The lignins used in the present work originate from spruce or birch sawdust, and while any specific

means of extraction would alter the native structures, they are also different from the start. Work
done by [https://www.sciencedirect.com/science/article/pii/S0958166918301289?
casa_token=aFGHJ_ZrVDcAAAAA:B_qrr8g4JpBK4xlrKtGjhpRr9fYcYAZeXEibQq8ygyeyiAELmatwoo2ye
Hyqvi2Y3Hvr7rf9] summarizes the main differences in lignin structures based on which lignocellulosic
biomass they come from. In short, softwoods such as spruce lignins, are dominated by the guaiacyl
monomer, being derived from the coniferyl alcohol precursor. The interunit linkages vary both in
structure and content. While the β-O-4’ linkage is present to the largest extent (approx. 60%), also
the dibenzodioxocin (5-5’/4-O-β’), phenylcoumaran (β-5’) and pinoresinol (β-β’) are present to
approximately 10, 10, and 5%. Apart from these structures, structures such as spirodienone (β-1’)
and biphenyl ether (4-O-5’) are present at lower contents (around 1%). In hardwoods such as birch,
the situation is different. In general, the monomeric distribution is dominated by syringyl units (S:G
ratio around 65:35), while the amount of β-ether units (β-O-4’) is often higher than in softwoods
(approx. 80%). Else, the content of β-5’, β-β’, and 5-5/4-O-5’ as in general lower, with contents
approximately equal to 5, 5, and 3%. Structures occurring at lower contents (1-2%) in softwoods,
namely the spirodienone (β-1’) and biphenyl ether (4-O-5’) the content is about the same as in
hardwoods.
Now, while it is of great importance to have some sort of idea as on what chemical structures are
present in the raw material set for hydrothermal liquefaction, the process prior, namely the
extraction which in this case is organosolv, will modify the distribution of the mentioned chemical
characteristics. There are few papers exploring the chemical nature of lignins extracted through
organosolv of the raw materials for the conditions relevant in this work. However,
[https://pubs.acs.org/doi/abs/10.1021/acsomega.0c05719] recently elucidated the chemistry of
lignins extracted from birch at conditions similar to the two used to extract the lignins applied in the
present work, and they will be mentioned at a later stage in this section. For spruce, similar
characterization has been performed on lignins extracted upon similar conditions as those used for
the lignins employed in this work [CITE SPRUCE PAPER]. For these lignins, employing acid does not
seem to promote Cα ethoxylation, but instead reduce the extent of this. In fact, employing acid seem
to reduce native lignin structures such as β-O-4’, and β-5’, while β-β’ and β-1’ contents are fairly
unchanged. Interestingly though, despite the decrease in native inter-unit motifs, the molecular
weight (both Mn and Mw) increases, and as mentioned in the referred paper, this is likely related to
incorporation of sugar dehydration products (and thus rich content of oxygenated species).
 Mass Distribution
Oil direct extract Char Losses
100%
90%
80%
70%
Mass Yield

60%
50%
40%
30%
20%
10%
0%
90 92 128 129 88 93 127 127_2
Reaction

Figure 1:

Bio-oil mass Distribution

HO LO
60%
50%
40%
Mass Yield

30%
20%
10%
0%
90 92 128 129 88 93 127 127_2
Reaction

Figure 2:
 Mass Distribution Monomers
14%

12%

10%
Monomers in LO
8% Monomers in HO

6%

4%

2%

0%
90 92 129 128 88 93 127 127_2

Figure 3:

Comparing the LO and HO yields obtained from treatment 90 and 92, applying acid during organosolv
extraction (92) of lignins from spruce seem to generate similar LO yields while increasing the HO
fraction (Figure 2). Considering the results obtained through GC/MS (Table 1) the monomers being
kept out of the LO fraction of 92 are guaiacol species (guaiacol, 4-ethylguaiacol and 4-
methylguaiacol ), this while guaiacylacetone is enriched in the LO of 92. What is interesting is that the
guaiacol species present in the LO fraction of oil produced from acid extracted spruce lignin, is the
one with oxidized Cα side-chain
[https://www.sciencedirect.com/science/article/pii/S1364032121009965], while the remaining
guaiacol structures either have no residuals from the side-chain present on the aromatic ring, a
methyl or ethyl group. This is likely related to the fact that the lignins extracted without acid have
greater content of intact side-chain structures (β-O-4’) [Ref spruce results] which can undergo both
C-C bond cleavage under subcritical conditions
[https://www.sciencedirect.com/science/article/pii/S1364032121009965], while an oxidized Cα is
less prone for C-C bond cleavage in the side chain. Thus no acid and intact aryl ether structures are
more likely to first have C-C bond cleavage in the side-chain, then experience dehydration forming
the alkylated guaiacol structures. Indeed, as mentioned by the latest reference, the C α-Cβ bond often
experiences greater dissociation energy when the Cα is oxidized to a ketone, which could be why the
acetone motif is intact (side-chain is cleaved further out). Now, 92, have far fewer of these
structures, and instead have aliphatic condensed side-chain motifs, alongside elevated contents of
oxidized Cα, which would explain the presence of only guaiacylacetone in the LO fraction. Obviously,
an oxidized side-chain (Cα) would likely keep the lignin from undergoing formation of alliphatic
condensed structures, which would mean that these lignins instead display a larger content of
linkages more unlikely to depolymerize, and would thus not form LO components thereby explaining
the higher yield of HO.
Guaiacol has been found to generate catechol and phenol, where catechol is considered a precursor
upon repolymerization reactions and for the formation of bio-chars, this while phenol is considered
stable [https://www.sciencedirect.com/science/article/abs/pii/S1364032113004589]. As mentioned
by [https://www.sciencedirect.com/science/article/pii/S096195341630294X], especially formation of
reactive intermediates such as guaiacol seem to promote repolymerization processes, this while
alkylated phenolics (Ar with a OH group) are simple phenols are less reactive. Now, looking at
treatment 128 and comparing this against 92, it would give an indication of what an increase in
substrate would affect the process. As can be seen in Figure 3, it enhances the amount of monomers
in the LO fraction, this while the overall LO yield (Figure 2) is effectively unaffected while the HO
yields decreases. Thus, it would seem as HO components are degraded giving rise to more monomers
in the LO, while inter-unit/side-chain structures likely are lost through volatiles keeping the LO yield
unaletered. Considering the side-chain of the lignin polymer, it has been considered as an important
structure for formation of organic acids in the hydrothermal product
[https://www.sciencedirect.com/science/article/abs/pii/S1364032113004589]. Meanwhile, there is
also suddenly formation of char, which could be due to formation of reactive intermediates
originating from the greater content of monomers. This was not the case for treatment 90 and 92, so
the appearance of new monomers, or the disappearance of any would be of interest. From the
GC/MS (Table 1), certain new structures appear in the LO fraction of 128. Among these are creosol,
catechol, 5-formylguaiacol but also syringaldehyde, among these catechol has been suggested to
form char https://www.sciencedirect.com/science/article/abs/pii/S1364032113004589], as have
guaiacol [https://link.springer.com/article/10.1007/s13399-016-0228-4] which is present at higher
contents in 128 LO.
Moving forward and looking at the continuous process (129; t = 20 min; 180 m(lignin)/g), conversion
of lignin into LO seem highly efficient, this as the HO and char yields are low (Figure 2 and Figure 1),
while the overall mass distribution of monomers in the LO is fairly high (Figure 3). Now, this is
peculiar, because a shorter treatment time and higher masses set for treatment generates a higher
content of LO. Expecting that LO originates from HO, and further can be converted into char is thus
not obvious anymore. Now, looking at the monomers present in the LO fraction of 129, it can be seen
that the overall fraction of monomers in the LO is 27%, this opposed to 72% for treatment 92 (30
min; 6 m(lignin)/g), so it seems as while LO components are formed, they are still not sufficiently
depolymerized to generate a high content of monomers thus explaining the low char formation. This
said, treatment 129 still generates char, and as with 128, they both present contents of catechol,
creosol, 5-formylguaiacol, which are not present in the non-char-forming treatments of 90 and 92.
Still, it would seem as potentially this treatment for some reason is more ‘severe’ as a lot of the
original lignin is converted into LO. Seemingly efficient conversion of HO into LO has been observed
before [https://www.mdpi.com/1996-1073/12/4/723] where increased severity causes steady
decrease of HO yield while the LO increases. Now, this could originate from the increased presence
of hydrophobic surfaces into the liquid which are preferably solvated by the hydroxide ion
[https://pubs.acs.org/doi/abs/10.1021/jp906978v], and lignin treated upon neutral hydrothermal
conditions have shown a preference to undergo base-catalyzed elimination/hydrolysis reactions
[https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cssc.201700528]. Thus while,
the presence of greater lignin contents may experience enhanced depolymerization,
repolymerization in the bulk might not be preferred. Worth mentioning on what seems to be a fairly
stable LO fraction, 5-tert-butylpyrogallol appears only for this spruce fraction, and alkylated
phenolics are reported stable
[https://www.sciencedirect.com/science/article/pii/S096195341630294X].

Table 1:

SSD1 (90) SSD2 (92) SSD2 (129) SSD2 (129) SSD2 (128) SSD2 (128)
CONT CONT Pre-S-U Pre-S-U
(5)LO (6)LO (10)LO (10)HO (9)LO (9)HO
(monomers (monomers (monomers (monomers (monomers (monomers
) mg/g ) mg/g ) mg/g ) mg/g ) mg/g ) mg/g
2-
Methyltetrahydrofura
n
Furfural
Diacetone alcohol
Phenol
p-Cresol 15.5 19.4
Guaiacol 274.2 212.9 106 238
4-methylguaiacol 113.3 73.7
Catechol 19 66
Creosol 21 80
p-hydroxyphenol
3-methoxycatechol 35.1 3 20
3-methoxycatechol (or
other isomer)
4-ethylguaiachol 48.4 34.6 17 36
4-methylcatechol 16
2,6-dimethoxy phenol 62.5 17.4 10
4-propylguaiacol 18.5 14.5
1,3,4- 16.7
dimethoxyphenol
Vanilin 76 56.9
Phenol, 4-methoxy-3-
(methoxymethyl)
5-Formylguaiacol 42 74
Phenol, 2,6- 28.6
dimethoxy-4-methyl
Ethanone,1-(2,3,4-
trihydroxyphenyl)
Acetoguaiacone 35.8 26.4 10 43
2,6-Di-tert-butyl-p-
cresol
5-tert-butylpyrogallol 41
Guaiacylacetone 58.1 110.9 169
2-butanone,4-(4-
hydroxyphenyl)
p-cresol, 2,6-di-tert- 10 2
butyl
Escaline 11
Syringaldehyde 2 79
Gallacetophenone-4’- 5
methylether
Benzenepropanol, 4- 57.4 60.60
hydroxy-3-methoxy-
Syringaldehyde 19.1
Gallacetophenone-4’-
methylether
Phenol (unknown)
Acetosyringone
Syringylacetone
5-(3-hydroxypropyl)- 26.2 18.2
2,3-dimethoxyphenol
Hexadecanoic acid or
likewise
Phenol (unknown) 20.40
Phenol (unknown)
Phenol (unknown) 16.8 17.4

Sum of quantified 885.5 716 272 10 820 2

mass
Monomers in fraction 89% 72% 27% 1% 82% 0%

Yield LO 7% 7% 33% 9%
Yield HO 36% 47% 7.1% 43%
Yield char 0% 0% 1.0% 1.3%

Yield monomers in LO 6.2% 5.0% 8.9% 7.3%

Yield monomers in HO 0.1%
As with spruce, there will be differences in birch lignin chemistry due to the application of acid or not
during lignin extraction. As elucidated by [https://pubs.acs.org/doi/abs/10.1021/acsomega.0c05719],
employing acid reduces the number and weight average molecular weight, increases the content of
condensed structures in the aliphatic side chain (β-5’, β-β’, β-1’, 5-5’, 4-O-5’) slightly enhances the
content of S units in the extract, and promotes the cleavage of β-O-4’ linkages (especially those
coupled to the S units). In addition to this, the application of acid also enhances ethoxylation in C α
position.

For birch, employing acid during the production of the organosolv lignins also seem to hamper
formation of any chars when subsequent performing hydrothermal liquefaction of the lignins (88 and
93; Figure 1), and instead increases the overall mass yield of bio-oils (Figure 1).

Now, as seen with spruce lignins, this could also originate from the fact that the structures present in
the lignin substrate are locked up in ‘less depolymerizable motifs’, which hinders formation of
reactive monomer intermediates during the hydrothermal liquefaction. Interestingly, this is also seen
alongside increased HO yields, while there is a slight decrease in the LO yield (Figure 2), which indeed
indicates that the majority of structures are to a greater extent locked up in less viable ‘stronger’
structures. This said, a greater content of the LO of 93 is constituted by monomers (Figure 3)
suggesting that some of the structures are still susceptible for depolymerization under the applied
conditions. Evaluating the monomers actually present in the LO of 88 and 93. As mentioned in the
discussion on bio-oils originating from spruce, catechol has been reported as a precursor for char,
this while alkylated aromatics and substituted phenols on the other hand are suggested more stable,
the formation of char for 88 and the potential involvement of catechol could be seen through its
absence in 88 LO, while 4-methylcatechol and 4-ethylcatechol are present at approx. 6-fold contents
in the 93 LO where no char formation is detected. Thus it would seem plausible that the involvement
of the catechol precursor in certain lignin motifs prior to bio-oil production would favor the
subsequent formation of either alkylated or non-alkylated intermediates, which has different affinity
for involvement in re-polymerization reactions. Their exact chemical destiny in the char is probably
complex, but phenols are reported to disappear from the light oil fraction upon elevated
hydrothermal treatment temperatures of lignin
[https://www.sciencedirect.com/science/article/pii/S0196890419309616].

Now one scenario for which LO components end up in the char is through the formation of PAHs,
which could have relatively high solubility due to ‘clusters of delocalized π* electrons’ experiencing
elevated interaction with the solvent at elevated temperatures, but upon cooling aggregate forming
the char [https://www.sciencedirect.com/science/article/pii/S1385894711007029].
Interestingly, when scaling up 93 into 127, the bio-oil yield is enhanced, but this is associated with
formation of char (Fig. 1), which is observed with an increase in both HO and LO yields (Fig. 2). The
system (127) seem to be enriched with monomers in the LO fraction (Figure 3; which then upon
extraction of the LO would generate some residue which is left in the HO fraction). As seen earlier,
elevated contents of monomers seem to be involved with char formation
[https://www.sciencedirect.com/science/article/abs/pii/S1364032113004589]
[https://pubs.rsc.org/en/content/articlehtml/2017/se/c7se00357a]
[https://www.sciencedirect.com/science/article/pii/S096195341630294X]. For structures detected
through GC/MS, and absent in the other birch LO fractions, in the LO fraction of 127 contain
monomers such as catechol and creosol, as is the highest contents of guaiacol (also reported to be
reactive and precursor for char) from this raw material.

Also needing mentioning is the presence of S units in the birch lignin. The fact that these are present
could affect to what extent both depolymerization and re-polymerization occurs. As recently
reviewed by [https://www.sciencedirect.com/science/article/pii/S0960852421013778], the bonds
set to undergo alteration when treating lignins at hydrothermal conditions are through hydrolysis of
ether bonds (β-O-4’, α-O-4’, α-O-γ’, 4-O-5’, etc.) and C-C bonds (β-β’, β-5’, 5-5’, etc.) to yield phenolic
compounds. As temperature is further enhanced, the products are further altered through
hydrolysis, demethoxylation, and alkylation resulting in stable alkylated aromatics and further to
aromatic oligomers through re-polymerization. Interestingly, as mentioned by
[https://www.sciencedirect.com/science/article/pii/S0960852421013778], it has been found that the
presence of methoxy functional groups on the aromatic ring allows for easier cleavage of any aryl
ether linkages in the lignin side chain, likely due to a more flexible distribution of charge allowing for
stabilization of any partially charged intermediate species. Thus, one would expect S units of lignin to
potentially experience easier depolymerization as these experience greater stability as independent
structures. However, this would also allow for stabilization of any ionic intermediate states as
charges as easily displaced. Interestingly, while treatment 93 is the one displaying absence of char, it
also presents the highest combined content of syringaldehyde, acetosyringone, and syringaldehyde
(S/G ratio for lignins extracted at similar conditions such as 88 and 93 are reported to be fairly similar
[https://pubs.acs.org/doi/full/10.1021/acsomega.0c05719]), which thus could indicate that the
syringyl-based structures partake in formation of char. Also noteworthy, the acid extracted birch
lignin seem to have a preference of forming S-based monomers where the Cβ carries the ketone
functional group (syringylacetone; Table2), likely due to the fact that the Cβ ether is already
hydrolyzed during extraction.
To evaluate the stability of the heavy oil fraction produced from the scale-up trial, the HO fraction of
127 was reprocessed under condition else being similar to that of 90 and 93. Interestingly, this gave
rise to no char formation, this despite the fact that the HO of 127 displayed a content of monomers
(Figure 3). The monomers initially present in the HO fraction of 127 where of the highly substituted
phenolic type, such as 2,6-dimethoxy phenol, 1,2,4-dimethoxyphenol, 4-methoxy-3-
(methoxymethyl)phenol, 4-tert-butylpyrogallol, and 2,6-di-tert-butyl-p-cresol, thus indicating that
these structures are not significantly contributing to any significant formation of char. Instead, the
slight decrease in both HO and LO yield seen for 127_2 as compared to the mass distribution of HO of
127 (Fig. 2), indicate that some loss is still experiences upon reprocessing, likely due to the loss of
certain aliphatic and/or oxidized species substituted onto the aromatic rings of certain structures.

Table 2:

BSD1 (88) BSD2 (93) BSD2(127) BSD2(127)

S-U S-U
(4)LO (7)LO (8)LO (8)HO
(monomers) mg/g (monomers) mg/g (monomers) mg/g (monomers) mg/g
2-
Methyltetrahydrofura
n
Furfural
Diacetone alcohol
Phenol
p-Cresol
Guaiacol 60.1 66.3 152
4-methylguaiacol 24.1 45.1
Catechol 55
Creosol 39
p-hydroxyphenol
3-methoxycatechol 89.3 57.4 217
3-methoxycatechol (or
other isomer)
4-ethylguaiachol 21.4 96.7 21
4-methylcatechol 28.4
2,6-dimethoxy phenol 251 215.9 316 18
4-propylguaiacol 18 23.4
1,3,4- 20.2 32 1
dimethoxyphenol
Vanilin 40.5
Phenol, 4-methoxy-3- 16 1
(methoxymethyl)
5-Formylguaiacol 21.8 21
Phenol, 2,6- 65.5 61.2 103 8
dimethoxy-4-methyl
Ethanone,1-(2,3,4- 5
trihydroxyphenyl)
Acetoguaiacone 15.7 22.4 23
2,6-Di-tert-butyl-p- 23.2 25.4
cresol
5-tert-butylpyrogallol 33.5 4
Guaiacylacetone 20.4 80.3 94
2-butanone,4-(4- 30.4 7
hydroxyphenyl)
p-cresol, 2,6-di-tert- 9
butyl
Escaline 18.3
Syringaldehyde 23.3 40
Gallacetophenone-4’- 17.3 31
methylether
Benzenepropanol, 4- 23
hydroxy-3-methoxy-
Syringaldehyde 39.8 52.2
Gallacetophenone-4’- 17.1
methylether
Phenol (unknown) 24.4
Acetosyringone 28.7 56.2 32
Syringylacetone 42 121 108
5-(3-hydroxypropyl)-
2,3-dimethoxyphenol
Hexadecanoic acid or
likewise
Phenol (unknown)
Phenol (unknown) 18.1
Phenol (unknown)

Sum of quantified 796 1143 1312 41

mass
Monomers in fraction 80% 114% 131% 4%

Yield LO 7% 4% 8%
Yield HO 24% 28% 50%
Yield char 1% 0% 1.5%

Yield monomers in LO 5.6% 4.0% 8.3%

Yield monomers in HO 2.1%