Effects of Pore CaCO3 Form Agencies on Dissolution

Mechanisms of Amoxicillin Drugs Encapsulated in Hydrogels

Full-IPN Chitosan N-Vinyl Caprolactam

Emil Budianto1,a) and Maghfira Fauzia1,b)

1
Polymer Research Group, Department of Chemistry, Faculty of Mathematics and Natural Science,
Universitas Indonesia. Jalan Prof. Dr. Sudjono D Pusponegoro, Kampus UI Depok 16424, Depok

a)
Corresponding author: emilb@ui.ac.id
b)
maghfira.fauzia@ui.ac.id

Abstract. The administration of amoxicillin trihydrate in Helicobacter pylori infection is not effective enough because
the conventional preparations used have a short retention time in the stomach. To overcome this problem, amoxicillin
trihydrate was encapsulated into the floating drug delivery matrix-matrix. In this study, the full-ipn acetaldehyde
crosslinked hydrogel (N-vinyl caprolactam) was synthesized with a 10% CaCO3 pore forming agent and then
encapsulated on amoxicillin trihydrate and studied the mechanism of drug dissolution with its kinetic kinetics approach.
The K-PNVCL Hydrogel produces optimal properties which are then loaded with amoxicillin trihydrate in situ and
post loading. In this research, we have got the percentage of swelling, floating time, the efficiency of in situ and post
loading 873%; 3.15 minutes; 99.8% and 99.4%. The dissolution test was performed on amoxicillin trihydrate which
had been encapsulated K-PNVCL hydrogel in vitro at pH 1.2 resulting in 94.5% for in situ loading and 98.5% for post
loading. Results of the kinetics of drug release for post loading and in situ loading methods tend to follow the Higuchi
model kinetics. The drug release mechanism occurs by Fickian diffusion. Proof of drug release mechanism from K-
PNVCL hydrogel matrix is further done by Scanning Electron Microscope (SEM) instrument.

Keywords: full-IPN, floating drug delivery system, dissolution test, poly (N-vinylcaprolactam), release mechanism.

INTRODUCTION
Helicobacter pylori bacteria cause chronic gastritis and peptic ulcer1, to cure bacteria-infected patients
used oral administration of amoxicillin. The amoxicillin tablets have a short retention time in the stomach,
making it difficult to achieve optimum absorption conditions2. Therefore, a floating system is used as drug
delivery to prolong the absorption time in the stomach3. CaCO3 pore-forming agents are required to form
FDDS-based drugs (Floating Drugs Delivery System). The matrix used in this FDDS is a hydrogel 4, its
specification is poly (N-vinyl caprolactam) or KPNVCL. In this research, the mechanism of drug
dissolution from the hydrogel matrix using kinetics approach will be done 5,6.
Chitosan-poly(N-vinyl caprolactam) hydrogel will be synthesized with full interpenetrating polymer
network (full-IPN) method as a matrix in the floating drug delivery system7. Study of capacity floating time
hydrogel will be evaluated by in vitro study. The ability of hydrogel as a matrix in the floating drug delivery
system will be evaluated by the encapsulation and dissolution of amoxicillin trihydrate by in vitro study8.
In this study, we investigated the dissolution mechanism of amoxicillin trihydrate from the chitosan
hydrogel matrix of chitosan-poly (N-vinyl caprolactam) using a pore-forming agent of CaCO3 in vitro in a
stomach fluid simulation solution (pH 1.2 artificially). Dissolution mechanism can be known by observing
surface morphology and hydrogel pore shape after drug release using SEM. The mechanism of drug
dissolution can also be predicted by the kinetic approach of the measurement data through the graph of the
relationship between the percent release of amoxicillin dissolution to the time which will then be
determined the order of reaction.

MATERIAL AND METHODS

The entire experiment of this study was conducted by in vitro study. The material used in this research
among others is amoxicillin trihydrate from PT. Kimia Farma, generic tablet and tablet patent (Amoxsan®)
of amoxicillin trihydrate, chitosan with DA 94.13% (medical grade) and 25.20 cps viscosity from PT.
Biotech Surindo, N-vinylcaprolactam from Sigma-Aldrich, ammonium persulfate (APS) from PT. Clariant,
glacial acetic acid pro analysis from Merck, N-N’-methylene bisacrylamide from Sigma-Aldrich,
acetaldehyde, N2 gasses from PT. Berkah, CaCO3 as a crosslinking agent.

Preparation of the Hydrogel

Acetaldehyde 0.1 M (2% w/w against the weight of chitosan) was added to 2 g of chitosan that has been
dissolved into 50 mL of 1% acetic acid in a three-neck flask. Then the three-neck flask was placed in an oil
bath, connected with a condenser, flow the nitrogen gasses and heat the three-neck flask. The NVCL
monomer and the initiator of polymerization (APS 3%) were added after system temperature reached 70
ºC. Then, 0.5% MBA was added and stirred in a nitrogen atmosphere for 2 hours. The solution of the
hydrogel was poured into a beaker glass, then added pore forming agent (with variation concentration) and
stirred them until homogenous. Finally, the solution was poured into a mold to form the hydrogel film and
dried it in an oven for 24 hours at a temperature of 60 ºC7.

Determination of Swelling Ratio

Swelling ratio was determined by immersing the hydrogel into water (aquabidest) for 30 minutes at
room temperature. Then the hydrogel surface was blotted using tissue paper. The swelling ratio of hydrogel
was determined by the equation:
Wk−Wa
Swelling ratio (%) = 𝑥 100% (1)
Wa

Wk is the weight of hydrogel after swelling at a predetermined time (t), and Wa is the weight of hydrogel
before immersion5.

Determination of Floating Time and Floating Lag Time

Floating lag time is the time required for the hydrogel to be raised to the surface of the medium and be
float while floating time is how long hydrogel floats on the surface of the medium. Determination of floating
time and floating lag time were measured in a vial containing 10 mL of pH 1.2 solution at 37 ± 0.5 °C
through visual observation8.

Encapsulation of Amoxicillin Trihydrate (In Situ Loading Method)

Encapsulation of amoxicillin trihydrate by in situ loading method was applied to optimum hydrogel
(produce the fastest floating lag time and floating time more than 3 hours). Preparation of optimum hydrogel
which loaded by amoxicillin trihydrate using the same procedures as in the preparation of hydrogel, but
there was the addition of amoxicillin trihydrate (500 mg) before the addition of 0.5 % MBA.

Encapsulation of Amoxicillin Trihydrate (Post Loading Method)

Dissolve 6.25 mg of amoxicillin trihydrate into 250 mL aquabidest. Then added to the hydrogel with
optimum swelling for 30 minutes, at room temperature. Post loading microcapsule then rinsed with
aquabidest.
Determination of Encapsulation Efficiency of Amoxicillin Trihydrate
The hydrogel (mass already known) was immersed into 5 mL of pH 1.0 solution and sonicated for 1
hour. 1 mL filtrate was taken and diluted to 10 mL; then the filtrate was analyzed using UV-Vis
spectrophotometer at the maximum wavelength of amoxicillin trihydrate 9.

Dissolution Test of Amoxicillin Trihydrate

The hydrogel (mass already known) was immersed into 10 mL of pH 1.2 solution and shook using
shaker (100 rpm at 37 ± 0.5 °C ). 1 mL filtrate was taken each interval (0, 5, 10, 20, 30, 60, 90, 120, 150
and 180) minutes, and diluted to 10 mL then analyzed the filtrate using UV-Vis spectrophotometer at the
maximum wavelength of amoxicillin trihydrate.
 Determination of Drug Release Kinetics
Kinetics of the release of the active substance of a modified release preparation can be obtained by
using the Higuchi equation, zero order, first order, and Korsmeyer-Peppas10.

Table 1. Calculation formula of Drug Kinetics

Equation y = a + bx
Zero orde Mt/Mo = k0.t
First orde Log Mt = log Mo – k1.t/2,303
Higuchi Mt/Mo = kH.t1/2
Korsmeyer-Peppas ln Mt/Mo = log k + n log t

Characterization of Hydrogel by FTIR Instrument

The hydrogel characterized by FTIR and compared to the others. A thin sheet of sample hydrogel placed
on a holder which contain KBr powder. The transmittance was measured at wavenumber 400-4000cm-1.

Characterization of Hydrogel by Scanning Electro Microscope

The morphology hydrogel was characterized by scanning electro microscope. A thin sheet of sample
hydrogel placed in a preparation container, then the focus and the intensity of light were manually to get
good results.

RESULTS AND DISCUSSIONS

FTIR Characterization of Chitosan-PNVCL Hydrogel with Addition of Pore

Forming Agent

The result of FTIR spectrum of chitosan-PNVCL hydrogel with a pore-forming agent is not much
different with FTIR spectrum of chitosan-PNVCL hydrogel without pore forming agent. The FTIR
spectrum of chitosan-PNVCL provides absorption at a wavelength of 684 cm-1 shows the bond between O-
C-N from PNVCL cross linked by MBA polymer network7. In the chitosan spectrum, there is an uptake at
3337 cm-1 indicating the presence of stretching hydroxy groups (O-H). This is the main characteristic of
chitosan structure. The 1150 cm-1 wave number indicates the shifting vibration of the C-O-C bridge. The
spectral shifts in the 1620 - 1650 cm-1 wave number occurring between the K-PNVCL hydrogel and the K-
PNVCL pore and hydrogel-forming agent without pore-forming agents are due to the increasing number of
carbonyl groups and the physical interactions between the carbonyl groups. The emergence of new
absorption bands at wave number 1150 cm-1 on hydrogel K-PNVCL with pore forming agent CaCO312. The
appearance of this band indicates that the pore-forming agent has succeeded in entering the hydrogel
matrix9,13.

Hidrogel K-PNVCL

C=O K-PNVCL/CaCO3

O-H C-O-C

FIGURE 1. FTIR spectra of chitosan-PNVCL hydrogel and chitosan-PNVCL hydrogel with addition of pore-
forming agent
 The Swelling Ratio of Chitosan-PNVCL Hydrogel with Addition of Pore Forming
Agent

The pore-forming agent which has been added will enter into a three-dimensional matrix of the hydrogel
and will fill the space between networks. The space should have been filled with a liquid medium that used
for the measurement of the swelling ratio14. The increase addition of pore-forming agent will cause the
space that can be filled by liquid medium in networks become fewer, causes the swelling ratio of the
hydrogel is also getting smaller. The weight ratio of aquadest adsorbed with dry hydrogel weight indicates
percent swelling ratio. In this research, the swelling ratio test with a pore-forming agent was performed.
Generated 873%, which means the hydrogel can absorb water as much as 873 times.

Floating Time Capacity of Chitosan-PNVCL Hydrogel with Addition of Pore

Forming Agent

The presence of the pore forming agent with a greater concentration would result in the number of pores
is formed to be a lot more because the more CO2 is trapped within the hydrogel layer and create a floating
lag time of hydrogel becomes faster. Carbon dioxide gasses formed during the reaction would be trapped
within the hydrogel network when carbon dioxide out of the hydrogel network then the gas will leave the
air cavities or pores within the hydrogel2. The addition of CaCO3 gives a floating time of 3.15 minutes
because of the floating mechanism that occurs, the CO2 formed pressing the hydrogel layer, so that the pore
or cavity formed this occurs due to the decrease in density of hydrogel material and the process of floating
(low-density system).

Encapsulation Efficiency of Chitosan-PNVCL Hydrogel as Matrix in Floating

Drug Delivery System

Pores formed in the hydrogel has a major influence on the efficiency of drug encapsulation. Pores are
formed too much lead the encapsulation efficiency of drugs to be small. It might be due to the appearance
of pores in the hydrogel makes the internal structure of the hydrogel becomes less dense, so it is not able to
maintain the drug in the matrix2,6. The increase addition of pore-forming agent in a hydrogel produce the
faster floating lag time of the hydrogel14,15.
The chitosan-PNVCL hydrogel with the addition of CaCO3 10% as a pore-forming agent was loaded
by drug with in situ loading method has floating lag time of 3 minute 15 seconds. The changes of floating
lag time of optimum chitosan-PNVCL hydrogel before and after drug loading occur because the increasing
burden of the drug was entrapped in the hydrogel matrix, caused the floating lag time of hydrogel become
longer16. From the data obtained, chitosan-PNVCL hydrogels with CaCO3 10% as pore forming agent has
the fast floating lag time and has the big encapsulation efficiency, for post loading is 99,4% and in situ
loading is 99,8%

Dissolution of Amoxicillin Trihydrate from Chitosan-PNVCL Hydrogel as Matrix,

Generic Tablet, and Tablet Patent

Figure 2. shows that the chitosan-PNVCL hydrogel was loaded by amoxicillin trihydrate with pore
forming agent CaCO3 with in situ loading and post loading method provides great dissolution results. The
concentration of amoxicillin trihydrate which released by chitosan-PNVCL hydrogel until 3 hours which
is almost 100% indicates that the dissolution of amoxicillin trihydrate encapsulated by chitosan-PNVCL
hydrogel was much better than the dissolution of generic tablet and tablet patents of amoxicillin trihydrate
The result of dissolution and the encapsulation of amoxicillin trihydrate by the chitosan-PNVCL hydrogel,
generic tablet, and tablet patent the highest is post loading 98%. So, we can conclude that the chitosan-
PNVCL hydrogel as a matrix in the floating drug delivery system can be used for encapsulation of
amoxicillin trihydrate which produces better dissolution of amoxicillin trihydrate than generic tablet and
tablet patents of amoxicillin trihydrate, wherein the best pore forming agent is CaCO3 10% with post
loading method. Meanwhile generic and tablet patent just 71% and 72%.
 Dissolution of Hydrogel K-PNVCL
100,0000

80,0000

%Dissolution
60,0000

40,0000

20,0000

0,0000
5 10 20 30 60 90 120 150 180
Time (minutes)

in situ post generik paten

FIGURE 2. The dissolution graph of amoxicillin trihydrate

Kinetic Model of Dissolution Results of Amoxicillin Drugs In Hydrogel K-PNVCL

Based on the result of drug dissolution test, it can be known the mechanism of drug release from
K-PNVCL hydrogel matrix by using kinetics approach of drug release. Kinetics of the release of the active
substance of a modified release preparation can be obtained by using the Higuchi equation, zero order, first
order, and Korsmeyer-Peppas10. The drug release data obtained is incorporated into the equation. Based on
the highest correlation coefficient (r2), it shows that the kinetics of drug release of Amoxicillin at post
loading tends to follow Higuchi's kinetics (r2 = 0.9712). And for in situ loading kinetics discharge same,
that is a kinetic release of Higuchi (r2 = 0,9679). Higuchi's release kinetics describes drug release from the
matrix depending on the time root based on Fickian diffusion. Based on the results obtained, it can be seen
that the mechanism of drug release amoxicillin trihydrate has a value of n <0.45 It identifies that the drug
release mechanism follows the Fickian diffusion mechanism. This is evidenced by SEM measurement
results, the results of morphological observation of hydrogels before and after the dissolution by using a
scanning electro microscope can be seen in Table 4.

TABLE 2. The results of morphological observation of hydrogels in situ and post loading in SEM

Time In situ loading Post loading

180 minutes

The observation of chitosan-PNVCL hydrogel matrix after dissolution with post loading and in
situ loading method in the 180-minute hydrogel matrix shows the number of amoxicillin trihydrate drug
molecules in the hydrogel matrix, thus causing pores in the many hydrogel matrixes to be covered by the
amoxicillin trihydrate drug molecules. The kinetics approximation concludes the K-PNVCL hydrogel
matrix encapsulated amoxicillin trihydrate based on the results of the Korsmeyer-Peppas equation with n
<0.45 only diffusion. However, with observations using SEM, the results are diffusion and erosion. This is
due to the morphology of its irregular shape indicating the matrix erosion occurs. The diffusion captured
from its kinetic equation, indicating that the kinetics equation can only calculate the release of amoxicillin
trihydrate from the surface of the matrix which is still remnant of amoxicillin trihydrate, not from within
its (transverse) matrix.

CONCLUSION
Full-IPN chitosan-poly(N-vinylcaprolactam) hydrogel has been successfully synthesized via crosslink
between chitosan-acetaldehyde and poly(N-vinylcaprolactam)-MBA. CaCO3 pore forming agent produces
a floating lag time of 3.15 minutes and floating time over 3 hours. In this research, efficiency test and 99.8%
of the in situ loading method were performed, and the biggest dissolution test was 98% of post loading.
Then for the kinetics profile of drug release amoxicillin trihydrate showed that the release of post loading
and in situ loading methods tended to follow the Higuchi model. The release mechanism of amoxicillin
trihydrate tends to follow the release mechanism of Fickian diffusion. The best drug loading method into
the hydrogel matrix is in situ loading method because it has a higher encapsulation efficiency than with
loading.

ACKNOWLEDGEMENT
The Authors would like to thank Directorate of Research and Community Engagement (DRPM)
Universitas Indonesia for supporting our research financially through Hibah Publikasi Internasional
Terindeks untuk Tugas Akhir (Hibah PITTA) 2017. We also would like to thank Department of Chemistry,
Universitas Indonesia for supporting us and providing the instrumental facilities for our research. Emil
Budianto supervised this research and designed the research pipeline, while Maghfira Fauzia conducted the
experimental details, analyzed the data, and wrote the final manuscript. Herewith, we would declare that
there is no conflict of interest regarding this manuscript.

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