Recent years have witnessed tremendous growth of nanotechnology based drug delivery system which reduces drug toxicity and side effects and increases the therapeutic index of the drug. Aim of the study is to develop a biodegradable, non-toxic nanoparticle, solely from natural polymers.  Zein – pectin nanoparticle comprising of a hydrophobic zein core and a hydrophilic pectin shell was developed by ultrasonication method. SEM images confirm the nanosize of the nanoparticle. UV- Visible and FT-IR spectroscopic results confirm the incorporation of zein, pectin and the encapsulation of the model drug quercetin in the nanoparticle. Zein is a prolamine class of protein found in wheat, maize etc and pectin is a polymer of galacturonic acid units found in plant cell wal

A T, Dhanya

K R, Haridas

N, Divia

S, Sudheesh

English

International Journal of Drug Delivery

  
 
International Journal of Drug Delivery 4 (2012) 147-152 
http://www.arjournals.org/index.php/ijdd/index 
 
Original Research Article 
             
Development of Zein-Pectin Nanoparticle as Drug carrier 
Dhanya AT1, Haridas KR1, Divia N1, Sudheesh S1* 
 
*Corresponding author: 
 
Sudheesh S 
   
1School of Chemical 
Sciences, Kannur 
University, Payyanur 
Campus, Kerala, India 
 
 
 
 
 
 
A b s t r a c t  
 
Recent years have witnessed tremendous growth of nanotechnology based drug delivery system 
which reduces drug toxicity and side effects and increases the therapeutic index of the drug. Aim of 
the study is to develop a biodegradable, non-toxic nanoparticle, solely from natural polymers. 
Zein – pectin nanoparticle comprising of a hydrophobic zein core and a hydrophilic pectin shell was 
developed by ultrasonication method. SEM images confirm the nanosize of the nanoparticle. UV- 
Visible and FT-IR spectroscopic results confirm the incorporation of zein, pectin and the 
encapsulation of the model drug quercetin in the nanoparticle. 
Zein is a prolamine class of protein found in wheat, maize etc and pectin is a polymer of galacturonic 
acid units found in plant cell wall 
Keywords: Zein, Pectin; Nanoparticle; cytotoxicity; quercetin; ultrasonication 
 
 
 
Introduction 
Improving delivery techniques that minimize toxicity of drug has a 
significant effect on its efficacy [1]. Drug delivery with the aid of 
nanoparticles enhances delivery or uptake by target cells and 
reduces the toxicity of the free drug to non-target organs and 
thereby results in an increase of therapeutic index [2]. The 
cytotoxicity of nanoparticles or their biodegradation products 
remains a major problem, and improvements in biocompatibility 
obviously are a main concern of future research [1]. The 
employment of natural polymers enables the achievement of 
materials with many interesting features such as low toxicity, 
biocompatibility and biodegradability and thereby allows their use 
as carriers. Several works have been published relating to the use 
of natural polymers such as zein, chitosan, alginate, pectin etc in 
drug carrier development [3-8]. 
Zein, a major prolamine class protein from corn is considered and 
generally recognized as safe (GRAS) and food grade ingredient by 
the Food and Drug Administration (FDA). It contains three quarter 
of lipophilic and one quarter of hydrophilic amino acid residues. 
Although zein is a water- insoluble protein, it remains soluble in 
aqueous solutions containing at least 70% alcohol. Because of its 
high hydrophobicity, zein has been successfully applied as a 
promising carrier for encapsulation and controlled release of fat 
soluble compounds [3, 8].  
Pectin is a natural, non-toxic and amorphous polysaccharide 
present in cell walls of plant tissues with considerable potential for 
uses in pharmaceutical formulations, chemistry, natural science 
and so forth [9, 10]. It has high molecular weight heteropolymers 
containing at least 65% (by weight) of D- galacturonic acid units 
which are joined to one another by means of α (1       4) glycosidic 
linkages [4]. Pectin macromolecules are able to bind with some 
organic or inorganic substances via molecular interactions. So, 
pectin can be used to construct matrices to absorb desired 
materials and deliver them in a controlled manner [9]. Incorporating 
low methoxy pectin in the polymer matrix is expected to increase 
the efficiency of encapsulation [4]. Moreover we have reported the 
hypolipidemic and hypoglycaemic activity of pectin earlier [11, 12]. 
Hence our study aims at developing a biodegradable, nontoxic 
nanoparticle by using a low methoxy pectin and zein with an 
average size most appropriate for drug delivery. The developed 
nanoparticle was encapsulated with a model drug quercetin. It was 
subjected to different characterization studies and confirmed the 
incorporation of all the components. 
Materials & Methods 
Zein was purchased from Sigma Aldrich Co, U.S.A. Pectin was 
isolated from locally available fruits of Coccinia indica (Ivy Gourd) 
by the modified method Mohamadzadeh et.al [13]. All other 
chemicals required for the study was purchased from Merck India 
Ltd. 
Estimation of Degree of esterification 
Degree of esterification of isolated pectin was calculated by 
estimating total uronic acid content and by HPLC method [14].  
 
ISSN: 0975-0215 
 
  
This work is licensed under a Creative Commons Attribution 3.0 License.  
Dhanya et al. International Journal of Drug Delivery 4 (2) 147-152 [2012] 
 
PAGE | 148 | 
 
 
 
Preparation of nanozein  
The nanozein was prepared by the modified method of Nicholas et. 
al [5] 
Preparation of drug encapsulated core-shell nanopectin 
The drug encapsulated core-shell nanopectin was prepared by the 
modified method of Nicholas et. al and Vinicius et.al [3,5]. To 
0.01% silicone oil under sonication, Zein and Quercetin (model 
drug) solution in 80% ethanol was added and sonicated until a 
single phase was formed. Pectin solution (1.04mg/ml) was added 
drop wise during sonication at a flow rate of 2.5ml/ min and 
continued sonication for 5 minutes. The solution was then freezed, 
lyophilized and stored. 
Characterization of nanozein and drug encapsulated 
core-shell nanopectin 
Morphology of nanozein and drug loaded core-shell nanopectin 
was investigated by Hitachi SU 6600 variable Pressure field 
Emission Scanning Microscope. The nanoparticle surfaces were 
vacuum sputter coated with gold for allowing the SEM visualization. 
The diameters of nanoparticles were determined.  
UV- Visible absorption spectra of the lyophilized samples of drug 
encapsulated zein nanoparticle, drug encapsulated zein-pectin 
nanoparticle, pure zein, pectin and quercetin (model drug) were 
recorded in the wavelength in the range of 200-500 nm with 
Shimadzu spectrophotometer by using water as reference standard 
and 80% ethanol and water as solvents.   The chemical structure of 
zein, pectin, quercetin, quercetin/zein nanoparticle, 
quercetin/zein/pectin nanoparticle was observed by FT-IR 
spectroscopy (Shimadzu).  
Results and Discussion 
Degree of esterification of pectin from Coccinia indica shows 1.9% 
and 0 .062% of degree of methylation and degree of acetylation 
respectively and confirms that it is low methoxy pectin.  It has been 
reported earlier that low methoxy pectin enhances the drug 
encapsulation. Hence pectin from Coccinia indica can be used for 
drug encapsulation. 
Morphological observation 
Morphological observation of zein nanoparticles and drug 
encapsulated zein- pectin nanoparticle by Scanning Electron 
Microscope are shown in figure 1 & 2.  Zein nanoparticles formed 
smooth surfaced nanospheres with an average diameter of 
approximately100nm. However the size ranged from 54nm to 
300nm. More homogenous and smaller nanospheres than zein 
nanospheres were formed when pectin solution was added. The 
approximate size was 93nm and the size ranged from 56nm 
to178nm. The reduced size of nanopectin may be due to the 
electrostatic interactions existing between quercetin/zein and 
pectin particles. 
The zein nanoparticle and drug encapsulated zein –pectin 
nanoparticle was characterized by means of UV- Visible and 
Fourier Transform Infra Red spectroscopic analysis. The analysis 
shows that the drug has been incorporated in the zein-pectin 
nanoparticle. 
Overlay spectrum (Fig.3) shows UV-Vis spectra of the zein, 
quercetin, nanozein, drug loaded zein-pectin nanoparticle (80% 
ehanol as solvent), pectin and drug loaded zein-pectin nanoparticle 
(water as solvent). From the UV- Visible spectrum it can be 
observed that λ max of pure pectin matches with the λ max of drug 
loaded core shell – nanopectin (water as solvent for both) which 
confirms the presence of pectin in the drug loaded zein-pectin 
nanoparticle. λ max’s of all other components were not visible since 
those are insoluble in water. When 80% ethanol was taken as 
solvent, λ max’s of pure zein and quercetin was also seen in drug 
loaded nanozein in which zein and quercetin are incorporated and 
in drug loaded zein-pectin nanoparticle in which pectin, zein and 
quercetin was incorporated. From these data it can be assumed 
that core- shell nanopectin is formed and drug has been 
incorporated in the nanoparticle. 
FT-IR spectrum of drug loaded zein-pectin nanoparticle (Fig.7) 
shows very prominent peaks at 3300, 1650, 1540 and 1450 cm-1. 
The peak at 3300 cm-1 which is assigned to – OH is found to be 
shifted from 3400 cm-1 range in pectin, zein and quercetin, this may 
be due to  interactions of –OH groups among them. The peak at 
1650 cm-1 may be due to the C-C multiple bonds stretching in tetra 
substituted alkene shifted towards a higher wave number. The 
peaks at 1540 and 1510 cm-1 may be assigned to – NH bending 
vibrations which is seen in pectin and zein as well. Another peak at 
1450 cm-1 can be due to C-C multiple bonds stretching of aromatic 
compounds which show the presence of quercetin in zein-pectin 
nanoparticle. A new band at 1748 cm-1 assigned to C=O of –
COOH confirms the presence of pectin in the nanoparticle 
developed. Another band observed at 1300 cm-1 which can be 
assigned to OH bending and C-O stretching vibrations of phenols 
confirms the presence of the model drug quercetin. 
  
 
 
 
 
Figure: 1 SEM image of Nanozein 
 
 
Figure: 2 SEM image of drug encapsulated                            
zein – pectin nanoparticle 
 
Dhanya et al. International Journal of Drug Delivery 4 (2) 147-152 [2012] 
 
PAGE | 149 | 
 
 
 
 
Figure: 3 Overlay spectrum of zein, Quercetin, pectin, nanozein and zein-pectin nanoparticle 
 
 
 
Figure: 4 FT- IR Spectrum of Pectin from Coccinia indica 
 
 
 
Dhanya et al. International Journal of Drug Delivery 4 (2) 147-152 [2012] 
 
PAGE | 150 | 
 
 
 
 
Figure: 5 FT-IR spectrum of Zein 
 
 
Figure: 6 FT-IR spectrum of Quercetin 
 
Figure: 7 FT-IR spectrum of Drug loaded  zein – pectin nanoparticle 
Dhanya et al. International Journal of Drug Delivery 4 (2) 147-152 [2012] 
 
PAGE | 151 | 
 
 
 
FT-IR spectrum of drug loaded zein-pectin nanoparticle (Fig.7) 
shows very prominent peaks at 3300, 1650, 1540 and 1450 cm-1. 
The peak at 3300 cm-1 which is assigned to – OH is found to be 
shifted from 3400 cm-1 range in pectin, zein and quercetin, this may 
be due to  interactions of –OH groups among them. The peak at 
1650 cm-1 may be due to the C-C multiple bonds stretching in tetra 
substituted alkene shifted towards a higher wave number. The 
peaks at 1540 and 1510 cm-1 may be assigned to – NH bending 
vibrations which is seen in pectin and zein as well. Another peak at 
1450 cm-1 can be due to C-C multiple bonds stretching of aromatic 
compounds which show the presence of quercetin in zein-pectin 
nanoparticle. A new band at 1748 cm-1 assigned to C=O of –
COOH confirms the presence of pectin in the nanoparticle 
developed. Another band observed at 1300 cm-1 which can be 
assigned to OH bending and C-O stretching vibrations of phenols 
confirms the presence of the model drug quercetin was taken as 
solvent, λ max’s of pure zein and quercetin was also seen in drug 
loaded nanozein in which zein and quercetin are incorporated and 
in drug loaded zein-pectin nanoparticle in which pectin, zein and 
quercetin was incorporated. From these data it can be assumed 
that core- shell nanopectin is formed and drug has been 
incorporated in the nanoparticle.  
FT–IR Spectroscopy 
Figures 4-7 shows the FT-IR spectra of pectin from Coccinia 
indica, zein, quercetin (model drug), and drug loaded zein – pectin 
nanoparticle respectively.  
FT – IR spectrum of pectin (Fig. 4) exhibits a broad intense peak at 
3425.1 cm-1 which may be assigned to O-H vibrational stretching. 
An intense peak at 1748 cm-1 is due to C=O of –COOH and the 
peak at 1642 cm-1 may be due to C-C multiple bond stretching. 
Another peak at 1531.5 cm-1 is due to N-H bending vibrations. The 
peaks 1236.4 cm-1 and 1153.5 cm-1 are assigned to O-H bending 
and C-O stretching of 10 and 20 alcohols respectively. The peak 
seen at 1019.4 cm-1 may be due to the aliphatic C- N vibration. 
FT – IR spectrum of zein (Fig. 5) exhibits an -OH peak at 3400 cm-
1 and another peak at 1637 cm-1 may be due to C-C multiple bond 
stretching. The peak at 1475 cm-1 is due to N-H bending vibrations 
of amine salts. Less intense peaks for -OH bending and stretching 
vibrations of 10, 20 and 30 alcohol are observed at 1287.5 cm-1 
to1175 cm-1. Two bands at 1225 cm-1 and  1037.5 cm-1 may be 
assigned to C- N vibrations of aliphatic and aromatic moieties. 
FT-IR spectrum of quercetin (Fig. 6) shows intense peaks at 3411 
cm-1 and 3311 cm-1 which may be due to the presence of –OH 
group. Another peak observed at 3125 cm-1 may be assigned to - 
OH stretching of the chelate compounds due to the polymeric 
association. A peak observed at 1687.5 cm-1 may be due to the 
ketone stretching. The peaks observed at 1512 cm-1 and 1450 cm-1 
are due to the presence of aromatic C-C multiple bond stretching. 
The peak observed at 1300 cm-1 may be assigned to OH bending 
and C-O stretching vibrations of phenols. 
Overlay spectrum (Fig.3) shows UV-Vis spectra of the zein, 
quercetin, nanozein, drug loaded zein-pectin nanoparticle (80% 
ehanol as solvent), pectin and drug loaded zein-pectin nanoparticle 
(water as solvent). From the UV- Visible spectrum it can be 
observed that λ max of pure pectin matches with the λ max of drug 
loaded core shell – nanopectin (water as solvent for both) which 
confirms the presence of pectin in the drug loaded zein-pectin 
nanoparticle. λ max’s of all other components were not visible since 
those are insoluble in water. When 80% ethanol  
Conclusion 
A natural core-shell nanoparticle comprising of zein and pectin was 
successfully developed and the model drug quercetin was 
encapsulated within.    
From overall results of morphological and physical 
characterization, it can be concluded that the zein-pectin 
nanoparticle developed can be used as a drug carrier. A further 
study on temperature stability, drug release and drug targeting is 
under progress. 
Author’s Contributions 
DAT carried out extraction of pectin from Coccinia indica, 
estimation of degree of esterification, preparation of nanozein and 
zein – pectin nanoparticle and preparations for SEM, UV-Visible, 
FT-IR studies etc.SS guided the study by designing and 
coordinating the work and helped in drafting the manuscript. 
HKR participated in interpreting the FT – IR spectral data. 
DN performed UV- Visible analysis of the samples and interpreted 
the overlay spectrum 
Conflict of Interest 
There is no conflict of interest 
 
Acknowledgment 
The authors are thankful to National Institute for Technology, 
Kozhikode, Kerala, for SEM analysis and Sree Chitra Tirunal 
Institute for Medical Sciences & Technology, Thiruvananthapuram, 
Kerala, for HPLC analysis.The financial support provided by Kerala 
Biotechnology Commission (KSCSTE) for conducting this research 
is also gratefully acknowledged. 
.
 
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Development of Zein-Pectin Nanoparticle as Drug carrier

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Development of Zein-Pectin Nanoparticle as Drug carrier

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