Content uploaded by Ahmed Abbas Hussein
Author content
All content in this area was uploaded by Ahmed Abbas Hussein on Oct 22, 2024
Content may be subject to copyright.
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
DOI: https://doi.org/10.31351/vol31iss1pp119-129
119
Investigation of Lipid Polymer Hybrid Nanocarriers for Oral Felodipine
Delivery: Formulation, Method, In-vitro and Ex-vivo Evaluation
Hayder Kadhim Drais1,*, Ahmed Abbas Hussein**
*Ministry of Health and Environment, Babil Health Directorate, Babil, Iraq.
**Baghdad College of Medical Sciences, Baghdad, Iraq.
Abstract
The antihypertensive felodipine is a calcium-channel blocking agent. It is practically insoluble in water
and shows low oral bioavailability (15%-20%). This investigation aims to formulate and characterize felodipine
lipid polymer hybrid nanocarriers (LPHNs) to be given orally by two nanovesicles formulating methods and make
comparative analysis through characterization process and in vitro and ex vivo intestinal permeation evaluation.
The felodipine LPHNs formulations (HF1-HF3) were prepared by the new microwave-based method and that
felodipine LPHNs formulations (HF4-HF6) were prepared by a single emulsification solvent evaporation technique
(SESET). All formulations (HF1-HF6) enter the characterization process. The felodipine LPHNs formulations
(HF1-HF6) were prepared successfully and undergo different characterization processes to make a comparative
study between formulations prepared by different methods. It was found that formulas prepare by a microwave-
based method are most superior to the SESET. The felodipine LPHNs formulations HF1-HF3 has lower particle
size, lower PDI and higher zeta potential, significantly higher (p< 0.05) dissolution rate, and significantly higher
(p< 0.05) intestinal permeation study than the felodipine LPHNs formulations HF4-HF6. The microwave-based
method is a very successful technique in preparing felodipine LPHNs formulations (HF1-HF3) and prepotent to
the SESET. All the felodipine LPHNs formulations (HF1-HF6) show extended drug release nanosystem.
Keywords: Lipid-polymer hybrid nanocarriers, Felodipine , Microwave based method, SESET.
ةصخلا
LPHNs
HF1-HF3LPHNs HF4-HF6
SESETHF1-HF6LPHNsHF1-HF6
LPHNs HF1-HF3
PDI p p LPHNsHF4-HF6 LPHNs HF1- HF3SESETLPHNs HF1-HF6
Introduction
It became clear that nanotechnology has
entered into the development of most areas of life,
including the development and manufacture of
medicines (1). Despite the great discoveries of drugs,
they remain ineffective until a safe way is found to
deliver them to the circulatory system or site
targeting. Felodipine is the calcium-channel
antagonist. Its use in the treatment of hypertension
and angina pectoris.
It is practically insoluble in water and
undergoes a hepatic first-pass effect that leads to low
oral bioavailability (15%-20%) (2). The oral route of
drug administration is most prevalent where and has
many merits such as its safe, easy to administer
therapeutic agent, not associated with pain as in the
intravascular route. But the oral route has some
demerits such as intestinal and hepatic first-pass
effect and is associated with low bioavailability
deera2020@gmail.com :mail-Corresponding author E
1
Received: 12/6/2021
Accepted: 5/ 9/2021 Iraqi Journal of Pharmaceutical Science
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
120
for drugs that have low solubilities such as
felodipine and low permeability(2). In addition to
oral route obstacles, the conventional oral delivery
system has many demerits such as poor patient
amenability, increased opportunity of dose missing
of a drug with a short half-life for which frequent
taking is required and presence of typical peak and
valley in the blood concentration/time curve lead to
fluctuations in therapeutic agent level this will create
adverse effects particularly in therapeutic agent with
low therapeutic index (3). Nanoparticles played a
major role in carrying therapeutic agents and
delivering them to the place of effectiveness. The
presence of nanoparticles led to an increase in
solubility, an increase in the surface area for drug
release, and thus increased absorption and
bioavailability (4). There are many nanoparticles
used in drug delivery, but they are associated with
some obstacles that interfere with drug efficacy. The
most important obstacles are low drug loading, drug
expulsion, and drug instability after administration
and in long-term storage. The lipid polymer hybrid
nanocarriers (LPHNs) system is a nanoparticulate
system for drug delivery. It protects the
encapsulated drug from obstacles associated with
another nanoparticle system(5-7). The LPHNs consist
of lipid content and polymeric ingredient (8). The
lipid content enhances the solubility of hydrophobic
drugs, improves membrane permeability leads to
increase bioavailability. The presence of polymer
within the structure of LPHNs provide more control
to release of loaded drug. The hybridization between
lipid and polymeric ingredients creates a
nanoparticulate system which is LPHNs that
characterized mainly by toughness, robust
nanoparticles, provide extended-release drug
delivery, higher drug payload, and high stability in
the human circulatory system and during
formulation storage(9). There are many methods of
LPHNs preparation with some limitations such as:
the presence of impurities in final products, costly,
require high time that delays research field, and low
stability. The newly microwave-based method is
employed with great success in the formulation of a
highly advanced nano system which is LPHNs and
characterized by economical, absence of impurities
associated with other nanoparticle preparation
methods, inexpensive, high stable of final
formulation (10-12). The single emulsification solvent
evaporation technique (SESET) is widely used in the
preparation of nanocarriers. The SESET has been
successfully used to loading a variety of
hydrophobic therapeutic agents with good
reproducibility, high yield, and ease of scaling up
(13).
This study aims to prepare and characterize oral
felodipine lipid polymer hybrid nanocarriers
(LPHNs) by two nanoparticle preparing methods to
make a comparative study through the
characterization process and in vitro and ex vivo
intestinal permeation evaluation.
Material and Methods
Materials
The Labrasol, PEG laurate, and PEG
oleate were purchased from Beijing Yibai
Biotechnology Co., Ltd. China. The methanol,
ethanol, KCl, HCl, KH2PO4, and Na2HPO4 Grin land
chemical comp. The U.K. The felodipine, lauric
acid, polysorbate 80, polysorbate 20, span 80,
propylene glycol, and NaOH were purchased from
Nanjing Duly Biotech Co., Ltd. China. The aniseed
oil, argan oil, and cardamom oil were purchased
from Hemani international KEPZ, Karachi,
Pakistan. The Olibanum oil was purchased from AI-
Emad for plant oil products. Iraq. The fenugreek oil
was purchased from BAR-SUR-LOUP GRASSE
(A.M).
Methods
The microwave-based method
The hydrophobic blend was prepared under
a magnetic stirrer device at 1000 rpm for 5 minutes.
It contains felodipine, lauric acid and chitosan that
was dissolved in cardamom oil : PEG-laurate. The
hydrophilic blend contains distilled water,
polysorbate 80 and propylene glycol related to
optimized amounts. By the application of
microwave device for less than 15 seconds to the
mixture of the two blends and under magnetic stirrer
device at 1000 rpm for adequate time (seconds to
minutes according to a final volume of dosage
form), a colloidal dispersion system of felodipine
LPHNs will be prepared. The optimized felodipine
LPHNs formulations (HF1-HF3) as shown in Table
(1), are immediately used for the study or
lyophilized to be filled in hard gelatin capsules. In
the lyophilization process (freeze-drying), the
samples were first frozen at -20 °C for 2 h, then
transferred at -80 oC for 22 h., and then lyophilized
at 0.001 mbar at -104 °C for 24 h using lactose
10%(w/w) as a cryoprotectant (10,14).
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
121
Single emulsification solvent evaporation
technique (SESET)
In this method, the felodipine and the
chitosan are dissolved in organic solvent and mix
with the lipophilic phase, then it was added into an
aqueous phase containing PEG laurate: polysorbate
80: propylene glycol. Under magnetic stirrer device
and probe or ultra-sonication process result in the
formation oil in water (o/w) emulsion. The organic
solvent is removed by a rotary evaporator, yielding
the felodipine lipid polymer hybrid nanocarriers.
The optimized felodipine LPHNs formulations
(HF4-HF6) as shown in Table (1), are immediately
used for investigation or pass through freeze drying
to fill in hard gelatin capsules. In freeze-drying, the
samples were first frozen at -20 °C for 2 h, then
transferred at -80 oC for 22 h., and then lyophilized
at 0.001 mbar at -104 °C for 24 h using lactose 10%
as a cryoprotectant (15,16).
Table 1. The selected felodipine LPHNs formulations (HF1-HF6) for characterization and optimization.
Formulation
code
Method of
preparation
Felodipine
% (w/w)
Cardamon
oil
% (w/w)
Lauric
acid
%
(w/w)
Chitosan
% (w/w)
PEG-(400) laurate
:Polysorbate
80: Propylene
glycol % (w/w)
Distilled
water
% (w/w)
up to
HF1
Microwave
based
method
1
8
2
0.2
17.5:8.75:8.75
100
HF2
Microwave
based
method
1
8
2
0.25
20:10:10
100
HF3
Microwave
based
method
1
8
2
0.35
22.5:11.25:11.25
100
HF4
SESET
1
8
2
0.2
17.5:8.75:8.75
100
HF5
SESET
1
8
2
0.25
20:10:10
100
HF6
SESET
1
8
2
0.35
22.5:11.25:11.25
100
Characterization of the felodipine LPHNs
formulations(HF1-HF6)
Globule size determination
The globule size is determined by the
nanoparticle analyzer model SZ-100 - nanopartica
series instruments from Horiba scientific company.
The photo correlation spectroscopy (PCS), is a
technique that has been used to determine the
particle size of felodipine LPHNs formulation. The
experiments were performed in triplicate (17).
Polydispersity Index (PDI) determination
The uniformity of the nanosystem of
colloidal attributes is determined by the
Polydispersity Index (PDI). It is measured by PCS
technique. As PDI value increases, the uniformity
of nanocarriers of the felodipine LPHNs
formulations will decrease. The experiments were
performed in triplicate (17).
Zeta Potential (ZP) measurement
The ZP is an index to determine the
stability of the colloidal dispersion system that is
affected by DLVO forces. It is measured by PCS
technique. The ZP determines the surface charge
which can develop around nanocarriers in a
dispersion medium. The experiments were achieved
in triplicate (17).
Entrapment efficiency (EE) and drug loading (DL)
determination
The entrapment efficiency of felodipine
LPHNs formulation is determined by the indirect
method through, 0.1 mL of freshly prepared
felodipine LPHNs formulation was taken and add
to it 9.9 mL ethanol for dilution. The obtained
colloidal dispersion was centrifuged for 15 minutes
at 10000 rpm. The supernatant layer was removed
and filtered through a 0.45 μm filter. The filtrated
liquid was diluted in a sufficient amount of ethanol
and analyzed by an ultraviolet (UV)
spectrophotometer at 361.5 nm. This study was
performed in three trials (18).
The encapsulation efficiency (EE) can be indirectly
determined by the following equation (1):
EE (%)= [(Weight of felodipine in LPHNs)/𝑇𝑜𝑡𝑎𝑙
felodipine amount)] ×100 ………..… (1)
The drug loading (DL) expressed in percentage (%)
is the quantity of active pharmaceutical agents
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
122
present in the nanocarriers divided by the total
quantity of lipid present in the nanosystem. It is
measured by the equation(2):
DL (%)=[(Weight of felodipine in LPHNs)/Weight
of LPHNs] × 100 ………..… (2)
The pH determination
It has a great effect on the solubility of the
therapeutic agents, the formulation attributes,
tolerability, formulation stability, and the
therapeutic agent's activity. The pH determination is
an important factor in felodipine LPHNs
formulations due to its relation to the stability and
activity of pharmaceutical nanocarriers. The
alteration of pH may be related to chemical
interactions that can affect product quality. The
digital pH meter employs to measure the pH of the
felodipine LPHNs formulations. The study was
achieved in triplicate (19).
Percent of light transmittance measurement
It is an important parameter that ascertains
colloidal attributes of felodipine LPHNs
formulations. The percent of light transmittance was
determined by a UV-Visible spectrophotometer to
preserve double distilled water as blank at 600 nm.
Results were being taken in triplicate (20).
In vitro felodipine release experiment
The experiment was achieved for
felodipine LPHNs (HF1-HF3) prepared by
microwave-based method and felodipine LPHNs
(HF4-HF6) that is prepared by single emulsification
solvent evaporation technique and compare it with
drug dissolution from pure drug suspension using
the combination method of (USP dissolving type I
apparatus - dialysis bag technique).Two dissolution
media have been used which are HCl buffer pH 1.2
+ 0.3 % polysorbate 80 solution and phosphate
buffer pH 6.8 + 0.3 % polysorbate 80 solution. The
dissolution medium volume is 900 mL for each
experiment at 37 ± 0.5 o C with constant stirring at
50 rpm. The drug amount in each of felodipine
LPHNs (HF1-HF6) and pure drug suspension was
5mg of felodipine. Samples were withdrawn at
predetermined intervals of time (5, 10, 15, 30, 60
minutes, and 2,4,8,12,24,36 hours) and filtered by
microfilter paper of 0.45 µm pore size and
compensate by equal withdrawn volume. At 361.5
nm, the felodipine concentration determines by
spectrophotometrically(21,22). The study was
performed in triplicate and the results were analyzed
using ANOVA statistical test at level p< 0.05. The
drug liberation kinetics study performed by fit the
obtained kinetic data into the various kinetic
equations. The value of regression coefficients (R2)
will explain the resultant model. The mechanism of
felodipine release was obtained from the slope of the
Korsmeyer Peppas equation (23,24).
Ex-vivo intestinal permeation study
The study of ex-vivo intestinal permeation
was performed using the non-everted sac technique
(25,26). The fasted male sheep weighing about 16 kg
was slain and anatomized under license university of
Baghdad/ College of Pharmacy. The small intestine
is a tested region that isolated and mesentery residue
was removed and washed with normal saline
solution. Several pieces of 5 cm in length and 2 cm
diameter of the small intestine was produced after
the cutting process. Insert in each piece that ligates
from one end one capsule of (felodipine LPHNs
HF1-HF6 ) and (felodipine drug suspension) where
all capsules contain 5mg of felodipine and add 4.5 g
of phosphate buffer pH 6.8 solution and tied the
other end. Insert the tested segments in 900 ml of
liquid which is phosphate buffer pH 7.4
solution+0.3% polysorbate 80mg using apparatus 1
rotating basket (Biobase Meihua Trading Co.,
Ltd.). At predetermined time intervals
(5,10,15,30,60,90,120,150,180,210, 240 minutes)
samples (5 ml) were taken and filtered by microfilter
paper(0.45 µm) then find the felodipine
concentration by UV spectrophotometer where the
wavelength is 361.5 nm. The replenishment with an
equal volume of withdrawn sample by diffusion
liquid at once. The study was achieved in three trials
and the outcome was analyzed statistically by
ANOVA test at p< 0.05.
The effective membrane permeability was achieved
by equation (3).
M = Peff S Cd tres ………..… (3)
where,
M = therapeutic agent amount that absorbed
Peff = permeability coefficient (effective membrane
permeability).
Cd = drug concentration at donor compartment
tres = residence time of drug in GI lumen.
S = surface area available for absorption
Statistical analysis
The research experimental data
documented as mean ± SD (n=3). A statistical study
was performed by analysis of variance (ANOVA)
where a p-value less than 0.05 indicates a significant
outcome(36).
Results and Discussion
Preparation of felodipine lipid polymer hybrid
nanocarriers
The felodipine LPHNs formulations (HF1-
HF3) were prepared by the new microwave-based
method according to specified concentrations of
components as shown in Table (1). The microwave's
role in the formulation process was according to the
following mechanism: when microwave passes
through the ingredient of felodipine LPHNs
formulation, the hydrogen bond, Vander walls
bonds, electrostatic interactions, and hydrophobic
forces between the molecules undergo temporary
breakage. The microwave causes dipolar rotation
and ionic conduction that cause molecular
oscillation and molecular collisions that raise the
thermal energy of the system lead to weak bonds
disruption. The increased thermal energy for the
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
123
system create tamed molecules that is more
favorable and faster to get self-assembly by the
magnetic stirrer agitation to produce felodipine
LPHNs formulation(10,14). The felodipine LPHNs
formulations (HF4-HF6) were prepared by single
emulsification solvent evaporation technique
(SESET) according to specified concentrations of
components as shown in Table (1). The mechanism
of SESET in felodipine LPHNs formulation depends
on the formation of oil/water emulsion that finally
produces hybrid nanocarriers through a single
formulation step(15,16).
Characterization of the prepared felodipine
LPHNs formulations
The following tests were utilized to characterize the
prepared felodipine LPHNs systems:
Globule size determination
The results were HF1 (146.2 nm); HF2
(94.2 nm); HF3 (75.1 nm); HF4 (743.2 nm); HF5
(169.7) and HF6 (179.2 nm). The outcomes indicate
that all felodipine LPHNs formulations( HF1-HF6)
have nanoscale particle size and colloidal dispersion
attributes. The analysis of variance indicates that
there is a significant (p<0.05) decrease in particle
size as the increase the concentration of PEG laurate:
polysorbate 80: propylene glycol blend at constant
lipid content.
Polydispersity Index (PDI) determination
PDI is a homogeneity parameter that
determines the size distribution of nanocarriers
within a colloidal dispersion system. Its value ranges
from 0 to 1. The smaller values near zero indicate a
more homogenous globule size distribution while
large values that approach 1 indicate a wider globule
distribution (27). The results of felodipine LPHNs
formulations (HF1-HF6) are HF1 (0.547); HF2
(0.54); HF3 (0.335); HF4 (0.888); HF5 (0.551) and
HF6 (0.411). The outcome explains that there is a
significant (p<0.05) decrease in PDI as the increase
the concentration of PEG laurate: polysorbate 80:
propylene glycol blend at constant lipid
concentration.
Zeta Potential (ZP) measurement
The zeta potential is a parameter employ to
measure the surface charge of felodipine LPHNs. It
explains the physical stability of some colloidal
dispersion systems. The results of felodipine
LPHNs formulations (HF1-HF6) are HF1 (2.3 mV);
HF2 (10.3 mV); HF3 (10.2 mV); HF4 (0.6mV); HF5
(-2.8mV) and HF6 (1.3mV). The felodipine LPHNs
formulations(HF1-HF6), depend on non DLVO
forces which are steric forces and hydration forces
to stabilize the nanovesicles. Therefore the low
value of zeta potential of LPHNs formulations(HF1-
HF6) do not affect the physical stability and remain
to withstand the process of nanocarrier aggregation
because the stabilization process performed by non
DLVO forces (28) The ANOVA explain there is a
nonsignificant (p 0.05) correlation between
independent variables and zeta potential factor.
Entrapment efficiency (EE) and drug loading (DL)
determination
The entrapment efficiency and felodipine
loading is an important factor that employs for the
evaluation and characterization of felodipine
LPHNs formulations. The EE outcome of
felodipine LPHNs formulations (HF1-HF6) are HF1
(85.443 % w/w ); HF2 (84.81w/w); HF3 (84.177%
w/w); HF4 (84.2% w/w); HF5 (83.4% w/w ) and
HF6 (82.8% w/w). It was found that the preparations
(HF1-HF3) are more EE than preparations (HF4-
HF6). The ANOVA indicates there is a significant
(p 0.05) correlation between independent variables
and EE parameter.The DL outcome of felodipine
LPHNs formulations (HF1-HF6) are HF1 (8.544%
w/w ); HF2 (8.481% w/w); HF3 (8.418% w/w); HF4
(7.7% w/w); HF5 (7.5% w/w ) and HF6 (7.4% w/w).
It was found that the preparations (HF1-HF3) are
more DL than preparations (HF4-HF6) due to lower
particle size and lower lipid content. The ANOVA
indicates there is a significant (p 0.05) correlation
between independent variables and the DL factor.
The pH determination
The pH outcome of felodipine LPHNs
formulations (HF1-HF6) are HF1 (4.2); HF2 (4.1);
HF3 (4.3); HF4 (4.1); HF5 (4.1 ) and HF6 (4.2). The
result shows that the felodipine LPHNs formulations
(HF1-HF6) had a suitable acidic pH value in the
range of (4.1 – 4.3) that is better for oral
administration(29). Also that the preparations (HF1-
HF3) are nearly similar pH values of preparations
(HF4-HF6). The analysis of variance shows a
significant relationship (p<0.05) between
independent variables and pH parameter.
Percent of light transmittance measurement
The percent (%) of light transmittance is an
attractive parameter to explain physically the
colloidal properties of felodipine LPHNs
formulations (HF1-HF6).The results of light
transmittance percentage of felodipine LPHNs
formulations (HF1-HF6) are HF1 (94.3%); HF2
(95.2%); HF3 (92.3%); HF4 (89.2%); HF5 (90.1%)
and HF6 (90.8%).The outcomes show that the
preparations (HF1-HF3) are more transparent than
preparations (HF4-HF6) and all felodipine LPHNs
formulations (HF1-HF6) gave features of colloidal
dispersion (30). The analysis of variance indicated a
significant relationship between independent
variables and light transmittance percentage at a
level (p<0.05).
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
124
In vitro felodipine release experiment
The experiment was done using the
combinational method which is (USP type I (Basket)
- dialysis bag technique in two dissolution media
which are HCl buffer pH 1.2 + 0.3 % polysorbate 80
solution and phosphate buffer pH 6.8 + 0.3 %
polysorbate 80 solutions. The outcome ascertains
there is no burst release of the therapeutic agent from
all felodipine LPHNs formulations (HF1-HF6) and
there was an extended-release process over 36 hours
from all felodipine LPHNs formulations (HF1-HF6)
(46).
In dissolution medium of HCl buffer pH 1.2 + 0.3 %
polysorbate 80 solutions as shown in Figures (1,2,3),
the felodipine release profile was significantly
higher (p-value <0.05) in dissolution rate for HF3
and was significantly lower (p-value < 0.05) in
dissolution rate of the pure drug. The comparability
profile of felodipine release for formulas have
similar concentration with different preparation
method was explained as following: HF1> HF4,
HF2> HF5 and HF3> HF6 while the comparability
profile of the felodipine release from LPHNs
formulation (HF1-HF6) and the pure drug
suspension explains in the following descending
order: HF3 > HF 6 > HF 2 > HF 5 > HF 1 > HF 4 >
pure drug suspension. The result indicates that
formulas were prepared by microwave-based
method to provide a higher dissolution rate in
comparison to formulas were prepared by SESET.
This is due to the microwave-based method was
prepared formulas (HF1-HF3) has lower particle
size than formulas (HF4-HF6) which prepared by
SESET, which provide higher surface area for
exposure to the dissolution medium lead to increase
the rate of felodipine dissolution.
In dissolution medium of phosphate buffer pH 6.8 +
0.3 % polysorbate 80 solution as shown in Figures
(4,5,6), the felodipine release profile was
significantly higher (p-value <0.05) in dissolution
rate for HF2 and was significantly lower (p-value <
0.05) in dissolution rate of pure drug suspension.
The comparability profile of felodipine release for
formulas have similar concentration with a different
preparation method was explained as following:
HF1> HF4, HF2> HF5 and HF3> HF6 while the
comparability profile of the felodipine release from
felodipine LPHNs formulations (HF1-HF6) and the
pure drug suspension explains in the following
descending order: HF2 > HF 3 > HF 1 > HF 6 > HF
5 > HF 4 > pure drug suspension. The result
indicates that formulas were prepared by a
microwave-based method to provide a higher
dissolution rate in comparison to formulas were
prepared by SESET. This is due to the microwave-
based method was prepared formulas (HF1-HF3)
has lower particle size than formulas (HF4-HF6)
which prepared by SESET, which provide higher
surface area for exposure to the dissolution medium
lead to an increased rate of felodipine dissolution.
In the dissolution rate of felodipine profile, it was
observed that the pure drug suspension gives a lower
release rate of the therapeutic agent in comparison
to all optimized LPHNs formulations (HF1-HF6)
because that LPHNs formulations (HF1-HF6) have
a large surface area that exposes to the dissolution
medium and this will allow a higher interaction with
the dissolution medium that increases dissolution
rate (47,48). The analysis of variance indicated a
significant (p<0.05) relationship between
independent variables and felodipine release.
Figure 1. In vitro felodipine release profile from
LPHNs formulations HF1,HF4 and the pure
drug at HCl buffer pH 1.2 + 0.3 % polysorbate
80 solutions, the values of mean ±SD (n=3).
Figure 2. In vitro felodipine release profile from
LPHNs formulations HF2,HF5 and the pure
drug at HCl buffer pH 1.2 + 0.3 % polysorbate
80 solutions, the values of mean ±SD (n=3).
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
125
Figure 3. In vitro felodipine release profile from
LPHNs formulations HF3,HF6 and the pure
drug at HCl buffer pH 1.2 + 0.3 % polysorbate
80 solutions, the values of mean ±SD (n=3).
Figure 4. In vitro felodipine release profile from
LPHNs formulations HF1,HF4 and the pure
drug suspension at phosphate buffer pH 6.8 + 0.3
% polysorbate 80 solutions the values of mean
±SD (n=3).
Figure 5. In vitro felodipine release profile from
LPHNs formulations HF2,HF5 and the pure
drug suspension at phosphate buffer pH 6.8 + 0.3
% polysorbate 80 solutions.
Figure 6. In vitro felodipine release profile from
LPHNs formulations HF3,HF6 and the pure
drug suspension at phosphate buffer pH 6.8 + 0.3
% polysorbate 80 solutions.
Kinetic analysis of release drug
The kinetic data were summarized in Table
(3) and Table (4), were obtained from in vitro release
of felodipine LPHNs formulations (HF1-HF6) and
the pure drug suspension at HCl buffer pH 1.2 + 0.3
% polysorbate 80 solution and phosphate buffer pH
6.8 + 0.3 % polysorbate 80 solutions. It was fitted to
different models that determine the mechanism of
therapeutic agent release. The kinetic models that
employ in the investigation were zero-order, first-
order kinetic, Higuchi model, and Korsmeyer-
Peppas model. The outcome indicates Higuchi's
model was obtained due to the higher regression
coefficient (R2) is obtained for it. This indicates
felodipine was release from the monolithic system.
The felodipine liberation from LPHNs formulations
(HF1-HF6) following non Fickian/anomalous
dissolution (diffusion and erosion) due to the drug
liberation exponent were significantly higher
(p<0.05) than 0.43(23).
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
126
Table (2). Summary of characterization results of felodipine LPHNs formulations (HF1-HF6).
code
Globule size
(nm)*
PDI*
Zeta
potential
(mV)*
Entrapmen
t efficiency
% (w/w)*
Drug
loading
% (w/w)*
pH*
Percent of light
transmittance*
Release percent of
felodipine in HCl
buffer pH 1.2 + 0.3
% polysorbate 80
solutions
at 24 hours*
Release percent of
felodipine in
phosphate buffer
pH 6.8 + 0.3 %
polysorbate 80
solutions at 24
hours*
HF1
146.2±10.095
0.547±
0.0502
2.3± 0.264
85.443±5.0
58
8.544±0.478
4.2±0.435
94.3± 0.8
74.21± 0.7
89.882± 1.268
HF2
94.21± 1.587
0.54± 0.078
10.3±0.721
84.81±
4.013
8.481±0.354
4.1±0.264
95.2± 0.964
77.682± 0.532
90.575± 0.238
HF3
75.1±2.816
0.335± 0.005
10.2±0.754
84.177±2.5
61
8.418±0.256
4.3± 0.36
92.3± 0.888
81.154± 0.872
90.2± 0.435
HF4
743.2±8.861
0.888± 0.007
0.6± 0.264
84.2± 4.529
7.7± 0.608
4.1±0.132
89.2± 0.869
69.21± 0.642
80.6± 1.587
HF5
169.7± 9.553
0.551± 0.005
-2.8±1.081
83.4± 3.984
7.5± 0.529
4.1±0.264
90.1± 1.058
74.98± 1.048
85.3± 0.264
HF6
179.2± 8.827
0.411± 0.011
1.3± 0.2
82.8± 3.862
7.4± 0.435
4.2±0.264
90.8± 0.964
78.1± 1.228
87.1± 0.754
*Values are expressed as mean ± SD (n=3).
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
127
Table 3. The correlation coefficient (R2) and release exponent (n) of different kinetic models of felodipine
LPHNs (HF1-HF6) and the pure drug released in HCl buffer pH 1.2 + 0.3 % polysorbate 80 solutions.
Table 4. The correlation coefficient (R2) and release exponent (n) of different kinetic models of felodipine
LPHNs (HF1-HF6) and the pure drug released in phosphate buffer pH 6.8 + 0.3 % polysorbate 80 solutions.
Ex-vivo intestinal permeation study
The permeability coefficient (cm/min) was
calculated after obtaining felodipine flux (μg/mL)
as shown in Table (5). The comparative study was
performed on the felodipine LPHNs formulations
(HF1-HF3) that prepare by microwave-based
method and felodipine LPHNs formulations (HF4-
HF6) that prepare by SESET. It was found that the
permeability coefficient (cm/min) has the following
descending order HF1 > HF4, HF2 > HF5, HF3>
HF6. This indicates that formulas (HF1-HF3) were
prepared by microwave-based method provide a
higher permeation rate across the intestinal
membrane in comparison to formulas (HF4-HF6)
were prepared by SESET.
The outcomes of the ex vivo intestinal permeation
experiment indicate that the permeability coefficient
(cm/min) of felodipine was significantly higher (p-
value <0.05) for F3 and was significantly lower ( p-
value < 0.05) for pure drug suspension. The
comparability profile of the felodipine release from
LPHNs formulation (HF1-HF6) and the pure drug
suspension explains in the following descending
order: HF3 > HF 6 > HF 2 > HF 5 > HF 1 > HF 4 >
pure drug suspension as shown in Figures (7,8,9).
It was noted that the pure drug suspension gives a
lower intestinal flux in comparison to all felodipine
LPHNs formulations(HF1-HF6) due to increase
solubility and intestinal permeation with felodipine
LPHNs as lipid-based technology. In addition, the
nanosize of these lipid-based carriers provide a large
surface area for drug release and rapid absorption to
the systemic circulation (25,26). The analysis of
variance indicated a significant(p-value <0.05)
relationship between independent variables and ex
vivo intestinal permeation factor.
Table 5. The slope and permeation coefficient for felodipine from LPHNs formulations (F1-F9) and pure
drug suspension through non-everted sheep intestine.
Formulation code
Flux (μg/mL)
Permeability coefficient (cm/min)
HF1
0.0211
0.000559
HF2
0.0213
0.000564
HF3
0.0226
0.000599
HF4
0.0199
0.000527
HF5
0.0211
0.000559
HF6
0.022
0.000583
Pure drug suspension
0.006
0.000159
Korsemeyer-peppas model
Higuchi model
First Order
model
Zero Order
model
Formulation code
n
R2
R2
R2
R2
0.5064
0.948
0.982
0.866
0.971
HF1
0.496
0.934
0.979
0.8821
0.968
HF2
0.4914
0.919
0.984
0.906
0.958
HF3
0.516
0.960
0.983
0.756
0.969
HF4
0.504
0.942
0.992
0.870
0.972
HF5
0.494
0.929
0.984
0.885
0.968
HF6
0.4133
0.908
0.965
0.89
0.843
Pure drug
suspension
Korsemeyer-peppas
model
Higuchi model
First Order
model
Zero
Order
model
Formulation code
n
R2
R2
R2
R2
0.7304
0.924
0.989
0.965
0.895
HF1
0.6774
0.964
0.988
0.968
0.891
HF2
0.6356
0.9794
0.9884
0.9563
0.917
HF3
0.649
0.968
0.996
0.909
0.936
HF4
0.645
0.98
0.993
0.937
0.911
HF5
0.622
0.983
0.992
0.944
0.905
HF6
0.4795
0.9318
0.9755
0.958
0.939
Pure drug suspension
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
128
Figure 7. Permeation of felodipine from LPHNs
formulations HF1, HF4 and pure drug
suspension through non-everted sheep intestine,
the values of mean ±SD (n=3).
Figure 8. Permeation of felodipine from LPHNs
formulations HF2, HF5 and pure drug
suspension through non-everted sheep intestine,
the values of mean ±SD (n=3).
Figure 9. Permeation of felodipine from LPHNs
formulations HF3, HF6 and pure drug
suspension through non-everted sheep intestine,
the values of mean ±SD (n=3).
Conclusion
The microwave-based method is a very
effective and reproducible technique in preparing
felodipine LPHNs formulations (HF1-HF3) and
prepotent to the SESET that prepare felodipine
LPHNs formulations (HF4-HF6). All the felodipine
LPHNs formulations (HF1-HF6) show colloidal
dispersion properties but The (HF1-HF3) superior to
that (HF4-HF6) according to the research
characterization process. All the felodipine LPHNs
formulations (HF1-HF6) show an extended drug
release nanosystem that makes it an advanced
system for control therapeutic agent delivery to
improve the patient's commitment to taking
treatment on time.
References
1. Keskinbora KH, Jameel MA. Nanotechnology
Applications and Approaches in Medicine: A
Review. J Nanosci Nanotechnol Res. 2018; 2:
2:6
2. Vishwakarma N, Jain A, Sharma R, et al. Lipid-
Based Nanocarriers for Lymphatic
Transportation. AAPS Pharm SciTech. 2019
Jan;20(2):83. DOI: 10.1208/s12249-019-1293-
3.
3. Mandhar Piyush, G. Joshi. "Development of
Sustained Release Drug Delivery System: A
Review.” 2015.
4. Siddartha Venkata Talluri, Gowthamarajan
Kuppusamy, Veera VenkataSatyanarayana
Reddy Karri, Shashank Tummala, SubbaRao V.
Madhunapantula. Lipid-based nanocarriers for
breast cancer treatment – a comprehensive
review. Drug Delivery. 2016; 23:4. 1291-1305,
DOI: 10.3109/10717544.2015.1092183
5. Westesen K, Siekmann B. Biodegradable
colloidal drug carrier systems based on solid
lipids. Drugs Pharmaceut Sci.1996; 73:213–58.
6. Yaghmur A, Glatter O. Characterization and
potential applications of nanostructured
aqueous dispersions. Adv Colloid Interface Sci.
2009; 147:333–42.
7. Cheow W S, Hadinoto K. Factors affecting drug
encapsulation and stability of lipid–polymer
hybrid nanoparticles. Colloids Surf. B
Biointerfaces. 2011; 85: 214–20.
8. Wu X Y. Strategies for optimizing polymer–
lipid hybrid nanoparticle-mediated drug
delivery. Expert Opin. Drug Deliv. 2016; 5:
609–12.
9. Hadinoto K, Sundaresan A, Cheow WS. Lipid–
polymer hybrid nanoparticles as a new
generation therapeutic delivery platform: a
review. Eur J Pharm Biopharm. 2013;.85:427–
43.
10. Drais H K, Hussein A A. Design and
Preparation Lipid Polymer Hybrid Nanocarrier
as Pulmonary Dispersion System Using a Novel
Microwave Method. Research Journal of
Pharmacy and Technology.2021; 14:1233-7.
11. Zhi W, Wang L, Hu X. Recent advances in the
effects of microwave radiation on brains.
Military Med Res. 2017; 4: 29.
12. Roer T.G. Microwave Electronic Devices.
Springer Science and Business Media. 2012; 1–
12.
13. 13.Nava-Arzaluz, M. G. et al. “Single
emulsion-solvent evaporation technique and
modifications for the preparation of
pharmaceutical polymeric nanoparticles.”
Recent patents on drug delivery & formulation.
2012; 6(3): 209-23.
Iraqi J Pharm Sci, Vol.31(1) 2022 Felodipine LPHNs
129
14. Drais Hayder Kadhim and Hussein Ahmed
Abbas. Formulation and evaluation lipid
polymer hybrid nanocarriers using a new
innovative microwaves-based method.
International Journal of Pharmaceutical
Research. 2020;12 (4):1264-69.
15. Bershteyn A, Chaparro J, Yau R, et al. Polymer-
supported lipid shells, onions, and flowers. Soft
Matter. 2008;4(9):1787–91.
16. Mukherjee A, Waters A. K, Kalyan P, Achrol A
S, Kesari, S, Yenugonda V M. Lipid-polymer
hybrid nanoparticles as a next-generation drug
delivery platform: state of the art, emerging
technologies, and perspectives. International
journal of nanomedicine. 2019; 14: 1937– 52.
https://doi.org/10.2147/IJN.S198353
17. Dave Vivek, Tak Kajal, Sohgaura Amit, Gupta
Ashish, Sadhu Veera, Reddy Kakarla. Lipid-
polymer hybrid nanoparticles: Synthesis
strategies and biomedical applications. Journal
of Microbiological Methods. 2019.
18. Zhou J, Zhou D. Improvement of oral
bioavailability of lovastatin by using
nanostructured lipid carriers. Drug Design,
Development and Therapy. 2015; 9: 5269 - 75.
19. Vázquez-Blanco S, González-Freire L, Dávila-
Pousa MC, Crespo-Diz C. pH determination as
a quality standard for the elaboration of oral
liquid compounding formula. Farm Hosp. 2018;
42(6):221- 7. doi: 10.7399/fh.10932. PMID:
30381041.
20. Srilatha R, Aparna C, Srinivas Prathima,
Sadanandam M. Formulation evaluation and
characterization of glipizide nanoemulsion.
Asian journal of pharmaceutical and clinical
research. 2013;6(2).66-71.
21. D'Souza Susan. A Review of In Vitro Drug
Release Test Methods for Nano-Sized Dosage
Forms. Advances in Pharmaceutics. 2014: 1-12.
22. Joshi M, Pathak S, Sharma S, Patravale V.
Design and in vivo pharmacodynamic
evaluation of nanostructured lipid carriers for
parenteral delivery of artemether: Nanoject.
International journal of pharmaceutics. 2008;
364 (1): 119-26 .
23. Mohima Tania, Dewan Irin, Islam S. Mashraful,
Rana Sohel, Hossain Alamgir. Encapsulation of
zidovudine in different cellulosic acrylic and
compatibility studies. International Journal of
Pharmacy and Pharmaceutical Sciences. 2015;
7(1):487-95.
24. Ramakrishna S, Mihira V, Vyshnavi K Raja,
Ranjith V. Design and evaluation of drug
release kinetics of meloxicam matrix tablet.
International Journal of Current Pharmaceutical
Research. 2012;4(1):90-9.
25. Attari Z, Bhandari A, Jagadish P C, Lewis S.
Enhanced ex vivo intestinal absorption of
olmesartan medoxomil nanosuspension:
Preparation by combinative technology, Saudi
Pharmaceutical Journal 2015. do i. org / 10
.1016 / j .jsps .2015 .03.008
26. Abed Haithem N, Hussein H. “Ex-vivo Ex-
Vivo Absorption Study of a Novel Dabigatran
Etexilate Loaded Nanostructured Lipid Carrier
Using Non-Everted Intestinal Sac Model. Iraqi
Journal of Pharmaceutical Sciences.
2019;28(2):37-45.
27. Anarjan N, Jafarizadeh-Malmiri H, Nehdi IA,
Sbihi HM, Al-Resayes SI, Tan CP. Effects of
homogenization process parameters on
physicochemical properties of astaxanthin
nanodispersions prepared using a solvent-
diffusion technique. Int J Nanomedicine.
2015;10:1109-18. doi:10.2147/IJN.S72835
28. Sawant K, Dodiya S. Recent Advances and
Patents on solid lipid nanoparticles. Recent Pat
Drug Deliv Formul. 2008; 2:120- 35.
29. Klang M, McLymont V, Ng N. Osmolality, pH,
and compatibility of selected oral liquid
medications with an enteral nutrition product.
JPEN J Parenter Enteral Nutr. 2013;37(5):689-
94. doi:10.1177/0148607112471560
30. Bali V, Ali M, Ali J. Study of surfactant
combinations and development of a novel
nanoemulsion for minimizing variations in
bioavailability of ezetimibe. Colloids Surf B
Biointerfaces 2010;76:410-20.
31. Mandal B, Mittal NK, Balabathula P, et al.
Development and in vitro evaluation of core-
shell type lipid–polymer hybrid nanoparticles
for the delivery of erlotinib in non-small cell
lung cancer. Eur J Pharm Sci . 2016; 81:162–
71.
32. Chaudhary Amity, Nagaich Upendra, Gulati
Neha, Khosa V K, R L Enhancement of
solubilization and bioavailability of poorly
soluble drugs by physical and chemical
modifications. Journal of Advanced Pharmacy
Education & Research. 2012;2(1):32-67.
33. Khan MM, Madni A, Torchilin V, et al. Lipid-
chitosan hybrid nanoparticles for controlled
delivery of cisplatin. Drug Deliv. 2019;
26(1):765-72. doi: 10. 1080 /10717544 .2019
.1642420
This work is licensed under a Creative Commons Attribution 4.0 International License.
