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Acta Poloniae Pharmaceutica ñ Drug Research, Vol. 67 No. 4 pp.

407ñ415, 2010 ISSN 0001-6837


Polish Pharmaceutical Society

PHARMACEUTICAL TECHNOLOGY

DEVELOPMENT AND CHARACTERIZATION OF EUDRAGIT


RS 100 LOADED MICROSPONGES AND ITS COLONIC DELIVERY
USING NATURAL POLYSACCHARIDES
VIKAS JAIN* and RANJIT SINGH

School of Pharmaceutical Sciences, Shobhit University, Meerut, Uttar Pradesh, 250110, India

Abstract: In the present work, paracetamol loaded eudragit based microsponges were prepared using quasi-
emulsion solvent diffusion method. The compatibility of the drug with various formulation components was
established. Process parameters were analyzed in order to optimize the formulation. Shape and surface mor-
phology of the microsponges were examined using scanning electron microscopy. The formulations were sub-
jected to in vitro release studies and the results were evaluated kinetically and statically. The in vitro release data
showed a bi-phasic pattern with an initial burst effect. In the first hour drug release from microsponges was
found to be between 17ñ30%. The cumulative percent release at the end of 8th hour was noted to be between
54-83%. The release kinetics showed that the data followed Higuchi model and the main mechanism of drug
release was diffusion.The colon specific tablets were prepared by compressing the microsponges followed by
coating with pectin:hydroxypropylmethylcellulose (HPMC) mixture. In vitro release studies exhibited that com-
pression coated colon specific tablet formulations started releasing the drug at 6th hour corresponding to the
arrival time at proximal colon. The study presents a new approach for colon specific drug delivery.

Keywords: microsponge, colonic delivery, pectin, diffusion

The requirement for an oral colonic drug deliv- pH-dependent systems, because of large variations
ery system is to reduce the drug release to a mini- in the pH of the gastrointestinal tract, is very well
mum prior to the cecum (1). Colon as a site offers documented. The site-specificity of timed-release
distinct advantages on account of a near neutral pH, dosage forms is considered poor because of large
a much longer transit time, reduced digestive enzy- variations in gastric emptying times and passage
matic activity and a much greater responsiveness to across the ileo-cecal junction (6). However,
absorption enhancers (2). These criteria favor this microflora-activated systems formulated making
distal part of the gastrointestinal tract (GIT) as a site use of non-starch polysaccharides are highly prom-
for the delivery of vermicides, colonic diagnostic ising because the polysaccharide remain undigested
agents and sustained release of drugs in treatment of in the stomach and the small intestine and can only
nectural asthma, angina and arthritis (3). A colon- be degraded by the vast anaerobic microflora of the
specific drug delivery system should prevent drug colon. Furthermore, this strategy exploiting the
release in the stomach and small intestine and affect abrupt increase of the bacteria population and corre-
an abrupt onset of drug release upon entry into the sponding enzyme activities will also accomplish
colon (4). greater site specificity of initial drug release (7). The
Various approaches have been used for deliv- polysaccharides for colonic drug delivery are also
ery of drugs to the colon via oral route, which inexpensive, naturally occurring and abundantly
include coating with pH-dependent polymers, available (8).
design of time-release dosage forms and the utilisa- Single unit colon targeted drug delivery sys-
tion of carriers that are degraded exclusively by the tems may suffer from the disadvantage of uninten-
colonic bacteria (5). Every system has advantages as tional disintegration of the formulation due to man-
well as disadvantages. The poor site-specificity of ufacturing deficiency or unusual gastric physiology

* Corresponding author: phone: +919219610427, fax: +911212575724 e-mail: vikasjain11118059@rediffmail.com, vikasjain111180@


gmail.com

407
408 VIKAS JAIN and RANJIT SINGH

Table 1. Composition of various microsponge formulations.

Formulations
Components
FPRS1 FPRS2 FPRS3 FPRS4
Paracetamol (mg) 600 1200 1800 2400
Eudragit RS-100 (mg) 200 200 200 200
Triethylcitrate (% w/v) 1 1 1 1
Dichloromethane (mL) 5 5 5 5
PVA (% w/v) 0.5 0.5 0.5 0.5

that may lead to drastically compromised systemic Methods


drug bioavailability or loss of local therapeutic Paracetamol loaded microsponge preparation
action in the colon. Recently, much emphasis is Paracetamol microsponges were prepared by
being laid on the development of multiparticulate quasi-emulsion solvent diffusion method. The inter-
dosage forms in comparison to single unit systems nal phase consisted of Eudragit RS-100 (200 mg)
because of their potential benefits like increased and triethylcitrate (1% w/v) dissolved in 5 mL of
bioavailability, reduced risk of local irritation and dichloromethane. Triethylcitrate (TEC) was used as
predictable gastric emptying (9). plasticizer. This was followed by addition of drug
Microsponges are polymeric delivery systems with gradual stirring (500 rpm). The internal phase
composed of porous microspheres. They are tiny was then poured into polyvinyl alcohol
sponge like spherical particles that consist of myri- 30,000ñ70,000 (PVA) solution in water, the external
ad of interconnecting voids within a non-collapsible phase. After 8 h of stirring, the microsponges were
structure with large porous surface (10). Moreover, formed due to removal of dichloromethane from the
they may enhance stability, reduce side effect and system. The microsponges were filtered and dried at
modify drug release favorably (7). 40OC for 12 h. The composition of microsponge for-
Paracetamol (PCM), an antipyretic and anal- mulations are given in Table 1.
gesic drug, which has been widely used in clinical
practice, was selected as a model drug. It has a short Fourier transform infrared (FTIR) analysis
half life in plasma about 1ñ4 hours. FTIR spectra of the drug, physical mixture of
The present study is aimed at developing drug and Eudragit RS-100, formulations
microsponge based novel colon specific drug deliv- FPRS1ñFPRS4 were recorded in potassium bromide
ery system containing PCM. The microsponges of disc using a Shimadzu Model 8400 FTIR spectrom-
PCM were prepared and characterized. They were eter to ascertain compatibility.
formulated as colon specific tablets and subjected to
in vitro characterization for various attributes. Differential scanning calorimetric (DSC) analysis
Thermal analysis using DSC was carried out
EXPERIMENTAL on drug, physical mixture of the drug and Eudragit
RS-100, and formulations FPRS1ñFPRS4
Materials (Shimadzu DSC-60 Thermal Analyzer). Accurately
Paracetamol was purchased from Jackson weighed samples were loaded into aluminum pans
Laboratories Pvt. Ltd. Amritsar (India). Eudragit and sealed. All samples were run at a heating rate of
RS-100 was kindly gifted by Evonic India Pvt. Ltd. 20OC/min. over a temperature range 40ñ430OC.
Mumbai (India). Polyvinyl alcohol 30,000ñ70,000
(PVA), triethylcitrate, and HPMC (100,000 cps) Morphology
were purchased from Sigma-Aldrich (USA). The morphology and surface characteristics of
Pectinex Ultra SP-L (26,000 FDU/mL), pectin (from the microsponges were studied using scanning elec-
citrus fruits, methoxy content 9.4%) and sodium car- tron microscopy (SEM). All the samples were coat-
boxymethylcellulose (Na-CMC) were procured ed with goldñpalladium alloy under vacuum. Coated
from Sigma (USA). All chemicals used for analysis samples were then examined using LEO 430 SEM
were of analytical grade. analyzer.
Development and characterization of Eudragit RS 100 loaded microsponges... 409

Actual drug content and encapsulation efficiency punches on an eight station tablet punch machine
The weighed amount of drug loaded (Cambart, D-8) using 1500 kgf/cm2 compression
microsponges (100 mg) was kept in 100 mL of pressure. Core tablet formulations are given in Table
phosphate buffer pH 6.8 for 12 h with continuous 2.
stirring. The samples were filtered using 0.45 µm Pectin:HPMC (80 : 20) mixture was used as
membrane filter and the samples were analyzed at outer shell for compression coating. The coating
256 nm against blank using UV spectrophotometer material used was 400 mg. Fifty percent of coating
(UV 1700, Shimadzu, Japan). The drug content and material was placed in the die cavity and the core
encapsulation efficiency were calculated using the tablet was placed in centre followed by addition of
following formulas (7): the remainder of the coating material. The coating
Actual drug content (%) = Mact/Mms ◊ 100 material was compressed around the core tablet at an
Encapsulation efficiency (%) = Mact/Mthe ◊ 100 applied pressure of 2500 kgf/cm2 using round flat
where Mact is the actual drug content in punches (14 mm) on the same tabletting machine.
microsponges, Mms is the total amount of the
microsponges and Mthe is the amount of drug added In vitro drug release studies of colon specific for-
to the microsponges. All analyses were carried out mulations
in triplicate. The drug release studies were done with the
same method used for microsponges and core
In-vitro drug release studies of microsponge for- tablets. Additionally, Pectinex Ultra SP-L was
mulations added to the dissolution medium at 6th hour in order
The microsponges containing 250 mg of parac- to simulate the enzymatic action of the colonic bac-
etamol ware subjected to in vitro drug release stud- teria. Samples were withdrawn periodically and
ies. In vitro release studies were carried out in USP compensated with an equal amount of fresh dissolu-
basket apparatus with stirring rate of 50 rpm at 37 ± tion media. The samples were analyzed for drug
0.5OC. Initial drug release was carried out in 900 content by measuring absorbance at 256 nm using
mL. of 0.1 M HCl for 2 h, followed by phosphate UV spectrophotometer.
buffer pH 6.8 for next 6 h. Samples were withdrawn
at regular intervals of time. The sink conditions were RESULTS AND DISCUSSION
maintained by adding equal amount of dissolution
medium. The samples were analyzed spectrophoto- Quasi-emulsion solvent diffusion method was
metrically (Shimadzu UV-1700) at a wavelength of used for preparation of microsponges because of its
256 nm. Dissolution tests were performed in tripli- simplicity and reproducibility. Moreover, it has
cate for each sample. advantage of avoiding solvent toxicity. The drug
and polymer in the ratios 3 : 1, 6 : 1, 9 : 1, and 12 :
Preparation of colon specific tablet formulations 1 were taken to prepare different microsponge for-
The core tablets consisting of microsponges mulations namely FPRS1, FPRS2, FPRS3, and
containing 250 mg drug, Na-CMC and magnesium FPRS4, respectively. In each formulation, the
stearate were prepared by direct compression amounts of polymer (200 mg), dichloromethane (5
method. All tablet constituents were weighed and mL), PVA (0.5% w/v) were kept constant. The
mixed in motor passel for 15 min. Final powder microsponge formulations were prepared using
mixture was compressed using 10 mm round flat mechanical stirrer (Remi RQ1217-D) at a stirring

Table 2. Core tablet formulations of PCM microsponges.

Core tablet Microsponges formulations (mg) Na-CMC Magnesium


formulation codes FPRS1 FPRS2 FPRS3 FPRS4 (mg) stearate (mg)

CPRS1 350.0 - - - 42 8
CPRS2 - 300 - - 92 8
CPRS3 - - 290 - 102 8
CPRS4 - - - 280 112 8
410 VIKAS JAIN and RANJIT SINGH

rate of 500 rpm for 8 h. The composition of various crystals on the surface. Figure 1 shows that drug-
microsponge formulations are presented in Table 1. polymer ratio has considerable effect on the mor-
The effect of various variables like drug/poly- phology and size of microsponges. It was observed
mer ratio, stirring rate, volume of internal phase, that as the ratio of drug to polymer was increased,
amount of emulsifying agent on the nature of the particle size decreased. This could probably be
microsponges was studied. due to the fact that in high drug to polymer ratios,
the amount of polymer available per microsponge
Effect of drug-polymer ratio on microsponges was comparatively lower. Probably in high drug-
The morphology of the microsponges was polymer ratios less polymer amount surrounds the
studied by scanning electron microscopy (SEM). drug and microsponges with smaller size were
The representative photographs of the microsponges obtained (11).
are shown in Figure 1. The microsponges were Production yield, actual drug content, encapsu-
observed to be spherical and uniform with no drug lation efficiency, and mean particle size of formula-

Figure 1(a-h). SEM photograph of microsponge formulations (drug: Eudragit RS-100). The photograph coded ëAí represents whole
image, ëBí represents surface photograps.
Development and characterization of Eudragit RS 100 loaded microsponges... 411

Figure 2. SEM photograph of drug: Eudragit RS-100 microsponges prepared at different stirring rates of (a) 300 rpm; (b)
400 rpm; (c) 500 rpm

Table 3. Production yield, actual drug content, encapsulation efficacy, and mean particle size of various microsponges formulations (n = 3).

Production Theoretical Actual drug Encapsulation Mean particle


Drug:Polymer
Formulation yield drug content content efficiency size
ratio
(% ± S.D.) (%) (% ± SD) (% ± SD) (µm ± SD)
FPRS1 3:1 72 ± 0.43 75.00 73.56 ± 0.09 98.08 ± 0.89 62.34 ± 6.89
FPRS2 6:1 74.12 ± 0.34 85.71 84.32 ± 0.04 98.37 ± 0.56 54.67 ± 5.39
FPRS3 9:1 76.23 ± 0.36 90.00 88.32 ± 0.45 98.13 ± 0.09 48.23 ± 7.24
FPRS4 12 : 1 75.02 ± 0.60 92.30 90.81 ± 0.34 98.38 ± 0.67 41.45 ± 5.34

tions FPRS1-FPRS4 are presented in Table 3. The was found to be dependant on the agitation speed.
production yield, actual drug content, and encapsu- As the speed was increased, the size of
lation efficiency of FPRS1-FPRS4 formulations was microsponges was reduced and the microsponges
found to be between 72ñ75%, 73ñ91% and 98ñ99%, were found to be spherical and uniform (12). When
respectively. The data obtained for various formula- the rate of stirring was increased up to 500 rpm the
tions in respect to production yield, actual drug con- spherical microsponges were formed with mean par-
tent, and encapsulation efficiency were subjected to ticle size of 62.34 ± 6.89 µm.
t-test at 95% level of significance. No significant It was noted that at higher stirring rate the pro-
difference in relation to these parameters was duction yield was decreased. Possibly, at the higher
observed amongst various formulation at p < 0.05. stirring rates the polymer adhered to paddle due to
the turbulence created within the external phase, and
Effect of stirring rate on the morphology and hence production yield decreased (13).
yield of microsponges
The effect of stirring rate on the morphology of Effect of volume of internal phase on the produc-
microsponges is shown in Figure 2. The formulation tion of microsponges
with the lower drug to polymer ratio (i.e., 3:1) was It was observed that on increasing the volume
chosen to investigate the effect of stirring rate on the of internal phase from 5 to 10 mL microsponges
morphology of microsponges. The stirring rate was were not formed. This may be due to the decrease in
varied in the range of 300 to 500 rpm. The disper- viscosity of internal phase (14). As the amount of
sion of the drug and polymer into the aqueous phase dichloromethane was increased, the finely dispersed
412 VIKAS JAIN and RANJIT SINGH

Figure 3. DSC thermogram of paracetamol physical mixture of drug & Eudragit RS-100 and FPRS1-FPRS4 microsponges
formulations

spherical quasi-emulsion droplets were seen in sol- resulted in larger microsponges. The dispersion of
vent under the agitation, but as the stirring was dis- the solution of the drug and polymer into droplets
continued emulsion droplets adhered together and was effected by the concentration of polyvinyl alco-
coalesce. Consequently, no microsponges could be hol in the external phase. When the concentration of
formed. The result suggests that the amount of PVA was increased in dispersion phase, the size of
dichloromethane need to be controlled within an microsponges was found to be decreased (18). The
appropriate range to effect not only the formation of results of the significant effect of emulsifying agent
quasi-emulsion droplets at the initial stage but also on production yield and mean particle size are
the solidification of drug and polymer in the shown in Table 4. An increased amount of emulsi-
droplets. The good microsponges were produced fying agent decreased the production yield from 72
when 3 to 5 mL of dichloromethane were used. to 67% and increased the mean particle size from 62
to 66 µm.
Effect of amount of emulsifying agent on the pro-
duction yield and particle size of microsponges Characterization of microsponges
An increase in amount of emulsifying agent DSC studies were carried out to confirm com-
resulted in decreased production yield and increased patibility (15). The thermal behavior of drugs, phys-
mean particle size, as the emulsifier was non-ionic ical mixture of drug and polymer, and formulations
in nature and possibly formed some hydrophobic FPRS1ñFPRS4 were studied. In the thermogram,
region which dissolved some of the drug and poly- the drug showed a sharp endothermic peak (at
mer. The molecules might have associated away 174.23OC) which corresponds to the melting point of
from the oil-water interface at higher concentrations drug in the crystalline form. In the DSC curve of
resulting in alternative hydrophobic region, which physical mixture of drug and polymer, and formula-
dissolved some portion of drug resulting in a reduc- tions FPRS1ñFPRS4, the characteristic peaks of
tion in production yield of microsponges (13). On drug(s) were observed. The result showed that there
the other hand, an increase in the amount of emulsi- is no incompatibility between the drug and poly-
fying agent resulted in increased larger mers. Microsponge production process did not
microsponges. This could be due to the increased change the nature of drug in microsponges. The
viscosity wherein larger emulsion droplets formed thermal behavior of the drug, physical mixture of
Development and characterization of Eudragit RS 100 loaded microsponges... 413

Table 4. The effect of emulsifying agent on microsponges formulation.

Internal Phase External phase Mean


Formulation Yield diameter
code PCM Polymer Dichloromethane Water PVA (%)
(mg) (mg) (mL) (mL) (% w/v) µm ± SD

FPRS1 (a) 600 200 5 100 0.5 72 ± 0.43 62.34 ± 6.89


FPRS1 (b) 600 200 5 100 1.0 67.35 ± 2.56 66.12 ± 3.15

Table 5. In vitro drug release models for different microsponges formulations.

Zero order First order Higuchi model Korsmeyer-Peppas model


Code R K R K R K R ëní
(mg/h) (h-1) (mg/h-1/2)
FPRS1 0.9729 7.7948 0.9846 0.1255 0.9848 24.347 0.9558 0.7939
FPRS2 0.9710 8.3785 0.9869 0.1497 0.9895 26.535 0.9622 0.6829
FPRS3 0.9805 9.4463 0.9941 0.2057 0.9954 29.678 0.9849 0.6323
FPRS4 0.9865 11.296 0.9934 0.3473 0.9957 35.083 0.9951 0.6908

Figure 4. FTIR spectra of paracetamol physical mixture of drug & Eudragit RS-100 and FPRS1-FPRS4 microsponges for-
mulations

drug and Eudragit RS-100 and formulations bonyl stretching band at 1654 cm-1 were seen.
FPRS1ñFPRS4 are presented in Figure 3. Eudragit RS 100 showed an ester C=O stretching
FTIR spectra were recorded to assess the com- peak around 1726.17 cm-1. All characteristic peaks
patibility of the drug and excipients (16). FTIR of paracetamol were observed in the FTIR spectra of
spectra of drug, physical mixture of drug and FPRS1ñFPRS4 formulations. The results showed
eudragit RS-100 and formulations FPRS1ñFPRS4 that no chemical interaction or changes took place
were examined. In FTIR spectra of paracetamol during preparation of the formulations and the drug
powder, characteristic N-H stretching band at 3325 was found to be stable in all the formulations. The
cm-1, O-H stretching band at 3161.11 cm-1, and car- FTIR spectra of the drug, physical mixture of drug
414 VIKAS JAIN and RANJIT SINGH

Figure 5. In vitro drug release profile of different formula- Figure 6. In vitro drug release profile of drug from differ-
tions of microsponges (FPRS1-FPRS4) ent colon specific formulations (CPRS1-CPRS4)

and Eudragit RS-100 and formulations FPRS1ñ value for Peppas model was found to be between
FPRS4 are presented in Figure 4. 0.5ñ1 indicative of non-fickian diffusion.
The in vitro dissolution data were subjected to
In vitro release studies of the microsponge formu- statistical analysis using ANOVA. The p value was
lations found to be 0.5207 indicating no significant differ-
The microsponge formulations were subjected ence in the release behavior (p > 0.05).
to in vitro release studies using USP XX1V dissolu-
tion assembly at the stirring rate at 50 rpm and tem- In vitro dissolution studies of the colon specific
perature at 37 ± 0.5OC. Initially, drug release was tablet formulations
carried out in of 0.1 M hydrochloric acid for 2 h fol- In order to prepare the compression coated
lowed by phosphate buffer pH 6.8 for the next 6 h. tablet formulations, core tablets were prepared as
The release profiles obtained for the formulations the first step. The homogenous granular characteris-
FPRS1-FPRS4 are presented in Figure 5. It was tic of microsponges is due to their highly porous
observed that the drug release decreased with an structure and in these means, microsponges have the
increase in the amount of polymer for each formula- compressibility to produce strong tablets and
tion. This may be due to the fact that the release of 1000ñ2000 kgf/cm2 pressure did not cause the struc-
drug from the polymer matrix takes place after com- ture deformation of microsponges (19). In vitro drug
plete swelling of the polymer and as the amount of release studies of the colon specific tablet formula-
polymer in the formulation increases the time tions were carried out using USP basket apparatus
required to swell also increases. The release showed with stirring rate 50 rpm at 37 ± 0.5OC. The release
a bi-phasic pattern with an initial burst effect. In the profiles obtained for the formulations CPRS1-
first hour drug release was found to be 17ñ30%. CPRS4 are presented in Figure 6. No drug was
This may be attributed to the drug present in the released in the first 6 h. After this lag time, the drug
pores of the microsponges or improper entrapment release started at the beginning of 7th hour due to
of drug (17). The cumulative percent release for the addition of the Pectinex Ultra SP-L and contin-
FPRS1-FPRS4 at the end of 8 h was found to be ued up to 14th hour for CPRS1 (68.65%), 14th hour
54ñ83%. The microsponge formulations were sub- for CPRS2 (88.23%), 13th hour for CPRS3
jected to in vitro dissolution studies and the data (92.45%) and 12th hour for CPRS4 (95.76%).
were analyzed using various mathematical models The results of in vitro drug release showed that
(Table 5). Based on highest regression value, the the ratio of Pectin: HPMC (80 : 20) protected the
best fit was observed for Higuchi matrix. The n cores up to 6th hour corresponding to the time to
Development and characterization of Eudragit RS 100 loaded microsponges... 415

reach the colon and after that under the influence of 4. Varshosaz J., Dehkordi A.J., Golanfshan S.: J.
the enzyme, the system could be degraded faster and Microencapsul. 23, 329 (2006).
deliver the drug to the proximal colon that forms the 5. Srimornsak P., Nunthanid J., Wanchana S.,
main site of bacterial carbohydrate metabolism. So, Luangtana-Anan M.: Pharm. Dev. Technol. 8,
the results were in accordance with the triggering 311 (2003).
mechanism due to the very active metabolism in the 6. Krishnaiah Y.S.R.: J. Control. Release 55, 245
proximal part compared with the distal part of colon (1998).
and pectin could find the appropriate environment to 7. Orlu M., Cevher E., Araman A.: Int. J. Pharm.
be degraded. 318, 103 (2006).
8. Vandamme T.H., Lenourry A., Charroeau C.,
CONCLUSION Chaumeil J.C.: Carbohydr. Polym. 48, 219
(2002).
This study presents new approach for the 9. Asghar L.F.A., Chandran S.: J. Pharm. Pharm.
preparation of modified microsponges. The pre- Sci. 9, 327 (2006).
pared microsponges exhibited characteristics of an 10. Nokhodchi A., Jelvehgari. M., Siahi M.R.,
ideal delivery system for colon targeting. The Mozafari M.R.: Micron 38, 834 (2007).
unique compressibility of microsponges offers a 11. Chourasia M.K., Jain S.K.: Drug Deliv. 11, 201
new alternative for producing mechanically strong (2004).
tablets. Further colon specific tablets based on 12. Perumal D.: Int. J. Pharm. 218, 1 (2001).
microsponges could provide effective local action as 13. Jelvehgari M., Siahi-Shadbad M.R., Azarmi S.,
microsponges may selectively be taken up by the Martin G.P., Nokhodchi A.: Int. J. Pharm. 308,
macrophages present in colon. 124 (2006).
14. Yang M., Cui F., You B., Fan Y.: Int. J. Pharm.
Acknowledgment 259, 103 (2003).
15. Ceschel G.C., Badiello R., Ronchi C., Maffei
The authors are thankful to the Director, P.: J. Pharm. Biomed. Anal. 32, 1067 (2003).
School of Pharmaceutical Sciences, Shobhit 16. Mukherjeea B., Mahapatraa S., Guptab R.,
University, Meerut for providing necessary facili- Patraa B., Tiwarib A., Arora P.: Eur. J. Pharm.
ties. Biopharm. 59, 475 (2005).
17. Mastiholimath V.S., Dandagi P.M., Jain S.S.,
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