or CTMP-pulped (Fig. 6b) –
methoxyl structures of lignin in the fibre samples, with the chemically show clear dissimilarities in intensities of lignin-derived 13C peaks (δC:
pulped fi bres having lower intensities of lignin-derived 13C resonances 105–155 ppm). This is a reflection of the difference in their lignin
composition, where hardwood lignin contains syringyl (S), guaiacyl (G)
(negative peaks in the aromatic spectral region 110–160 ppm and
and p-hydroxyphenyl (H) monomers while softwood lignin contains
negative methoxy peak at 56 ppm). The hemicellulose peak at 101
almost exclusively G monomers [34]. The main bond formed between
ppm and the acetyl peak at 21.5 ppm are also negative, indicating the lignin monomers is the relatively weak aryl-ether bond (i.e. β-O-4
more effective removal of hemicellulose in the chemically pulped bond), which makes up 35–60% and 50–70% of the bonds in softwood
fibres. and hardwood, respectively [35,36]. The lower occurrence of this bond
The second component in Fig. 4 (i.e. t2) separates the softwood in softwood is coupled to the ability of G-type lignin to form the more
(Sw) samples from the hardwood (Hw) samples, highlighting the chemically stable C–C bonds between the G units, whereas C–C bonds
differences in organic matter properties of the fibres in relation to the cannot be formed between S units [26,35]. For this reason, hardwood
raw material used. However, this only accounts for 6% of the variation fibre lignin is more likely to be affected by the CTMP process, and the
in NMR spectral data among the samples (Fig. 4). A detailed analysis accessibility of the hemicellulose and cellulose for degradation is
of the loadings of the second principal component was not attempted improved. In terms of AD, however, previous lab-scale BMP tests on
as it showed a complex pattern (not shown). However, a positive peak ground wood samples revealed no clear degradation of lignin despite
degradation times of more than 440 days [37,38]. This suggests that
was observed at 146 ppm, originating from guaiacyl (G), reflecting the
degradation of lignin itself is not the explanation for the observed dif
 ference in methane production between hardwood and softwood. has a higher content of mannose [40,41]. In a study by De la Cruz et al.
In another study, AD of ground hardwood (sycamore) showed a [38], the xylose content in hardwood during AD was decreased by 30%
faster methane production than from softwood (pine) in BMP tests in BMP tests, whereas the sugar content of the softwood sample
carried out for 27 days, and at the same time a higher removal of xylan (mainly
[39]. Generally, hardwood has a higher content of xylose and softwood

5
E.-M. Ekstrand et al.
VS− 1⋅day− 1, Table 3). One possible explanation for the slower degra
dation could be the presence of residual bleaching chemicals or wood
compounds that have been released during bleaching. This is
supported by findings from another mill using the same bleaching
chemical
Biomass and Bioenergy 143 (2020) 105824

Fig. 3. 13C CPMAS NMR spectra on raw fibres and

fibres from three different pulping processes. For ab
breviations, see Table 2. C refers to cellulose, with the
numbers referring to the different carbon positions in
each structure. Ac and C=O refers to methyl and
carbonyl resonances of acetyl groups in hemicellulose
and MeO- to the methoxy groups in lignin. C4cr and
C4am are the peaks from carbon 4 in crystalline and
amorphous cellulose, respectively. Cellulose assign
ment adapted from VanderHart and Atalla [30].

mannose) was unaffected. This indicates that xylose is more easily

degraded and may explain why hardwood fibres produced methane
more quickly than did softwood fibres in this study.
Furthermore, hardwood hemicellulose is acetylated to a larger
extent compared to softwood hemicellulose (reviewed by Pawar et al.
[42]). The presence of acetyl groups in xylans and mannans may
(peroxide). Here CTMP wastewater inhibited methane formation during
hinder enzymatic hydrolysis of polysaccharides [43], and selective
BMP tests of wastewater generated during the production of bleached
deacetyla
pulp, whereas the corresponding wastewater for unbleached pulp
tion has been shown to increase enzymatic (cellulase, cellobiase) di
showed no such effect (Appendix D).
gestibility of wood [44,45]. The decrease in intensity of the acetyl peak
The hemicellulose composition of the CTMP softwood fibres was
observed in this study (Fig. 6) for hardwood – but not for softwood –
altered by bleaching as evident by the different appearance of the
after pulping may therefore have facilitated hydrolysis and partly ex
hemicellulose C1 region in the 13C CPMAS spectra (δC: 95–102 ppm);
plains the higher methane potential of hardwood CTMP fibres
compared to softwood CTMP fibres during AD. see Fig. 8a. The dominant hemicellulose in softwood is
galactoglucomannan and a significant portion of this hemicellulose is
present even after bleaching, as evidenced by the presence of the C1
3.3. Effect of bleaching signal from mannose at δC: 102 ppm (Fig. 8a). However, some of the
hemicellulose has been removed, observed in the reduction in 13C
Like the results for raw material, bleaching had no significant effect
on the methane potentials of the fibres from kraft and sulphite pulping CPMAS signal intensity around 98 ppm as well as in several other
carbohydrate peaks between 60 and 91 ppm. Exactly how the
(Fig. 7). Further, there was no difference in kmax for kraft fibres, whereas
hemicellulose is modified is difficult to deduct but a plausible
for sulphate fibres, bleaching had a positive effect, particularly for explanation may be the removal of the galactose units attached to the
hardwood, with a kmax of 190 compared to 150 Nml g VS− 1⋅day− 1. main chain of galactoglucomannan. Deacetylation during bleaching is
There was a significant difference (p < 0.01) in methane potential for also apparent as the acetyl signal (21 ppm) completely disappeared
unbleached and bleached softwood CTMP fibres (160 ± 8 and 210 ± 12 (Fig. 8). Interestingly, bleaching did not change the lignin content of
Nml CH4 g VS− 1, respectively), and the estimated kmax indicated a CTMP softwood fibres (Appendix C), yet a higher methane yield was
slightly faster degradation of bleached softwood CTMP fibres (8 Nml g achieved for this sample compared to unbleached CTMP fibres. This
VS− 1⋅day− 1 compared to 5 Nml g VS− 1⋅day− 1, Table 3). The BMP suggests that the removal of acetyl groups and the altered
tests for these two samples were, however, not continued until hemicellulose structure made the softwood fibres more suscep tible to
methane production ceased completely, meaning that the total degradation, but the mechanistic details are not clear.
methane produced could eventually be more similar. In the context of The lignin content of hardwood CTMP fibres decreased from 25% to
using the fibres for biogas production, it is not feasible to allow the 18% (Appendix C), which is also visible in Fig. 8b as a decrease in the
BMP test to continue longer than 90 days. peak representing the methoxy group in lignin (δC 56 ppm). Further
For hardwood CTMP fibres, bleaching had no significant effect on more, the acetyl groups that remained after pulping were removed by
methane potential, with 290 ± 8 and 300 ± 10 Nml CH4 g VS− 1 for bleaching, whereas the decrease in hemicelluloses observed for soft
wood was not apparent for hardwood (Fig. 8b). Despite the decrease in
unbleached and bleached, respectively, whereas kmax was higher for
lignin and acetyl groups, neither the rate of degradation nor the
the unbleached fibres (30 Nml g VS− 1⋅day− 1
compared to 20 Nml g methane potential of the bleached hardwood sample was improved;
 instead kmax was lower than for the unbleached fibres (20 compared to the degradability.
−1 −1 In a study by Larsson et al. investigating the methane potential of
30 Nml g VS ⋅day , Table 3). Possibly, the CTMP pulping of
bleached CTMP wastewaters, the concentrations of total organic
hardwood fibres and the removal of acetyl groups improved the
accessibility/degrad ability of the fibres to such an extent that additional carbon (TOC) and acetic acid were about twice as high in wastewaters
modifications to the hemicellulose by bleaching did not further improve from

6
E.-M. Ekstrand et al. Biomass and Bioenergy 143 (2020) 105824

Fig. 4. a) Scores plot of the PCA model with all solid-state 13C CPMAS NMR data included, and b) loading plot of the variables, where positive peaks correlate
positively with the samples that are positive along t1 in Fig. 4a, and negative peaks correlate positively with the samples that are negative along t1. See Table 2 for
abbreviations and Fig. 3 for peak assignments.
bleaching, a large number of aliphatic peaks still persist, whereas the
hardwood pulping as raw material (3800–4600 mg TOC L− 1, 60–70 peaks from the double bonds are not present. Furthermore, aromatic
structures not associated with lignin appear in the HSQC spectrum of
mmol acetic acid L− 1) compared to softwood (2100–2800 mg TOC L− 1,
unbleached TMP fibres, but those peaks are not present after
20–30 mmol acetic acid L− 1) [47]. This corresponds well to the disap
pearance of the acetyl peak in Fig. 8a and b, and the proportionally bleaching. A previous chemical analysis of the bleaching wastewater at
higher intensity of the acetyl peak in hardwood before bleaching. Still, the TMP mill showed the presence of extractives such as resin acids
the higher TOC in the hardwood CTMP wastewater indicates that other and tri glycerides (50 mg l− 1 and 72 mg l− 1, respectively). As the TMP
organic structures are removed from the wood as well. For TMP, the fibres were collected from the wastewaters, these compounds were
fact that unbleached fibres produced less methane than the inoculum likely present in the sample and could be the cause of the inhibition
control (as evident by the negative methane potential) suggests that observed during the BMP test; the release of these compounds into the
AD was inhibited. Similarly, inhibition of the AD process was observed wastewater during bleaching explains why the bleached TMP fibre
during the initial days of BMP tests of both pulping and bleaching sample did not give rise to any apparent inhibition.
wastewater from the same mill, confirming that inhibitory compounds
are released during these processes. Bleached TMP fibres, however,
did not demonstrate an inhibition of degradation, resulting in a 3.4. Pulp and paper mill primary sludge
−1
methane potential of 130 ml CH4 g VS (Fig. 7) and an estimated kmax
An important consideration is that this study was based on pulp fi bres
of 20 Nml g VS− 1⋅day− 1 (Table 3). The 2D HSQC spectrum (Fig. 9)
sampled from the product line or the pulp washing. The primary sludge
indicated the presence of unsaturated aliphatic CH bonds in the un
from the primary clarification at the mill would likely contain residue
bleached fibre sample in addition to a large number of peaks in the
compounds from the different process steps, such as CaCO3 (caused
aliphatic region in both bleached and unbleached TMP fibres. After
 by poor chemical recovery) or fillers from the paper production [48]. primary sludge per ton of TS. Within this study, BMP tests of primary
The inorganic chemicals would reduce the proportion of VS in the sludge from three different kraft mills were performed, demonstrating a
primary sludge and thereby also lower the methane potential of the considerable variability in both VS content and methane potential

7
E.-M. Ekstrand et al. Biomass and Bioenergy 143 (2020) 105824

Fig. 5. Accumulation of methane minus control in BMP tests of hardwood (Hw) and softwood (Sw) from different pulping processes (before bleaching). CTMP =
chemical thermo-mechanical pulping.

Fig. 6. 13C CPMAS NMR spectra on hardwood (Hw; black) and softwood (Sw; grey) in a) raw fibres and b) fibres produced with CTMP. N = 3 for both CTMP
samples. S and G and refers to syringyl (S) and guaiacyl (G) units in lignin, with numbers referring to the different carbon positions in each structure. See Fig. 3 for
assignment of the non-lignin peaks. Lignin assignment adapted from Almendros et al. [46].
 8
E.-M. Ekstrand et al. Biomass and Bioenergy 143 (2020) 105824

Fig. 7. Accumulation of methane minus control in BMP tests on unbleached/bleached fibres from different pulping processes. See Fig. 1 for abbreviations. All
samples are softwood, unless denoted hardwood (Hw).

Fig. 8. 13C CPMAS spectra of bleached (black) and unbleached (grey) fibres produced by CTMP using a) softwood (Sw) or b) hardwood (Hw). The region where
hemicellulosic C1 carbons appear is marked with an asterisk [29].

9
E.-M. Ekstrand et al. Biomass and Bioenergy 143 (2020) 105824

Fig. 9. 2D 1H–13C HSQC spectra from unbleached (black) and bleached (cyan) thermo-mechanically pulped (TMP) softwood (Sw) fibres. (For interpretation of the
references to colour in this figure legend, the reader is referred to the Web version of this article.)
(54% if including recovered fibre pulp [49]). CTMP-pulped fibres would,
however, give rise to a substantial variation in methane pro duction
Table 4 during shifts in raw material and between unbleached/bleached
Methane potential, organic content (volatile solids (VS) as per cent of total qualities of pulp. Here the methane potentials were 300 ± 10 and 160 ±
solids (TS)) and the approximate composition of the primary sludge from three
8 Nml CH4 g VS− 1 for CTMP hardwood and softwood pulp, respectively.
kraft pulp and paper mills.
This means that the application of CTMP fibres for methane production
CH4 (Nml g VS− 1) VS (% of TS) Composition
would vary from site to site, depending on the ratio of hardwood to
310 ± 4 90 100% kraft 170 ± 6 85 80% kraft, 20% NSSC 200 ± 13 70 100% kraft softwood that is used at each mill and how often the mill shifts between
these different raw materials during pulp production. The mechanism
behind the higher degradability in hardwood was likely related to a
(Table 4). Evidently, the higher VS content of the first sample resulted
lower lignin content with more β-O-4 bonds that are easier to break
in a higher methane potential in the sludge compared to the primary
compared to softwood. Furthermore, the hemicellulose was more
sludge from the other two mills. Furthermore, the second mill also
affected by pulping, possibly by a higher degree of deacetylation of the
produced 20% NSSC pulp, which, in line with the results presented for
hardwood xylan, which made it more susceptible to degradation. In
pulp fibres above, reduced the methane potential of the primary
addition, bleaching had a positive effect on the methane potential for
sludge. Thus, site-specific conditions may affect the amount of
softwood CTMP fibres, where the acetyl groups of softwood CTMP
methane that can be extracted from the available primary sludge,
fibres were removed upon bleaching and the composition of the
which should be consid
hemicellulose was affected, likely improving fibre degradability. For
ered prior to any full-scale implementation.
TMP fibres, the presence of extractives originating from the wood
inhibited biogas production, while bleaching alleviated the inhibitory
4. Conclusions
effects of these compounds, and enabled the conversion of fibres to
biogas during AD.
In summary, fibres from kraft and sulphite pulping would be suitable
substrates for AD, demonstrating high methane potentials (400 ± 17
Acknowledgements
and 390 ± 6 Nml CH4 g VS− 1, respectively). These fibres also had the
− 1 − 1
highest rate of methane production (130–200 Nml g VS ⋅day ), The authors wish to thank the personnel at SCA and
most likely due to the efficient removal of lignin during the pulping
BillerudKorsnas ¨ and Xu-Bin Truong at Scandinavian Biogas Fuels AB
processes. Furthermore, as the methane potential for these fibres did
for their assistance within this study. The authors would also like to
not differ during shifts in raw materials used or shifts in production
between un thank NMR for Life, a project funded by the Knut and Alice Wallenberg
bleached and bleached pulp, the methane production at a full-scale AD foundation. The study was funded by the Swedish Energy Agency
plant would likely remain stable despite frequent variations in these (Grant No. 32802-2), Scan
operational conditions. Thus, our results strongly promotes the appli dinavian Biogas Fuels AB, Poyry ¨ AB, BillerudKorsn¨as AB, SCA,
cation of biogas production from kraft and sulphite primary sludge, and Fiskeby Board AB and Purac AB. It was also supported by the
would result in a considerable methane production worldwide as Swedish Research Council Formas [Grant number: 2016-01054] and
chemical pulping represents 82% of the world’s virgin pulp production Biogas Research

10
E.-M. Ekstrand et al.
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