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these disorders are directly or indirectly connected with lipid metabolism. Inulin
has shown its efcacy in mitigating these disorders by altering lipid metabolism
(Chen etal. 2023). Studies on diabetic mice model with abnormal lipid metabolism
as well as gut microbiota dysfunction have shown that consumption of Jerusalem
artichoke inulin improves the biochemical parameters, including blood glucose,
total cholesterol, and pro-inammatory cytokine reduction (Li etal. 2022). Another
study conducted to determine the effect of inulin administration in pre-weaning
period in piglets has shown that supplementing inulin has a signicant effect in
lipid-metabolizing gene of the ileum (Schroyen etal. 2021). Furthermore, in obsess
ob/ob mice, inulin has shown ameliorative effect on metabolic disorder by remodel-
ing gut microbiota and enhancing the production of short-chain fatty acid level
through the expression of angiopoietin-like protein 4 (Guo etal. 2022).
2.7.1.4 Stimulator oftheImmune System
Metabolism of inulin occurs at the colon part of GIT which led to the production of
metabolites such as short-chain fatty acids. These fatty acids provide energy to
intestinal cells and further act in differentiation of immune cells (Sheng etal. 2023).
Research has shown that consuming inulin in diet alters the epithelium layer of the
colon by intestinal stem cell proliferation which has resulted in deeper crypts and
elongated colons (Corrêa etal. 2023). Furthermore, a study done on goats stated
that a low-carbon and high-inulin diet led to improvement in intestinal mucosal bar-
rier function and provided immunity against infection (Yuan etal. 2023).
2.7.1.5 Absorption Enhancer
Inulin has a signicant effect on the absorption of several elements such as calcium,
iron, and many more. It has been found to increase calcium absorption by passive
diffusion, through bolstering via ion exchange and expanding the surface for absorp-
tion by stimulating colon cell growth. Furthermore, it enhances calcium solubility
and stimulates the level of calcium-binding proteins expressions (Bakirhan and
Karabudak 2023). Furthermore, inulin has also shown effect on iron absorption. In
a study it was found that adding high-performance inulin, oligofructose, and syn-
ergy1in diet led to increase in intestinal absorption of iron in rats (de Cássia Freitas
etal. 2012). Another study stated that supplementing 4% of inulin in the diet of
piglets led to enhanced utilization of intrinsic iron in soya bean and corn diet
(Yasuda etal. 2006).
2.8 Pharmaceutical Benefits
2.8.1 Coating ofDrug
Inulin has a notable characteristic that withstands in the acidic environment of the
stomach and hydrolyzes only in the intestine by microbiota. Digestive enzymes
have no role in its metabolism and absorption. Therefore, it can be utilized as a
delivery vehicle for acid-liable drugs as well as for providing a sustained released
M. Vishwakarma et al.
37
action (Akhgari etal. 2009). Many researchers are unitizing this feature of inulin in
development of novel drug delivery vehicles which provides a protective layer to
drug and helps to overcome the drug’s limitations. In a research, inulin in combina-
tion with eudragit RS was used in the coating of 5-aminosalicylic acid (5-ASA) for
the treatment of inammatory bowel disease (IBD). The results have shown that the
formulations have the ability to withstand in the acidic pH and a sustained release
of pattern of 5-ASA was achieved in small and large intestines (Imran etal. 2012).
Similarly, the same combination was used for the delivery of budesonide for the
treatment of IBD (Soltani etal. 2023). Furthermore, in another study inulin was
modied to impart it with a positive charge by using glycidyl trimethyl ammonium
chloride for using it as a shell for the core of lecithin sodium acetate for the con-
struction of core-shell lipid nanoparticle for the co-delivery of curcumin and pacli-
taxel in the case of breast cancer. The developed formulation has shown enhanced
cytotoxic effect on MDA-MB-231 cell lines (Vatansever etal. 2023).
2.8.2 As Lyoprotectant andCryoprotectant
Several strategies have been developed by the researchers for protecting the vesicu-
lar drug from degradation and drug leakage during storage. Lyophilization is one of
the efcient methods generally used for removing the bounded and unbounded
aqueous content of the component in order to enhance their long-term stability.
Lyoprotectant plays a major role in lyophilization as it helps to prevent the compo-
nent damage. In several studies the efcacy of inulin as lyoprotectant has been ana-
lyzed. Inulin has been reported to enhance the stability of curcumin-loaded
liposomes as lyoprotectant (Jiang etal. 2023). Whereas, for the freeze-drying of
lactobacillus strain, it has also shown its efcacy as lyoprotectant for long-term stor-
age (Bodzen etal. 2021). Furthermore, in preservation of mammalian cell culture,
it has also shown its activity as a cryoprotectant (Tornacı 2022). Another research
has also shown that inulin can improve the cryopreservation of red blood cells by
promoting the vitrication and inhibiting the ice recrystallization (Hu etal. 2024).
2.8.3 In PEGylation
Systemically administered formulation should possess long circulatory and colloi-
dal stability in order to provide its therapeutic efcacy. PEGylation of formulation
with polyethylene glycol (PEG) was the gold standard for the development of long
circulator novel drug delivery system. But some cases that have been reported state
the development of PEG-directed immunoglobulins after rst administration of for-
mulation. Researchers have worked for the identication of another option for
PEGylation. Inulin has been explored for PEGylation of nanocarriers as an alterna-
tive of PEG.The developed formulation has shown its ability to replace PEG (Sardo
et al. 2022). Furthermore, a mucoadhesive PEGylated inulin-based nanoparticle
formulation that was developed by prima et al. for the enhancement of the
2 Extraction andPurication ofInulin
38
permeation of corticosteroids for transcorneal delivery has also shown its efcacy
(Di Prima etal. 2019).
2.8.4 Used asAdjuvant inVaccine
Apart from drug delivery, inulin has also shown its efcacy in developing an immune
response by working as adjuvant in vaccine. A multi-epitope subunit vaccine devel-
oped by inulin acetate microparticles decorated with mannan has shown immuniza-
tion efcacy in foot and mouth diseases (Yoon et al. 2020). Furthermore,
polycaprolactone- and inulin-based particles were developed to improve the hepati-
tis B (HBV) vaccination strategy. The formulation in the concentration of 4μg/mL
has shown its efcacy to produce Interleukin 12 and 10 (Duarte 2021). Furthermore,
delta inulin has shown its activity in intrapulmonary vaccination by stimulating
nonpolarized signaling chemotactic for combatting respiratory pathogens (Ferrell
etal. 2021). In another research delta inulin was used in mucosal immunization of
mice against SARS-CoV-2 which showed that developed vaccine could elicit lung
resident immune memory and provide protection (Stewart etal. 2022), whereas,
inulin has also shown immune enhancement ability with ptfA gene DNA vaccine
for chicken for Pasteurella multocida (Gong etal. 2020).
2.8.5 As Gel
Inulin has been reported to form gel with water when its concentration raised to
10–15/100g. Providing thermal treatment and shear force to inulin water suspen-
sion led to formation of a gel-like network. This property of inulin has been used in
many food and pharmaceutical industries (Florowska et al. 2020). Inulin oral
administration has reported to show antitumor immunity by modulating gut micro-
biota (Han et al. 2021). Furthermore, for the effective treatment of acute colitis,
orally administrated inulin gel containing Bacillus subtilis and antioxidant atomic
nanocatalyst has shown its therapeutic efcacy (Lu etal. 2024). In another study,
Jerusalem artichoke inulin was used as emulsion gel for delaying the linoleic acid
oxidation due to its higher phenolic content and this study also stated that emulsion
gel with inulin could be a potential fat replacer (Li etal. 2020). Some of the research
on inulin-based drug delivery systems is given in Table2.2.
2.9 Diagnostic Benefits
With therapeutic and pharmaceutical benet, inulin is also used in diagnosis of
many diseases. It has been used as biomarker for determining the glamour ltration
rate in analysis of kidney functioning (Gu and Yang 2022; Wu etal. 2022). Apart
from this, inulin-coated iron oxide super-paramagnetic nanoparticles have been
used in MR imaging in case of hepatic failure. In vivo studies have shown that the
M. Vishwakarma et al.
39
Table 2.2 Application of inulin in drug delivery
Carrier
Drug Disease Outcome
Reference
Inulin stearic
acid
bioconjugate
nanoparticles
Genistein Colon cancer Developed formulations
have shown controlled
drug release pattern with
potential cytotoxicity
against HCT 116
colorectal cell
Jangid
etal.
(2022)
Chitosan inulin
aldehyde
hydrogel
Dopamine and
indomethacin
– Excellent
cytocompatibility against
L-929 broblast
Rahnama
etal.
(2021)
Inulin-based
glycovesicle
Levooxacin Inammatory
bowel disease
Developed formulations
have shown good
antibacterial activity
against salmonellosis
Xu etal.
(2024)
Honey-
stabilized inulin
nanoparticles
Irinotecan
hydrochloride
Colon cancer Formulations have shown
more cytotoxicity and
apoptosis than pure drug
Joseph
etal.
(2024)
Inulin and
β-cyclodextrin
conjugate
Curcumin Colon cancer Cytocompatibility of
developed formulation
with Caco-2 cells was
veried
Catenacci
etal.
(2020)
Sodium alginate
and inulin
hydrogels
Bovine serum
albumin
– Formulations have shown
high stability in acidic
medium with good release
efcacy
Najwa and
Solehah
(2020)
Inulin-modied
double-layered
nanoparticles
Paclitaxel Orthotopic
colon cancer
Formulations have shown
its safe drug delivery
property after oral
administration
Hou etal.
(2022)
developed nanocomposites have excellent contrast between the normal liver and the
injured one (Kermanian etal. 2021). Inulin has also been utilized as a model mole-
cule for analyzing the effect of intranasal dosing speed and volume of administra-
tion in nose-to-brain delivery (Fukuda etal. 2021).
2.10 Limitations ofExtraction andPurification ofInulin
The extraction process is often labor-intensive and time-consuming, involving mul-
tiple complex steps such as homogenization, enzymatic treatment, and precipitation
(Du etal. 2023). Additionally, variability in yield and purity poses challenges, as
these are inuenced by factors like plant source, environmental conditions, and
extraction methods. Economic feasibility and environmental impact are also critical
concerns, with the need for cost-effective and sustainable extraction techniques
Lastly, regulatory hurdles can complicate the commercialization of inulin-based
products (Fuso 2023).
2 Extraction andPurication ofInulin
40
2.10.1 Complexity ofExtraction Processes
The process of extracting inulin from plant sources entails several intricate proce-
dures, such as enzymatic treatment, precipitation, ultrasound-assisted extraction
(UAE), microwave-assisted extraction (MAE), and supercritical uid extraction
(SFE). These complexities can limit the scalability of the process for industrial
applications. In a study by Lingyun etal., the enzymatic extraction of inulin from
Jerusalem artichoke (Helianthus tuberosus) was investigated. The researchers uti-
lized a combination of cellulase and inulinase enzymes to enhance the extraction
yield. While the process improved inulin recovery, it required precise control of
enzymatic activity, pH, and temperature, making the process intricate and sensitive
to operational conditions. The study concluded that although enzymatic extraction
is effective, the need for enzyme optimization adds to the complexity and cost
(Lingyun etal. 2007). Petkova etal. explored the use of ultrasound-assisted extrac-
tion (UAE) for isolating inulin from dahlia tubers. The application of ultrasound
was found to disrupt cell walls, thereby enhancing the release of inulin. However,
the study noted that the effectiveness of UAE is highly dependent on factors such as
ultrasound frequency, power, and extraction time. Moreover, the scale-up of UAE
for industrial purposes poses technical challenges due to the need for specialized
equipment and energy considerations (Petkova etal. 2018). Alabadi etal. conducted
a study on the microwave-assisted extraction (MAE) of inulin from Jerusalem arti-
choke (Helianthus tuberosus). The use of microwaves signicantly reduced the
extraction time and improved yield. Nonetheless, the process requires careful opti-
mization of microwave power and exposure time to prevent degradation of inulin.
Additionally, the high initial cost of microwave extraction systems can be a barrier
for widespread adoption in industrial settings (Alabadi and Abood 2020). A study
by Zhu etal. examined the supercritical uid extraction (SFE) of inulin from dahlia
tubers (Dahlia pinnata). SFE, utilizing carbon dioxide as a solvent, provided a high-
purity product with minimal environmental impact. However, the process requires
high-pressure equipment and precise control of temperature and pressure condi-
tions, which complicates the operational protocol and increases costs (Zhu etal.
2016). The researchers highlighted the need for further development to make SFE
more cost-effective and scalable.
2.10.2 Yield Variability
The yield of inulin is highly dependent on the plant source and the extraction method
used. Variations in environmental conditions, plant maturity, and processing param-
eters can lead to signicant uctuations in yield (Gholami etal. 2018). This vari-
ability poses challenges in standardizing the extraction process to achieve consistent
product quality. A study by Kanakasabai etal., which investigated the extraction of
inulin with fructo-oligosaccharides (FOS) from plant sources, such as chicory roots
(Cichorium intybus L.), was optimized using specic extraction conditions.
Variables including volume of water (10–75 mL), time (10–30 min), and
M. Vishwakarma et al.
41
temperature (30–90°C) were investigated for their effects on inulin yield through
single regression modeling. Additionally, the inuence of water volume on the Brix
index and pH of inulin was examined. Results showed the highest inulin yield of
1.80% at 50 mL water volume, 1.86% at 20min. Heating time, and 1.83% at
90°C.A direct relationship between inulin yield and temperature was observed.
The highest Brix index of 9.25 was recorded at 15mL water volume (Kanakasabai
etal. 2023; Puangbut etal. 2012). A study conducted by Aly etal. examined the
impact of plant maturity on inulin yield from Jerusalem artichoke (Helianthus
tuberosus). The researchers observed that the stage of plant maturity at the time of
harvest played a crucial role in inulin content. Plants harvested early (at 90days)
had an inulin yield of 18.6% ± 1.4%, while those harvested at full maturity (at
150days) had a signicantly higher yield of 32.4%±2.0%. This nding suggests
that optimizing harvest times is critical for maximizing inulin yield, but it also intro-
duces variability based on the precise timing of harvests (Aly and Khalil 2017). In
a study by García Lopez etal., different extraction methods were evaluated for their
impact on inulin yield from agave (Agave tequilana). The researchers compared
ultrasound (U), microwave (M), simultaneous ultrasound-microwave (UM), and
heat treatment (HT), nding signicant differences in yields. The yield obtained by
UM (72%) was similar to the other methods; however, the extraction time was
shorter (5min). In U, M, and HT, the yields were 86, 76, and 85% in 35, 30, and
180min, respectively. These results demonstrate the superior efciency of ultra-
sound-assisted extraction in maximizing inulin yield. However, the study also noted
that the efciency of each method varied depending on the specic agave species
used, highlighting the inuence of both extraction technique and plant variety on
inulin yield (García-Villalba etal. 2023).
2.10.3 Purity Challenges
Achieving high purity levels of inulin is challenging due to the presence of other
polysaccharides and impurities that co-extract with inulin. Advanced purication
techniques such as ultraltration and chromatography, while effective, are costly
and may not be feasible for large-scale production. A study by Li etal. investigated
the use of ultraltration for purifying inulin extracted from Jerusalem artichoke
(Helianthus tuberosus). The study compared the purity levels achieved using mem-
branes with different molecular weight cutoffs. Results indicated that a membrane
with a 10kDa cutoff achieved an inulin purity of 85.6%±1.5%, whereas a 5kDa
cutoff membrane increased the purity to 92.3%±1.2%. However, the study high-
lighted that the cost of ultraltration increased signicantly with higher purity lev-
els, making it less feasible for large-scale production (Sağcan etal. 2024). Alexsandra
etal. explored the effectiveness of various chromatographic techniques, including
ion-exchange and size exclusion chromatography, for purifying inulin from. Agave
sisalana boles. Ion-exchange chromatography achieved an inulin purity of
88.4%±2.0%, while size exclusion chromatography reached a higher purity level
of 95.1%±1.3%. Despite the high purity, the study pointed out the high operational
2 Extraction andPurication ofInulin
42
costs and technical expertise required for these techniques, which limit their appli-
cability in industrial settings (Apolinário etal. 2017). A study by Olvera etal. inves-
tigated a combined approach of ultraltration followed by ethanol precipitation for
purifying inulin extracted from agave (Agave tequilana). The combined method
achieved an inulin purity of 93.8%±1.7%, signicantly higher than using ultral-
tration alone (87.2% ± 2.1%). The researchers noted that while the combined
method improved purity, the additional precipitation step increased both the com-
plexity and cost of the purication process, posing challenges for large-scale imple-
mentation (Olvera and Lopez-Munguia 2014).
2.10.4 Regulatory andSafety Issues
The regulatory framework for inulin products differs among various nations, posing
obstacles to the commercialization and worldwide distribution of products contain-
ing inulin. Ensuring adherence to food safety regulations and obtaining necessary
approvals can be a lengthy and complex process (McBurney etal. 2019). Anadón
etal. (Anadón etal. 2016) conducted a study examining the regulatory prerequisites
for inulin products in the European Union. It was discovered by the researchers that
inulin products are required to adhere to the Novel Food Regulation (EU) 2015/2283.
This regulation entails thorough safety evaluations and approval procedures. The
study emphasized a scenario in which a recently developed inulin product derived
from an innovative source encountered a lengthy approval process, resulting in a
substantial delay in its introduction to the market. Moreover, the substantial expense
associated with adherence, which is approximated at €250,000, presented a hin-
drance for small and medium-sized enterprises (Anadón etal. 2016). Tee etal. con-
ducted a case study that analyzed the challenges encountered by a Chinese inulin
producer in their efforts to export to several global markets. The study revealed that
the presence of varying regulatory standards between nations, such as the obligatory
labeling of genetically modied crops (GM crops) in the European Union but not in
China, resulted in substantial obstacles. The company had to modify its labeling and
product compositions to comply with the individual rules of each country, resulting
in extra expenses and logistical difculties. This case highlights the intricate nature
and challenges involved in the international trading of inulin products (Tee and
Chan 2022).
2.11 Future Scope
The extraction and purication of inulin have a promising future due to technologi-
cal improvements, a rising demand for natural and functional components, and a
growing focus on sustainable practices. Advancements in extraction technology
play a crucial role in improving the efciency and productivity of inulin. Novel
techniques such as pulsed electric eld (PEF) extraction, supercritical uid extrac-
tion (SFE), and the utilization of deep eutectic solvents (DES) have promising
M. Vishwakarma et al.
43
advantages in terms of increased productivity, decreased energy usage, and mini-
mized environmental harm (AlYammahi et al. 2023). However, these techniques
require further optimization and validation for industrial-scale applications.
The high expense of the available purication methods continues to be a major
obstacle to inulin’s widespread application. Future research should concentrate on
creating scalable, reasonably priced processes that preserve high purity levels.
Methods like adsorptive separation, membrane ltration, and integrated extraction-
purication processes could provide viable solutions (Huang etal. 2022). Growing
importance is being given to sustainability in the synthesis of bioactive substances.
Reducing the environmental impact of inulin production requires the adoption of
green technologies, such as the use of renewable solvents and energy-efcient pro-
cedures. Investigating the lifecycle assessment (LCA) of various extraction and
purication techniques will aid in determining the most environmentally friendly
procedures (Tsatsaragkou et al. 2023). A further option for raising yield is to
increase the inulin content of plant sources through genetic modication and better
agronomic techniques. Research on improving cultivation conditions and geneti-
cally engineering plants like chicory and Jerusalem artichokes to increase inulin
synthesis can have a big impact on the availability and cost of inulin (Marou 2010).
Addressing regulatory challenges through the harmonization of standards across
regions can facilitate the global trade of inulin products. Future work should aim at
simplifying compliance processes and reducing the time and cost associated with
regulatory approval. Additionally, expanding the market by exploring new applica-
tions of inulin in various industries will drive further growth (Ghosh etal. 2019).
Beyond its conventional uses as a prebiotic and dietary ber, inulin can be used
to explore and develop new applications that could lead to new market opportuni-
ties. Research into inulin’s role as a functional ingredient in pharmaceuticals, cos-
metics, and bioplastics, as well as its potential in biotechnological applications, will
be crucial (Shoaib etal. 2016; Anjuomo etal. 2021). Innovations in technology,
environmental concerns, and growing market demand are driving substantial prog-
ress in inulin extraction and purication. To fully realize the potential of inulin,
future research should concentrate on rening innovative extraction and purication
techniques, improving the sustainability of production processes, and investigating
novel applications for inulin. Collaboration between academia, industry, and regula-
tory bodies will be essential to overcome existing challenges and promote the
growth of the inulin market.
2.12 Conclusion
Inulin is a stored polysaccharide of natural origin but with huge potential in the eld
of application in food and pharmaceutical industries. Plants with the main sources
are Jerusalem artichokes, chicory, and dahlia. Having a conguration of a β-(2,1)-
glycosidic bond structure makes inulin undigestible by human enzymes in the small
intestine. It classies inulin as a soluble dietary ber with prominent prebiotic
effects because it is fermented by benecial gut ora in the large intestine to
2 Extraction andPurication ofInulin
44
produce SCFAs like butyrate, propionate, and acetate. These SCFAs have been
linked to a wide array of health advantages that range from gut health improvement
to protection against some diseases. Interest in inulin is growing, and techniques for
its extraction and characterization have increased. Some of the optimized methods
are HWE, UAE, MAE, and EME, which were developed for high yield, efciency,
and reduced impact on the environment. Each has distinct benets, but UAE and
MAE are especially noted for efciency and lesser energy use. Inulin, together with
its chemical modications, offers further applications in industry and pharmacy.
Esterication, etherication, and carboxymethylation improve its properties to act
as an emulsier, a stabilizer, and in the development of new biomaterials. Most
modications use eco-friendly solvents, aligned with increasing public emphasis on
sustainable practices in chemical processes.
These diverse biological activities, such as immunomodulation, antioxidant
properties, antitumor effects, hepatoprotection, hypoglycemic effects, and gastroin-
testinal protection presented by inulin, make it, therefore, a bioactive compound
with potential value. With the advancement of research, inulin is found to be a very
promising ingredient in functional food and pharmaceuticals for signicant contri-
butions to the enhancement of healthcare and biomedicine.
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2 Extraction andPurication ofInulin