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1 Agro-Wastes-Based Feedstock as a Source for Bionanomaterials Production…
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24h and centrifuged. The obtained powder was analyzed for its physical characteristics. UV adsorption spectra within the range of 350–450nm conrmed the formation of AgNPs, and they have signicant antibacterial activity against Salmonella
typhimurium, S. aureus, E. coli, and K. pneumoniae in comparison with standard
antibiotic, streptomycin. Also, free radical scavenging assay of AgNPs indicated the
enhancement of antioxidant activity from 40% to 88%, paving ways for several
biomedical applications (Karunakaran etal. 2016). Synthesis of ZnO nanoparticle
from Nyctanthes arbor-tristis (night owering jasmine) using zinc acetate dihydrate
resulted in crystalline-structured nanoparticles in the size range of 12–32nm, proving to be an efcient antifungal agent against plant pathogens—Alternaria alter-
nata, Aspergillus niger, Botrytis cinerea, Fusarium oxysporum, and Penicillium
expansum. Highest minimum inhibitory concentration values were observed at
128μg/mL for Botrytis cinereal and Penicillium expansum (Jamdagni etal. 2018).
Metallic nanocomposites in the eld of pharmacology is rarely studied. Spherical
Au nanomaterials, with the average diameter range of 15–45nm synthesized from
ower extract (aqueous) of Hibiscus sabdariffa, had a potency in controlling
myeloid leukemia effect in rodents. Induction of acute myeloid leukemia was done
in 75 mice, which were divided into 6 subgroups—HAucl4, Hibiscus sabdariffa,
AuNPs, daunorubicin, untreated, and control. AuNPs enhanced the antiinammatory cytokines parameters as compared to untreated mice. The signicant
increase in antioxidant potential of the Au-treated mice is in par with that of
daunorubicin- treated one, proving the potential of Au-NPs in the eld of drug discovery (Zangeneh and Zangeneh 2020).
13
1.7 Bionanomaterials fromSeed
Nanoproducts made out of seed extracts have several antimicrobial and anticytotoxic properties. Au and Ag nanomaterials have good antibacterial effect, like that
of Zn and copper nanomaterials. A study was conducted on Phoenix sylvestris (silver date palm) seed extract as a reducing agent for shaping AgNPs from AgNO3.
The total phenolic and avonoid content of the seed extract was assessed and free
radical scavenging activity was tested with DPPH-scavenging assay and bactericidal activity analysis was carried out in Broth microdilution and disk diffusion
methods. The higher amount of avonoid content in the seed extract contributed to
the higher antibacterial and antioxidant activity against Acne vulgaris, the acnecausing pathogen. Despite having high microbicidal activity, AgNPs as a cosmeceutical nanocomposite are not yet reported. Depending on their physical and
morphological characteristics, AgNPs might be an effective tool in drug discovery
(Qidwai etal. 2018). Unlike biological and fabrication applications certain nanomaterials from seed extracts have conducting applications. For example, Caesalpinia
bonducella (fever nut) seed extracts were used to reduce Cu (NO3)2·3H2O resulting
in the formation of rice-shaped CuONPs. CuONPs were incorporated in parafnimpregnated graphite electrode (Cu-PIG) to detect the presence of riboavin. The

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stability of Cu-PIG stands strong and satisfactory for 120days and makes it suitable
to apply in real samples such as egg yolk, milk powder, and B-complex tablets.
Recovery rates and detection sensitivity extend the nanofabricates into the eld of
electrochemical sensing (Sukumar etal. 2020).
P. Sankaranarayanan et al.
1.8 Nanoproducts fromSugarcane Bagasse
Sugarcane bagasse, a biomass fraction obtained after extraction of sugarcane
juice, is predominantly rich in cellulose, hemicellulose, and lignin. Almost 2–5%
of the pulp and paper industry uses lignocellulosic biomass (sugarcane bagasse,
straw) as its raw material. Silica from sugarcane bagasse can be modied with
polyethylene imine (PEI) to produce magnetic paper. Silica-PEI and Cobalt ferrite
nanoparticles were co-precipitated via the lumen loading process resulted in
agglomeration due to their interaction with cellulose bers to inculcate the magnetic ability. The magnetic property of the bers was analyzed through its physical
properties such as, the idea of replacing cobalt ferrite in place of magnetite results
in higher magnetization, enhanced magnetic coercivity, tensile strength, hardness,
and chemical stability. Toxicity analysis of these magnetic bers must be overlooked to be a potential magnetographic printing paper (Piñeres etal. 2020). The
sugarcane bers’ higher lignin and cellulose content contributes to the adsorption
property. Using bioadsorbents in dye removal was previously done using rice husk
ash and straw ash. Sugarcane bagasse was scrutinized for its adsorption properties
and showed a maximum adsorption capacity of 9.41mg/g on methylene blue dye
in wastewater. Chemical reaction kinetic studies describe pseudo-second-order
kinetics, and the process is endothermic, spontaneous, and feasible. High availability, excellent adsorption property, and eco-friendly properties of sugarcane
bagasse help it to be a better alternative source in wastewater treatment (Siqueira
etal. 2020).
1.9 Agro-Wastes inImproving Soil Fertility
Quality of plantation greatly depends on the cultivability and nutrient availability of
the soil. Synthetic fertilizer cannot maintain the fertility of the soil due to its toxicity, which leads to less nutrient uptake by plants. To enhance optimal production,
agricultural wastes should be used as a compost to increase the organic materials in
soil, providing long-term soil fertility. The reusability of plant wastes in agriculture
might make the food production unit self-sufcient. For example, the natural fertilizer prepared with mixture of sugarcane bagasse, cow manure, and lter mud
yielded maximum saplings and showed better growth and higher stem weight compared to basic inorganic NPK fertilizers (Rahmad etal. 2020). Another eld experiment study was conducted to investigate the effect of biochar, rice husk, and rice

1 Agro-Wastes-Based Feedstock as a Source for Bionanomaterials Production…
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straw on maize yield. The combination of biochar with NPK fertilizer yielded 7.5t/
ha grain yield of maize and improved the owering rate signicantly. Soil fertility
rates increased with the usage of higher proportion of agri-composting and lesser
quantity of inorganic fertilizers. This outcome may be suitable for smallholder
farmers to enhance the nutrient value of fragile lands with plantation wastes as an
integrated soil management approach (Dzomeku etal. 2018).
15
1.10 Metallic Nanoparticles asPesticides andInsecticides
Metallic nanoparticles and their role in bactericidal and fungicidal activities were
well studied. But the synergistic activity of nanoparticles with pesticides/antibiotics
was least evaluated. The usage of chemically synthesized AgNPs as pesticides and
bactericides is commercially available (Rahmad etal. 2020). But the development
of agricultural waste-based cidal agents was least focused. A nanocomposite developed by integrating AgNPs synthesized from Ligustrum lucidum (Glossy Privet)
leaf extracts with epoxiconazole was evaluated for its antifungal property against
Setosphaeria turcica, and its IC50 was found to be 170.20 μg/mL. Synthesized
AgNPs when combined with fungicide epoxiconazole at a ratio of 9:1 can be an
efcient fungistat. Further research regarding AgNPs properties such as surface
charge, acid-base property, aggregation behavior and its fate on the environment
must be carried out to make it a potent pesticide (Huang etal. 2020). Usage of copper and selenium nanoparticles against pests, insects, and larvae was extensively
studied. An eco-friendly approach of using metal-contaminated soil-based copper
nanoparticles against Tribolium castaneum excelled than those of synthetic pesticides (El-Saadony etal. 2020).
1.11 Limitations
Despite having several advantages of nanomaterials, there are debates regarding
their adverse effects on the environment. Initially, nanoparticles in land have the
potential to contaminate groundwater bodies and enter human body leading to pulmonary disorders. Nano TiO2 and ZnO used in cosmetics enter human skin, causing
skin allergies and organ toxicity. Though Au and AgNPs have efcient cytotoxic
activities, some amount of cellular damage is inevitable. This can be overcome by
the application of citrate-capped gold nanospheres, which have an effective cytotoxic effect, followed by its less toxic nature. Literature reviews suggest that many
of the surfactants used in the manufacture of nanospheres are toxic (Ray etal. 2009).
The choice of plants as precursors might be an alternative source for producing toxic
nanomaterials. Meanwhile, there are two types of aggregation of nanomaterials in
the environment—homo- and hetero-aggregation. Those that exist in waterdispersible colloidal form such as multi-walled carbon nanotubes have more

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surfactants, which get deposited in the soil for a long time, thereby affecting their
stability.
In case of nanofertilizers, the biochemical composition of nanoparticles greatly
impacts its fate on the environment. The use of “green” precursors was found to
be less polluting, as they compost easily. Most chemically synthesized inorganic
nanoparticles have toxic effects compared to organic nanoparticles. Since there
are very few studies of nanomaterials in soil fertility, impact of nanomaterials on
soil nutrition as well as with soil microbes is unclear. Progress in the development
of tailored nanomaterials has a precise control over their efciency.
Bionanomaterials and their role in bioremediation need to be assessed in order to
view their capability in integrated remediation. Meanwhile, nano-assisted farming
requires experimentation to be performed under controlled conditions, with a variety of parameters optimized. Once the investigation regarding nano-agriculture is
completed in small scale, eld study has to be performed to evaluate its toxicity
(Usman etal. 2020).
In recent times, the use of nanomaterials excelled in the eld of medicine especially in case of sensing and drug delivery applications, marking its breakthrough
in the management of several diseases. Still, several mysteries around the fate of
nanomedicine in body’s defence mechanism are not clear. But those therapies
dealing with gene alterations and pre-natal stage treatments were not permitted,
owing to their fatal outcomes. Similarly, workplace safety must be maintained
while working with nanomaterials as they are too small and might enter patients’
bodies, causing severe respiratory problems such as asthma and cold. Toxicity
evaluation must be carried out to work with nano-fabricated devices (Odiba
etal. 2017).
P. Sankaranarayanan et al.
1.12 Signicance ofUsing Bionanomaterials
fromAgro-Wastes
Nanomaterials produced from agro-wastes have lesser toxicity than that of the other
green synthesis routes, mainly because of the organic compounds in the plant
sources, such as proteins and avonoids. Despite having lesser applications of bionanomaterials from agro-wastes in terms of electronics, food safety, and energy, the
availability of literature on green synthesis makes it easier to produce nanocompounds at a cheaper rate (Odiba etal. 2017). The conversion of waste to wealth,
especially in nano-applications, helps reroute the waste into raw materials, making
it suitable for several applications. Agro-wastes-based bionanomaterials serve as a
feedstock for soil fertility applications in nanosensors, food safety, bionanopesticides, biocatalysis, and biostimulant for ower, fruit, and seed production (Ray
etal. 2009). Novel toxicity evaluation approaches for agro-based bionanoproducts
must be extensively carried out to determine the futuristic prospects of using agrowastes in nano-synthesis.

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17
1.13 Conclusion
This chapter has highlighted the outcomes of agro-based bionanomaterials and their
signicance in the eld of health care, electronics, agriculture, and engineering
applications. Metallic nanobiomaterials have a greater demand in the near future
due to their environmentally friendly synthesis and commercial viability.
Furthermore, effective techniques have to be adapted in making the synthesis in
simpler forms. The use of easily accessible raw materials such as agro-wastes, metal
contaminated soils, and pre-processed industrial wastes might be a forthcoming
energy source. Applications of bionanomaterials from agro-wastes are found to be
suitable for precision agriculture in different forms such as nanofertilizers, nanoinsecticides, and nanopesticides. To conclude, scrutiny of plant-based bionanomaterials in day-to-day applications has to be assessed. More research revelations from the
academic sector might add up more elite bionanomaterials for positive considerations in commercialization.
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P. Sankaranarayanan et al.

Chapter 2
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Application ofBiotechnology forRaw
Material Analysis
SomsubhraGhosh, SujayitaMazumder, RajeswarDas, andSandipanJana
Abstract Raw materials are the most important substance in the pharmaceutical
and biotechnology industries nowadays. These are obtained and produced from
different sources, by using several techniques. These are available in different
forms and phases, so the processing, storage, and analytical process of the raw
materials are very crucial and essential to maintain the highest standard of the
nished product. Several instruments and analytical processes are available and
used frequently in the industry for routine checkup of the raw materials.
Conventional analytical techniques like spectroscopy, chromatography, etc. are
mostly used among all other analytical techniques. Some the techniques are either
time-consuming or very costly for routine analysis. Nowadays, several biotechnological techniques or processes are being used as alternatives to conventional techniques. Biotechnology is one of the emerging elds in the healthcare and
pharmaceutical industry. It has several applications and uses, including patient
care, treatment of several diseases, and even for diagnostic purposes. Apart from
all those conventional uses, in the recent past, biotechnological techniques have
also been used for analyzing and testing raw materials. A detailed discussion is
provided systematically to highlight the application of biotechnology in the analysis of raw materials.
Keywords Biotechnology · Raw material · Analysis · 3D biological printing
S. Ghosh (*) · S. Mazumder · R. Das · S. Jana
School of Pharmacy, The Neotia University, Sarisa, West Bengal, India
e-mail: somsubhra.ghosh@tnu.in
Ltd. 2024
S. Bose et al. (eds.), Concepts in Pharmaceutical Biotechnology and Drug
Development, Interdisciplinary Biotechnological Advances,
https://doi.org/10.1007/978-981-97-1148-2_2
21© The Author(s), under exclusive license to Springer Nature Singapore Pte

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S. Ghosh et al.
2.1 Introduction
The rules and guidelines pertaining to biotechnology products do not provide a
uniform denition of a raw material. Regulations make reference to elemental substances, constituents, and in-process materials. A raw material is dened as “starting
materials, reagents, and solvents intended for the use in the production of intermediates or APIs” in the International Conference on Harmonisation (ICH) Q7. A monoclonal antibody used as a raw material in afnity chromatography is known as ICH
Q6B.As a result, the producer must dene a raw material’s meaning according to
its own standards on safety, quality, and adherence to the current Good Manufacturing
Practices (cGMPs). A thorough assessment of the factors that must be taken into
account for novel compounds is necessary to show that the raw material satises
acceptable quality standards for use in the production of biotechnology products
intended for human consumption. No specic threshold has been set for how much
a raw material must adhere to cGMP guidelines (Noh etal. 2018). Numerous biotechnology and pharmaceutical enterprises, such as those afliated with the Rx-360
Consortium, prioritize the oversight of supply quality, including raw materials. ICH
Q9 provides guidance on incorporating quality risk management into materials
management procedures. Since a biotechnology product may be manufactured
using a range of raw materials, certain considerations are made when introducing a
new raw material. Presently, the Chinese Pharmacopoeia (2020) uses two biotechnologies for the analysis of raw materials for animal and herbal medicine: polymerase chain reaction (PCR) and polymerase chain reaction-restriction fragment
length polymorphism (PCR-RFLP) (Li etal. 2016).
2.2 Molecular Methods toVerify Raw Material Authenticity
Progress in sequencing technology, ongoing cost reductions in sequencing, collection of genetic data from a diverse range of species, and creation of multiple databases enable the advancement of molecular identication technology and the
breeding of new varieties. Some DNA-based technologies that are capable of precisely identifying raw materials are barcoding, molecular markers, and fast
detection.
Molecular markers: Molecular markers use DNA fragments carrying genetic
information that differs between species to determine the principles governing gene
organization and the manifestation of phenotypic features. They are resistant to differences in tissue and organ composition, changes in the external environment, and
developmental phases. They are able to recognize distinct tissues, organs, and even
individual cells in various stages of development. The markers have found widespread use in the study and identication of herbal medicines due to their stable
qualities, broad dispersion, abundance, ease of selection, ease of operation, and
speedy detection. For example, nucleotide sequences that have been inserted or
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