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Synthesis and Characterization of Methanolic Root Extract of
Imperata Cylindrica and Its Nanoencapsulation with Chitosan
P. O. Okwuego
1
*, V. O. Offiah
1
, C. M. Okey-Nzekwe
1
Department of Pure and Industrial Chemistry, Chukwuemeka Odumegwu Ojukwu University,
Anambra State, Nigeria
*
Corresponding Author
DOI: https://doi.org/10.51583/IJLTEMAS.2026.15020000108
Received: 25 February 2026; Accepted: 02 March 2026; Published: 20 March 2026
ABSTRACT
The development of efficient nano-based drug delivery systems remains a critical strategy for improving the
stability, bioavailability, and therapeutic performance of plant-derived bioactive compounds. In this study,
methanolic root extract of Imperata cylindrica was synthesized, characterized, and nanoencapsulated using
chitosan as a biodegradable and biocompatible polymeric carrier, with the objective of enhancing its suitability
for wound-healing and hemostatic drug delivery applications. Qualitative and quantitative phytochemical
analyses revealed the presence of alkaloids (5.4%), tannins (5.7%), flavonoids (10.3%), saponins (2.25%), and
terpenoids (43.3%), which are compounds commonly associated with anti-inflammatory, antimicrobial, and
tissue-repair activities. Nanoencapsulation was achieved via chitosan-assisted precipitation, and the resulting
nanoparticles were characterized using Fourier transform infrared spectroscopy (FTIR), ultraviolet–visible (UV–
Vis) spectroscopy, transmission electron microscopy (TEM), and X-ray diffraction (XRD). FTIR spectra
confirmed the successful incorporation of the extract within the chitosan matrix through the presence of
characteristic O–H, N–H, and C=O functional groups without structural degradation. TEM analysis revealed
predominantly spherical, mesoporous nanoparticles with particle sizes ranging from 2 to 50 nm and average
diameters between 12.66 and 17.98 nm. UV–Vis spectroscopy demonstrated a hypsochromic shift in absorption
maxima from 550 nm for the free extract to 375 nm for the nanoencapsulated formulation, indicating improved
molecular dispersion and an encapsulation efficiency of 31.81%. XRD analysis revealed crystalline phases
containing mineral oxides relevant to biological and pharmaceutical applications. Overall, the chitosan-based
nanoencapsulation of Imperata cylindrica root extract demonstrates significant potential as a natural
product-derived drug delivery system for topical wound-healing and hemostatic applications, aligning with
current advances in polymeric nanoparticle-mediated drug delivery.
Keywords: Imperata cylindrica; chitosan nanoparticles; drug delivery system; nanoencapsulation; wound
healing; hemostatic agents
INTRODUCTION
Efficient drug delivery remains a major challenge in pharmaceutical science, particularly for bioactive
compounds derived from medicinal plants that often suffer from poor stability, low aqueous solubility, and
limited bioavailability. Nano-enabled drug delivery systems have emerged as effective tools for overcoming
these limitations by enhancing drug protection, controlled release, and targeted delivery to diseased tissues.
Polymeric nanoparticles, especially those derived from natural polymers, are of increasing interest due to their
favorable safety profiles and tunable physicochemical properties Agnihotri et al (2004).
Imperata cylindrica is a perennial rhizomatous grass widely distributed across tropical and subtropical regions
and extensively used in traditional medicine for wound healing, hemostatic activity, anti-inflammatory effects,
and treatment of liver disorders. Phytochemical studies have shown that the plant contains flavonoids,
terpenoids, alkaloids, tannins, and glycosides, which contribute to its pharmacological activities. However, the
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direct application of crude plant extracts is often limited by instability, rapid degradation, and uncontrolled
release at the site of action Alonso et al (2003); Cui et al (2012); Chen et al. (2016); Chen et al. (2015).
Chitosan, a cationic polysaccharide obtained through the deacetylation of chitin, has gained prominence in drug
delivery science due to its biocompatibility, biodegradability, low toxicity, mucoadhesive behavior, and inherent
antimicrobial properties. Chitosan-based nanoparticles are particularly attractive for topical and transdermal
drug delivery owing to their ability to enhance drug retention, promote tissue interaction, and facilitate sustained
release Ahmed et al (2016).
The present study focuses on the development of a chitosan-based nanoencapsulated formulation of methanolic
root extract of Imperata cylindrica. The work emphasizes nanoparticle synthesis, physicochemical
characterization, phytochemical profiling, and structural evaluation in the context of drug delivery science. By
integrating traditional medicinal knowledge with modern nanotechnology, this study aims to provide a
scientifically validated nano-drug delivery platform suitable for wound-healing and hemostatic applications Ali
et al (2018).
MATERIALS AND METHODS
Sample Collection and Preparation
Fresh roots of Imperata cylindrica were collected from Umuenechi Village, Nibo, Anambra State, Nigeria. The
plant was authenticated at the National Root Crops Research Institute Herbarium, Umudike, Abia State. The
roots were washed, air-dried at room temperature for seven days, pulverized, and stored in airtight containers
for subsequent analysis.
Extraction of Root Sample
One hundred and fifty grams (150 g) of the powdered root sample was macerated in 250 mL of methanol for 24
h. The extract was filtered using muslin cloth and concentrated to near dryness using a water bath at 70 °C. The
concentrated extract was cooled and stored under refrigeration.
Synthesis of Chitosan
Chitosan was prepared via alkaline deacetylation of chitin obtained from carbonated periwinkle shells. The shells
were activated using 0.5 M phosphoric acid, washed to neutral pH, oven-dried at 70 °C, and stored for
nanoparticle synthesis.
Preparation of Nanoencapsulated Extract
One gram (1 g) of the methanol extract was dissolved in 150 mL of deionized water and added dropwise to 250
mL of 0.5 M ferric nitrate under continuous stirring. Chitosan (10 g per 100 mL of mixture) was subsequently
introduced, and the mixture was stirred until gelation occurred. The product was concentrated at 65 °C,
oven-dried, and stored for characterization.
RESULTS AND DISCUSSION
Organoleptic and Formulation Stability Assessment
The organoleptic evaluation of Imperata cylindrica root, chitosan, and the encapsulated particles revealed
distinct differences in colour, texture, taste, and odour, reflecting their intrinsic properties and the effects of
nanoencapsulation (Table 1). The root of I. cylindrica exhibited a milky colour, fine texture, mildly sweet taste,
and mild odour, consistent with its natural plant composition and high phytochemical content. Chitosan appeared
ash-coloured, powdery, with a shelly taste and soapy odour, typical of its polysaccharide structure. The
encapsulated particles displayed a brown colour, prickly texture, sour taste, and pungent but non-choking odour,
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indicating successful integration of the extract into the polymer matrix. Importantly, the organoleptic properties
of the nanoencapsulated formulation showed no observable deterioration in colour, odour, texture, or consistency
throughout the analysis period. This stability highlights the protective role of the chitosan matrix, which
preserves the bioactive constituents, maintains uniform sensory attributes, and enhances formulation robustness
a critical factor for topical or oral drug delivery systems where consistency influences physicochemical stability
and patient compliance Nkachukwu. et al (2025); Mmuo et al (2024); Okwuego, et al (2025).
Table 1. Organoleptic Properties of Imperata cylindrica, Chitosan, and Encapsulated Particles
Parameter
Imperata cylindrica Root
Chitosan
Encapsulated Particles
Colour
Milk
Ash
Brown
Texture
Fine
Powdery
Prickly
Taste
Sugary
Shelly
Sour
Odour
Mild
Soapy
Pungent but not choking
Morphological Characteristics and Particle Size Distribution
Transmission electron microscopy (TEM) analysis demonstrated that the nanoencapsulated Imperata
ylindricaextract consisted predominantly of spherical nanoparticles with mesoporous structures and particle
sizes ranging from 2 to 50 nm. The average particle sizes (12.66–17.98 nm) fall within the optimal range reported
for polymeric nanoparticles intended for drug delivery applications Okwuego et al (2021); Ochie et al (2025);
Okwuego (2025). Nanoparticles within this size regime are known to exhibit enhanced surface area, improved
tissue interaction, and prolonged retention at the site of application, particularly in topical and wound-healing
formulations.
Figure 1 Transmission of Electron Microscope of Methanol Extract
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Figure 2 Transmission Electron Microscope of Chitosan
Figure 3 Transmission Electron Microscope of Encapsulated particle
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Compared with larger micro-scale carriers, the observed nanoscale dimensions are advantageous for facilitating
intimate contact with damaged tissue surfaces and for promoting uniform drug distribution.
Mild particle agglomeration observed in some regions is consistent with previous reports on chitosan-based
nanoparticles and may be attributed to intermolecular hydrogen bonding; however, the extent of agglomeration
observed is unlikely to compromise delivery efficiency.
FTIR Analysis and Encapsulation Confirmation
FTIR analysis of Imperata cylindrica methanol extract (Table 2), chitosan (Table 3), and the encapsulated
particles (Table 4) revealed the presence and preservation of key functional groups throughout the formulation
process. The extract exhibited characteristic O–H and N–H stretches (3218–3498 cm⁻¹), C=O stretches (1618–
1863 cm⁻¹), C–O stretches (1135–1295 cm⁻¹), C–H stretches (3013 cm⁻¹), and minor C≡N peaks (2016–
2798 cm⁻¹), consistent with alkaloids, flavonoids, tannins, and terpenoids. Chitosan displayed broad O–H and
amide N–H stretches (3245–3426 cm⁻¹), C=O stretches (1613–1884 cm⁻¹), and C–O stretches (1277–1415 cm⁻¹),
reflecting its polysaccharide structure capable of hydrogen bonding.
The encapsulated particles retained all major extract and polymer peaks, with slight shifts in O–H and N–H
bands (3141–3548 cm⁻¹) indicative of hydrogen bonding between the extract and chitosan matrix. Preservation
of C=O, C–O, and C–H vibrations confirms that the bioactive compounds remain chemically intact after
encapsulation Okwuego (2023); Ochie et al (2025); Okwuego et al (2021); Okwuego et al (2025)The FTIR
results demonstrate successful incorporation of the extract into chitosan particles without chemical degradation,
supporting the formulation’s potential as a stable, natural product-based drug delivery system Ochie et al (2025);
Okorie et al (2025)
Table 2. FTIR Spectral Data for Imperata cylindrica Methanol Extract
Functional Group / Assignment
C–O, C–H deformation bonds for alkyl and methyl groups
C–O, C–H deformation bonds for alkyl and methyl groups
C–H, C–O deformation bonds for alkyl groups
C–O deformation bonds for ketones and acids
C–O stretch for ketones and alcohols
C=O stretch for ketones, acids, and amides
C=O stretch for ketones, acids, and amides
C≡N stretch for nitriles
C≡N stretch for nitriles
C≡N stretch for nitriles
C≡N stretch for nitriles
C≡N stretch for nitriles
C–H stretch for alkenes
O–H stretch for alcohols, amides, and acids
N–H and O–H stretch for amines, alcohols, and phenols
O–H stretch for alcohols, acids, and amides
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Figure 4 FTIR Spectra of Methanol extract
Table 3. FTIR Spectral Data for Chitosan Extract
Functional Group / Assignment
C–O, C–H deformation bonds for alkyl and methyl groups
C–O, C–H deformation bonds for alkyl and methyl groups
C–H, C–O deformation bonds for alkyl groups and ketones
C–O stretch for ketones and alcohols
C–O stretch for ketones and alcohols
C=O stretch for ketones, acids, and amides
C=O stretch for ketones, acids, and amides
C=O stretch for ketones, acids, and amides
C≡N stretch for nitriles
C≡N stretch for nitriles
C≡N stretch for nitriles
O–H stretch for acids
O–H stretch for acids
C–H stretch for alkanes
C–H stretch for alkenes
O–H stretch for alcohols, acids, and amides
O–H stretch for alcohols, acids, and amides
O–H (unbound)
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Figure .5 FTIR Spectra of Chitosan
Table 4. FTIR Spectral Data of Encapsulated Particles
Frequency (cm⁻¹)
Functional Group / Assignment
839.80
C–O, C–H deformation bonds for alkyl and methyl groups
1016.40
C–O, C–H deformation bonds for alkyl groups and ketones
1400.32
C–O stretch for ketones and acids
1633.70
C=O stretch for ketones and amide groups
1871.38
C=O stretch for ketones, acids, and amides
2117.08
C≡N stretch for nitriles
2275.43
C≡N stretch for nitriles
2450.16
C≡N stretch for nitriles
2526.01
C≡N stretch for nitriles
2631.72
C–H stretch for alkenes and aromatic groups
2830.92
C–H stretch for alkenes and aromatic groups
2954.72
C–H stretch for alkanes, alkenes, acids, and aromatics
3045.69
C–H stretch for alkenes
3140.57
N–H and O–H stretch for amines, alcohols, and phenols
3261.19
O–H stretch for alcohols, amides, and acids
3548.20
O–H stretch for alcohols, acids, and amides
3698.69
O–H unbound for alcohols
3828.11
O–H unbound
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Figure .6 FTIR SPECTRA of Encapsulated particle.
Importantly, no disappearance or excessive shifting of major peaks was observed following encapsulation,
indicating that the structural integrity of the bioactive phytochemicals was preserved. This observation is
particularly significant for drug delivery systems, as chemical alteration of active compounds during formulation
can compromise therapeutic efficacy.
UV–Visible Spectroscopy and Encapsulation Efficiency
UV-Vis analysis of chitosan, the methanol extract of Imperata cylindrica, and the encapsulated extract (Table 5)
revealed distinct absorption patterns and structural interactions within the polymer matrix. Chitosan exhibited
low, nearly constant absorbance across 350–850 nm, consistent with its polysaccharide structure and absence of
strong chromophores. The methanol extract showed a pronounced peak at 500–600 nm, attributed to π→π*
transitions in conjugated systems such as flavonoids, terpenoids, and polyphenols. Upon nanoencapsulation, the
extract displayed substantially reduced absorbance across all wavelengths, with a hypsochromic shift of the
absorption maximum from 550 nm for the free extract to 375 nm in the encapsulated formulation. This blue shift
indicates reduced molecular aggregation and improved dispersion of the bioactive compounds within the
chitosan matrix Nkachukwu. et al (2025); Mmuo et al (2024); Okwuego, et al (2025). The calculated
encapsulation efficiency of 31.81% demonstrates effective incorporation of the extract, comparable to other
chitosan-based plant bioactive formulations. The diminished optical activity and similarity in absorption
behavior between chitosan and the encapsulated particles confirm successful entrapment and compatibility of
the polymer drug system. From a drug delivery perspective, these spectral features suggest enhanced stability,
controlled presentation of active constituents, and potential for improved bioavailability and sustained release at
the wound site.
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Table 5. Absorbance per Unit Wavelength of Chitosan, Methanol Extract, and Encapsulated Extract of
Imperata cylindrica
Wavelength (nm)
Chitosan
Methanol Extract of I. cylindrical
Encapsulated Extract of I. cylindrical
350
0.344
1.758
0.107
400
0.344
1.762
0.107
450
0.274
2.225
0.039
500
0.250
6.000
0.017
550
0.202
6.000
-0.002
600
0.182
6.000
-0.008
650
0.149
0.850
-0.011
700
0.133
0.535
-0.012
750
0.122
0.345
-0.014
800
0.110
0.241
-0.014
850
0.100
0.180
-0.015
Figure 7Absorption maxima of the various sample
3.5 Crystallinity and Mineral Composition Analysis by XRD
X-ray diffraction (XRD) analysis of the encapsulated nanoparticles (Tables 6 and 7)revealed a multi-phase
crystalline composition consisting of Barium Terbium Niobium Oxide (Ba₂TbNbO₆), Romarchite (SnO), Ziroite
(ZrO₂), Freibergite ((Cu, Ag, Zn)), and Osumilite (K–Na–Ca–Mg–Fe–Al–Si). Quantitative analysis showed that
Osumilite (32.1%) and Barium Terbium Niobium Oxide (31.2%) were the predominant phases, followed by
Freibergite (19.5%), Romarchite (11.4%), and Ziroite (5.8%).
The presence of sharp diffraction peaks indicate partial crystallinity, confirming the structural integrity of the
nanoparticles and suggesting stability during storage and application Okwuego, et al (2025); Nwankwo et al
(2025). Importantly, the identified mineral components may contribute physiologically relevant properties, such
as blood clotting, tissue regeneration, and enzymatic activity, which could synergize with the bioactive
phytochemicals from Imperata cylindrica. The XRD results demonstrate that the nanoencapsulation process
preserves the crystalline mineral phases while integrating them into a stable polymeric delivery system,
supporting both structural robustness and potential therapeutic functionality Ochie et al (2025).
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Table 6. Qualitative Analysis of Encapsulated Nanoparticles
Phase Name
Formula
Phase Details
Barium Terbium Niobium Oxide
Ba₂TbNbO₆
Minerals
Romarchite, syn
SnO
Minerals
Ziroite, syn
ZrO₂
Minerals
Freibergite, syn
(Cu, Ag, Zn)
Minerals
Osumilite
K–Na–Ca–Mg–Fe–Al–Si
Minerals
Table 7. Quantitative Analysis of Encapsulated Nanoparticles
Parameters
Weight Fraction (%)
Barium Terbium Niobium Oxide
31.2 (15)
Romarchite, syn
11.4 (7)
Ziroite, syn
5.80 (13)
Freibergite, syn
19.5 (4)
Osumilite
32.1 (7)
Figure 8 X –ray Diffraction spectral
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3.6 Phytochemical Profile and Therapeutic Relevance
Qualitative and quantitative phytochemical analyses confirmed that terpenoids (43.3%) were the dominant
constituents of the methanolic root extract of Imperata cylindrica, followed by flavonoids (10.3%), tannins
(5.7%), alkaloids (5.4%), and saponins (2.25%). These classes of secondary metabolites are well documented
for their antioxidant, anti-inflammatory, antimicrobial, and hemostatic activities, which are critical therapeutic
attributes for wound-healing applications Okwuego, et al (2025).
Table 8 Phytochemical screening results of Imperata cylindrica root extract
Phytochemical class
Test
Observation
Result
Alkaloids
Wagner’s reagent
White precipitate
++
Mayer’s reagent
Reddish-brown precipitate
++
Saponins
Frothing test
Persistent frothing
++
Emulsion test
Stable emulsion
++
Fehling’s solution test
Light reddish precipitate
+
Flavonoids
Ammonium test
Formation of green and light-green layers
++
NaOH/Acetic acid test
Formation of green and light-green layers
++
Tannins
Ferric chloride test
Greenish-black precipitate
++
Lead acetate test
Cream precipitate
++
Steroids and terpenoids
Ethanol–chloroform–H₂SO₄
test
Reddish-brown interface layer
++
Glycosides
Fehling’s test
Opaque brick-red precipitate
++
+, slightly present; ++, moderately present; +++, present; −, absent.
Table 9 Quantitative phytochemical composition of Imperata cylindrica root extract
Phytochemical parameter
Amount (mg/4 g sample)
Percentage (%)
Tannins
0.057 ± SD
5.7
Flavonoids
0.103 ± SD
10.3
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Alkaloids
0.054 ± SD
5.4
Saponins
0.023 ± SD
2.25
Terpenoids
0.433 ± SD
43.3
Values are expressed as mean ± standard deviation (SD), where applicable.
Phytochemical analysis of Imperata cylindrica root extract (Tables 8 and 9) revealed alkaloids, flavonoids,
tannins, saponins, steroids, terpenoids, and glycosides, with terpenoids being the most abundant (43.3%),
followed by flavonoids (10.3%), tannins (5.7%), alkaloids (5.4%), and saponins (2.25%). These findings
corroborate previous reports of high terpenoid and phenolic content, supporting the plant’s antioxidant and
pharmacological activities Okwuego, et al (2025); Mmuo et al (2024). The pronounced terpenoid content
suggests potential for inflammation modulation and oxidative stress reduction, while flavonoids and tannins may
contribute to antimicrobial activity and hemostasis. Alkaloids and saponins further enhance antimicrobial
defense and membrane interactions. Nanoencapsulation within chitosan is expected to protect these bioactive
constituents from degradation, allow controlled release, and preserve phytochemical integrity, demonstrating the
extract’s suitability as a stable, natural product-based drug delivery system with localized therapeutic efficacy.
CONCLUSION
The study successfully developed a chitosan-based nanoencapsulated formulation of Imperata cylindrica
methanolic root extract, demonstrating structural integrity, preserved bioactive constituents, and nanoscale
particle size. FTIR, UV-Vis, and XRD analyses confirmed retention of functional groups, improved
phytochemical dispersion, and partial crystallinity of mineral phases, indicating stability and structural
robustness. The encapsulated particles maintained acceptable organoleptic properties and exhibited potential for
controlled release, enhanced tissue interaction, and therapeutic activity, including anti-inflammatory,
antimicrobial, and antioxidant effects. These findings highlight the formulation as a promising natural product-
derived nanoparticle system for topical wound-healing and hemostatic applications, with potential for further in
vitro and in vivo validation.
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16. Okwuego, P.O, Offiah V.O, Nkachukwu, M.B, Ifeakor, C.O. (2025) Comprehensive Analysis of
Leachables and Extractables from Pharmaceutical Packaging: Investigating Ink and Adhesive Migration
in Selected Drug Products in Nigeria International Journal Of Research And Innovation In Applied
Science (IJRIIAS) ISSN NO. 2454-6194 DOI : https://doi.org/10.51584/IJRIASs.10040007
17. Okwuego, P. O, Momoh, E. R. (2025) Phytochemical, Antioxidant and Antimicrobial Evaluation of Oil
Palm Tree (Elaeis guineensis) Bark for Potential Nutraceutical and Pharmaceutical Applications
International Journal of Health and Pharmaceutical Research E-ISSN 2545-5737 P-ISSN 2695-2165 Vol.
10. No. 11 2025 www.iiardjournals.org
18. Okorie E.A, Offiah V,O, Oragwu P.I, Okwuego P.O (2025) Assessment of Heavy Metals and Polycyclic
Aromatic Hydrocarbons in Soil and Water in Selected Mining Areas of Ebonyi State, Nigeria. Journal of
Global Ecology and Enviroment. Doi: 10.56557/jogee/v21i29123.