This document discusses a study comparing the effect of chitosan from different sources on the in-vitro release of esomeprazole from chitosan/polylactic acid nanoparticles. It begins with an introduction to nanotechnology and chitosan as a natural polymer for drug delivery. It then states the research aim to evaluate the effect of chitosan from different sources (crayfish, snail shells, periwinkle shells, and shrimp) on the drug release. The methodology discusses extracting chitosan from the sources and preparing chitosan/polylactic acid nanoparticles loaded with esomeprazole. A literature review identifies research on chitosan nanoparticles and drug delivery and gaps such as further studies

1. A Comparative Study on Chitosan and it’s
Effect on the In-Vitro Release of Esomeprazole
From Chitosan/Polylactic Acid Nanoparticles
Ugochi E. Ewii
ACEFUELS/21/PHD/14000112
Qualification: B. Pharm
(Igbinedion) M. Pharm (Uniuyo)

2. • 1.1 General overview /key concepts
• The ability of nanotechnology to transverse across several aspects
of our daily lives and deliver top notch functions makes it quite
distinctive and desirable (Putheti et al., 2008).
• The role of nanotechnology to pharmaceutical science and
technology centers around the development of innovative drug
delivery systems to target specific sites in the body, maintain a
steady drug release and deliver profound therapeutic effects with
the reduction of systemic side effects, dose reduction and better
patients’ compliance (Douglas, 2020).
1. BACKGROUND/INTRODUCTION

3. • 1.1 General overview /key concepts
• Several nano-carriers have been utilized in nanomedicine. These
include organic and inorganic nanocarriers.
• Natural polymers are desirable nanoparticles for drug delivery
because they are not toxic and have very good biocompatibility
and low immunogenicity, and are biodegradable(Anwunobi &
Emeje, 2011).
• Chitosan is a natural polymer obtained from Chitin, found in the
shells of Crustaceans (Dev et al., 2010a).
1. BACKGROUND/INTRODUCTION
CONTD.

4. • 1.1 General overview /key concepts
 Several drugs can easily be transported for controlled drug delivery
following co-administration with Chitosan (van der Lubben et al.,
2003).
 Numerous benefits to using Chitosan are enhanced residence time, easy
chemical modification and its physicochemical properties (Thacharodi
& Panduranga Rao, 1996).
 Chitosan is often derived from Chitin through chemical or enzymatic
deacetylation. The main natural sources of Chitin are Shrimp and crab
shells (Elieh-Ali-Komi & Hamblin, 2016). It could also be extracted
from periwinkle and snail shells (Gbenebor et al., 2017).
1. BACKGROUND/INTRODUCTION
CONTD.

5. 1.2 PROBLEM STATEMENT/RESEARCH QUESTION/HYPOTHESIS
• One problem often faced with the utilization of nanoparticles especially for drug
delivery is the high cost of these nanomaterials. By exploring the natural polymers
from sources in our community and advocating it’s utilization, through tremendous
research and exportation, these sources would be a source of foreign exchange and
would lead to cheaper nanomedicines.
 HYPOTHESIS: Chitosan from different sources, formed into nanoparticles will create
suitable controlled drug delivery systems for invitro/invivo drug delivery, precisely for the
treatment of ulcer.

6.  This research is aimed at evaluating the effect of the Chitosan from different sources on the
in-vitro release of Esomeprazole from Chitosan/Polylactic acid nanoparticles

 Objectives

 1. Characterization of the Chitosan obtained from different sources.

 2. Preparation of the Chitosan/Polylactic acid Nanoparticles

 3. Loading of Esomeprazole into the Chitosan/Polylactic acid nanoparticles.

 4. Characterization of the formed nanoparticles

 5. Evaluation of the In-vitro drug release properties of the drug loaded nanoparticles.
1.3 RESEARCH AIM AND OBJECTIVES

7. 1.4 JUSTIFICATION & SCOPE
1. In-vitro release test is one of the preliminary tests that must be carried out on every drug formulation
prior to it’s use. It gives an idea on the release of the drug from its dosage form inside the body. Given
that Chitosan from different dosage forms have different physiological properties, it is expedient to
explore these sources, characterize them and evaluate their use in nano drug delivery.
3. Unique properties of polymeric nanoparticles like biocompatibility, biodegradability, non- toxicity,
stability and drug targeting makes the study of Chitosan/Polylactic acid nanoparticles for drug delivery
suitable.
4. This research will be carried out on Chitosan from 4 sources (crayfish, snail shells, periwinkle shells
and shrimps.Amongst the 3 strategies for loading drugs into nanoparticles, only encapsulation will be
utilized.
www.futo.edu.ng

8. 2. LITERATURE REVIEW
2.1 Tables with specific Literature
S/N Literature Findings Reference
1 Nanotechnology in
Pharmaceutical Science: A
Concise Review
The word Nanotechnology was actually coined by
Prof. Norio Taniguchi in 1974, and it simply involves
using technology at a nanoscale size
Popat
Jadhav et
al., 2020.
2 Nanotechnology: Applications,
techniques, approaches, & the
advancement in toxicology and
environmental impact of
engineered nanomaterials.
Since its inception, nanotechnology has been applied
in vast areas, these include energy, transportation,
agriculture, medicine, water treatment etc.
Zaib &
Iqbal,
2019.
3 Recent Applications of Natural
Polymers in Nanodrug Delivery
When applied to pharmaceutical science, its benefits
are immense and unquantifiable as new discoveries
are continuously being made.
Anwunobi
& Emeje,
2011.

9. 2. LITERATURE REVIEW CONTD.
S/N Literature Findings Reference
4 A review on status of
nanotechnology in
pharmaceutical sciences
Some of these benefits are targeted site-specific
delivery, biosensors, biomarkers, tissue engineering,
diagnosis, development of nanomedicines etc.
Suttee et
al., 2019.
5 Nanostructure-mediated drug
delivery.
Nanotechnology helps to enhance drugs
performance, safety and effectiveness as well as
patient’s adherence etc.
Hughes,
2005.
6 Pharmaceutical Nanotechnology:
A Therapeutic Revolution
Nano-carriers may be referred to as transporters of
the active pharmaceutical ingredient in the nanoscale
to a targeted site in the body.
Douglas,
2020.

10. 2. LITERATURE REVIEW CONTD.
S/N Literature Findings Reference
7. Colloidal nanocarriers: a review
on formulation technology, types
and applications toward targeted
drug delivery.
Nano-carriers are classified into two broad groups;
Organic (polymeric nano-carrier, dendrimer,
liposome, micelles and solid lipid nano-carrier) and
Inorganic Nano-carriers (mesoporous silica, quantum
dot, magnetic nanocarrier, gold nano-carrier and
carbon nanotube). Characteristics of nano-carriers are
enhanced biocompatibility, solubility, stability,
enhanced pharmacokinetics, low toxicity and
targeted/sustained drug delivery
Mishra et
al., 2010
8. Advances of Cancer Therapy by
Nanotechnology
Polymeric nanocarriers can be made from natural (e.g.,
Chitosan, gelatin, collagen, alginate etc.) and synthetic
polymers (Polyethylene glycol and polylactic acid).
Wang et
al., 2009
9. Cancer-cell-specific induction of
apoptosis using mesoporous silica
nanoparticles as drug-delivery
Advantages of polymeric nanocarriers are higher
stability, drug payload, improved half-life and
sustained drug release. Polymeric nano-carriers may
Rosenholm
et al., 2010.

11. 2. LITERATURE REVIEW CONTD.
S/N Literature Findings Reference
10. Chitosan-based drug
delivery systems
Chitosan is one unique polymer in the pharmaceutical industry
known for its distinctive properties. It is peculiar for its cationic
properties which could be ascribed to its primary amino groups
Bernkop-
Schnürch &
Dünnhaup
2012.
11. Comparison of the
mucoadhesive properties
of various polymers.
These amino groups are held responsible for its controlled drug
release, mucoadhesion permeation enhancement Insitu gelation
and transfection enhancing properties
Grabovac
al., 2005,
Schipper e
1996,
Sakloetsak
et al., 2009
Mao et al.,
2010.
12. Extraction and
characterization of chitin
and chitosan from
Chitosan drug delivery systems have been utilized in oral drug
delivery, nasal drug delivery, vaginal drug delivery, buccal drug
delivery and vaccine delivery Chitin can be derived from many
Bernkop-
Schnürch &
Dünnhaup

12. 2.2 SUMMARY AND IDENTIFIED GAPS
 1. Shangyong et al., 2022, studied the application of chitosan/alginate nanoparticle in oral drug delivery
systems, its prospects and challenges, however monitoring the pharmacological effect of the drugs
prepared with these nano-carriers is a huge knowledge gap.
 2. Mostafa et al., 2022 studied the Customizing nano-chitosan for sustainable drug delivery. Some
knowledge gaps that were identified. These include Cytotoxicity of Chitosan based systems (these needs
to be studied further in future works), Synthesis of low molecular weight chitosan should be studied for
better solubility and Safety and targeting studies should also be carried out.
 3. Casalini et al., 2019 studied a perspective on polylactic acid based polymers use for nanoparticles
synthesis and applications. Physical properties, chemical properties and applications were studied.
Cytotoxicity and hemolysis needs to be studied further.

13. 2.2 SUMMARY AND IDENTIFIED GAPS CONTD.
4. Oyekunle and Omoleye (2019) studied the new process for synthesizing chitosan from snail shells
through elimination of the drying stage. The formed chitosan were characterized showed suitable
properties and this process proved to be more economical. However further works need to be done on
the utilization of the formed chitosan in nano-drug delivery.
5. Abere et al., 2022 studied the derivation of composites of chitosan-nanoparticles from crustaceans
source for nanomedicine. They found out that indeed chitosan nanoparticles are promising
nanocarriers for encapsulating molecules in medications or active chemicals and that shellfish waste
could be a tremendous source of chitosan. Large scale greener methods needs to be sxplored for
chitosan extraction.

14. 3. RESEARCH DESIGN / METHODOLOGY
 3.1 Flow chart of Experimental design & approach:
 Steps in Extraction of Chitosan

15. 3.2 MATERIALS PREPARATION
 Chitosan (CS) from periwinkle, snails, crabs and crayfish, poly (lactic acid) (PLA),
dichloromethane, Pothylene oxide) (PEO), phosphate buffer saline (PBS) and acetic acid will
be obtained from Sigma–Aldrich, USA. Esomeprazole drug will be purchased from Sigma,
USA. All other chemicals will be of analytical grade.
 Preparation of PLA/CS nanoparticles
 About 100 mg of PLA will be dissolved in 10 ml of dichloromethane to form a fine
dispersion. This solution will rapidly be poured into10 ml of 1% acetic acid solution
containing 40 mg of CS and 200 mg of PEO. The mixture will then be sonicated for 15 min
to form an emulsion and vigorously stirred until the organic solvent evaporates. Finally, the
nanoparticles will be precipitated by adding water and then lyophilized.

16. 3.2 MATERIALS PREPARATION CONTD.
 Preparation of Esomeprazole loaded PLA/CS nanoparticles
 To encapsulate the drug, w/o/w emulsion technique will be applied. Aqueous drug solution
(2 ml) will be first poured into a polymer solution (100 mg of PLA dissolved in 10 ml
dichloromethane) to form a w/o emulsion. The w/o emulsion will then be rapidly poured into
10 ml of 1% acetic acid solution containing 40 mg of chitosan and 200 mg of PEO. The
mixture was sonicated for 15 min to form an emulsion, and then vigorously stirred. Stirring
will be continued until the organic solvent evaporates. The nanoparticles will be precipitated
by adding water and then be lyophilized.

17. 3.3 EXPERIMENTAL/COMPUTATIONAL PROCEDURE
 Characterization of Formed Nanoparticles.
 Several analysis will be done on the nanoparticles. These include Fourier transform infrared (FTIR)
spectroscopy, Scanning Electron Microscopy SEM, Dynamic Light Scattering and Thermogravimetric
analysis and X-Ray diffraction.
 Evaluation of Drug encapsulation efficiency
 The obtained nanoparticles will be frozen and lyophilized by a freeze dryer system to obtain a dried
nanoparticle product. The weighed product of nanoparticles will be washed with distilled water and then
centrifuged and the supernatant was collected. The solution will be measured by UV spectrophotometer
(UV-1700 Pharma Spec, Shimadzu) at the wavelength 270 nm (Fernandes, 2006) and the weight of drug
will be calculated by use of a calibration curve.
 Encapsulation efficiency = (weight of Esomeprazole drug in nanoparticles)/(weight of Esomeprazole
drug initially) * 100%

18. 3.3 EXPERIMENTAL/COMPUTATIONAL PROCEDURE CONTD.
 In-vitro drug release studies
 The in-vitro drug release tests will be carried out on all formulations (3% and 6% drug loaded samples).
Fifty milligrams of each sample will be suspended in 100 ml of PBS buffer at various pH at 37°C and
will be placed in an incubated shaker at 120 rpm. At predetermined time intervals, 3 ml of aliquots will
be withdrawn and the concentration of drug released will be monitored by UV spectrophotometer (UV-
1700 Pharma Spec, Shimadzu) at 270 nm. The dissolution medium will be replaced with fresh buffer to
maintain the total volume.
 The drug release percent can be determined by the following equation: Drug release {%] = C(t) / C (0) ꭓ
100 where C (0) and C(t) represents the amount of drug loaded and amount of drug released at a time t,
respectively. All studies will be done in triplicate.

19. 3.4 DATA PROCESSING, PRESENTATION AND ANALYSIS
 Raw data collected will be analyzed using SPSS and Microsoft excel software etc.

Polymer % of
Drug
loaded
w/w
Concentration of
Drug loaded (mg)
Encapsulation
Efficiency EE +-
SD
Particle
Size nm
+-SD
Time of
Drug
release
(mins)
Concentration of
Drug Released
(mg)
Chitosan from
Periwinkle Cp
3 - - - 30
60
90
120
240
360
-
-
-
-
-
-
6 - - - 30
60
90
-
-
-

20. 3.5 WORK PLAN (GANTT CHART)

21. 4. EXPECTED RESULTS
 4.1 Expected Results (linked to objectives)
 It is expected that Chitosan from different sources will yield nanoparticles for controlled
drug delivery. The physio-chemical properties will be evaluated and the one which yields
desirable properties will be recommended.
 4.2 Expected outcomes (linked to problem statement)
 It is expected that cost effective nanoparticles suitable for controlled drug delivery, precisely
for the treatment of Duodenal ulcer will be derived from Chitosan obtained from our local
sources like crayfish and periwinkle shells. These nanoparticles will then be utilized for
foreign exchange.

22. 4. EXPECTED RESULTS CONTD.
 4.3 Key beneficiaries of Project
 The community at large stands to benefit from this project,
beginning from the ACE-FUELS center, to the prestigious people of
Imo state to Nigeria, foreign countries and even our future
generations. The success of this project would proffer a
breakthrough in the treatment of Duodenal ulcer.
 4.4. Relevant Sustainable Development Goals (SDG)
 Out of the 17 sustainable development goals, the one most relevant
to this research is the third one ‘Good health and Well being’.
Because the success of one goal enhances the rest, all the
sustainable development goals will be enhanced by this research.