Environmental Research 188 (2020) 109828

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Environmental Research
journal homepage: www.elsevier.com/locate/envres

Hydrochar production from high-ash low-lipid microalgal biomass via T

hydrothermal carbonization: Effects of operational parameters and products
characterization
Choon Gek Khooa, Man Kee Lamb,c, Abdul Rahman Mohameda, Keat Teong Leea,∗
a
School of Chemical Engineering, Universiti Sains Malaysia, Engineering Campus, 14300, Nibong Tebal, Pulau Pinang, Malaysia
b
Chemical Engineering Department, Universiti Teknologi PETRONAS, 32610, Seri Iskandar, Perak, Malaysia
c
HICoE-Centre for Biofuel and Biochemical Research, Institute of Self-Sustainable Building, Universiti Teknologi PETRONAS, 32610, Seri Iskandar, Perak, Malaysia

ARTICLE INFO ABSTRACT

Keywords: This study aims to produce hydrochar from high-ash low-lipid Chlorella vulgaris biomass via hydrothermal
Microalgal biomass carbonization (HTC) process. The effects of hydrothermal temperature and retention time with respect to the
Hydrothermal carbonization physicochemical properties of hydrochar were studied in the range of 180–250 °C and 0.5–4 h, respectively. It
Energy densification was found that the hydrothermal temperature had resulted in a significant reduction of hydrochar yield as
Hydrochar
compared to the retention time. The raw microalgal biomass was successfully converted into an energy densified
Nutrient recycling
hydrochar via an optimized HTC reaction, with higher heating value (HHV) of 24.51 kJ/g, which was ap-
proximately two-times higher than that of raw biomass. In addition, the overall carbon recovery rate and energy
yield were in the range of 53.2–86.4% and 46.9–76.6%, respectively. The high quality of the produced hy-
drochar was further supported by the plot of van Krevelen diagram and combustion behaviour analysis. Besides,
the aqueous phase collected from HTC process could be further used as nutrients source to cultivate C. vulgaris, in
which up to 70% of the biomass yield could be attained as compared to the control cultivation condition. The
reusability of the aqueous phase collected from HTC process as an alternative nutrients source to cultivate
microalgal indicated the feasibility and positive integration of HTC process in microalgal biofuel processing
chain.

1. Introduction power generation (Brennan and Owende, 2010). However, extraction of

In the past few years, microalgal biomass has attracted huge at-
tention from researchers as the third-generation feedstock for biofuels
production. This is because certain microalgal species (e.g. Chlorella sp.
and Scenedesmus sp.) contain high quantity of lipid and carbohydrate,
which can be converted to biodiesel and bioethanol, respectively
(Sugunya et al., 2016). Besides, microalgal can be cultivated on non-
arable area that helps to minimize the competition with agricultural
land; at the same time being able to bio-fix CO2 from flue gases and
bioremediate wastewater that is rich in nitrogen and phosphorus con-
tents (Pires et al., 2012).
In an effort to drive the commercialization of microalgal biofuels
production, intensive researches have been conducted especially on the
photobioreactor designs, optimization of cultivation conditions, cells
harvesting and biofuels conversion. Among all the biofuels that can be
derived from microalgal biomass, biodiesel have received the highest
attention as it can be directly used in the existing engine and boiler for
the lipid using chemical solvents from the microalgal biomass requires
extensive pre-treatment steps to disrupt the microalgal cell wall to
enhance the lipid extraction efficiency (Arenas et al., 2017). In addi-
tion, drying process is needed to dry the wet-paste microalgal before
lipid extraction to avoid complication in the subsequent solvent re-
covery process. Besides, some microalgal species that are able to grow
fast but with low lipid content are not feasible to be cultivated for
biodiesel production.
The introduction of hydrothermal technology, which is a thermo-
chemical technique that converts wet biomass feedstocks into hydro-
char via thermal disintegration in hot compressed aqueous medium; is
an alternative way to produce microalgal biofuels (Kumar, 2014). The
advantages of hydrothermal process included: (i) rapid reaction rate
(Garcia Alba et al., 2012), (ii) replacement of acid/bases with green
solvent as reaction medium (Peterson et al., 2008), (iii) do not require
the addition and recovery of chemicals from water (Rizzo et al., 2013),
(iv) minimized corrosion issue in hydrothermal reactor (Elliott et al.,

∗
Corresponding author.
E-mail address: ktlee@usm.my (K.T. Lee).

https://doi.org/10.1016/j.envres.2020.109828
Received 31 March 2020; Received in revised form 8 June 2020; Accepted 10 June 2020
Available online 21 June 2020
0013-9351/ © 2020 Elsevier Inc. All rights reserved.
C.G. Khoo, et al.
2013), and (v) simple and economical operation (Elliott et al., 2014).
The microalgal biomass which contains lower sulphur content than
lignocellulosic biomass minimize corrosion problem in HTC reactor and
thus making it a better feedstock (Thakkar et al., 2019). Through hy-
drothermal process, the entire microalgal cells (e.g. lipid, protein and
carbohydrate) are converted into hydrochar with high energy densifi-
cation value. In addition, intensive drying process of microalgal bio-
mass can be avoided as the reaction medium in the hydrothermal
process is water-based. However, study on the hydrochar production
from microalgal biomass is still lacking in the literature and more un-
derstanding are required to enhance the commercialization potential of
this solid fuel.
Several challenges are associated with hydrothermal processing of
biomass including the discharging of aqueous phase by-products. The
microalgal slurry that was being investigated was ranged from 5 to
50 wt%, where higher solid content can be achieved by removing ex-
cessive water through energy-intensive drying process (Jazrawi et al.,
2013). To address this concern, the approach of using dilute microalgal
slurry (i.e. with concentration obtained from harvesting process – bio-
mass-to-water ratio at 1-to-100 or 1 wt% of biomass content) for hy-
drothermal treatment was investigated in the present study. Besides,
the aqueous phase collected after hydrothermal processing could be
further used as nutrients source for microalgal cultivation to improve
the water footprint of hydrothermal process (Biller et al., 2012; López
Barreiro et al., 2015; Leng et al., 2018). This would make the hydro-
thermal process more viable with lower environmental impact.
Thus, the aim of the present work is to investigate the effects of
hydrothermal carbonization (HTC) operating conditions towards the
quality of hydrochar produced from microalgal biomass. The HTC
process was reported to only require mild reaction conditions that
normally range between 180 and 250 °C and 10–40 bar, with retention
time ranging from 0 to 5 h (Peterson et al., 2008; Heilmann et al.,
2010). Hence, the hydrothermal temperature, T and retention time, t,
were studied in the range of 180–250 °C and 0.5–4 h, respectively. The
quality of the hydrochars was characterized by using proximate ana-
lysis, elemental analysis, energetic assessment, functional groups
identification, and combustion behaviour. In addition, the aqueous
phase produced from HTC process was further analysed for the possi-
bility to be used as an alternative nutrient source for microalgal culti-
vation.
2. Materials and methods
2.1. Biomass preparation
This existing study is a continuation work from our previous study
that had been published elsewhere (Khoo et al., 2016). In the previous
publication, the influence of CO2 mass transfer and hydrodynamics
effect exerted on microalgal in a bubble column photobioreactor cul-
tivation system were investigated (Khoo et al., 2016). In that study, a
robust microalga strain was required and hence, the classical species –
Chlorella vulgaris was selected, in which this microalgal species has a
wide range of temperature tolerance and able to grow under harsh
environment (Safi et al., 2014). Subsequently, this study aims to study
the downstream application of the selected microalgal strain. C. vulgaris
that was cultivated in a pilot-scale bubble column photobioreactor (BC-
PBR) was used as the feedstock for hydrothermal carbonization (HTC)
process. The optimum cultivation conditions to grow the microalgae are
reported in our previous publication (Khoo et al., 2016). After growing
the microalgae to matured phase, the microalgae were harvested from
the BC-PBR through flocculation method using Alum (QRëC™, AR
grade, Malaysia) as flocculant. The highest flocculation efficiency
(> 99%) was attained with 500 mg/L dosage and 20 min of floccula-
tion time. The wet paste microalgae were further dried in oven
(memmert, UFP600TS, German) at 70 °C overnight for the ease of
storage.
2
Environmental Research 188 (2020) 109828
2.2. Characterization of biomass
2.2.1. Biochemical analysis
The biochemical analysis conducted on the microalgal biomass (in
dry ash free weight basis, d. a.f. wt%) included total lipid, total car-
bohydrate, and crude protein. The total lipid extraction was performed
by modified Bligh-Dyer method (Bligh and Dyer, 1959; Lam et al.,
2014). Initially, 5 g of dried microalgal biomass was weighed, followed
by adding 150 mL of chloroform-methanol mixture, CHCl3/MeOH (1:2
v/v) into a 500 mL conical flask. The mixture was stirred at 1000 rpm
for 4 h under ambient conditions. Then, the mixture was filtered using
Whatman Grade 1 filter paper. The conical flask was further rinsed with
50 mL of CHCl3 to ensure all the solid residue were transferred to the
filter paper. Thereafter, the filtrate was evaporated via a rotary eva-
porator at 55 °C under vacuum condition. The remaining microalgal
lipid was collected and weighted. The total lipid content in the mi-
croalgal biomass was calculated by Eq. (1):
m2
Total lipid(d.a. f.wt%) = × 100%
m1 (1)
where m1 is the weight of microalgal biomass sample (g) and m2 is the
weight of dried lipid collected (g).
The total carbohydrate content was determined via two-step sul-
phuric acid (H2SO4) hydrolysis method (Wychen and Laurens, 2015).
Initially, 25 mg of dried microalgal biomass was added into a 10 mL test
tube. Then, 250 μL of 72 wt% H2SO4 was added into the glass tube with
biomass sample and vortexed thoroughly. The vortexed sample was
then incubated at 30 °C in water bath for 1 h. The test tube was then
removed from water bath and 7 mL of 18.2 MΩ deionized water was
added into the test tube to neutralize the H2SO4 concentration to 4 wt
%. A rubber stopper was used to enclose the top of the test tube. The
tube was then inverted for complete mixing and autoclaved at 121 °C
for 15 min. Then, the hydrolysed sample was vortexed and the hydro-
lysate (i.e. separated from solids) was removed for neutralization. The
neutralization process was conducted by adding calcium carbonate,
CaCO3 into the hydrolysate. The neutralization pH was ranged between
6 and 8. Thereafter, the neutralized sample was filtered through a
0.2 μm nylon filter for subsequent high-performance liquid chromato-
graphy (HPLC) analysis. The total carbohydrate content was re-
presented by the summation of the concentration of monomers sugar
(mg/mL) calculated from the HPLC calibration curves. The total car-
bohydrate content in the microalgal biomass was calculated through
Eq. (2):
M1 V1
Total carbohydrate(d.a. f.wt%) = × 100%
m (2)
where M1 is the total carbohydrate (i.e. sugar monomers) concentration
(mg/mL), V1 is the injection sample volume (mL), and m is the weight
of microalgal biomass sample (mg).
According to Andersen (Andersen et al., 2013), the biochemical
composition of ash-free microalgal biomass consists of carbohydrate,
lipid, and protein. Hence, the crude protein was represented by the
differences of total carbohydrate and total lipid contents from the raw
biomass, which was represented by Eq. (3).
Crude protein (d.a.f. wt%) = 100 – Total lipid – Total carbohydrate
(3)
2.3. Proximate analysis
The proximate analysis composition of the raw microalgal biomass
was examined by using thermogravimetric analysis (TGA), in which the
sample was heated to constant weight under ASTM specified conditions
(Donahue and Rais, 2009). Proximate analysis includes the determi-
nation of moisture, volatile matter, fixed carbon, and ash content of the
sample, in term of dry weight basis (wt%). The proximate analysis
C.G. Khoo, et al.
composition was determined by TGA, which was conducted by ther-
mogravimetric analyzer (SDTQ600 TGA, TA Instruments, U.S.), which
measured the weight loss of sample that undergoes heating up to 900 °C
under purified nitrogen, N2 (inert) environment, and then held at
900 °C by switching to purified air environment. The moisture content
was determined by the weight loss of sample when heated to 110 °C due
to evaporation of water content. Meanwhile, the volatile matter content
was corresponded to the volatile products that evolved (weight loss)
between 110 and 900 °C. Then, the fixed carbon content was re-
presented by the combustion of solid material when the sample was
held at 900 °C by switching to purified air environment. Lastly, the
residue after combustion process was the ash content of biomass.
2.3.1. Elemental analysis
The elemental analysis of raw microalgal biomass in terms of carbon
(C), hydrogen (H), nitrogen (N) and sulphur (S) was conducted via an
Elemental Analyzer (PerkinElmer, 2400 Series II CHNS/O, U.S.). The
result for CHNS was obtained in mass percentage (wt%). The remaining
on mass balance was referred to mass percentage of oxygen (O) atom,
which was calculated via Eq. (4).
O(wt %) = 100– C H N S (4)
2.3.2. Functional group analysis
Fourier transform infrared (FT-IR) spectrometer (Shimadzu,
IRPrestige-21, Japan) was used to determine the functional groups of
the raw microalgal biomass. FT-IR spectrum was scanned between 4000
and 600 cm−1 at ambient temperature. A total of 20 scans were taken
for each interferogram at 2 cm−1 resolution.
2.3.3. Combustion behaviour
The combustion behaviour of the raw microalgal biomass was de-
termined by using thermogravimetric analysis (TGA). The TGA was
conducted by using a thermogravimetric analyzer (SDTQ600 TGA, TA
Instruments, U.S.) from 30 °C to 900 °C with a ramping rate of 10 °C/
min under purified air atmosphere. Non-isothermal combustion was
conducted in the furnace of TGA system at atmospheric pressure. The
weight loss and its respective derivative weight loss rate of the raw
microalgal biomass were continuously recorded.
2.4. HTC reaction
The schematic diagram of the hydrothermal carbonization reaction
rig is shown in Fig. 1. The hydrothermal carbonization (HTC) was
conducted in a batch autoclave reactor system with a volume capacity
of 200 mL. For HTC process, initially the microalgal feed slurry was
prepared by rehydrating the raw microalgal biomass with deionized
water in the HTC reactor vessel, to a biomass to water ratio of 1:100.
The microalgae slurry was kept in the HTC reactor vessel and mixed
with a stirring rod to obtain uniform biomass-to-water ratio throughout
the experiments. After the HTC reactor was sealed tightly, purified N2
gas was flowed into the reactor to purge out the residual air through the
gas outlet. The reactor was then heated from ambient temperature at
heating rate of 10 °C min−1 to the designated reaction temperatures
(180–250 °C) and held at the pre-determined residence times (0.5–4 h).
Upon completion of HTC reaction, the reactor was cooled down to
ambient temperature. Then, the solid and aqueous phase products were
separated through filtration, and the hydrochar was rinsed with deio-
nized water thoroughly. The hydrochar was weighted separately and
stored for further analysis.
2.5. HTC products analysis
2.5.1. Hydrochar
The hydrochar was characterized according to the properties of
solid fuel, which included proximate and elemental analyses, energetic
3
Environmental Research 188 (2020) 109828
assessment, functional groups, and combustion behaviour. The prox-
imate and elemental analyses were conducted according to the standard
methods as mentioned in previous sections for the raw microalgal
biomass. More details on hydrochars characterization results are given
in Supplementary data.
The calorific energy content of hydrochar samples was determined
by a bomb calorimeter (IKA, C200 auto, China). Hydrochar samples
were combusted in an oxygen-rich environment inside a bomb that was
contained within a water bath. The increment of temperature in the
water bath was used to determine the gross higher heating value (HHV)
through a comparison to a calorific-grade benzoic acid standard. The
solid yield (Yhydrochar), energy densification (DE), and energy yield (YE)
were calculated based on the following equations, respectively.
mhydrochar
Yhydrochar = × 100%
mbiomass (5)
HHVhydrochar
DE =
HHVbiomass (6)
YE = Yhydrochar × DE (7)
where mhydrochar is the mass of hydrochar, mbiomass is the mass of raw
microalgal biomass, HHVhydrochar is the HHV of hydrochar, and
HHVbiomass is the HHV of raw microalgal biomass.
The functional groups analysis of hydrochar samples was de-
termined by FT-IR spectrum, meanwhile the combustion behaviour of
hydrochar samples was determined by using a thermogravimetric
analysis (TGA). The details protocols for both FT-IR and TGA analysis
were same as per reported for microalgal biomass in previous sections.
2.5.2. Aqueous phase
After the separation of aqueous phase from hydrochar, the pH value
of the aqueous phase was measured immediately using a pH meter
(Fisher Scientific, AB15, U.S.). Meanwhile, further analyses such as
total organic carbon (TOC), total phosphorus (TP), total nitrogen (TN)
and chemical oxygen demand (COD) were conducted in batches after
several days of refrigerated storage. The analyses for TN, TP and COD
were performed spectrophotometrically through test kits supplied by
HACH. It was noted that through screening analysis, there was no major
degradation on the aqueous phase for the products of interest occurring
during refrigerated storage.
The TOC content was determined by using a PC-controlled Total
Organic Carbon Analyzer (Shimadzu, TOC-VWP, Japan). A 50 μL
sample was treated with 2 M HCl and sparged with air for 1.5 min
before introduction into the catalytic combustion chamber at 680 °C.
The oxidation products were passed through the non-dispersive in-
frared (NDIR) detector, where CO2 was quantified by comparing the
peak areas of samples to the three-point calibration curve generated
from analysis of potassium hydrogen phthalate (KHP) at the beginning
of each run. Replication runs were required if standard error of peak
area > 0.2%. TOC was calculated by the difference between the mea-
sured total carbon (TC) and inorganic carbon (IC). Details aqueous
phase characterization results are available in Supplementary Data.
The nutrient concentration analysis for aqueous phase products
included TP and TN. The concentrations of TP and TN were measured
through HACH method 8190 (AHPA 4500-P, Persulfate Digestion
Method (America Public Health Ass, 1999a)) and HACH method 10,071
(AHPA 4500-N, Persulfate Digestion Method (America Public Health
Ass, 1999b)); in both cases using a digester unit (HACH, DRB200, U.S.)
and a spectrophotometer (HACH, DR2800, U.S.) were required. The
HACH reagent vials with pre-mixed chemicals was digested at various
temperature and time, which are: (1) TP: 150 °C for 30 min, and (2) TN:
105 °C for 30 min. The digested reagent vials were then analysed with
the pre-set programs in spectrophotometer accordingly. All TP and TN
concentrations were expressed as mg/L. Details aqueous phase char-
acterization results are available in Supplementary data.
C.G. Khoo, et al.
Fig. 1. Schematic diagram of hydrothermal carbonization reactor.
The COD in aqueous phase product was determined by HACH test
kit through HACH method 8000 (AHPA 5220, Dichromate oxygen de-
mand method (America Public Health Ass, 1999c)). A digester unit
(HACH, DRB200, U.S.) and a spectrophotometer (HACH, DR 2800,
U.S.) were required for the COD analysis. The HACH reagent vials with
pre-mixed chemicals was digested at 150 °C for 120 min. The digested
reagent vials were then analysed with the pre-set programs in the
spectrophotometer. The concentration of COD was expressed as mg/L.
Details aqueous phase characterization results are available in Supple-
mentary data.
2.6. Cultivation of microalgae
The C. vulgaris used in this study was obtained from School of
Biological Science, Universiti Sains Malaysia. The C. vulgaris was in-
oculated into 5 L photobioreactor (PBR) with tap water containing
100 mL of organic fertilizer medium (Serbajadi, Japanese Humus 27,
Malaysia), which contains the following components: 1323.2 mg/L TN,
634.4 mg/L TP, 634.4 mg/L potassium (K), 269.9 mg/L calcium (Ca),
54.5 mg/L magnesium (Mg), 1.0 mg/L manganese (Mn), 4.1 mg/L
boron (B), 1.3 mg/L iron (Fe), 576.0 mg/L BOD5, and 1729.9 mg/L
COD. When reaching nearly maximum density, the microalgae seed
were transfer to the 56 L pilot-scale bubble column photobioreactor
(BC-PBR) containing 1 L of organic fertilizer medium as control sample.
Meanwhile, the aqueous phase produced from HTC reactions were
pooled together and used as nutrient medium for microalgae cultiva-
tion. The microalgae growth performances by using these two types of
nutrient sources were compared under optimum growth conditions as
per reported in our previous publication (control sample): ~0.3 g L−1
inoculum concentration, 0.16 vvm compressed air aeration rate, pH 3,
and 24 h illumination with an fluorescent light intensity of
60–70 μmol m−2 s−1 (Khoo et al., 2016).
The microalgal biomass concentration, N (in dry weight basis) of
cultures was measured according to the method reported previously
(Khoo et al., 2016). The regression equation of the relationship between
optical density and biomass concentration was established and shown
4
Environmental Research 188 (2020) 109828
in Eq. (8):
N (g L 1) = 0.5419 OD540 , R2 = 0.9924 (8)
−1
where N is biomass concentration (g L ) and OD540 is optical density
measured at a wavelength of 540 nm.
The optical density of microalgae culture was measured by the
UV–vis spectrophotometer (Shimadzu, UV mini-1240, Japan).
Meanwhile, the relative microalgal biomass concentration was mea-
sured gravimetrically. A 10 mL of microalgae culture sample was cen-
trifuged at 4400 rpm for 10 min to decant the supernatant. The mi-
croalgae culture was then dried in oven at 105 °C until the weight was
invariant.
2.7. Reproducibility of results
At least duplicate independent runs were conducted under identical
conditions to ensure the reproducibility of the experimental results.
Results reported herein represent the mean values from each in-
dependent trial.
3. Results and discussion
3.1. Characterization of microalgal biomass
Table 1 shows the biochemical analysis of microalgal biomass (on
dry ash free basis, d. a.f.), which consists of 57.0 d. a.f. wt% of crude
protein, 19.4 d. a.f. wt% of total lipid, and 23.6 d. a.f. wt% of carbo-
hydrate, respectively. The distribution of biochemical compositions is
in good agreement as per reported by Safi et al. 24, in which the con-
tents of protein, lipid and carbohydrate of C. vulgaris biomass are within
the range of 42–58 wt%, 5 to 40 wt% and 12 to 55 wt%, respectively.
Generally, the moisture content of dry weight basis biomass is
highly dependent on the types of biomass and their structural storage.
From Table 1, the raw microalgal biomass consists of 2.5 wt% moisture
content, 29.0 wt% volatile matter, 43.8 wt% fixed carbon, and 24.7 wt
% ash content. The low moisture content in biomass was contributed by
C.G. Khoo, et al. Environmental Research 188 (2020) 109828

Table 1 Table 2
Characterization of Chlorella vulgaris biomass. Biomass band regions for biochemical compositions of Chlorella vulgaris biomass
in FTIR spectrum. Adapted from Driver (Driver et al., 2015).
Properties Value
Composition Wavelength (cm−1) Functional group(s)
a
Biochemical analysis (d.a.f. wt%) Crude protein 57.0
Total lipid 19.4 Lipid 3566–3000 –OH of carboxylic acid
Total carbohydrate 23.6 3000–2877 C–H & C]CH– chains of lipid
Proximate analysis (wt%) Moisture 2.5 –CH2 & –CH3 from fatty acids
Volatile matter 29.0 Protein 1640–1412 C=O & N–H of amide
Fixed carbona 43.8 Carbohydrate 1113–1007 –C–O–R in aliphatic ethers
Ash 24.7 –C–O in alcohol
Elemental analysis (wt%) Carbon 27.4
Hydrogen 7.0
Nitrogen 5.5
calorific value of microalgal biomass (Demirbas, 2004).
Sulphur 0.4
Oxygena 59.7 The FT-IR spectrum in Fig. 2 illustrates the functional groups of raw
Calorific value (kJ/g) 12.58 microalgal biomass. According to Driver (Driver et al., 2015), the bio-
Empirical formula (on ash free dry basis) C183H560O299N32S chemical compositions such as protein, carbohydrate and lipid can be
H/C molar ratio (on ash free dry basis) 3.07
represented by various functional groups as listed in Table 2. The lipid
O/C molar ratio (on ash free dry basis) 1.63
content of raw microalgal biomass were represented by the band re-
a
Value calculated by difference. gions ranging from 3000 to 2877 cm−1 and 3566–3000 cm−1 (i.e., first
and second segregated regions), which indicated the presence of C–H &
the moisture content from humid environment, especially in Malaysia. C]CH– and –CH2, & –CH3 chains from lipid and fatty acids, in asso-
The ash content of raw microalgal biomass in this study was higher than ciated with the carboxylic acid –OH group. Generally, microalgal lipids
other types of biomass (i.e. basically 2–7 wt%). This high ash content can be categorized into nonpolar lipids (e.g. acyglycerols, free fatty
could give a negative effect on the biomass combustible behaviour due acids (FFA), sterols and steryl esters) and polar lipids (phosphoglycer-
to fouling problems, whereas the amount of fixed carbon and volatile ides, glycosylglycerides and sphingolipids) (Halim et al., 2012). The
matter in biomass could strongly affect its thermal decomposition detection of carboxylic acid –OH group indicated clearly the presence of
pathways (Biller and Ross, 2011). Hence, the raw microalgal biomass free fatty acid. This finding indicated that the raw microalgal biomass
should be further upgraded to meet the standard of biofuel combustion. consist of high FFA content and remain unreacted during the lipid ex-
The elemental contents of carbon, hydrogen, oxygen, nitrogen and traction process. A broader protein region (i.e., third region ranges from
sulphur in the raw microalgal biomass was found as 27.4, 7.0, 59.7, 5.5 1600 to 1400 cm−1) indicates the stretching vibration of C]O and N–H
and 0.4 wt%, respectively (Table 1). Hence, the empirical formula of functional groups of amides. The high intensity peaks at the fourth
the raw microalgal biomass was calculated as C183H560O299N32S. Be- regions (i.e. 1113 and 1007 cm−1) indicate the stretching of –C–O–R
sides, the H/C and O/C molar ratios for raw microalgal biomass were and –C–O in aliphatic ethers and alcohol, respectively from carbohy-
calculated from elemental composition as 3.07 and 1.63, respectively drates in biomass.
(on dry basis). The calorific value for C. vulgaris biomass was found to The thermogravimetry-derivative thermogravimetry (TG-DTG)
be 12.58 kJ/g. This calorific value is lower than the reported calorific curves for C. vulgaris biomass in purified air environment is shown in
values that is between 18 and 21 kJ/g (Scragg et al., 2002). This might Fig. 3. There are four stages of weight loss during the combustion of raw
be due to the low lipid content in the raw microalgal biomass, in which microalgal biomass. Stage A corresponded to the initial 7.75 wt%
the heating value is proportionally correlated to the content of lipid weight loss from the evaporation of physically absorbed water in the
(Illman et al., 2000). In addition, raw microalgal biomass with higher biomass at temperatures ranging from 30 to 140 °C (Rizzo et al., 2013).
ash content and lower carbon content could also contributed to the low This was followed by continuous decrease in raw biomass weight in

Fig. 2. FT-IR spectrum of Chlorella vulgaris biomass.

5
C.G. Khoo, et al.
Fig. 3. The TG-DTG profile of Chlorella vulgaris biomass in an air atmosphere. Note: Stage A represented weight loss from evaporation of water, Stage B represented
the release of volatiles and combustion, Stage C represented the combustion of volatiles, and Stage D represented the passive combustion reaction zone.
Stage B and Stage C at temperature ranging from 190 to 375 °C and
375–635 °C, respectively. Stage B represented the release of volatiles,
while Stage C represented the combustion of volatiles. Stage B and C
were associated to the main biomass decomposition regions, that con-
tributed to 38.5 and 32.5 wt% of biomass weight loss, respectively.
Hence, Stage B and C were denoted as the active combustion reactions
zone, where the combustion of primary biochemical compositions in
the raw microalgal biomass (i.e. lipid, carbohydrate, and protein) took
place (Gai et al., 2015a). Besides, the weight loss at Stage B of the DTG
thermogram indicated that devolatilization of primary components
during the active combustion reactions region. This observation re-
flected the devolatilization of volatile components from primary com-
ponents of raw microalgal biomass, which released a large amount of
heat at Stage B.
The peak of DTG thermogram at Stage C represented the raw bio-
mass combustion after devolatilization process. Meanwhile, Stage D
represented the passive combustion reaction zone, occurring at tem-
perature ranging from 635 to 720 °C, where slower biomass weight loss
(i.e. 2.25 wt%) was observed at the last stage. The decomposition step
at Stage D was attributed by the decomposition of inorganic material/
minerals, which were in good agreement with the high ash content of
raw microalgal biomass (López-González et al., 2014). Previous study
by Gai (Gai et al., 2015a) demonstrated that high ash content has sig-
nificant influence on the combustion process. This is because high ash
content (i.e. ash content for biomass feedstock was approximately
2–7 wt%) could increase the biomass degradation rate, which in turn
increased the raw microalgal biomass oxidation for higher hydrochar
production yield. Hence, based on the combustion behaviour of C.
vulgaris biomass, solid biofuel production is a preferable biofuel product
from this raw microalgal biomass.
3.2. Effects of HTC temperature and retention time
This section will report the effects of hydrothermal carbonization
(HTC) temperature and reaction time for the conversion of the raw
microalgal biomass to hydrochar at fixed biomass to water ratio
(B:W = 1:100). As shown in Fig. 4, the hydrochar yield was 41.85 wt%
at 180 °C and 30 min. The yield decreased to 26.32 wt% at 250 °C for
4 h reaction time. Overall, the hydrochar yield decreased when the
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Environmental Research 188 (2020) 109828
hydrothermal temperature and retention time were increased from 180
to 250 °C and from 0.5 to 4 h, respectively. The maximum difference of
hydrochar yield across the hydrothermal temperature investigated was
34.46% (i.e., at 4 h), whereas only 17.52% (i.e., at 220 °C) across the
retention time. It was obvious that the decrease in hydrochar yield was
strongly affected by hydrothermal temperature instead of reaction time.
This observation could be due to the following reasons: (1) macro-
molecules in the biomass underwent severe degradation and formed
liquid oil and gases under high temperature (Biller and Ross, 2012); and
(2) high hydrothermal temperature (i.e., above 200 °C for prolonged
retention time) caused secondary degradation of hydrochar during the
process of carbonization (Nizamuddin et al., 2017).
In addition, drastic decreased in hydrochar yield was observed at
hydrothermal temperature above 200 °C, especially for longer HTC
retention time. This is due to the decomposition of macromolecules in
biomass into liquid phase products, instead of retaining in solid phase.
According to Gai (Gai et al., 2015b), these changes were attributed by
the decomposition of cyclic oxygenates from reducing sugars (i.e., at
200 and 210 °C) and decomposition of N&O-heterocyclic compounds
from hydrolysis of protein (i.e., at 240–250 °C) into bio-crude oil. Apart
from that, slow decreasing rate in hydrochar yield was also observed at
high hydrothermal temperatures, which was attributed by the sec-
ondary degradation of hydrochar (i.e., lower conversion rate) under
subcritical water conditions (Mazaheri et al., 2010). These observations
indicated that hydrochar yield from HTC reaction showed an inverse
relationship for both hydrothermal temperature and retention time.
However, the rate of hydrochar yield decreased with the increasing
retention time was slower than that of hydrothermal temperature. This
finding is in good agreement with other well-established literatures
reported by [Román (Román et al., 2012), Benavente (Benavente et al.,
2015), Basso (Basso et al., 2016)].
3.3. Quantitative analysis on hydrochar
3.3.1. Composition analysis
Elemental analysis (carbon, hydrogen, nitrogen, sulphur, and
oxygen contents) and proximate analysis (moisture content, ash con-
tent, volatile matter and fixed carbon) for raw microalgal biomass and
its derived hydrochar, along with mass yield and carbon recovery rate
C.G. Khoo, et al. Environmental Research 188 (2020) 109828

Fig. 4. Hydrochar yield from HTC reaction of Chlorella vulgaris biomass at different hydrothermal temperatures and retention times with constant biomass to water
ratio (B:W = 1:100).

are shown in Supplementary data. Also included in Supplementary data and N: 5.7 wt%). A negligible amount of sulphur content (0.1–0.2 wt%)
are the carbon recovery rate, higher heating value (HHV) and energy was detected in hydrochars, with raw microalgal biomass showing
yield (EY) of the hydrochar obtained from various HTC operating 0.4 wt%. Besides, carbon content increased when the hydrothermal
conditions. temperature was elevated from 180 to 250 °C and the retention time
Generally, the quality of hydrochar is evaluated by proximate increased from 0.5 to 4 h. It was found that HTC was able to double the
analysis and heating value. Volatile matter refers to the low boiling densification of carbon as shown in Fig. 5. The main reactions that were
point organic compounds that usually condense into oils and tars responsible for carbon enrichment in hydrochars were condensation
during cooling stage, in which higher volatile content will produce and aromatization that took place in subcritical water during HTC
hydrochar with lower quality (Kaeg et al., 2000). The overall volatile (Funke and Ziegler, 2010). These carbon enrichment hydrochars are
matter in hydrochar was lower than that of raw microalgal biomass, beneficial for carbon sequestration process. In contrast, the oxygen
which indicated that hydrochar demonstrated a better quality as com- content shows an opposite trend compared to that of carbon with a
bustion fuels (Supplementary data). On the other hand, the fixed carbon lower range from 25.3 to 39.8 wt%, as compared to the raw microalgal
represents non-degradable carbon for ground burial that contribute to biomass (i.e., O: 59.7 wt%). However, the carbon, hydrogen and oxygen
carbon credits in carbon cycle (Zhu et al., 2015). Based on Supple- content at retention time of 1 and 4 h remained almost the same. This
mentary data, for sample 180_0.5 (i.e. HTC conducted at 180 °C for
0.5 h), it is estimated that for every 100 kg of raw microalgal biomass,
HTC could recover 84.6 kg of C in hydrochar form (i.e., carbon recovery
rate) for ground burial, in which 58.1 kg of carbon is in stable form (i.e.,
fixed carbon). In comparison with raw microalgal biomass, the ash
content of hydrochar increased from 3.1 to 7.7 wt% at higher hydro-
thermal temperatures (230–250 °C) and longer retention time (4 h).
Meanwhile, there was insignificant changes in the ash content ( ± 1 wt
%) for samples carbonized at hydrothermal temperature of
180 °C–210 °C with retention time of 2 h. This was due to the HTC
treatments as most of the organic and inorganic elements (especially
Na+ and K+) were removed from raw microalgal biomass, especially
when the HTC was conducted at severe operating conditions (Reza
et al., 2015). Moreover, HTC at higher temperature and prolonged re-
tention time could contribute to major loss of organic matters in hy-
drochars to the aqueous phase, resulting to increment of the ash frac-
tion in the hydrochars (Libra et al., 2011).
The elemental analysis from Supplementary data shows that HTC
has insignificant effect on the hydrogen and nitrogen contents of hy- Fig. 5. Carbon densification of hydrochar produced from HTC of Chlorella
drochars that ranged between 6.6 to 9.5 wt% and 0.8–5.5 wt%, re- vulgaris biomass at different hydrothermal temperatures and retention times
spectively, in comparison with raw microalgal biomass (i.e., H: 7.0 wt% with constant biomass to water ratio (B:W = 1:100).

7
C.G. Khoo, et al.
Fig. 6. The van Krevelen diagram of hydrochar produced from Chlorella vulgaris biomass under various HTC processing conditions.
observation showed that prolonged retention time would not improve
the overall hydrochars quality.
On the other hand, the changes in H/C and O/C atomic ratios with
respect to the dehydration and decarboxylation reactions during HTC
can be visualized by the van Krevelen diagram (Fig. 6). The atomic
ratios of H/C (1.48–2.04) and O/C (0.30–0.59) of hydrochars shows a
reduction in comparison to microalgal biomass (H/C: 3.07 & O/C:
1.63). This finding indicated that the reduction of atomic ratios of H/C
and O/C of hydrochars occurred along with the increased in hydro-
thermal temperature and prolonged retention time, in which extreme
HTC reactions caused significant changes in the structural element
composition. In terms of elemental analysis through ven Krevelen dia-
gram, it was found that the dehydration and decarboxylation reactions
contributed to the reduction of H and O contents in hydrochars (Funke
and Ziegler, 2010; Titirici et al., 2012). Notably, the hydrochars derived
from raw microalgal biomass displayed much higher ratios of H/C and
O/C than those of the others solid fuels such as coal, which reflected
high quality of microalgal hydrochars due to reduction in the con-
densed aromatic structure (Hrnčič et al., 2016).
The HHVs of hydochars obtained in the present work ranged from
21.00 to 25.30 kJ/g (Supplementary data), which were comparable to
the calorific value of sub-bituminous coals (20.83–21.32 kJ/g)
(Engineering Toolbox and 2010, 2018). By taking the average value of
HHVs, it was approximately 1.84-time higher than the raw microalgal
biomass sample (12.58 kJ/g). Overall, the HHVs were highly dependent
on the hydrothermal temperature, in which the highest energy densi-
fication of hydrochar was produced at 210 °C. Notably, the energy yield
(EY) calculated varies significantly with the yield of hydrochars. Based
on the results obtained, the maximum EY (i.e., 76.6%) was attained
under optimum HTC operating conditions (i.e., T = 210 °C, t = 0.5 h).
3.4. Functional groups analysis
The FT-IR spectra of raw microalgal biomass and selected hydrochar
samples (i.e., T = 200, 210, and 220 °C; t = 0.5 h) are presented in
Fig. 7. From the figure, the stretching of –OH bonds in hydroxyl or
carboxylic acid were observed between the bandwidth of 3600 to
3000 cm−1. Under this range of bandwidth, the intensity of peaks for
hydrochars samples were reduced when hydrothermal temperature was
measured from 200 to 220 °C. The intensities of peaks were lower
compared to raw microalgal biomass, which indicated dehydration of
biomass during HTC reaction. Nonetheless, the sharp peaks between the
band regions of 2850–2950 cm−1 showed the presence of hydrocarbon
chains such as –CH2 and –CH3 in the hydrochars samples produced
8
Environmental Research 188 (2020) 109828
under higher HTC temperature. This observation reflected better
quality of hydrochars as solid fuel (i.e. with higher C/H atomic ratio).
Interestingly, the hydrochar produced at 210 °C showed the most in-
tense band region for bandwidth 1700–1450 cm−1, indicating higher
nitrogen content in the hydrochar and thus suitable for soil amendment
(Reza et al., 2014). On the other hand, the bandwidth of
1150–950 cm−1 was less intense in hydrochars compared to raw mi-
croalgal biomass. This is due to the decomposition of C–O–R and C–O
bonds from ethers and polysaccharides during HTC reactions (Gai et al.,
2015b).
3.5. Combustion behaviour
The thermogravimetric analysis (TG-DTG profiles) of raw micro-
algal biomass and selected hydrochar samples (i.e., T = 200, 210, and
220 °C; t = 0.5 h are illustrated in Fig. 8. Overall, the TG-DTG de-
composition trends of hydrochar samples were similar to the raw mi-
croalgal biomass. The combustion behaviors of the samples were fur-
ther divided into several combustion stages based on the rate of weight
loss at different temperature zones in TG curves (Fig. 8(a)) and DTG
curves (Fig. 8(b)). Stage A, B, C, and D represented different combus-
tion stages for raw biomass that has been discussed in previous section.
Meanwhile, Stage A1, B1, and C1 represented the combustion stages for
hydrochar samples.
For all the hydrochar samples, Stage A1 represents evaporation of
moisture content at temperature ranging from 30 to 120 °C, resulted to
a weight loss of 3–4.5 wt%. The combustion of hydrochars took place in
two stages aside from the weight loss during moisture evaporation.
Firstly, the release of volatiles took place at Stage B1, followed by the
combustion of hydrochars occurring at the second stage (Stage C1),
which corresponds to the devolatization and combustion of the volatiles
and hydrochars (Parshetti et al., 2013). The major hydrochar weight
loss of hydrochars at Stage B1 and C1 (35–40.5 wt% and 31 to 37 wt%
respectively) occurred at temperature ranging from 195 to 373 °C and
373–680 °C, respectively. When compared the DTG curves of hydro-
chars to raw biomass, it was found that area of the weight loss peaks
displayed by hydrochars was larger than raw microalgal biomass. This
corresponds to higher fixed carbon contents in the hydrochar and thus,
indicating that intensified devolatilization and combustion reactions
contributed to larger weight loss. Lastly, the residual weight of each
sample was lower than their original ash content, each sample was
lower than their original ash content, indicating a successful ash car-
bonization during combustion of sample.
The reaction mechanism of hydrochars during combustion can be
C.G. Khoo, et al.
Fig. 7. FTIR spectra of Chlorella vulgaris biomass and selected hydrochars processing at hydrothermal temperatures of 200, 210 and 220 °C, for a retention time of
0.5 h. Note for regions: 1: OH; 2: CH2 & –CH3; 3: CH2 & –CH3; 4: C]O & N–H; 5: C]O & N–H; 6: C–O–R; and 7: C–O.
detected through DTG curves. During devolatilization and char com-
bustion stage, the hydrochars samples illustrated three obvious peaks in
DTG profile (i.e., from 220 to 540 °C). The peak occurred at 240 °C
could be contributed by the depolymerization reactions of low ther-
mally stable substances such as volatile matter in hydrochars, mean-
while further degradation of thermally stable substances such as fixed
carbon content in hydrochars was continued in the char combustion
stage at 300 °C (Chen et al., 2014). The combustion of raw microalgal
biomass (i.e. at Stages C and D) resulted to a more higher intensity
peaks as compared to hydrochar samples. This is mainly due to the loop
reactions of polymerization when the raw microalgal biomass was hy-
drothermally carbonized under subcritical water condition (Kambo and
Dutta, 2015). The biochemical components in the raw microalgal bio-
mass could be converted into different types of compounds through
various reactions routes such as hydrolysis, dehydration, decarboxyla-
tion, polymerization, and aromatization (Funke and Ziegler, 2010). The
compounds produced during HTC reactions could also be recombined
to form new substances for the subsequent polymerization reactions
(Demirbas, 2004). Hence, this has caused the detection of more peaks in
the DTG thermogram for hydrochars compared to raw microalgal bio-
mass.
Apart from that, the combustion behaviour of biofuels can be de-
scribed by ignition and burnout temperatures, which is an indicator for
the safety of storage and delivery of fuel, and for its respective com-
bustor design (Du et al., 2007). The ignition temperature (Ti) is the
minimum temperature for the fuel to ignite spontaneously while
burnout temperature (Tb) is the temperature when the fuel is depleting.
Tm indicates the temperature of maximum mass loss at the maximum
peak. The values of Ti, Tm, and Tb for both raw microalgal biomass and
selected hydrochar samples are tabulated in Table 3.
All the selected hydrochar samples attained higher Ti (approxi-
mately 250–257 °C) than the raw microalgal biomass (170 °C). This is
due to the higher fixed carbon content in hydrochars that required
higher Ti. Also, the Ti for hydrochars samples remained constant for
different HTC temperatures due to the devolatization peaks in DTG
curves were located at similar temperature region. This observation
indicated that increment in hydrothermal temperature has insignificant
influence towards coal fuel properties of hydrochars. Besides, the Ti of
hydrochars were in good agreement with the biochar produced from
duckweed samples (255.5–286.9 °C) along with wet torrefaction tem-
perature ranging from 130 to 250 °C (Zhang et al., 2016). As a result,
9
Environmental Research 188 (2020) 109828
this moderate Ti indicated that the produced hydrochar had a good coal
fuel property. It should be noted that Ti value that is too low could
cause safety issue in storage (i.e., easily ignited) whereas Ti that is too
high may require more energy to ignite the reactions. This finding in-
dicated that the coal fuel properties of hydrochar has improved in terms
of storage and delivery safety compared to the raw microalgal biomass.
Besides that, it was found that the Tm of hydrochar samples were
55 °C–70 °C higher than that of biomass, indicating that longer HTC
retention time was required in order to achieve maximum HTC reaction
under the same heating rate during combustion reaction. In addition,
the Tb of hydrochars were generally lower than the raw microalgal
biomass, indicating the important of burnouts performances of the
hydrochars, which were more thermally stable (i.e. more resistance
towards decomposition at high temperatures) compared to the raw
microalgal biomass. Thus, a lower Tb value indicated shorter burnout
time of fuel and lower combustible matter content in ash.
3.6. Application of aqueous phase produced from HTC
3.6.1. Characterization of aqueous phase
The aqueous phase produced from HTC reaction was characterized
to determine the concentration of carbon (i.e. through COD and TOC
analyses), total nitrogen and total phosphorus. In general, the aqueous
phase was amber in colour and released foul odour due to volatile
components such as short chain fatty acids (Levine et al., 2013). The
characterization results of aqueous phase produced from HTC of C.
vulgaris are tabulated in Supplementary data.
Fig. 9 illustrates the distribution of pH in the aqueous phase after
HTC reactions ranging from 2.95 to 4.50. The pH is suitable to be used
for microalgae cultivation as based on our previous study the optimum
pH is around 3 (Khoo et al., 2016). The pH of the aqueous phase de-
monstrated a trend of ascending order when the hydrothermal tem-
peratures increased. The most acidic pH attained at the lowest HTC
temperature (180 °C), indicating that the HTC reaction favours the
formation of organic acids at low temperature; meanwhile at higher
HTC temperature, decarboxylation of organic acid was favoured and
caused the increment in pH values (Gai et al., 2015b).
Besides that, the TOC in aqueous phase was about 43–83% of that in
C. vulgaris culture medium (2512.20 mg/L), and the concentration de-
creased with increasing hydrothermal temperature. This showed that
the organic acid components produced during HTC reactions continued
C.G. Khoo, et al. Environmental Research 188 (2020) 109828

Fig. 9. The pH of the aqueous phase separated after HTC of Chlorella vulgaris
biomass at different hydrothermal temperatures and retention times with
constant biomass to water ratio (B:W = 1:100).

Fig. 8. TG-DTG thermogram for combustion of Chlorella vulgaris biomass and

selected hydrochar samples (T = 200, 210 & 220 °C) under HTC retention times Fig. 10. The COD/TOC ratios of aqueous phase separated after HTC of Chlorella
of 0.5 h as data representative: (a) TG profile and (b) DTG profile. Note:Stage A vulgaris biomass at different hydrothermal temperatures and retention times
& A1 represented weight loss from evaporation of water, Stage B & B1 re- with constant biomass to water ratio (B:W = 1:100).
presented the release of volatiles and combustion, Stage C & C1 represented the
combustion of volatiles, and Stage D represented the passive combustion re-
total organic compounds that can be retained in solid phase. Among all
action zone.
the investigated hydrothermal temperatures, the temperature with
maximum energy yield, EY (210 °C) elucidated a nearly constant
Table 3 change of COD/TOC ratios under various HTC retention time. Overall,
Characteristic temperature of Chlorella vulgaris biomass and selected hydrochar the COD/TOC ratios were relatively low (i.e. less than one) and in-
samples produced at hydrothermal temperatures 200, 210 And 220 °C for re- dicated low concentration of oxidable organic substances (e.g. sugars,
tention time of 0.5 h as data representative.
formate) presence in the aqueous phase. In addition to that, the aqu-
Sample Characteristic temperature (oC) eous phase contained trace amount of total phosphorus (i.e.,
1.37–7.52 mg/L). Significant amount of total nitrogen
Ti Tm Tb
(67.10–122.70 mg/L) was also detected, indicating the possibility of
Biomass 170.0 250.0 700.0 recycling the aqueous phase as alternative nutrient source for micro-
T200_t0.5 250.0 320.0 635.0 algae cultivation.
T210_t0.5 255.5 305.0 600.0
T220_t0.5 257.0 312.5 600.0

3.7. Cultivation of microalgae with aqueous phase

to react to form carboxyl under severe conditions (Gai et al., 2015b).
Analysis on the aqueous phase produced from HTC showed that it
Hence, by taking the ratio of COD to TOC, the sensitivity of the organic
contained high concentration of nutrient contents such as TN and TP.
substances in aqueous phase to be oxidized can be evaluated. Fig. 10
Therefore, the aqueous phase was tested if it is possible to be used as
shows the COD/TOC ratio of the aqueous phase from HTC and its values
alternative nutrient source to cultivate C. vulgaris. The aqueous phase
ranged from 0.25 to 0.40. The distribution of the COD/TOC ratios il-
obtained during optimum HTC reactions condition (hydrothermal
lustrated a nearly constant change across hydrothermal temperatures,
temperature and retention time at 210 °C for 0.5 h, respectively) was
except for 250 °C. This is due to the secondary degradation of organic
used for this part of study. The comparison of the major nutrient
compounds in hydrochars produced at 250 °C, causing the reduction of
compositions between the control nutrient (i.e. organic fertilizer) and

10
C.G. Khoo, et al.
Table 4
Distribution of nutrient compositions sourced from organic fertilizer and aqu-
eous phase produced from optimized HTC reaction.
Nutrient source Control Aqueous phase produced from HTC
COD (mg/L) 34.6 600.0
Total nitrogen (mg/L) 26.5 78.2
Total phosphorus (mg/L) 4.27 2.66
TOC (mg/L) 25.7 2027.5
Fig. 11. Comparison of growth performances of Chlorella vulgaris that utilized
HTC produced aqueous phase as nutrients source with the control sample for 15
days. Other cultivation condition: inoculum concentration ≈0.3 g L−1, illu-
mination = 24 h with light intensity of 60–70 μmol m−2 s−1, compressed-air
aeration rate = 0.16 vvm, pH 3 culture medium, and volume of
nutrients = 1 L.
aqueous phase produced from HTC are tabulated in Table 4. It was
found that the aqueous phase produced from HTC contained higher
nutrients concentration than the organic fertilizer medium (control),
indicating the possibility to be used as nutrients source to grow mi-
croalgae.
Fig. 11 shows the comparison of C. vulgaris growth performance in
the pilot-scale BC-PBR cultivation system with control conditions (i.e.
organic fertilizer as nutrients source) and aqueous phase produced from
HTC as nutrients source. From the nutrients composition analysis, it
was found that the aqueous phase produced from HTC contained sig-
nificant amount of nutrients, especially excess in TOC that could acted
as additional carbon source supplied to the cultivation system. From
Fig. 11, the growth of C. vulgaris was not affected by using aqueous
phase produced from HTC as nutrient source, and able to produce sa-
tisfactory quantity of biomass concentration within 15 days of culti-
vation period. The results showed that the microalgae growth with
aqueous phase produced from HTC was able to retain up to 92% of the
biomass concentration as compared to the control sample. The reduc-
tion in biomass concentration could be due to the high COD value and
low total phosphorus concentration in the HTC produced aqueous
phase, which limits the growth of microalgae cells. However, it is still
feasible to utilize the aqueous phase produced from HTC as an alter-
native nutrient source since it is still able to significantly support the
growth of microalgae.
4. Conclusions
The present study has successfully demonstrated the feasibility to
produce hydrochar from low-lipid and high-ash microalgal biomass
through HTC process. It was found that the HHV value of hydrochar
11
Environmental Research 188 (2020) 109828
(24.51 kJ/g) produced under optimized reaction condition was ap-
proximately two-times higher than that of raw microalgal biomass
(12.59 kJ/g). Moreover, the combustion behaviour study performed on
the hydrochar had significant higher ignition and lower burnout tem-
perature than the raw microalgal biomass. This result indicated that the
produced hydrochar through HTC process can be used as a safe solid
fuel. In addition, the aqueous phase produced from HTC process could
be further utilized to re-cultivate C. vulgaris with promising growth
performance. This finding indicated the promising opportunity of HTC
to be integrated in the upstream microalgal cultivation process and to
enhance the techno-economic feasibility of microalgal hydrochar pro-
duction.
Credit author statement
Choon Gek Khoo: Conceptualization, Methodology, Formal analysis,
Investigation, Writing - original draft. Man Kee Lam: Supervision,
Writing - review & editing. Abdul Rahman Mohamed: Supervision. Keat
Teong Lee: Supervision, Writing - review & editing, Funding acquisi-
tion.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgements
This work was supported by the Ministry of Higher Education
(MOHE) Malaysia [Fundamental Research Grant Scheme (FRGS) –
Malaysia's Rising Star Award (MRSA) 2016 (203/PJKIMIA/6071362)]
and the Universiti Sains Malaysia [Short Term Grant (304/PJKIMIA/
6315016)].
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.envres.2020.109828.
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