Biomass Conversion and Biorefinery

https://doi.org/10.1007/s13399-024-05573-7

ORIGINAL ARTICLE

High quality hydro‑char production through co‑hydrothermal

treatment of sewage sludge and wheat straw: novel properties
Qing An1,2 · Dezhen Chen1,2 · Mengxi Han1,2 · Yuheng Feng1,2

Received: 16 August 2023 / Revised: 6 December 2023 / Accepted: 8 December 2023

© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2024

Abstract
Co-hydrothermal carbonization (Co-HTC) of sewage sludge (SS) with wheat straw (WS) has emerged as a promising method
for producing high quality biochar products through the utilization of low-cost biomass resources. The objective of this study
was to investigate the physico-chemical properties of hydro-char derived from the Co-HTC treatment of the mixture of SS
and WS at different Co-HTC temperatures (200, 240, and 280 °C). The research findings indicated that the inclusion of WS
in the Co-HTC process positively influenced the properties of the hydro-char and higher Co-HTC temperatures enhanced
this trend. It is revealed that the hydro-char produced at 280 °C exhibits the biggest surface area, abundance of micropore
structures, and the highest electron accepting capacity (EAC = 0.4389 ­mol−/ghydor-char). Moreover, when increasing the Co-
HTC temperature from 200 to 240 °C, the higher degree of aromatization (H/C = 1.1170 (atomic ratio)) can be achieved,
which correlates with the electron donating capacity (EDC = 0.7358 ­mol−/ghydor-char). For the secondary dissolution of
organic substances leaching from hydro-chars, the blending of SS and of WS can significantly enhance the content of aro-
matic protein II, with marginal impact on humic acid. Furthermore, the Co-HTC process proved effective in immobilizing
heavy metals during the treatment. All of the improved properties facilitate the SW hydro-chars to be used as promoters in
anerobic digestion system.

Keywords Co-hydrothermal carbonization (Co-HTC) · Sewage sludge · Wheat straw · Hydro-char · Physico-chemical
properties

1 Introduction disposal [4]. However, challenges persist, such as the genera-

Sewage sludge (SS) is a by-product of wastewater treat-
ment plants, and its production continues to escalate due
to global economic development and construction of more
wastewater plants for water pollution control [1]. Improper
handling of sludge can lead to severe environmental risks
due to the presence of harmful organic substances, including
pathogens, drugs, hormones, heavy metals, and polycyclic
aromatic hydrocarbons etc. in SS [2, 3]. Presently, landfill,
composting, and incineration are the primary methods for SS
* Dezhen Chen
chendezhen@tongji.edu.cn
1
Thermal and Environmental Engineering Institute,
Mechanical Engineering College, Tongji University,
Shanghai 201804, China
2
Shanghai Engineering Research Center of Multi-Source
Solid Wastes Co-Processing and Energy Utilization,
Shanghai 201804, China
tion of secondary pollutants, wastage of land resources, and
operational complexities [5]. Consequently, finding an effec-
tive and sustainable sludge treatment technology remains a
pressing issue.
Carbonization is a thermal treatment process that con-
verts heterogeneous organic biomass into char under oxy-
gen-deficient environment, comprising hydrothermal car-
bonization (HTC) and pyrolysis methods [6, 7]. The strong
adhesion of water molecules through hydrogen bonding
on the surface of SS results in high moisture content and
poor dehydration ability, leading to operational difficul-
ties and increased energy consumption during pyrolysis
[8]. In comparison, HTC treatment of SS can generate
high pressure with low energy consumption, improving
sludge dehydration, reducing volume, degrading organic
pollutants, and lowering heavy metal content in the result-
ing hydro-char. This method proves to be effective for
safe sludge disposal and carbon material production [9].
Specifically, hydro-char produced from Co-HTC process

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 Biomass Conversion and Biorefinery

carries rich functional groups, although it has relatively
small specific surface area, low level of porosity, and
higher ash content, which restrict its applications [10, 11].
As a solution, a new technique known as Co-HTC has been
developed.
Co-HTC treatment involves processing two or more
feedstocks in the same Co-HTC system [12]. This
approach enables the efficient amalgamation of advan-
tageous characteristics from various feedstocks such as
sawdust, corncobs, cornstalks, and rice straws [13]. Lev-
eraging the synergistic effects of two types of solid wastes,
they can be repurposed and recycled as hydro-char solid
fuel with superior quality.[14, 15]. For instance, He et al.
(2019) found that Co-HTC treatment of SS with fruit
and agricultural waste promotes decarboxylation dur-
ing the process, thereby improving the fuel properties of
the resulting hydro-char [16]. Similarly, Lu et al. (2021)
reported improved fuel properties of hydro-char from Co-
HTC treatment of SS and woody biomass [17]. Addition-
ally, Tong et al. (2021) observed an increasing trend in
specific surface area and pore size of hydro-char with the
increasing addition of rice straw during Co-HTC treatment
with SS [18]. Furthermore, significant synergistic effects
were evident when SS was blended with pinewood sawdust
for Co-HTC conducted at 220 °C. This led to the produc-
tion of high-quality, nutrient-rich hydro-char [19]. How-
ever, studies on the synergetic effect of Co-HTC on the
hydro-char properties are lacking. Especially, hydro-char
from SS is often concerned with its heavy metal content
when applied as soil amendment or as additive/absorbents.
On the other hand, wheat straw is a widely available agri-
culture waste, but wide application of its hydro-char is
currently limited by low char yield, lack of some special
adsorption ability relevant with N content [20], and the
refractory hydrothermal liquid.
SS is rich with protein and N content [21]. Co-HTC of
SS and WS could produce hydro-char rich in N element
with satisfied yield. Based on the knowledge from previ-
ous researches, this study utilized WS, an available agricul-
tural waste, as a cheap auxiliary material added to SS in a
hydrothermal reactor. Different hydro-chars were obtained
by varying the Co-HTC temperature, and an in-depth evalu-
ation of the properties of the hydro-char was conducted. This
research contributes to the development of a reasonable and
effective SS treatment technology, exploring the potential of
Co-HTC approaches to enhance the properties of hydro-char
for various applications.
2 Materials and methods
2.1 Raw materials
The SS utilized in this study was sampled from Zhuyuan
Sewage Treatment Plant in China, while the WS was
obtained from Chongming Island, China. The raw SS and
WS were subjected to drying at 105 °C for 12 h, followed by
milling and sifting through a 100-mesh sieve. Subsequently,
the dried SS and WS were mixed by 1:1 ratio and designated
as SW. Table 1 presents the characteristics of the raw materi-
als used in this study.
2.2 Preparation methods
The milled SS and SW were subjected to hydrothermal treat-
ment to produce hydro-chars using the methods described

Table 1  Physicochemical characteristics of raw materials and prepared hydro-chars

Sample Proximate analysis/ (wt%, db 1) Ultimate analysis/ (wt%, db) BET surface Pore volume/ Average pore size/ HHV/
2 3 4
area/ ­(m2g-1) ­(cm3g-1) (nm) ­(MJkg-1)
Ash VM FC C H O N S

SS 45.43 48.01 6.56 25.12 4.34 20.48 4.09 0.54 4.28 0.02 20.95 10.86
WS 5.38 58.33 36.29 45.37 5.76 42.91 0.28 0.30 — — — 18.10
H-SS-200 58.38 34.95 6.67 22.21 3.23 12.89 2.94 0.35 14.70 0.13 36.17 9.03
H-SW-200 38.87 48.87 12.26 33.48 3.19 21.86 2.53 0.07 28.00 0.05 1.63 12.37
H-SS-240 62.25 28.31 9.44 21.69 2.75 10.49 2.46 0.36 12.93 0.11 34.17 8.45
H-SW-240 43.25 41.70 15.05 32.66 3.04 18.49 2.49 0.07 36.19 0.05 1.49 12.16
H-SS-280 65.46 24.27 10.27 20.77 2.57 8.60 2.26 0.34 10.30 0.07 28.28 8.04
H-SW-280 50.32 29.54 20.14 31.04 2.81 13.42 2.35 0.06 24.31 0.04 1.48 11.70
1
db: dry basis
2
VM: volatile matter
3
FC: fixed carbon
4
O: calculated by difference
Biomass Conversion and Biorefinery

Table 2  Preparation methods of Sample SS:WS Dry weight/ Volume/ Temperature/ Retention time/ Hydrothermal
hydro-chars (g) (mL) (°C) (h) liquid pH

H-SS-200 1:0 20 200 200 0.5 7.48

H-WS-200 0:1 20 200 200 0.5 3.51
H-SW-200 1:1 20 200 200 0.5 4.17
H-SS-240 1:0 20 200 240 0.5 7.74
H-WS-240 0:1 20 200 240 0.5 3.58
H-SW-240 1:1 20 200 240 0.5 4.19
H-SS-280 1:0 20 200 280 0.5 8.03
H-WS-280 0:1 20 200 280 0.5 4.11
H-SW-280 1:1 20 200 280· 0.5 7.24

in Table 2. The Co-HTC reactor operated at a water to dry
mass ratio of 4:1. Deionized water with an ohmic resistance
of 18 MΩ cm was used as the reaction medium. The heating
rate of 10 °C/min was adopted, after the target temperature
reached (i.e., 200, 240, and 280 °C), another reaction time
of 30 min was maintained, during which the reactor was
in a saturated state at the corresponding temperature, and
then the reactor stayed overnight to be cooled down to room
temperature. The hydro-char was obtained by separating the
solid, and it was dried at 105 °C for 12 h. The resulted hydro-
chars were ground into powder on a disk-rotating mill and
passed through a 100-mesh sieve for analysis. The hydro-
char samples produced at different Co-HTC temperatures
were named using the format of “raw materials + hydrother-
mal temperature”, as shown in Table 2. The yield of hydro-
char was calculated using the following equation:
Whydro−char
Yhydro−char (%) = × 100 (1)
Wraw
where ­Whydro-char was the mass of hydro-char solid after dry-
ing, and Wraw was the mass of raw materials of SS, WS, and
WS mixture before Co-HTC.
2.3 Analytical methods
The proximate analysis was conducted to measure the total
solid and volatile solid of the raw materials and hydro-chars,
following ASTM D-5142 standard methods. The Brunauer
Emmett and Teller (BET) method was used to analyze the
specific surface area and pore volume. The Vario Micro ana-
lyzer (Elementar Analysensysteme, Hanau, Germany) was
utilized to determine the elemental composition (N, H, O,
N, S) of the raw materials and hydro-chars. The high heating
value (HHV) was calculated based on the modified Dulong
formula, considering the hydro-chars’ element composition
[22]. To identify the functional groups on the surface of the
hydro-char, Fourier-transform infrared (FTIR) spectra was
obtained using an infinity Avatar 360 FTIR spectrometer
with a diffuse fluctuance cell (Version 8.0, Nicolet Instru-
ments Corporation, USA). The surface morphology of the
hydro-char was observed using a scanning electron micro-
scope (SEM) (Quanta 200 FEG, FEI Company, Czech
Republic). The soluble organic matter leaching experiments
of hydro-chars were based on the research of Jamieson et al.
(2014) [23]: 0.5 g hydro-char was added into 5 mL deionized
water to shake at 150 rpm for 8 h, and then centrifuged at
8000 rpm for 5 min to get the supernatant. The fluorescent
substance in the supernatant was analyzed using 3D exci-
tation emission matrix (EEM) fluorescence spectroscopy
(Cary Eclipse, Varian Co., USA), the excitation wavelengths
ranging from 220 to 550 nm and emission wavelengths rang-
ing from 220 to 620 nm, with a step length of 2 nm and slit
width at 12,000 nm/s scanning speed and 400 V of voltage
intensity. Meanwhile, to measure the leaching heavy met-
als concentration in the hydro-chars, the leaching tests of
hydro-char samples were conducted based on the Chinese
standards HJ/T 299–2007 and HJ/T 300–2007 (Table S1)
[24]. All the chemicals utilized in the leaching process were
of analytical purity and procured from Sigma-Aldrich. The
metals including As, Cr, Zn, Pb, Cu, Cd, Ni, Ca, Fe, and
Mn in the leachate solutions were analyzed with the help of
an inductively coupled plasma emission spectrometry (ICP-
OES) (Agilent 720 ES, USA). The electron accepting capac-
ity (EAC) and electron donating capacity (EDC) were meas-
ured using an electrochemical station (CORRTEST, China)
based on methods described in previous publications [25].
3 Results and analysis
3.1 Hydro‑char yield
Figure 1 shows the yields of the hydro-chars obtained from
the Co-HTC processes for SS, WS, and Co-HTC process
for SW. As illustrated in Fig. 1, the hydro-char yields for
H-SS-200, H-SS-240, and H-SS-280 were 66.27, 64.02, and
60.57 wt.%, respectively. For H-WS-200, H-WS-240, and

Fig. 1  Hydro-char yields from HTC of SS, WS, and SW
H-WS-280, the hydro-char yields were only 40.75, 37.31,
and 32.72 wt.%, respectively. For H-SW-200, H-SW-240,
and H-SW-280, their yields were 56.74, 46.88, and 41.15
wt.%, respectively. It is evident that the hydro-char yields
decreased as the Co-HTC temperature increased. This trend
can be attributed to the depolymerization and decomposi-
tion of organic substances, along with the removal of most
alkali metals during the Co-HTC treatment process [26, 27].
Furthermore, in comparison to the SS hydro-char samples,
the hydro-char yields were lower for the WS and SW hydro-
char samples. This divergence can be possibly ascribed to
the fact that the lower ash content and higher volatile matter
content from WS (Table 1).
3.2 Characteristics of hydro‑chars
3.2.1 Element composition and fuel characteristics
The proximate analysis, ultimate analysis, and HHV of
hydro-char from Co-HTC of SS and SW can been seen in
Table 1.
It can be observed that the SW hydro-char exhibited lower
ash content, higher fixed carbon (FC) content, and higher
volatile matter (VM) content compared to the SS hydro-
char. This can be attributed to the fact that WS has a lower
ash content and higher FC content; in addition, ash dissolu-
tion could happen during Co-HTC process caused by the
relatively lower pH values than SS alone. For SS hydro-
char, its ash and FC contents showed an increasing trend
with the increase in Co-HTC temperature, while the VM
content showed an opposite trend. A similar changing trend
was observed in the proximate analysis of the SW hydro-
char. These results are caused by the degradation of most
Biomass Conversion and Biorefinery
hemicellulose and amorphous cellulose, accompanied by
leaching of water-soluble inorganic species with increas-
ing Co-HTC temperature [28]. In addition, the dissolution
of inorganic species was more serious at lower temperature
(with lower pH value) than at higher temperature.
It was also found that blending WS resulted in an increase
in C content compared to SS hydro-char. This can be
attributed to the high C content and thus high FC content
contributed by WS. Moreover, Co-HTC temperature also
influenced the C content of hydro-chars; it decreased from
22.21 to 20.77% with temperature increased from 200 to
280 °C for SS hydro-char and from 33.48 to 31.04% for SW
hydro-chars. Furthermore, blending WS enhanced the trans-
formation of organic nitrogen-containing compounds into
the liquid phase and the leaching of organic and inorganic
sulfur in aqueous phase during the Co-HTC process, which
significantly reduced N and S concentrations in hydro-chars
[29, 30]. However, Co-HTC temperature only had a slight
influence on N and S concentrations in this study, and N
contents in SW hydro-chars are much higher than that in
WS material.
During the Co-HTC process, blending WS also resulted
in an increase in HHV compared to SS hydro-char, due
to the decrease in ash content and an increase in C con-
tent in hydro-char. The HHV value ranges within 12.37 to
11.70 MJ/kg, meaning that WS hydro-chars are not good
fuels. Furthermore, an increase in Co-HTC temperature has
a negative impact on HHV. This finding is consistent with
previous research which reported that raising the tempera-
ture was harmful to increasing HHV of hydro-char when
Co-HTC temperature exceeds 180 °C [31].
3.2.2 Carbonization degree
Figure 2 shows the Van Krevelen diagram of the raw materi-
als and hydro-char products. The value of H/C can serve as
an indicator of carbonization degree; it decreased slightly
when blending WS and increasing the Co-HTC temperature,
indicating improved aromatization degree. It can be seen that
blending WS under same temperature led to lower H/C and
higher O/C, implying that addition of WS can increase the
aromatization, and higher O/C means the higher hydropho-
bicity [32]. The H/C and O/C ratios of the hydro-char also
affect the electron transfer ability of the hydro-char. It was
reported that the carbon matrix of the char can be the main
electron transport medium only when the H/C and O/C ratios
are less than 0.35 and 0.09, respectively; otherwise the redox
functional groups on the surface of the char are the main
electron transport medium [33]. In this research the H/C and
O/C ratios are greater than 0.35 and 0.09, respectively, so
the main electron transport medium is the redox functional
groups (quinoid functional groups and phenolic moieties)
on the char’s surface.
Biomass Conversion and Biorefinery
Fig. 2  The Van Krevelen diagram for the hydro-chars
3.2.3 Porosity characteristics
Figure 3 shows the specific surface area and pore size dis-
tribution of the hydro-chars in nitrogen adsorption and des-
orption isotherms. According to the standard of the Interna-
tional Union of Pure and Applied Chemistry (IUPAC), the
absorption ranges of the SS, the H-SS-200, the H-SS-240,
and the H-SS-280 have obvious hysteresis loops in the range
of 0.5 < P/P0 < 1.0, where P is the pressure of ­N2 and P0 is
the saturated vapor pressure of ­N2. The absorption ranges
of the SS, the H-SS-200, the H-SS-240, and the H-SS-280
belong to the type IV isotherm and ­H1 hysteresis loops,
which indicate the existence of mesoporous. However, the
absorption ranges of the H-SW-200, the H-SW-240, and the
H-SW-280 belong to the V isotherm and H ­ 3 hysteresis loops,
implying the existence of micropore and the porous structure
is irregular.
Figure 3a, b depicts the micropore size distribution
and specific surface area of the hydro-char samples. It is
observed that all SS hydro-char samples, including the SS,
the H-SS-200, the H-SS-240, and the H-SS-280, possess
rich micropores. However, the H-SW-200, the H-SW-240,
and H-SW-280 have rich micropores and mesopores. Based
on the results presented in Table 1, it is inferred that blend-
ing WS has a positive effect on the specific surface area of
hydro-char. Under the HTC temperatures of 200 °C, 240 °C,
and 280 °C, the specific surface area increased from 14.70
to 28.00 ­m2 ­g−1, from 12.93 to 36.19 ­m2 ­g−1, and 10.03 to
24.31 ­m2 ­g−1, respectively when blending WS. However, an
increase in HTC temperature is detrimental to the specific
surface area of the SS hydro-char. It is reported that SS with
high moisture content and poor dewaterability has difficulty
in forming a large specific surface area during the Co-HTC
process [8, 34]. However, blending WS obstructs the for-
mation of large pore volumes and diameters, leading to the
presence of more micropores in SW hydro-char (Fig. 3).
This is also evident from the SEM images (Figure S1).
Furthermore, high temperature can result in the collapse
of some pores, leading to a reduction in pore volume. The
surface properties of hydro-char have a significant impact
on electron transfer between microorganisms because of
their large surface area and porous structure, which pro-
mote microorganism adhesion [35]. Studies have shown that
hydro-char with high specific surface area and increased
N content can be used as a promoter for electron transfer
between microorganisms during the anaerobic digestion
(AD) process [36]. Therefore, the results obtained here sug-
gest that hydro-chars prepared from SW are suitable to be
promoters in an AD process.
3.3 Surface functional groups and protein structure
Figure 4 shows the FTIR spectra of the hydro-chars produced
under different Co-HTC conditions. The peak at 3340 ­cm−1
corresponds to the alcoholic and phenolic hydroxyl groups
that are associated with intermolecular hydrogen bonding.
It can be observed from Fig. 4 that the amount of hydroxyl
groups in both SS and SW hydro-char decreases with
increasing HTC temperature, which can be attributed to the
loss of chemically bound water and hydroxyl groups from
SS and SW during the Co-HTC process [37, 38]. However,
the SW hydro-char exhibits a higher content of hydroxyl
groups than the SS hydro-char, suggesting that blending
WS is beneficial in retaining hydroxyl groups on the surface
of hydro-char. The peaks at 2981 ­cm−1 and 2850 ­cm−1 are
attributed to the vibration of aliphatic ­CHX groups. For both
SS and SW hydro-char, the C ­ HX content shows an increasing
trend compared to the raw SS. This indicates that the Co-
HTC process above 200 °C mainly causes the degradation of
most hemicellulose and amorphous cellulose, accompanied
by the leaching of aliphatic ­CHX [38]. The HTC temperature
(above 200 °C) and blending WS have a slight influence on
the ­CHX content. The peak at 1033 ­cm−1 indicates the vibra-
tion of aliphatic amine or the –C–O– of polyphenols, while
that at 600–800 ­cm−1 is due to the aromatic structure and
heterocyclic atom. As shown in Fig. 4, the HTC tempera-
ture has no influence on the aliphatic amine (the –C–O– of
polyphenols), aromatic structure, and heterocyclic atom.
However, blending WS causes a higher content of aliphatic
amine (the –C–O– of polyphenols), aromatic structure, and
heterocyclic atom.
In Fig. 4, it is evident that protein is the primary macro-
molecular organic compound in hydro-char. A higher pro-
tein content in hydro-char enables its use as promoter by
releasing essential nutrients (N) for microorganisms during
the anaerobic digestion process [39]. The increase in HTC

Fig. 3  Nitrogen adsorption and
desorption isotherms and pore
size distributions of hydro-chars
temperature for both SS and SW hydro-char, as depicted in
Fig. 4, leads to an increase in N–H, C–H, and C–N contents;
these are the products of protein hydrolysis. To investigate
the thermostability of the secondary structure of proteins in
hydro-char, Fig. 5 and Table S2 provide the necessary data.
The band at 1700–1600 ­cm−1 arises from the amide I band,
from which the secondary structural information of protein
is obtained by peak differentiation-imitating of the FTIR
spectra of hydro-chars. The primary secondary structures
of proteins are β-sheets and α-helix, which are located at
1640–1630 ­cm−1 and 1657–1648 ­cm−1, respectively [40].
Hydrothermal reaction and blending WS help maintain the
β-sheets and reduce the α-helix compared to the SS hydro-
char. However, the HTC temperature has no effect on either
the β-sheets or the α-helix. This suggests that the β-sheets
Biomass Conversion and Biorefinery
and α-helix exhibit good thermostability when the HTC
temperature exceeds 200 °C. The band at 1695–1680 ­cm−1
indicates the antiparallel β-sheets. Figure 5 shows that the
hydrothermal reaction contributes to the antiparallel β-sheets
content of hydro-char, while blending WS inhibits the reten-
tion of antiparallel β-sheets in the hydro-char. For the SW
hydro-char, 200 °C is the optimal temperature to produce
antiparallel β-sheets. However, for the SS hydro-char, the
content of antiparallel β-sheets decreases with an increase
in HTC temperature. In addition, the secondary structure
includes both β-turn (at 1666–1659 ­cm−1) and the aggre-
gated band (at 1625–1610 ­cm−1). Blending WS and increas-
ing the HTC temperature have a positive influence on the
aggregated band content. However, blending WS has a
negative effect on the β-turn content. The impact of HTC
Biomass Conversion and Biorefinery
Fig. 3  (continued)
temperature is material-dependent, as evident in the case
of SS hydro-char, where the β-turn content increases with
rising HTC temperatures, while the SW hydro-char exhibits
the opposite trend.
3.4 Mediated electrochemical analysis
of hydro‑char
The utilization of carbon material as an electron conduit
to facilitate direct interspecies electron transfer (DIET) has
been a conventional approach [41]. Previous studies have
demonstrated that carbon materials functionalized with spe-
cific groups (e.g., hydroquinone, quinone, and phenolic moi-
eties) could significantly accelerate the electron exchange
process between electron-donating bacteria and electron-
accepting methanogens [42]. Additionally, extracellular
electron transfer between carbon material and electroactive
microorganisms could be accelerated by biological elec-
tron components such as cytochromes and electron shut-
tles. Therefore, it is crucial to quantify the electron transfer
to evaluate the potential of carbon materials in enhancing
the efficiency of anaerobic digestion. In this study, electron
donating capacity (EDC) and electron accepting capacity
(EAC) of hydro-chars are described in Fig. 6.
As depicted in Fig. 6, the utilization of both SS and WS
blending (SW) strategies and elevated Co-HTC temperatures
led to an increase in both EAC and EDC levels. For the
SS hydro-char, the EAC content does not change too much
when HTC temperature increases, with values varying from
0.032 to 0.0305 ­mol−/ghydro-char; in contrast, the EDC content
increased from 0.2235 to 0.2846 ­mol−/ghydro-char with the rise
in Co-HTC temperature from 200 to 280 °C. Concerning
the SW hydro-char, the EAC/EDC contents exhibited more
pronounced increases, with EAC ranging from 0.2268 to

Fig. 4  FTIR spectra of hydro-char produced under different Co-HTC
conditions
0.4389 ­mol−/ghydro-char within 200–280 °C, and EDC increas-
ing from 0.6349 to 0.7358 ­mol−/ghydro-char as the Co-HTC
temperature increased from 200 to 240 °C, but dropping to
0.7058 ­mol−/ghydro-char at 280 °C. As a comprehensive result,
blending WS can significantly enhance the electron trans-
fer capacity, primarily due to the increase of aromatization
degree (as shown in Fig. 2).
Specifically, H-SW-280 displayed the highest EAC con-
tent (0.4389 ­mol−/ghydor-char). In line with earlier research,
the increase in EAC is linked to the augmentation of quinoid
functional group content. This observation strongly indicates
that H-SW-280 possesses a notable concentration of quinoid
functional groups, with high temperatures and elevated pres-
sure conditions promoting the formation of these valuable
functional groups. Simultaneously, H-SW-240 demonstrated
the most substantial EDC content (0.7358 ­mol−/ghydor-char),
corresponding to the lower H/C ratio. This result implies that
a higher degree of aromatization resulted in the higher EDC
levels in this study. Based on prior research, it is evident that
a higher concentration of phenolic moieties in the hydro-
char corresponds to an increased amount of EDC. This find-
ing underscores that both the incorporation of WS blending
and exposure to elevated temperatures can contribute to an
increase in the degree of aromatization and an augmentation
of phenolic moieties in hydro-chars. However, it is important
to note that an excessive increase in HTC temperature may
Biomass Conversion and Biorefinery
not be beneficial for EDC in this study. Based on the previ-
ous study, a positive linear correlation between the EDC
content of carbon material and ­CH4 yield has been estab-
lished in the carbon-amended anaerobic digestion system
[43]. Notably, In this study, the EDC content of SW hydro-
chars are much higher than SS hydro-char. These findings
suggest that SW hydro-chars show significant potential for
enhancing anaerobic digestion processes when compared to
SS hydro-chars.
3.5 Heavy metals evaluation
According to previous studies, a variety of heavy metals such
as Arsenic (As), Cadmium (Cd), Chromium (Cr), Nickel
(Ni), Lead (Pb), Zinc (Zn), Mercury (Mn), and Copper (Cu)
are commonly found in SS [44], which poses high biotoxic-
ity and cannot be removed or immobilized through anaero-
bic digestion [45]. Therefore, immobilizing heavy metals
in hydro-char during the Co-HTC process is necessary to
enhance the safety of anaerobic digestion when hydro-char
is used as an additive. In this study, the leaching concentra-
tion of heavy metals in both raw SS and prepared hydro-char
samples is presented in Table 3. Results show an obvious
decreasing trend in leaching heavy metal concentration in
hydro-char samples compared to raw SS, especially when
increasing the HTC temperature and blending WS. This
decrease can be attributed to several factors, such as higher
pressure in the hydrothermal reactor at higher temperatures
leading to the dissolution of heavy metals, low heavy metal
content in WS, and the dilution effect derived from mixing
WS and SS [18]. Additionally, poisonous elements such as
Cd, Cr, Mn, and Pb that are harmful to microorganisms can
be eliminated by the Co-HTC process, thus improving the
efficiency of anaerobic digestion. However, increasing the
HTC temperature and blending WS have no significant effect
on As content when the HTC temperature is below 280 °C,
and the hydro-char prepared from SW at 280 °C exhibits the
lowest As content. The Fe content shows a slight increasing
trend, while the Ni content declines with the increase in HTC
temperature and blending WS. Ca is an essential composi-
tion element for microorganisms. Results indicate that the Ca
content remains higher under lower HTC temperatures, while
blending WS significantly decreases the Ca content.
3.6 Secondary dissolution organic substance
Figure 7 delineates crucial information regarding the sec-
ondary dissolution of organic substances in hydro-chars
based on 3D-EEM results (Figure S2). Notably, it has been
reported that such organic substances are considered the
most readily assimilable carbon source for microorgan-
isms, as compared to particulate organic matter, during the
Biomass Conversion and Biorefinery

Fig. 5  The FTIR spectra of

protein secondary structures
in hydro-chars produced under
different HTC conditions

anaerobic digestion process [46]. With respect to the exci-
tation wavelength at 250 nm, the emission wavelengths at
325 nm indicate the presence of aromatic protein II. Previous
studies have established that aromatic protein II plays a piv-
otal role in dissolved organic nitrogen, which is responsible
for methane production during anaerobic digestion [47].
Additionally, for the excitation wavelength at 325 nm, the
emission wavelength at 400 nm is found to correspond to
humic acid. The influence of humic acid on promoting elec-
tron transport of direct interspecific and electron transfer
 Biomass Conversion and Biorefinery

535.15 ± 3.82
20.76 ± 0.26
3.02 ± 0.01
1.24 + 0.05
2.07 ± 0.14
1.58 ± 0.04
0.95 ± 0.09
Zn

129.95 ± 2.71
Pb

0
0
0
0
0
0
65.84 ± 1.25
2.08 ± 0.13
1.79 ± 0.02
1.32 ± 0.04

1.17 ± 0.01
1.07 ± 0.04
1.2 ± 0.02
Ni

275.27 ± 7.12

1.3 ± 0.17
Fig. 6  Electron donating capacity (EDC) and electron accepting
capacity (EAC) of hydro-chars

Mn

0
0
0
0
mechanism has been extensively reported in literatures [48,

20,386 ± 128.32
49].

21.49 ± 0.27
26.96 ± 1.22
9.17 ± 0.86
11.5 ± 1.07
5.44 ± 1.05
7.92 ± 0.96
As is evident from Fig. 7, both temperature and blend-
ing WS exert a notable influence on the content of organic

Fe
dissolution substances in hydro-chars. Among the various
hydro-chars, H-WS-200 and H-WS-280 display the highest
content of aromatic protein II. This can be attributed to the

148.98 ± 5.27
0.97 ± 0.025

0.44 ± 0.05
0.65 ± 0.01
hydrolysis of proteins in the SW into amino acids and their

2.6 ± 0.02
subsequent conversion into aromatic protein II [10], signify-
Table 3  The leaching heavy metals content of raw material (SS) and prepared hydro-chars

Cu

ing their potential to enhance large-scale methane produc-

0
0
tion. Moreover, it is worth noting that the hydro-char pre-
pared from the mixture of SS and WS demonstrates a higher
97.2 ± 2.51

content of aromatic protein II as compared to the hydro-char

prepared solely from SS. Regarding humic acid content, it
Cr

0
0
0
0
0
may be attributed to the hydrolysis of lipids [10]. In this 0
study, H-SS-240 attains the highest value, while H-WS-200
1.9 ± 0.05

ranks second in this regard. However, the content of humic

acid is found to be relatively similar across the other hydro-
Cd

0
0
0
0
0
0

chars, indicating that the influence of temperature and blend-

ing WS on humic acid content is marginal. It can be inferred
5277.00 ± 56.12
563.06 ± 19.95
622.44 ± 18.83
606.98 ± 15.75
472.15 ± 16.08

that blending WS can significantly enhance the content of

279.33 ± 6.07
143.57 ± 4.03

aromatic protein II, showing hydro-char’s property improve-

Concentration /(mg.kg-1)

ment when used as promoter for anaerobic digestion system.

Ca

4 Conclusions
31.39 ± 0.32

5.09 ± 0.06
5.2 ± 0.085
4.57 ± 0.21
4.97 ± 0.29
2.79 ± 0.30
5.8 ± 0.06

This research investigated SW hydro-char properties, includ-

As

ing elemental composition change, specific area, aromatiza-

tion degree, secondary dissolution organic substances, and
H-SW-200

H-SW-240

H-SW-280

the behavior of heavy metals etc. The following conclusions

H-SS-200

H-SS-240

H-SS-280
Sample

can be drawn:
SS
Biomass Conversion and Biorefinery
Fig. 7  The secondary organic
dissolution substance of hydro-
chars: a the spectra of 3D EEM
fluorescence at 250 nm of
excitation wavelength (aromatic
protein II); and b the spectra
of 3D EEM fluorescence at
325 nm of excitation wave-
length (humic acid)
1) Increasing the HTC temperature and incorporating
WS into the Co-HTC process positively influenced the
properties of the resultant hydro-chars. Notably, these
improvements encompassed an increase in specific sur-
face area, aromatization degree, secondary dissolution
of organic substances, and electron transfer capacity of
the hydro-chars.
2) The inclusion of WS in the Co-HTC process promoted
the formation of aromatic protein II in the hydro-chars
thus led to a higher content of aromatic protein II in the
hydro-char leaching solution.
3) The Co-HTC process effectively immobilized heavy
metals present in the SS and WS, mitigating the risk of
releasing these harmful elements into the environment.
Overall, this study demonstrates the Co-HTC of sewage
sludge and wheat straw at varying HTC temperatures can
produce hydro-chars with improved properties, especially
the hydro-chars can be used as promoters in an AD system.
The results paved the way for the development of carbon
products and their value-added applications.
The hydrothermal reaction involves many factors that
can significantly impact the products. In addition to factors
discussed in this study, other factors, such as reaction time,
precursor composition, and blending proportions, are the
potential directions in the future.
Supplementary information The online version contains supplemen-
tary material available at https://d​ oi.o​ rg/1​ 0.1​ 007/s​ 13399-0​ 24-0​ 5573-7.
Author contribution Qing An: conceptualization, methodology, inves-
tigation, data curation, writing—original draft. Dezhen Chen: concep-
tualization, writing—review and editing, resources, supervision, fund-
ing acquisition. Mengxi Han: investigation, resources. Yuheng Feng:
investigation, resources.
Funding This research work was financially supported by Shang-
hai Municipal Science and Technology Commission (Grant No.
20230712900) and Shanghai Science and Technology Innovation
Action Plan (Grant No.22dz1208200).
Data availability Not applicable.
Declarations
Ethical approval Not applicable.
Competing interests The authors declare no competing interests.
References
1. Yang G, Zhang GM, Wang HC (2015) Current state of sludge
production, management, treatment and disposal in China. Water
Res 78:60–73. https://​doi.​org/​10.​1016/j.​watres.​2015.​04.​002
2. Verlicchi P, Zambello E (2015) Pharmaceuticals and personal care
products in untreated and treated sewage sludge: Occurrence and
environmental risk in the case of application on soil—a critical
review. Sci Total Environ 538:750–767. https://​doi.​org/​10.​1016/j.​
scito​tenv.​2015.​08.​108
3. Fang W, Wei YH, Liu JG (2016) Comparative characterization of
sewage sludge compost and soil: heavy metal leaching character-
istics. J Hazard Mater 310:1–10. https://​doi.​org/​10.​1016/j.​jhazm​
at.​2016.​02.​025
4. Zabaniotou DF (2008) Utilization of sewage sludge in EU applica-
tion of old and new methods—a review. Renew Sust Energ Rev
12(1):116–140. https://​doi.​org/​10.​1016/j.​rser.​2006.​05.​014
5. Smith KM, Fowler GD, Pullket S, Graham N (2009) Sewage
sludge-based adsorbents: a review of their production, properties
and use in water treatment applications. Water Res 43(10):2569–
2594. https://​doi.​org/​10.​1016/j.​watres.​2009.​02.​038
6. Zhao PT, Shen YF, Ge SF, Yoshikawa K (2014) Energy recycling
from sewage sludge by producing solid biofuel with hydrothermal
carbonization. Energ Convers Manage 78:815–821. https://​doi.​
org/​10.​1016/j.​encon​man.​2013.​11.​026

7. Fonts I, Gea G, Azuara M, Abrego J, Arauzo J (2012) Sewage
sludge pyrolysis for liquid production: a review. Renew Sust Energ
Rev 16(5):2781–2805. https://​doi.​org/​10.​1016/j.​rser.​2012.​02.​070
8. Wzorek M (2012) Characterisation of the properties of alternative
fuels containing sewage sludge. Fuel Process Technol 104:80–89.
https://​doi.​org/​10.​1016/j.​fuproc.​2012.​04.​023
9. Chen C, Liu GM, An Q, Lin L, Shang Y, Wan CL (2020) From
wasted sludge to valuable biochar by low temperature hydrother-
mal carbonization treatment: insight into the surface characteris-
tics. J Clean Prod 263:121600. https://​doi.​org/​10.​1016/j.​jclep​ro.​
2020.​121600
10. Wang LP, Chang YZ, Li A (2019) Hydrothermal carbonization
for energy-efficient processing of sewage sludge: a review. Renew
Sust Energ Rev 108:423–440. https://​doi.o​ rg/​10.​1016/j.​rser.​2019.​
04.​011
11. Wang RK, Wang CB, Zhao ZH, Jia JD, Jin QZ (2019) Energy
recovery from high-ash municipal sewage sludge by hydrothermal
carbonization: fuel characteristics of biosolid products. Energy
186:115848. https://​doi.​org/​10.​1016/j.​energy.​2019.​07.​178
12. Ahmed M, Hameed B (2020) Insight into the co-pyrolysis of dif-
ferent blended feedstocks to biochar for the adsorption of organic
and inorganic pollutants: a review. J Clean Prod 265:121762.
https://​doi.​org/​10.​1016/j.​jclep​ro.​2020.​121762
13. Mahata S, Periyavaram SR, Akkupalli NK, Srivastava S, Matli C
(2023) A review on Co-Hydrothermal carbonization of sludge:
effect of process parameters, reaction path way, and pollutant
transport. J Energy Inst 110:101340. https://​doi.​org/​10.​1016/j.​
joei.​2023.​101340
14. Fan SS, Li H, Wang Y, Wang Z, Tang J, Tang J, Li XD (2018)
Cadmium removal from aqueous solution by biochar obtained by
co-pyrolysis of sewage sludge with tea waste. Res Chem Interme-
diat 44:135–154. https://​doi.​org/​10.​1007/​s11164-​017-​3094-1
15. Zhai YB, Peng C, Xu BB, Wang TF, Li CT, Zeng GM, Zhu Y
(2017) Hydrothermal carbonisation of sewage sludge for char
production with different waste biomass: effects of reaction tem-
perature and energy recycling. Energy 127:167–174. https://​doi.​
org/​10.​1016/j.​energy.​2017.​03.​116
16. He C, Zhang Z, Ge C, Liu W, Tang Y, Zhuang X, Qiu R (2019)
Synergistic effect of hydrothermal co-carbonization of sewage
sludge with fruit and agricultural wastes on hydrochar fuel qual-
ity and combustion behavior. Waste Manage 100:171–181. https://​
doi.​org/​10.​1016/j.​wasman.​2019.​09.​018
17. Lu XL, Ma XQ, Chen XF (2021) Co-hydrothermal carboniza-
tion of sewage sludge and lignocellulosic biomass: fuel properties
and heavy metal transformation behaviour of hydrochars. Energy
221:119896. https://​doi.​org/​10.​1016/j.​energy.​2021.​119896
18. Tong SR, Zhang SP, Yin HX, Wang JX, Chen MZ (2021) Study on
co-hydrothermal treatment combined with pyrolysis of rice straw/
sewage sludge: biochar properties and heavy metals behavior. J
Anal Appl Pyrol 155:105074. https://d​ oi.o​ rg/1​ 0.1​ 016/j.j​ aap.2​ 021.​
105074
19. Zhang XJ, Zhang L, Li AM (2017) Hydrothermal co-carboni-
zation of sewage sludge and pinewood sawdust for nutrient-rich
hydrochar production: synergistic effects and products characteri-
zation. J Environ Manage 201:52–62. https://​doi.​org/​10.​1016/j.​
jenvm​an.​2017.​06.​018
20. Reza MT, Rottler E, Herklotz L, Wirth B (2015) Hydrothermal
carbonization (HTC) of wheat straw: influence of feedwater pH
prepared by acetic acid and potassium hydroxide. Bioresource
Technol 182:336–344. https://d​ oi.o​ rg/1​ 0.1​ 016/j.b​ iorte​ ch.2​ 015.0​ 2.​
024
21. Cao JP, Li LY, Morishita K, Xiao XB, Zhao XY, Wei XY,
Takarada T (2013) Nitrogen transformations during fast pyroly-
sis of sewage sludge. Fuel 104:1–6. https://​doi.​org/​10.​1016/j.​fuel.​
2010.​08.​015
Biomass Conversion and Biorefinery
22. Kang SM, Li XL, Fan J, Chang J (2012) Characterization of
hydrochars produced by hydrothermal carbonization of lignin, cel-
lulose, d-xylose, and wood meal. I Ind Eng Chem Res 51:9023–
9031. https://​doi.​org/​10.​1021/​ie300​565d
23. Jamieson T, Sager E, Gueguen C (2014) Characterization of bio-
char-derived dissolved organic matter using UV-visible absorp-
tion and excitation-emission fluorescence spectroscopies. Che-
mosphere 103:197–204. https://​doi.​org/​10.​1016/j.​chemo​sphere.​
2013.​11.​066
24. MoEPo C (2007) HJ/T300-2007: Solid waste-extraction procedure
for leaching toxicity-acetic acid buffer solution method. China
Environmental Science Press, Beijing, China
25. Kluepfel L, Keiluweit M, Kleber M (2014) Sander, Redox proper-
ties of plant biomass-derived black carbon (biochar). Environ Sci
Technol 48:5601–5611. https://​doi.​org/​10.​1021/​es500​906d
26. Xu ZX, Song H, Zhang S, Tong SQ, Hu ZX, Wang Q, Li Q, Hu X
(2019) Co-hydrothermal carbonization of digested sewage sludge
and cow dung biogas residue: investigation of the reaction char-
acteristics. Energy 187:115972. https://​doi.​org/​10.​1016/j.​energy.​
2019.​115972
27. Safar M, Lin BJ, Chen WH, Langauer D, Chang JS, Raclavska
H, Pétrissans A, Rousset P, Pétrissans M (2019) Catalytic effects
of potassium on biomass pyrolysis, combustion and torrefaction.
Appl Energ 235:346–355. https://​doi.​org/​10.​1016/j.​apene​rgy.​
2018.​10.​065
28. Zhang SP, Chen T, Xiong YQ, Dong Q (2017) Effects of wet tor-
refaction on the physicochemical properties and pyrolysis prod-
uct properties of rice husk. Energ Convers Manage 141:403–409.
https://​doi.​org/​10.​1016/j.​encon​man.​2016.​10.​002
29. Lin YS, Ma XQ, Peng XW, Hu SC, Yu ZS, Fang SW (2015)
Effect of hydrothermal carbonization temperature on combustion
behavior of hydrochar fuel from paper sludge. Appl Therm Eng
91:574–582. https://​doi.​org/​10.​1016/j.​applt​herma​leng.​2015.​08.​
064
30. Liu TT, Guo YC, Peng NN, Lang QQ, Xia Y, Gai C, Liu ZG
(2017) Nitrogen transformation among char, tar and gas during
pyrolysis of sewage sludge and corresponding hydrochar. J Anal
Appl Pyrol 126:298–306. https://​doi.​org/​10.​1016/j.​jaap.​2017.​05.​
017
31. Wang LP, Li AM, Chang YZ (2016) Hydrothermal treatment
coupled with mechanical expression at increased temperature for
excess sludge dewatering: heavy metals, volatile organic com-
pounds and combustion characteristics of hydrochar. Chem Eng
J 297:1–10. https://​doi.​org/​10.​1016/j.​cej.​2016.​03.​131
32. Kim D, Lee K, Park KY (2014) Hydrothermal carbonization of
anaerobically digested sludge for solid fuel production and energy
recovery. Fuel 130:120–125. https://​doi.​org/​10.​1016/j.​fuel.​2014.​
04.​030
33. Sun TR, Levin BD, Guzman JJ, Enders A, Muller DA, Ange-
nent LT, Lehmann J (2017) Rapid electron transfer by the car-
bon matrix in natural pyrogenic carbon. Nat Commun 8(1):1–12.
https://​doi.​org/​10.​1038/​ncomm​s14873
34. Syed-Hassan SSA, Wang Y, Hu S, Su S, Xiang J (2017) Thermo-
chemical processing of sewage sludge to energy and fuel: funda-
mentals, challenges and considerations. Renew Sust Energ Rev
80:888–913. https://​doi.​org/​10.​1016/j.​rser.​2017.​05.​262
35. Jaafar NM, Clode PL, Abbott LK (2015) Soil microbial responses
to biochars varying in particle size, surface and pore properties.
Pedosphere 25(5):770–780. https:// ​ d oi. ​ o rg/ ​ 1 0. ​ 1 016/ ​ S 1002-​
0160(15)​30058-8
36. An Q, Chen DZ, Chen H, Yue X, Wang L (2022) Modification
of hydro-chars by non-thermal plasma to enhance co-anaerobic
digestion and degradation of sewage sludge pyrolysis oil. J Envi-
ron Manage 307:114531. https://d​ oi.o​ rg/1​ 0.1​ 016/j.j​ envma​ n.2​ 022.​
114531
Biomass Conversion and Biorefinery
37. Hossain MK, Strezov V, Chan KY, Ziolkowski A, Nelson PF
(2011) Influence of pyrolysis temperature on production and nutri-
ent properties of wastewater sludge biochar. J Environ Manage
92(1):223–228. https://​doi.​org/​10.​1016/j.​jenvm​an.​2010.​09.​008
38. Lu HL, Zhang WH, Wang SZ, Zhuang LW, Yang YX, Qiu RL
(2013) Characterization of sewage sludge-derived biochars from
different feedstocks and pyrolysis temperatures. J Anal Appl Pyrol
102:137–143. https://​doi.​org/​10.​1016/j.​jaap.​2013.​03.​004
39. Uchimiya M, Lima IM, Klasson KT, Wartelle LH (2010) Contam-
inant immobilization and nutrient release by biochar soil amend-
ment: roles of natural organic matter. Chemosphere 80(8):935–
940. https://​doi.​org/​10.​1016/j.​chemo​sphere.​2010.​05.​020
40. Yin CQ, Meng FG, Chen GH (2015) Spectroscopic characteriza-
tion of extracellular polymeric substances from a mixed culture
dominated by ammonia-oxidizing bacteria. Water Res 68:740–
749. https://​doi.​org/​10.​1016/j.​watres.​2014.​10.​046
41. Chen SS, Rotaru A-E, Shrestha PM, Malvankar NS, Liu FH, Fan
W, Nevin KP, Lovley DR (2014) Promoting interspecies electron
transfer with biochar. Sci Rep 4:5019. https://​doi.​org/​10.​1038/​
srep0​5019
42. Wang JF, Zhao ZQ, Zhang YB (2021) Enhancing anaerobic diges-
tion of kitchen wastes with biochar: link between different proper-
ties and critical mechanisms of promoting interspecies electron
transfer. Renew Energ 167:791–799. https://​doi.​org/​10.​1016/j.​
renene.​2020.​11.​153
43. Shen YW, Yu YM, Zhang Y, Urgun-Demirtas M, Yuan HP, Zhu
NW, Dai XH (2021) Role of redox-active biochar with distinc-
tive electrochemical properties to promote methane production
in anaerobic digestion of waste activated sludge. J Clean Prod
278:123212. https://​doi.​org/​10.​1016/j.​jclep​ro.​2020.​123212
44. Lang QQ, Guo YC, Zheng QF, Liu ZG, Gai C (2018) Co-hydro-
thermal carbonization of lignocellulosic biomass and swine
manure: hydrochar properties and heavy metal transformation
behavior. Bioresource Technol 266:242–248. https://​doi.​org/​10.​
1016/j.​biort​ech.​2018.​06.​084
45. Anjum R, Grohmann E, Krakat N (2017) Anaerobic digestion
of nitrogen rich poultry manure: impact of thermophilic biogas
process on metal release and microbial resistances. Chemoshere
168:1637–1647. https://​doi.​org/​10.​1016/j.​chemo​sphere.​2016.​11.​
132
46. Lu D, Xiao KK, Chen Y, Soh YNA, Zhou Y (2018) Transforma-
tion of dissolved organic matters produced from alkaline-ultra-
sonic sludge pretreatment in anaerobic digestion: From macro to
micro. Water Res 142:138–146. https://​doi.​org/​10.​1016/j.​watres.​
2018.​05.​044
47. Xiao KK, Yu ZC, Pei KY, Sun M, Zhu YW, Liang S, Hou HJ,
Liu BC, Hu JP, Yang JP (2022) Anaerobic digestion of sludge
by different pretreatments: changes of amino acids and microbial
community. Front Env Sci Eng 16:1–15. https://​doi.​org/​10.​1007/​
s11783-​021-​1458-7
48. Zhao C, Liu HB, Liu H, Zhan XM, Zhang XD, Fu B, Shi MZ, Lu
SL, Huang F, Cui MH, Li YJ (2023) Influence of thermal hydrol-
ysis on sludge anaerobic digestion: release of humic acid pro-
motes electron transport of methanogen. Chem Eng J 451:138455.
https://​doi.​org/​10.​1016/j.​cej.​2022.​138455
49. Wang XQ, Lyu T, Dong RJ (2022) Wu SB (2022) Revealing the
link between evolution of electron transfer capacity of humic acid
and key enzyme activities during anaerobic digestion. J Environ
Manage 301:113914. https://​doi.​org/​10.​1016/j.​jenvm​an.​2021.​
113914
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