Journal of Bioresources and Bioproducts 9 (2024) 310–321

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Journal of Bioresources and Bioproducts

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Research Article

Exploring hemp seed hull biomass for an integrated C-5

biorefinery: Xylose and activated carbon
Sreesha Malayil a,c, Luke Loughran b, Frederik Mendoza Ulken a,
Jagannadh Satyavolu a,b,∗
a Conn Center for Renewable Energy Research, University of Louisville, Kentucky 40292, United States
b Bioproducts LLC, Louisville, Kentucky 40206, United States
c
RTI International, Research Triangle Park, NC 27709, United States

a r t i c l e i n f o a b s t r a c t

Keywords: Large quantities of hemp hulls can be completely utilized for creation of value-added products
Xylose (cost effective biofuels and biochemicals) through a biorefinery approach. A sustainable approach
C-5 biorefinery in making xylose, a low calorie sweetener and high surface area activated carbons (AC) for su-
Methane sulphonic acid
per capacitors, attracts interest. The AC when leveraged as a co-product from biorefinery process
Delignification
makes it more cost effective and, in this paper, we discuss the production of xylose and AC from
Activated carbon
Dilute acid hydrolysis hemp seed hull with methane sulphonic acid (MSA) hydrolysis. Xylose recovery with MSA hy-
drolysis was 25.15 g/L when compared to the traditional sulphuric acid (SA) hydrolysis of 19.96
g/L at the same acid loading of 1.8 %. The scanning electron microscope (SEM) images and
Fourier transform infrared (FT-IR) spectra indicate partial delignification along with hemicellu-
lose hydrolysis responsible for high xylose recovery. Post hydrolysis fibers were KOH activated
and carbonized to make AC. The MSA hydrolyzed and KOH activated fiber produced pure, fluffier
and finer particle AC with a drastic increase in surface area 1 452 m2 /g when compared to SA
hydrolyzed of 977 m2 /g. These results indicate the potential of MSA in dilute acid hydrolysis of
biomass for xylose recovery and production of high surface area activated carbon. From a pro-
duction standpoint this can lead to increased use of sustainable low-cost agricultural biomass for
making high surface area AC as components in supercapacitors.

1. Introduction

Hemp containing 𝛿-tetrahydrocannabinol below legal limits (< 0.3 %) commonly referred as industrial hemp is an annual cash
crop gaining attention in the United States and European countries (Adhikary et al., 2021; Aloo et al., 2022). Hemp industry has grown
exponentially over the last few decades in the United States since it is a sustainable crop requiring low water and fertilizer with no
requirement of herbicide, pesticide etc., while nourishing soil, performing as an excellent break crop in crop rotation (Ranalli and
Venturi, 2004; Adesina et al., 2020). The global industrial hemp market is expected to grow from 4.6 billion yuan (US dollar) in 2019
to 26.6 billion yuan (US dollar) by 2025 (Burton et al., 2022; Mishra et al., 2022). Earlier, industrial hemp was grown for textile
application and enormous research has been conducted on developing techniques and processes for extraction of hemp fibers. Today
industrial hemp has been attracting growing interest worldwide for its oil, protein and seed (Poniatowska et al., 2019).
Hemp seed typically contains one third oil, one third protein and one third hull (Leonard et al., 2022). Hemp oil has been positioned
as a highly valuable product in food, pharmaceutical, nutraceutical and cosmetic industry, due to its uniqueness in composition,

∗
Corresponding author.
E-mail address: [email protected] (J. Satyavolu).

https://doi.org/10.1016/j.jobab.2024.01.002

Available online 24 January 2024

2369-9698/© 2024 The Authors. Publishing Services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd. This is an open access article
under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
S. Malayil, L. Loughran, F.M. Ulken et al. Journal of Bioresources and Bioproducts 9 (2024) 310–321

justifying hemp seed processing even if its oil yields are lower than the conventional oil seeds. The hemp seed protein estedin is of
high biological value due to its structural similarity to serum globulins and its amino acid composition (Malomo et al., 2014). Unlike
soy, which contains allergens and a combination of low and high molecular weight proteins hemp protein mainly consisting of easily
digestible low molecular weight protein making it a super food (Malomo et al., 2014). Even though hemp can be grown as a dual
crop for food and industrial markets, cultivation systems usually concentrate on producing either fiber or seed and generally not both
(Burton et al., 2022). Hemp cultivation systems concentrated on hemp seeds yields short fibers finding application in paper making
and are unsuitable for textile application. Therefore, hemp seed production system typically targets three segments of the global
industrial market-food and beverage, personal care products and paper (Rheay et al., 2021). With all this said and an expected boom
in hemp industry, it is anticipated that large quantities of waste such as hemp hulls would be produced in the coming years. Hemp
being a niche crop, still shows low economics with just oil and protein extraction, requiring investigation of novel use for hemp hulls
with the objective of improving the overall economics.
Earlier studies in this lab on seed hulls has showed its potential as substrate for C-5 biorefinery due to its low lignin and high hemi-
cellulose content (Malayil et al., 2022; Tadimeti et al., 2022). This is because in nature hemicellulose is found interlinked to lignin
which makes extraction difficult (Chanakya and Sreesha, 2012). Various seed hulls such as soy, corn and almond have been success-
fully converted to various bioproducts through a C-5 biorefinery concept (Fonseca et al., 2014; Herde et al., 2020; Satyavolu et al.,
2021; Malayil et al., 2022; Tadimeti et al., 2022). Economic models developed to predict cost effective production of fuels and
chemicals from biomass have concluded that the cost of substrate and handling to be the major share of the overall production cost
(Chandel et al., 2018). Therefore, the choice of appropriate lignocellulosic material for C-5 biorefinery is important for cost effective
production of fuels, chemicals and bioproducts. The C-5/hemicellulose based biorefinery mainly focuses on extracting hemicellulose
with various pre-treatment techniques and recovering it to produce xylose, chemicals and fuels. Hemicellulose being the second most
abundant component of lignocellulosic biomass after cellulose is about 15 %–30 % of biomass composition (Yousuf et al., 2020). The
amorphous and highly branched nature of hemicellulose makes it more unstable in comparison to cellulose and lignin and therefore
requires less harsh pre-treatment conditions for its degradation. The utilization of the hemicellulose fraction helps in justifying the
overall economics of the production of biofuels and bio-chemicals from lignocellulosic biomass. A typical C5 biorefinery focuses
on extraction of xylose by hydrolyzing biomass using dilute acids and converting xylose to xylitol and other chemical precursors
(Fonseca et al., 2014; Gori et al., 2015; Malayil et al., 2022; Tadimeti et al., 2022). The remaining xylose stripped biomass can
be saccharified with cellulases and other treatments for bioethanol production or can be converted to other value-added products
such as activated carbon, natural fiber composites etc. The economics of hemp seed industry can be multiplied several folds with an
integrated C-5 biorefinery.
For a C-5 biomass biorefinery process, the choice of acid and the hydrolysis conditions is an important factor (Almashhadani et al.,
2022). Commonly preferred acid for dilute acid hydrolysis is sulphuric acid (SA) due to its high reaction rates, mass transfer efficiencies
and low cost. The SA hydrolysis for C-5 sugar recovery is performed at lower acid concentrations 0.1 %–1 % and milder conditions
(time and temperature). However, SA pre-treatment/hydrolysis requires downstream processing in the form of acid neutralization
which is expensive and leads to generation of sulphate wastes (Chandel et al., 2012). Therefore, it is important to discover new “green
agents” which can address some of these challenges by facilitating higher xylose selectivity, separation of xylose from the hydrolysis
liquor, reusability and milder operating conditions. Methane sulphonic acid (MSA) is one such “green agent” which can be reused
without the need for neutralization and earlier studies conducted in this lab on MSA for dilute acid hydrolysis of almond waste such
as skin and shell has shown higher xylose selectivity and yields along with enhanced delignification resulting in separation of the
cellulosic fiber bundles suitable for downstream processing (Malayil et al., 2022). In this article we attempt to extend this knowledge
and test its action not only on xylose selectivity and yields but also on the other products of the C-5 biorefinery represented in Fig. 1
such as activated carbon (AC) from hemp seed hull.
The objective of this study is to evaluate a novel C-5 biorefinery process for hemp seed hull where a green acid such as MSA would
be tested for dilute acid hydrolysis, for simultaneous recovery of xylose (C-5 sugar) and high surface area activated carbon (use in
supercapacitors).

2. Materials and Methods

2.1. Substrate

Hemp seed hulls used in this study were provided by Victory hemp foods, Louisville, Kentucky, USA.

2.2. Substrate composition analysis

Hemp hulls were dried at 110 °C for 8 h, milled and sieved (4 mm) for homogenization. Proximate analysis (moisture, ash,
volatile matter and fixed carbon) was conducted using a thermogravimetric analyzer SDT TA Q600 in accord to the ASTM protocol
(Malayil et al., 2022). Elemental composition (C, H, N and O) was estimated using a TrueSpec CHN LECO elemental analyzer as
per the ASTM D5373 protocol (USA). The higher heat value (HHV) and proteins were calculated from the elemental composition as
described in (Malayil et al., 2022).

2.3. Hydrolysis

The effects of single stage dilute acid hydrolysis of hemp seed hulls were compared for two acids (methane sulphonic acid and
sulphuric acid). The analysis was performed separately in triplicates in a 100 mL glass pressure vessel (30 mm diameter × 190 mm

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Fig. 1. Proposed C-5 biorefinery for hemp seed hull.

Table 1
Hydrolysis condition for hemp seed hull.

Acid loading (dry weight basis) Hydrolysis temperature Solid to liquid ratio Concentration of acid in liquor (w/w)

4% 140 1꞉5 0.8 %

5% 140 1꞉5 1.0 %
6% 140 1꞉5 1.2 %
7% 140 1꞉5 1.4 %
8% 140 1꞉5 1.6 %
9% 140 1꞉5 1.8 %
10 % 140 1꞉5 2.0 %

length) with polytetrafluoroethylene (PTFE) bushings as outlined in (Malayil et al., 2022). The conditions chosen for acid hydrolysis
is shown in Table 1.
The proposed C-5 biorefinery process for hemp hull under investigation is illustrated in Fig. 1. A dilute acid hydrolysis was
tested on hemp seed hull with two different acids (sulphuric acid-SA and methane sulphonic acid-MSA) for optimizing xylose yields.
The conditions used for selective xylose hydrolysis are listed in Table 1. The hydrolysate collected (rich in xylose) was used for
xylose recovery employing a patented process developed at University of Louisville (US Patent # 10, 407, 453) (Gori et al., 2015;
Tadimeti et al., 2022). Once the xylose is removed, the remaining hydrolysate rich in glucose and other monomers from hemicellulose
and cellulose hydrolysis can be used for producing bioethanol/biogas (not discussed in this article). The hydrolyzed hemp hulls were
collected separately for two acid treatments (SA and MSA) and subjected to carbonization with and without activation agent to study
the impact. The activation agent used in this study is potassium hydroxide (KOH). Such a biorefinery process for hemp seed hull has
the potential to increase the overall economics for hemp seed industry and provide additional income to farmers.

2.4. Hydrolysate analysis

The hydrolysate from both the acid treatments were analyzed for glucose, arabinose and xylose and degradation products such as
acetic acid, 5-Hydroxy methyl furfural and furfural with a Waters 600E HPLC system (Waters corporation Milford, MA, USA) with an
Agilent 1260 infinity refractive index detector and Agilent Hi-Plex H column (300 mm × 7.7 mm × 8 mm) as outline in (Gori et al.,
2015; Malayil et al., 2022; Tadimeti et al., 2022). The combined severity factor was calculated using the procedure outlined in
Tadimeti et al. (2022).

2.5. Post hydrolysis fiber characterization

Hemp seed hull after hydrolysis were characterized for surface, chemical and mechanical properties using Fourier transform
infrared (FT-IR) spectroscopy and scanning electron microscope (SEM) for its potential in activated carbon making.

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Table 2
Elemental composition and proximate analysis of hemp seed hull.

Composition Hemp shell

Moisture (%) 19.52 ± 0.44

Volatile matter (%) 62.60 ± 1.23
Ash (%) 0.86 ± 0.09
Fixed carbon (%) 17.02 ± 0.36
Higher heating value (MJ/kg) 19.75
C (%) 49.40 ± 0.29
H (%) 5.94 ± 0.11
N (%) 1.93 ± 0.08
S (%) 4.3 ± 0.12
O (%) 38.00 ± 0.85
O/C ratio 0.76
Proteins (%) 12.06
Polar extractives (%) 23.90 ± 1.80
Cellulose (%) 54.50 ± 1.21
Hemicellulose (%) 16.71 ± 0.89
Lignin (%) 4.00 ± 0.21
Xylan:arabinan (%) 1:00

2.5.1. The FT-IR

For determining the changes in the chemical composition before and after hydrolysis MSA and SA treated hemp seed hulls and AC
were subjected to FT-IR analysis using an attenuated total reflection (ATR)-enabled Perkin Elmer Spectrum 100 FT-IR spectrometer
as outlined in Malayil et al. (2022), Tadimeti et al. (2022) and Yang et al. (2014).

2.5.2. The SEM

Changes in morphology of hemp seed hulls with acid treatments were analyzed using SEM (TESCAN USA, Inc.) at a voltage of 20
kV (Balla et al., 2021; Malayil et al., 2022; Tadimeti et al., 2022).

2.6. Chemical activation

The hemp seed hulls after dilute acid hydrolysis were further processed to produce activated carbon. A KOH activation process was
used wherein 100 g of hydrolyzed hemp seed hulls was mixed with KOH in a 1꞉1 ratio as previously described in Herde et al. (2020).

2.7. Carbonization

The KOH activated and as is samples (hemp hull as is, SA and MSA hydrolyzed) were then carbonized in a horizontal tube
furnace under a Nitrogen blanket at 950 °C for one hour as previously described in Herde et al. (2020). The AC samples obtained was
thoroughly washed with deionized water to remove KOH and air dried for further analysis.

2.8. Brunauer-Emmett-Teller (BET)

The prepared activated carbons (AC’s) were analyzed for surface area and other surface properties using a BET surface area
analyzer (Micromeritics, TriStar 3000, USA). This was done using Nitrogen gas as the adsorbed gas at 77.3 K. The samples were
degassed before the analysis at 160 °C for two and a half hours (Herde et al., 2020).

3. Results and discussion

3.1. Composition of hemp seed hull and hydrolysis

The composition of hemp seed hulls as proximate (moisture content, volatile matter, ash, fixed carbon, higher heating value),
ultimate (C, H, N, O) and fiber analysis (extractives, proteins, cellulose, hemicellulose and lignin) is reported for the first time and
presented in Table 2. Hemp seed hull was found to have high volatile fraction and fixed carbon with low ash and moisture content
making it a suitable substrate for both thermochemical and biological conversion to fuels (Yang et al., 2017). The other indicator
for potential conversion to fuels and chemicals is O/C ratio and higher C %, suggesting a clear possibility for C5 biorefinery process
(Cherubini and Strømman, 2011). Similarly, hemp hull is found to possess large quantities of water soluble pectic materials and low
lignin content making it an appropriate material for various conversion processes to biofuels, bioproducts and chemicals. Hemp hull
containing high volatile, fixed carbon fraction and low moisture and ash would make an ideal substrate for biorefinery processes.
Unlike other seed hulls studied earlier in this lab such as soy (Tadimeti et al., 2022), corn (Fonseca et al., 2014) and almond hull
(Malayil et al., 2022), hemp seed hull does not contain arabinose. Experience on previous research conducted in this lab suggests
that under dilute acid conditions, especially SA has better selectivity for arabinose than xylose suggesting the need for a two-stage

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Fig. 2. Xylose recovery and hydrolysis byproducts.

hydrolysis where in stage 1 with milder acid and process conditions leads to extraction of arabinose and stage 2 with slightly higher
acid concentrations and processing conditions leads to solubilization of xylose and removal of hemicellulose structures (Tadimeti et al.,
2022). In case of hemp seed hull, the hemicellulose structure contains only xylose suggesting the need for a single stage acid hydrolysis
for maximum xylose recovery leading to better process economics.

3.2. Kinetics of hydrolysis

The first and foremost step in creating a seed hull biorefinery process is to optimize the hydrolysis conditions (type of acid,
substrate to acid ratio, temperature and other process conditions) (Basar et al., 2021). Based on the previous studies conducted in
this lab on substrates such as almond industry waste (Malayil et al., 2022), soy hull (Tadimeti et al., 2022) and dry distiller’s grains
(DDGE) (Fonseca et al., 2014) the time for hydrolysis was kept constant at 60 min and the other varying parameters chosen for this
study are listed in Table 1. The release of xylose from hemp hull at various acid concentrations chosen for the study along with the
release of degradation products such as furfural, 5-hydroxymethyl furfural (5-HMF) and acetic acid is shown in Fig. 2 for two acids
(SA and MSA). Most of the seed hulls are found to have low lignin content and high hemicellulose (Saura-Calixto et al., 1983) making
them more suitable for C5 biorefinery and further processing of the hemicellulose free biomass to various value-added products.
Amorphous hemicellulose is readily hydrolyzed at dilute acid and milder processing conditions whereas the crystalline cellulose
structure requires > 1 % acid concentration, higher temperature and longer hydrolysis time (Negahdar et al., 2016). Severe treatments
also have adverse effects such as increase in degradation products like furfural and 5-HMF and reduced strength of hydrolyzed fibers
which is not satisfactory when the hemicellulose free biomass is considered for various other downstream processes. Therefore, an
attempt is made to study the effect of SA and MSA hydrolysis on hemp hull and optimize the process for maximum xylose release
and selectivity with minimal degradation products.
Even though the ideal acid concentration for most of the seed hull studied earlier in this lab for optimal xylose recovery was < 1 %,
in case of hemp hull the observation was that < 1 % acid (SA and MSA) lead to low recovery of xylose (< 2 g/L, Fig. 1). Maximum
xylose recovery was achieved at 1.8 % acid concentration for both SA and MSA. The MSA was found to yield more xylose (26.15
g/L) when compared to SA (19.96 g/L). The MSA being a stronger acid than sulphuric acid is expected to hydrolyze more xylose.
Similar observation of increased selectivity and recovery of xylose with MSA was observed with almond seed skin (Malayil et al.,

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Fig. 3. Combined severity factor (combined severity factor, CSF) and xylose yields.
SA: sulphuric acid; MAS: methane sulphonic acid.

2022). An interesting observation made in this study was the drastic increase in xylose release when the acid concentration was
increased from 1.2 % to 1.4 %. For most of the substrates studied earlier in this lab for xylose recovery and that reported in literature
suggest a gradual increase followed by a decrease with increasing degradations products (Fonseca et al., 2014; Malayil et al., 2022;
Tadimeti et al., 2022). However, this was not the case with hemp seed hull which requires further investigation. A possible reason
for this transition could be due to delignification. In biomass structures lignin is found bonded with hemicellulose and this forms a
covering over cellulosic microfibrils (Chanakya and Sreesha, 2012). Therefore, for optimal release of xylose from the hemicellulose
partial delignification is required. Literature has reported MSA to possess delignification properties (Gierer, 1985; Malayil et al.,
2022; Wu et al., 2023) and can be the reasons for higher xylose recovery which requires further study. Increasing acid concentrations
beyond 1.4 % lead to increased formation of degradation products: acetic acid, furfural and 5-HMF (Fig. 2).
The dependency of xylose yields in the hydrolysate on combined severity factor (CSF) is shown in Fig. 3. The varying severity
factors such as time, temperature and acid concentration combined as single entity and expressed as CSF is an excellent tool in
understanding the combined effect on xylose recovery and buildup of degradation products (Goh et al., 2011; Pappas et al., 2014;
Kresnowati et al., 2021; Tadimeti et al., 2022). From Fig. 2 it is evident that among the two acids MSA is preferred due to its high
xylose yields and low degradation products. Acetic acid is a degradation product and 5-HMF and furfural is a transformation product
of xylose at severe conditions. High severity factors contributed to loss of xylose (Fig. 3) in case of SA whereas in case of MSA there
was no substantial loss. With increasing CSF there is loss of xylose and buildup of degradation and transformation products in case
of SA whereas it was lower in case of MSA. A similar response with SA in case of soy hull was reported by (Tadimeti et al., 2022)
wherein the xylose recovery decreased with increasing CSF followed by buildup of degradation and transformation products where
a two stage SA hydrolysis was optimized for xylose recovery. Cassales et al. (2011) have reported significant reduction in operation
time with higher acid loading rates with 3–4 times higher degradation and transformation product formation when compared to
current study with SA which significantly impacts the xylose recovery process suggesting better performance of MSA at higher acid
loading rates. The xylose rich hydrolysate obtained would be used as a starting material for xylose isolation and separation process
(US Patent # 10 407 453) developed by biomass and biofuels group at University of Louisville. For optimal xylose recovery using
this process, it is very important to have low byproducts such as acetic acid, furfural and 5-HMF in the hydrolysate (Satyavolu et al.,
2021). Also, for the downstream processing of the hydrolyzed hemp fibers for making activated carbon, it is important to maintain
the structure of cellulose and avoid losses since at higher CSF cellulose is depolymerized.

3.3. Morphological and structural changes with hydrolysis

The morphological changes in hemp seed hull with dilute acid hydrolysis (SA and MSA) was studied using SEM imaging. Hemp
hull was found to have well defined three-layered structures (epidermis, sub hypodermis and hypodermis, Fig. 4a) like other seed
hulls studied earlier in this lab (rice, soy and almond hull). The surface of hemp hull was smooth (Fig. 4a) like soy hull but unlike rice
hulls which had well defined micro bumps and almond hull having reticulate-verrucate sculpture with convex and rugose anticlinal
walls and rugose periclinal walls (Xiao et al., 2014). Literature has reported disruption in the 3-layer seed hull structure for most of
the seed hulls studied for C5 and C6 biorefinery process with SA concentrations ranging from 0.1 % to 2 % (Malayil et al., 2022;
Tadimeti et al., 2022). However, in case of hemp seed hull the three-layered structure remained intact with 1.8 % acid (SA, Fig. 4b)
which requires further investigation. Such high acid concentration and processing conditions is expected to remove hemicellulose,
cellulose and lignin partially leading to separation of the cellulose microfibrils. But this was not the case observed with hemp seed
hull and the cellulose microfibrils was found intact. Previous studies on almond hull (Malayil et al., 2022) have reported rupturing

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Fig. 4. Scanning electron microscope (SEM) images of (a) as is (as received), (b) SA hydrolyzed and (c) MSA hydrolyzed.

of three-layered structure at 0.6 % MSA loading separating cellulosic fibers which was quite different from hemp seed hull requiring
further study. With MSA hydrolysis there was limited disruption in the hemp seed hull structure but did not lead to total collapse of
three-layered structure and separation of the cellulose fibers. With 1.8 % MSA there was disruption in the epidermis and hypodermis
layer (Fig. 4c). The MSA hydrolysis also led to loosening of the cellulose fiber bundles (Fig. 4c) and was different from SA. This
can be attributed to the delignification properties of MSA. Lignin is found interlinked with hemicellulose and this is helically wound
on cellulose microfibril (Chanakya and Sreesha, 2012) and therefore, removal of hemicellulose alone cannot lead to loosening of
cellulose microfibril and requires partial delignification.

3.4. The FT-IR

The FT-IR spectra (Fig. 5) represents the functional group and changes occurring, translating to the effect of chemical changes with
various treatments was used to understand the chemical compositional changes in hemp hull with hydrolysis. The analysis provides a
qualitative comparison of the changes occurring in functional groups among different hydrolysis treatments. All samples showed OH
(3291 cm−1 ), C–O (1042 cm−1 ) and C=O (1744 cm−1 ) and amide (1516 cm−1 ) groups indicating the hydrophilicity of hemp hull along
with C=C (1629 cm−1 ) and C–H bending (1454 cm−1 ) and C–H stretching (2853 cm−1 ) (Malayil et al., 2022; Tadimeti et al., 2022).
Reduction in peak intensity band corresponding to the –OH groups was observed due to hemicellulose removal. The MSA hydrolyzed
hemp seed hull had increased removal of –OH groups when compared to SA correlating with the increased xylose recovery (Fig. 5).

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Fig. 5. Fourier transform infrared (FT-IR) spectra of hemp hulls before and after hydrolysis.

The –CH stretching peak of cellulose and hemicellulose can be identified at peak 2925 and 2853 cm–1 in hemp seed hull. With acid
treatments there was flattening/reduction in peak intensities of –CH stretching indication loss of hemicellulose. Peak intensity at
1744 cm–1 can be assigned to the –C=O stretching from carbonyl and uronic acid ester groups of hemicellulose. Both SA and MSA
treatments were responsible in reducing this peak intensity suggesting removal of hemicellulose with maximum removal observed in
case of MSA hydrolysis. Amide peak (1516 cm−1 ) can be assigned to protein content of feedstock and reduction in amide group with
hydrolysis suggest removal of soluble proteins. With MSA treatment there was further flattening of this peak suggesting complete
removal of proteins when compared to SA and similar results were observed with almond industry waste such as skin and hull when
subjected to MSA hydrolysis (Malayil et al., 2022). Along with higher xylose yields, MSA hydrolysis could make hemp seed hull more
hydrophobic which is considered beneficial for further down streaming of this hydrolyzed biomass to various products such as natural
fiber composites, biocoal, activated carbon and carbon nano fibers.
The FI-IR spectra region specific to lignin in biomass is between 1800 and 800 cm–1 and can be used as an important tool in
understanding the type of lignin present with changes occurring due to hydrolysis (Faix, 1991). Drastic reduction in peak intensities
in this region especially with MSA suggest delignification along with hemicellulose hydrolysis. Expected lignin subunits in hemp are
syringyl (S) and hydroxy phenyl propane (H) with very few guaiacyl (G) since it belongs to the angiosperm family (Kiyoto et al.,
2018). Reduction in peak intensity at 1744 cm−1 with both SA and MSA suggest changes occurring in the acetate carbonyl group
of hemp lignin. Peak intensities assigned for S subunit are 1629 and 1516 cm−1 and reduced peak intensity indicate reduction in S
subunits. From Fig. 5, it can be observed that with MSA treatment there was drastic reduction in these peak intensities especially
at 1516 cm−1 suggesting removal of both hemicellulose and lignin with MSA. Flattening of peak at 1454 cm−1 with SA and MSA
indicate deformations in CH3 and CH2 groups of lignin moieties. No change in peak intensities were observed at 1331 cm−1 with SA
and MSA indicting no loss or structural deformation in the G subunit of lignin. Flattening of peak intensity at 1146 cm−1 with MSA
indicate deformation in the C–H in plane bending and C=O stretching of the aromatic structures. Similar reduction in peak intensity
with MSA at 1042 cm−1 suggest changes in the C–O stretching of lignin structures. The FT-IR results clearly indicate the activity of
delignification by MSA when compared to SA and similar results were observed when almond industry waste such as shell and skin
were subjected to hydrolysis with MSA (Malayil et al., 2022). This delignification property of MSA can be further tweaked to extract
pure cellulose microfibrils for various downstream applications.

3.5. Activated carbon (AC)

Hemp seed hull after hydrolysis was subjected to carbonization for AC making. Successful portable energy storage systems such
as Li-ion batteries, Li-S batteries supercapacitors, hydrogen storage mechanisms solar devices etc. require renewable, sustainable
and low-cost electrode material and high surface area AC’s can meet this demand (Lobato-Peralta et al., 2021). One of the major
challenges associated with production of AC’s is the access to low-cost substrate suitable for making high surface area AC (Baig and
Gul, 2021). Earlier research piloted in this lab has reported AC’s made from acid hydrolyzed biomass to have better surface area and
properties suitable for super capacitors (Herde et al., 2018). Herde et al. (2020) have demonstrated hemicellulose extracted biomass
fibers to produce high surface area AC’s. Hemicellulose removal from substrates like dry distiller’s grain (DDG) and soy hulls with
SA hydrolysis was found to increase the surface area of AC multifold (Herde et al., 2020). In this study, our aim is to understand the
effect of methane sulphonic acid in hemicellulose removal from hemp seed hull and its effect on AC properties and compare it to SA.
The AC’s were produces with and without activation process for comparison and the activation agent used is KOH.

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Table 3
Properties of SA and MSA hydrolyzed hemp shell activated carbon.

Sample Yield (%) Micropore volume (cm3 /g) Micropore area (m2 /g) BET surface area (m2 /g) External surface area (m2 /g)

As is (As received) 29.78 ± 1.02 0.05 ± 0.015 90 ± 2.22 108 ± 0.99 18 ± 0.69
As is + KOH 15.33 ± 0.75 0.33 ± 0.021 647 ± 4.35 706 ± 0.32 59 ± 1.12
SA 27.58 ± 1.36 0.05 ± 0.011 100 ± 1.09 123 ± 1.53 23 ± 0.45
SA + KOH 12.33 ± 0.90 0.51 ± 0.025 990 ± 5.21 977 ± 3.21 87 ± 1.88
MSA 25.94 ± 1s.18 0.05 ± 0.006 101 ± 2.01 143 ± 1.28 48 ± 2.11
MSA + KOH 10.50 ± 0.67 0.67 ± 0.021 1244 ± 3.33 1452 ± 3.54 118 ± 0.93

Note: as is, as received; SA, sulphuric acid; MAS, methane sulphonic acid; BET, Brunauer-Emmett-Teller.

Fig. 6. The FT-IR analysis of activated carbon (AC) made from hemp hull.

3.5.1. Yield and surface area

The surface area of AC’s made pre and post hydrolysis, with and without KOH activation is compared in Table 3. As shown in
Table 3, the surface area of pre hydrolysis and post hydrolysis materials are drastically different. The AC made from pre hydrolysis
hemp seed hull is 108 m2 /g and with KOH activation it goes up to 706 m2 /g whereas post hydrolysis with SA the surface area
further increases to 123 m2 /g and with KOH activation it goes upto 977 m2 /g (Table 3). The KOH activation is an intricate procedure
involving multiple reactions occurring on the surface of the biomass (Chen et al., 2020; Herde et al., 2020). The stage of physical
activation involves the enlargement of biomass pores with gas/volatiles released during heating of the biomass and involves three
major changes happening on the surface of the biomass. The first major surface change occurs due to various redox reactions occurring
between potassium and the free oxygen available in the hemp fiber resulting in increased surface area. As heating of the KOH activated
hemp shell progresses increasing amount of water vapor and carbon dioxide is released from the fiber trapping them in the pores
leading to increased pore volumes forming the second important surface modification. The third and final major change occurring on
the surface is due to the deposition of potassium forming a layer on the fiber surface leading to the expansion of the material causing
increased surface area.
The MSA hydrolysis further increased the surface area to 143 m2 /g and with KOH activation there is a potential to increase the
surface area to 1452 m2 /g (Table 3). The MSA is supposed to dehydrate the fiber and remove lignin along with hemicellulose (sup-
ported by the FT-IR data, Fig. 5) (Malayil et al., 2022) creating cavities in the hemp fiber structure for better penetration of potassium
leading to expanded carbon structures and higher surface area AC’s. Lignin is found interlinked to hemicellulose in biomass structure
and earlier studies in this lab on dilute acid hydrolysis of substrates such as almond skin with MSA has shown its potential in sepa-
rating the cellulose microfibrils due to delignification along with hemicellulose removal (Malayil et al., 2022). Partial delignification
along with hemicellulose removal has increased the surface area and micropore volume dramatically (Table 3). Use of MSA in dilute
acid hydrolysis for xylose recovery is relatively new and barely researched topic with minimal information available in literature for
comparison. Some interesting aspects for future study would be the mechanism of action in removal of lignin by MSA, affinity of
MSA towards type of lignin (S, G and H unit), its effect on physical and chemical structure of biomass and downstream application
of hydrolyzed fibers in making high surface area AC’s, nano cellulose, bioethanol, chemicals etc.

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Fig. 7. The SEM images of activated carbon made from (a) as is (As received), (b) as is with KOH activation, (c) SA hydrolyzed, (d) SA hydrolyzed
with KOH activation, (e) MSA hydrolyzed and (f) MSA hydrolyzed with KOH activation.

The yield of AC obtained from hemp as is and after dilute acid hydrolysis with and without KOH activation is shown in Table 3.
Maximum achievable yields of AC’s with surface area of > 1000 m2 /g from biomass is usually between 5 % and 12 % (Danish and
Ahmad, 2018) and MSA hydrolyzed hemp seed hull with KOH gave a reasonable good yield of 10.50 % (Table 3). The yields of
MSA+KOH AC’s was 10.5 % lower than SA+KOH and could be attributed to the additional removal of hemicellulose and lignin by
MSA. A similar loss in yields (6.2 %) without activation SA (27.58 %) and MSA (25.94 %) can be observed.

3.5.2. The FT-IR analysis of AC

The FT-IR analysis of AC made from dilute acid hydrolyzed hemp hull with and without activation providing information on
the chemical structural changes is shown in Fig. 6. Fig. 6 indicates a flat line with no peaks for the MSA hydrolyzed KOH activated
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AC suggesting pure carbon structure. The peak assigned to –OH stretching vibration of hydroxy functional groups usually present at
3430 cm–1 is absent for all the AC’s. The peak at 2921 cm–1 assigned to symmetric and asymmetric vibrations of methyl and methylene
groups is present is most of AC’s except the MSA hydrolyzed and KOH activated AC. Similarly, peaks assigned at 1744 cm–1 for C=O in
acetyl group and carboxylic group is present in AC made from MSA hydrolyzed hemp is absent in KOH activated MSA hydrolyzed hemp
hulls. Peaks at 1366 cm–1 assigned for C–H stretching and 1218 cm–1 for –OH in plane bending were all absent in MSA hydrolyzed
and KOH activated AC. Partial removal of lignin along with hemicellulose with MSA hydrolysis has a great positive impact on the AC
wherein pure hydrophobic uniform carbon structures can be obtained.

3.5.3. The SEM images of AC

The SEM images exhibiting changes in the surface properties of carbonized hemp seed hull with hydrolysis (SA/MSA), and with
and without KOH activation is shown in Fig. 7. Carbonized fibers made from post hydrolysis (SA and MSA) hemp seed hull exhibited
increased surface area (Fig. 7c and e). This is due to removal of hemicellulose causing increased acid penetration acting as an activation
agent. An interesting observation made in this study is that the AC made with MSA hydrolyzed hemp hull exhibited unique surface
modifications where in the material had minute pore like structures on the surface (Fig. 7e) along with better exposed cellulose
fiber bundles and can be due to the delignification properties of MSA needing further study. The AC’s made from SA hydrolyzed
hemp seed hull had smooth obsidian like surface, unlike MSA hydrolyzed substrate. The KOH activation made the surface of the
material more porous like pumice stone with increased surface area. Unhydrolyzed hemp seed hull with KOH activation had more
large particulate matter whereas both SA and MSA hydrolyzed, and KOH activated hemp seed hull AC was fluffier and finer particle
with porous structure. Even though MSA hydrolyzed, and KOH activated AC exhibited higher surface area the particle size and
surface morphology were like SA hydrolyzed and KOH activated AC. Similar surface morphology was observed for AC made from SA
hydrolyzed corn fiber with KOH activation (Fonseca et al., 2014; Herde et al., 2020).

4. Conclusions

The present work shows the effectiveness of methane sulphonic acid (MSA) in hemp seed hull hydrolysis for extracting xylose
and subsequent conversion of the hydrolyzed biomass to high surface area activated carbon (AC) for a C-5 biorefinery process. Dilute
acid hydrolysis of hemp seed hull with MSA at an acid loading of 1.8 % was found to yield 26.15 g/L of xylose which was 31 %
higher yields when compared to traditional dilute acid hydrolysis with sulphuric acid (SA) at the same acid loading rate. The residual
hemp hull fibers after hydrolysis were activated with KOH and carbonized to produce AC’s. Our work shows that AC produced from
MSA hydrolyzed and KOH activated hemp seed hull delivered an order of magnitude higher surface area when compared to SA
hydrolyzed and KOH activated AC. The surface area of pre-hydrolyzed and KOH activated hemp seed hull was only 706 m2 /g when
compared to SA hydrolyzed and KOH activated AC (977 m2 /g). However, the surface area increased dramatically when hydrolyzed
with MSA and activated with KOH to 1452 m2 /g with increased micropore volumes. The FT-IR analysis of the AC indicates that
the MSA hydrolyzed, and KOH activated AC had pure uniform hydrophobic carbon structures and SEM images of the same indicate
fine fluffier carbon structure. The MSA is found to have delignification properties and therefore removal of hemicellulose along with
lignin leads to formation of pure cellulose structures and this property has a great positive impact on high surface area AC formation.
These results indicate the potential of MSA in dilute acid hydrolysis of biomass for xylose recovery and production of high surface
area activated carbon. From a production standpoint this can lead to increased use of sustainable low-cost agricultural biomass for
making high surface area AC as components in supercapacitors.

Declaration of competing interest

The authors declare there is no conflict of interests.

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