INTRODUCTION (DNA) (Gombotz & Wee, 1998). However, there is

INTRODUCTION

This
paper will present study on the preparation and characterization of
nanoparticles based on hydrophobic alginate derivative as carriers for
sustained release of vitamin D3 and there is no organic solvent will
be used during the preparation of sample.

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1.1  Background of study

Nanoparticle
is one of the products from nanotechnology which is involving science,
engineering and technology which conducted material at nano scale from 1 to 100
nanometres (An introduction to nanotechnology, 2007-2016). The size range of
nanomaterial is finer than the human hair and requires a specific microscope to
observe the materials. Nowadays, revolution of nanotechnology brings many
benefit and development in various fields including in electronic,
construction, medicine and food industry. Nanotechnology gives us advantage in
control the physical and chemical characteristic of the particles to improve
the properties of the material. For example, most medicine nowadays consists of
nanoparticles in order to increase its effectiveness in the body.   

In
this study, the nanoparticle used is from the alginate which is sources from
the brown algae polysaccharides (M. D. Guiry, 2014). Alginate is natural linear
polysaccharides from brown algae that consist of two kinds of hexurunic acids
which are arranged in repeating block MM (GG) or alternating MG block (Qian Li, 2011). Alginate had been choosing
in this study because it has several properties such as biocompatibility, bio-adhesiveness,
and pH sensitivity (Qian Li, 2011) . For example,
hydrogen alginate particle which is produce in aqueous medium by cross-linking
sodium alginate with divalent cations were suggested for the encapsulation of
cells, proteins and oligonucleotides (DNA) (Gombotz & Wee, 1998). However, there is a
weakness in it cross-linking structure. These cross-linking structures are
easily to be destroyed in biological buffer which consist of chelators of
calcium ions or monovalent electrolytes (Qian Li, 2011) . To overcome the
problems, various type of hydrophobic group will be introduced to the main
chain of alginate backbone by using a different method. The amphiphilic derivative
of alginate bearing hydrophobic groups was synthesized in the presence of
organic solvent (Qian Li, 2011). Example of organic
solvent used such as dichloromethane, dimethyl sulfoxide and chloroform. Unluckily,
if the organic solvent was present at high lever than the supported by safety
data, it will bring harmful to human body and to the environment (Liu & Hu).

Vitamin
D3 is known as cholecalciferol (What Is Vitamin D3 (Cholecalciferol)?, 1996-2017) and it is naturally
occurring form of vitamin D (Qian Li, 2011). Vitamin D play
parts of the importance roles in prevention of health and disease which it will
helps to maintain the health of the bone and teeth. It also can be consumed in
the food or supplement. For more information, vitamin D will help support the
health of immune system, brain, and nervous system.  However, vitamin D insufficiency is a wide
spread public health problem since it is a fat-soluble vitamin (Qian Li, 2011). Due to the lining
of the digestive tract is aqueous in nature, to be assimilated; vitamin D is
absorbed in a bile-fat complex which is a water soluble globule with a fatty
core (Williams & Schlenker, 2002). In addition, the
absorption of the vitamin D will be distracted if there is least amount of lipids
in the intestine. In order to improve its dissolvable properties, it is
important to enhance the water solubility. However, in this study, oleoyl
alginate ester nanoparticles act as a carrier for vitamin D3 while vitamin
D3 is a model nutraceutical. Nutraceutical is used to describe about
product which derived from food sources with
extra health benefits to the basic nutritional value found in foods. The term
“nutraceutical” is combination between the two words of “nutrient” which mean a
nourishing food component and “pharmaceutical” which means a medical drug (Dr Tomislav Meštrovi?, MD, PhD, 2015). The nanoparticle
will release the vitamin D3 at a sustain rate in gastrointestinal
fluid. 

 

 

 

1.2  Problem statement 

Nanoparticle
has a wide range of application in variety of fields. Nowadays, nanoparticles
based on food proteins such as lacto globulin and casein has been studied widely
for nutraceutical delivery (Chen, 2005). However, there
still lack of research on application of nanoparticles based on polysaccharides
in the food industry. Recently, researcher has been found that
cholesteryl-grafted sodium alginate was able to self-aggregate as nanoparticles
and to encapsulate the hydrophobic compound pyrene (Yang, Zhang, Wen, Liang, & Zhang, 2007). The hydrophobic
alginate derivative was obtained from modification of alginate by acid chloride
reaction using oleoyl chlorie with the presence of organic solvent (Qian Li, 2011). However, the presence
of the chloroform, dichloromethane and dimethyl sulfoxide as an organic solvent
at higher levels than safety data can bring harmful to humans and to the
environment.  

 

1.3  Expected outcomes

In
this study, OAE samples will be able to self-aggregate to form nanoparticles
when dissolve at low concentrations in aqueous solution such as distilled water
and the nanoparticles will retain their structural integrity in both simulated
gastric fluid (SGF) and simulated intestinal fluid (SIF). This happen may be
due to hydrophobic nutraceutical compound is trapped within the nanoparticles. This
study predicts as the concentration of vitamin D3 increase, the
loading capacity (LC) and the loading efficiency (LE) also will be increase.
Next, this study will prove it is possible to produce hydrophobic oleoyl
alginate ester without using organic solvent.       

 

 

 

1.4  Objective

The
objectives of this study are:

       
i.           
To investigate
the loading and release characteristics of nanoparticles based on oleoyl
alginate ester (OAE) by using vitamin D3 as a model nutraceutical.   

     
ii.           
To elaborate nanoparticles
from hydrophobically modified alginate to develop a carrier for oral
administration of vitamin D3.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MATERIALS AND METHODS

3.1
Materials

The
material will be used during conducting this research are oleoyl chloride, sodium
alginate, formic acids, pancreatin, pepsin, pyrene, and vitamin D3
which will be purchased from Sigma Chemicals and without using too high
purification. Meanwhile methanol which is in the HPLC grade will be purchased
from Merck Chemicals. The rest of the chemicals purification will be based on analytical
grade. The lower molecular weight of alginate sample will be obtained from acid
hydrolysis process and its relative molecular mass is 1.72 x 104 Da (Qian Li, 2011). The molecular mass
will be measured with high-permeation chromatography on an Agilent 1100 system
equipped with a refraction index detector with a column of TSKgel G3000 SWXL.    

3.2
Methods

The
preparation of oleoyl alginate ester (OAE) will be carried out through
esterification process in the present of formic acids, by reacting the low molecular
weight alginate with the oleoyl chloride. The reaction will be handled in a
three-neck flask equipped with a magnetic bar; the first tube is to introduce
the reactant through a peristaltic pump, second tube is for nitrogen bubbling and
the last tube is a cooling system. The flask will be placed in a homo-thermal
magnetic stirrer machine (GuoSheng Co., China) (Qian Li, 2011). The formlylation reaction will be
carried out by impregnating 1 gram of alginate with 10 mL of formic acid for
ten minutes under stirring condition at temperature of 25°C. Next, the sample
of OAE will be prepared for different degrees of substitution by controlling
the reaction between formatted alginate and oleoyl chloride. 8 mL of oleoyl
chloride will be added into the reaction drop wise through the peristaltic
pump.

 

 

Later,
the reaction mixture will be heated for 20 minutes with 50°C temperature. The
another two samples of OAE will be prepared with 16 mL of oleoyl chloride and
will be heated at 30°C and 50°C. A 100 mL of ethanol solution with (95%v/v)
will be added in order to terminate or stop the reaction process. Later, to
obtain the solid phase, the mixture must be filter to separate them. The solid
phase will be washed with ethanol (95%v/v) for several times and will be dry at
room temperature in the vacuum.  

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

RESEARCH FLOW CHART

Figure
3.1 shows the overall experimental process of this study from the raw materials
until the analysis and observation process.

 

 

 

 

GANTT CHART

Year

2017

2018

Project Activities

J

F

M

A

M

J

J

A

S

O

N

D

J

Selection of Project Tittle
a)      Discussion with supervisor
b)      Collect information

 

 

 

 

 

 

 

 

 

 

 

 

Literature Review
a)      Collect information from journal, books and
past thesis.

 

 

 

 

 

 

 

 

 

 

 

Writing Research Proposal
a)      Literature review
b)      Discuss with supervisor
c)      Gathering data information
d)      Submission proposal

 

 

 

 

 

 

 

 

 

 

Research Proposal Presentation
a)      Presentation to supervisor and examiner

 

 

 

 

 

 

 

 

 

 

 

 

 

Final Draft proposal Submission(Chapter1,2,3)

 

 

 

 

 

 

 

 

 

 

 

 

 

Conducting research work
a)      Get the sample in FRIM
b)      Sample preparation for lab
c)      Lab-Moisture content, basic density,
antioxidant and total phenolic compound

 

 

 

 

 

 

 

 

 

 

 

 

 

Data analysis

 

 

 

 

 

 

 

 

 

 

 

 

 

Final report writing
a)      Discuss with supervisor
b)      Analysis the data gathering
c)      Compare data obtained with previous study

 

 

 

 

 

 

 

 

 

 

 

 

 

Final presentation

 

 

 

 

 

 

 

 

 

 

 

 

 

Final report submission

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MILLESTONE

Project activities

Date

Completion of
Final Draft Proposal

May 2018

Completion of
Sample Preparation

August 2018

Completion of
Experiment

October 2018

Completion of
Report and Documentation

December 2018