Renewable Energy

Volume 103, April 2017, Pages 490-500

Comprehensive characterization of lignocellulosic biomass through proximate,

ultimate and compositional analysis for bioenergy production
Yengkhom Disco Singh a, Pinakeswar Mahanta a b, Utpal Bora a c d

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Abstract

We report here the characterization of five biomass samples (Impereta cylindrica, Eragrostis airoides, Typha angustifolia
L., Arundinella khasiana Nees ex Steud, and Echinochloa stagnina (Retz.) P. Beauv) based on the proximate, ultimate and
compositional analysis. The biomasses were examined physico-chemically and characterized to understand their
compositional and structural properties. The moisture content was found to be highest in Typha angustifolia (13.951%)
and lowest in Eragrostis airoides (8.275%). Ash content was seen to be maximum in Arundinella khasiana (8.12%) and
minimum in Eragrostis airoides (3.660%). Derivative Thermogravimetric (DTG) peak was observed below 120 °C
indicating the loss of water molecules from the biomass. Cellulose degradation occured between 350 °C to 500 °C. The
maximum carbon content was visible in Typha angustifolia (52.895%) and minimum in Eragrostis airoides (41.024%). The
FTIR spectra showed a range of peaks such as 3450 cm−1, 2860 cm−1, 1668 cm−1, 1175 cm−1, 1097 cm−1, 7872 cm−1,
625 cm−1, 554 cm−1 etc. The cellulose content was found maximum in Eragrostis airoides (43.17%) and minimum in
Echinochloa stagnina (24.90%). The results demonstrate that the collected lignocellulosic biomass could be potential
candidate for bioethanol production.

Introduction

Currently climate change and diminishing oil supplies are key issues for most countries in the world. The available data
shows that the global consumption of petroleum based fuel was 85.7 million barrels per day (BPD) in 2008, which is
estimated to increase to 97.6 million (BPD) in 2020 and 112.2 million (BPD) by 2035 [1]. The imbalance between the
increased demand and shortage for energy will be a global challenge in near future [2]. In this scenario, bioethanol is
considered to partly replace the fossil-derived fuels [2]. In developing countries, the use of lignocellulosic biomass
(LCB) could significantly increase the bioethanol production and seems to be a promising candidate for the future
bioenergy needs.

Lignocellulosic biomass includes agricultural residues, energy crops, grasses, wood residues, forest residues and
municipal paper waste. Most of the lignocellulosic biomass is composed of 35–55% cellulose, 20–40% hemicellulose,
and 10–25% lignin. The lignocellulosic biomass vary in composition [31] depending on the type of biomass, locality,
climatic conditions and the soil where it grows. The sugar (pentose and hexose) composition in the biomass decides the
theoretical ethanol yield. It is the interaction between all these parameters that enables the biomass to be considered
as economically efficient energy resources. The qualitative and quantitative characterization of biomass components
are necessary from its application perspectives. This could be achieved only when a fundamental characterization of
biomass is done prior to microbial fermentation [3]. Hence, systematic characterization of biomass is indispensable in
biofuel production.

The use of LCB as feedstock for biofuel production will decrease the harmful effects caused by emission of CO2 gasses
associated with traditional fuels. Biomass derived fuels do not contribute to increase of CO2 in the atmosphere as the
amount released upon burning of such fuels do not exceed that fixed during photosynthesis. Therefore, the net
production of carbon dioxide to the atmosphere is balanced, and this makes it a remarkable bioenergy source in the
present context of environmental pollution caused by greenhouse gasses [4].

The physical characteristics of biomass are determined by proximate analysis which gives an estimate of moisture,
volatile matter, ash, fixed carbon, total solid, calorific value, decomposition temperature by thermogravimetric analysis
(TGA) and crystallinity by X-ray diffraction (XRD) study. These parameters affect the combustion behavior of the fuel
[16]. On the other hand, chemical analysis of biomass samples consists of ultimate analysis, which gives information
about the elemental composition. Usually, the major elemental components such as carbon, hydrogen, nitrogen, oxygen
and trace amount of sulphur are analyzed in a biomass sample.

The North-Eastern region of India is ecologically enriched with innumerable unexplored microbes, plant and animal
species. The region is one of the well-known global biodiversity hotspots of India which comes under the Indo-Burma
(Myanmar) and Himalayan Biodiversity regions. The region which is home to various native species of plants and
animals have plenty of lignocellulosic biomass, which grows at high terrain regions. These biomasses remains un-
explored for bioethanol production. Here an attempt is made to explore such new feedstock for biofuel production. The
collection of species was done randomly by considering the abundance in a locality.

In brief, we have collected Impereta cylindrica, Eragrostis airoides, Typha angustifolia L., Arundinella khasiana Nees ex
Steud and Echinochloa stagnina (Retz.) P. Beauv. and characterized the lignocellulosic biomass with proximate, ultimate
and compositional analysis to know their potential for bioethanol production.

Section snippets

Biomass sample collection

In this experimental study, the biomasses were collected from two states (Assam and Manipur) of North-Eastern India.
The latitude and longitude of the collection sites were recorded. Photographs were taken for each biomass sample. The
amount of feedstock collected was between 3 and 5 kg in weight. The visible sand, soil, and other contaminants were
removed manually. The experimental data presented are the average of three readings. The general outline of the
experimental methods is given in …

Biomass sample collection

The five biomass species collected for the study were identified as Impereta cylindrica, Eragrostis airoides, Typha
angustifolia L., Arundinella khasiana Nees ex Steud, and Echinochloa stagnina (Retz.) P. Beauv. The systematic
classification of the selected biomass along with their respective site of collection is given in Table 1.…

Discussion
 REVIEW
published: 08 February 2021
doi: 10.3389/fphar.2020.602364

Cordyceps spp.: A Review on Its

Immune-Stimulatory and Other
Biological Potentials
Gitishree Das 1, Han-Seung Shin 2, Gerardo Leyva-Gómez 3, María L. Del Prado-Audelo 3,
Hernán Cortes 4, Yengkhom Disco Singh 5, Manasa Kumar Panda 6, Abhay Prakash Mishra 7,
Manisha Nigam 8, Sarla Saklani 9, Praveen Kumar Chaturi 8, Miquel Martorell 10,
Natália Cruz-Martins 11,12,13, Vineet Sharma 14, Neha Garg 15, Rohit Sharma 14 and
Jayanta Kumar Patra 1*
1
Research Institute of Biotechnology and Medical Converged Science, Dongguk University-Seoul, Goyangsi, South Korea,
2
Department of Food Science and Biotechnology, Dongguk University-Seoul, Goyangsi, South Korea, 3Departamento de
Farmacia, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad de México, Mexico, 4Laboratorio de
Medicina Genómica, Departamento de Genética, Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra, Ciudad de
México, Mexico, 5Department of Post-Harvest Technology, College of Horticulture and Forestry, Central Agricultural University,
Pasighat, India, 6Environment and Sustainability Department, CSIR-Institute of Minerals and Materials Technology, Bhubaneswar,
India, 7Adarsh Vijendra Institute of Pharmaceutical Sciences, Shobhit University, Saharanpur, India, 8Department of Biochemistry,
H. N. B. Garhwal University, Srinagar Garhwal, India, 9Department of Pharmaceutical Chemistry, H. N. B. Garhwal University,
Edited by:
Srinagar Garhwal, India, 10Department of Nutrition and Dietetics, Faculty of Pharmacy, and Centre for Healthy Living, University of
Cheorl-Ho Kim,
Concepción, Concepción, Chile, 11Faculty of Medicine, Alameda Prof. Hernani Monteiro, University of Porto, Porto, Portugal,
Sungkyunkwan University, 12
Institute for Research and Innovation in Health, University of Porto, Porto, Portugal, 13Laboratory of Neuropsychophysiology,
South Korea
Faculty of Psychology and Education Sciences, University of Porto, Porto, Portugal, 14Department of Rasa Shastra and
Reviewed by: Bhaishajya Kalpana, Faculty of Ayurveda, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India, 15Department
Young-Ji Shiao, of Medicinal Chemistry, Faculty of Ayurveda, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India
National Research Institute of Chinese
Medicine, Taiwan
Ulrike Lindequist, In recent decades, interest in the Cordyceps genus has amplified due to its
University of Greifswald, Germany
immunostimulatory potential. Cordyceps species, its extracts, and bioactive
*Correspondence:
constituents have been related with cytokine production such as interleukin (IL)-1β, IL-
Jayanta Kumar Patra
[email protected] 2, IL-6, IL-8, IL-10, IL-12, and tumor necrosis factor (TNF)-α, phagocytosis stimulation of
immune cells, nitric oxide production by increasing inducible nitric oxide synthase activity,
Specialty section: and stimulation of inflammatory response via mitogen-activated protein kinase pathway.
This article was submitted to
Ethnopharmacology,
Other pharmacological activities like antioxidant, anti-cancer, antihyperlipidemic, anti-
a section of the journal diabetic, anti-fatigue, anti-aging, hypocholesterolemic, hypotensive, vasorelaxation,
Frontiers in Pharmacology
anti-depressant, aphrodisiac, and kidney protection, has been reported in pre-clinical
Received: 03 September 2020
studies. These biological activities are correlated with the bioactive compounds present in
Accepted: 02 December 2020
Published: 08 February 2021 Cordyceps including nucleosides, sterols, flavonoids, cyclic peptides, phenolic,
Citation: bioxanthracenes, polyketides, and alkaloids, being the cyclic peptides compounds the
Das G, Shin H-S, Leyva-Gómez G, most studied. An organized review of the existing literature was executed by surveying
Prado-Audelo MLD, Cortes H,
Singh YD, Panda MK, Mishra AP,
several databanks like PubMed, Scopus, etc. using keywords like Cordyceps, cordycepin,
Nigam M, Saklani S, Chaturi PK, immune system, immunostimulation, immunomodulatory, pharmacology, anti-cancer,
Martorell M, Cruz-Martins N,
anti-viral, clinical trials, ethnomedicine, pharmacology, phytochemical analysis, and
Sharma V, Garg N, Sharma R and
Patra JK (2021) Cordyceps spp.: A different species names. This review collects and analyzes state-of-the-art about the
Review on Its Immune-Stimulatory and properties of Cordyceps species along with ethnopharmacological properties, application
Other Biological Potentials.
Front. Pharmacol. 11:602364.
in food, chemical compounds, extraction of bioactive compounds, and various
doi: 10.3389/fphar.2020.602364 pharmacological properties with a special focus on the stimulatory properties of immunity.

Frontiers in Pharmacology | www.frontiersin.org 1 February 2021 | Volume 11 | Article 602364

Das et al. Cordyceps Spp., An Immunity Stimulator

Keywords: ethnopharmacology, cordyceps, cordycepin, natural medicine, immune system, immunostimulation,

immunomodulatory, clinical studies

INTRODUCTION Direct, Scopus databases, Google search, etc., using keywords

and its combination like Cordyceps, cordycepin, natural medicine,
The fungus Cordyceps spp. belongs to Tibetan medicine and immune system, immunostimulation, immunomodulatory,
consumers describe it as an important source of energy. pharmacology, anti-cancer, anti-viral, clinical trials,
Cordyceps spp. belongs to Ascomycota, Pyrenomycetes, ethnomedicine, pharmacology, phytochemical analysis, and
Hypocreales, and Clavicepitaceae, and at least 700 species are different species names. Usually, the search was carried out in
known. The word Cordyceps originates from the Greek term “title, abstract, and keyword” fields. In each search, normally the
"kordyle", which means “club”, and the Latin etymon “ceps”, review articles were omitted, however, in some instances, some
which means “head” (Olatunji et al., 2018). Cordyceps species important review articles were also considered. Further only
invade insects, arthropods, other fungi, and evades the host articles published in the English language were considered.
immune system by harmonizing the life cycle of its host with Articles that were published with only basic ethnobotanical
the intention of survival and multiplication. Their interaction assessment reports which lack substantial proof of the claim
with the host will produce different secondary metabolites were not included in the study.
(Olatunji et al., 2018), such as cordycepin, adenosine,
guanosine, cordymin, γ-aminobutyric acid (GABA),
exopolysaccharides, cordysinin A-E, among others (Liu Y HABITAT, DISTRIBUTION, AND
et al., 2015). CHARACTERISTICS OF CORDYCEPS SPP.
The different species of Cordyceps have beneficial properties
such as anti-cancer, anti-proliferative, anti-angiogenic, anti- From the more than 700 species of mushrooms recognized on the
metastasis, apoptosis induction, anti-inflammatory, antioxidant, genus Cordyceps, around 20 species parasitize on the genus
anti-fibrotic, anti-arteriosclerosis, anti-hypertensive, anti- Elaphomyces, meanwhile the remaining species do on insects and
thrombotic, antimalarial, antifungal, hypolipidemic, antidiabetic, arthropods belonging to Arachnida, Hymenoptera, Isoptera,
hypoglycemic, anti-asthmatic, steroidogenesis, spermatogenic, Coleoptera, Hemiptera, and Lepidoptera classes. This diversity
anti-aging, and immunomodulatory effects (Liu Y et al., 2015). of species includes the C. sinensis (Ophiocordyceps sinensis
These properties are concentration-dependent, and in most cases, (Berk.) G.H.Sung, J.M.Sung, Hywel-Jones and Spatafora), C.
no adverse effects were reported, although the evaluation of ophioglossoides (Tolypocladium ophioglossoides (Ehrh.)
isolated compounds such as cordycepin is preferred. Quandt, Kepler & Spatafora), C. militaris (L.) Fr., C. gracilis
Interestingly, Cordyceps spp. contains different compounds (Grev.) Durieu & Mont., C. sobolifera (Hill ex Watson), C.
with the ability to strengthen the response of the immune system subsessilis Petch, C. gunnii (Berk.) Berk., C. cicadae S.Z. Shing,
and also to control its exacerbated response. Most of the C. tuberculate (Lebert) Maire, C. scarabaeicola Kobayasi, C.
information on the effect of Cordyceps on the immune minuta Kobayasi, C. myrmecophila Ces., C. canadensis Ellis &
system derives from studies in cancer. In particular, Everh., C. nutans Pat., C. agriota A. Kawam., C. ishikariensis M.
Cordyceps spp. increases the production of interleukin (IL)- Zang, D. Liu and R. Hu, C. sphecocephala (Berk.) Sacc, C.
1β, IL-2, IL-6, IL-8, IL-10, IL-12, and tumor necrosis factor konnoana Kobayasi & Shimizu, C. nigrella Kobayasi &
(TNF)-α, induces phagocytosis of macrophages, mononuclear Shimizu, C. pruinosa Petch, C. tricentri Yasuda, among others
cells, nitric oxide (NO) release, and stimulates the (Tuli et al., 2013a; Lo et al., 2013; Baral et al., 2015; Pal and
inflammatory response via the mitogen-activated protein Misra, 2018).
kinase (MAPK) route (Lee et al., 2006; Wang M et al., These species exhibit different characteristics such as
2012). Furthermore, it presents synergism with interferon pharmaceutical properties, making them attractive to
(INF)-γ in the production of cytokines. These properties are traditional Chinese medicine (TMC) since the nineties, and
attractive in the search for new applications where the being C. sinesis the most studied and applied. Their
stimulation in the immune system response is wanted. geographic distribution is mainly based on the host
Therefore, this review collects and analyzes the state-of-the- distribution; however, they can grow in high mountains at an
art about properties of Cordyceps spp. focused on the altitude of 3,600–4,000 m above the sea level. Thus, Cordyceps
stimulatory properties of immunity. spp. has been found in North America, Europe, and Asia, mostly
in countries such as China, Japan, Nepal, Bhutan, Vietnam,
Korea, and Thailand. In India, it is principally present in
METHODOLOGY subalpine regions such as Kumaun Himalaya and Garhwal
Himalaya (at higher altitudes) (Maity, 2013; Chakraborty
An organized review of the existing literature was executed by et al., 2014). Furthermore, it has been reported that species
surveying pertinent peer-reviewed research articles, review such as C. gunnii (Berk.) Berk. was found in Australia
articles, etc. from several available bibliographic databanks (Olatunji et al., 2018). The composition of their metabolite
such as PubMed, SpringerLink, Elsevier journal, Science makes them able to tolerate characteristic severe conditions at

Frontiers in Pharmacology | www.frontiersin.org 2 February 2021 | Volume 11 | Article 602364