International Journal of Applied Pharmaceutics

Vol 2 Issue 2, 2010

ResearchArticle
FORMULATIONANDEVALUATIONOFTRIMETAZIDINEHYDROCHLORIDELOADED
CHITOSANMICROSPHERES

CHINTAGUNTAPAVANVEENA*,KAVITHAK,ANILKUMARSN
PharmaceuticsDivision,BharathiCollegeofPharmacy,Bharathinagara,Maddur,Karnataka571422,India.Email:chpveena@gmail.com
Received:14Jan2010,RevisedandAccepted:18Feb2010
ABSTRACT
Trimetazidine hydrochlorideloaded chitosan microspheres wereprepared by theioniccrosslinking technique using TPP as crosslinking agent.
Theprocessinducedtheformationofmicrosphereswiththeincorporationefficiencyof47%to77%.Theeffectofchitosanconcentration,cross
linking agents and conditions was evaluated with respect to entrapment efficiency, particle size, surface characteristics and in vitro release
behaviors.Infraredspectroscopicstudyconfirmedtheabsenceofanydrugpolymerinteraction.Differentialscanningcolorimetricanalysisrevealed
thatthedrugwasmolecularlydispersedinthechitosanmicrospheresmatricesshowingroughsurface,whichwasconfirmedbyscanningelectron
microscopystudy.Themeanparticlesizeandentrapmentefficiencywerefoundtobevariedbychangingvariousformulationparameters.Thein
vitro release profile could be altered significantly by changing various formulation parameters to give a sustained release of drug from the
microspheres. The kinetic modeling of the release data indicate that trimetazidine hydrochloride release from the chitosan microspheres follow
anomaloustransportmechanismafteraninitiallagperiodwhenthedrugreleasemechanismwasfoundtobefickiandiffusioncontrolled.
Keywords:microspheres,drugdelivery,targeting,drugrelease,chitosan.

INTRODUCTION
Oral controlled release (CR) dosage forms (DFs) have been
developed over the past three decades due to their considerable
therapeutic advantages such as ease of administration, patient
compliance and flexibility in formulation. Microspheres carrier
systemsmadefromthenaturallyoccurringbiodegradablepolymers
haveattractedconsiderableattentionforseveralyearsinsustained
drugdelivery.Recently,dosageformsthatcanpreciselycontrolthe
releaseratesandtarget drugstoaspecificbodysitehavemadean
enormousimpactintheformulationanddevelopmentofnoveldrug
delivery systems. Microspheres form an important part of such
novel drug delivery systems13. They have varied applications and
are prepared using assorted polymers4. However, the success of
thesemicrospheresislimitedowingtotheirshortresidencetimeat
thesiteofabsorption.Itwould,therefore,beadvantageoustohave
meansforprovidinganintimatecontactofthedrugdeliverysystem
withtheabsorbingmembranes58.Thiscanbeachievedbycoupling
bioadhesion characteristics to microspheres and developing
bioadhesive microspheres. Bioadhesive microspheres have
advantagessuchasefficientabsorptionandenhancedbioavailability
of drugs owing to a high surfacetovolume ratio, a much more
intimate contact with the mucus layer, and specific targeting of
drugstotheabsorptionsite912.Chitosan(obtainedbydeacetylation
of chitin) is a cationic polymer that has been proposed for use in
microsphere systems by a number of authors1317. Chitosan was
selected as a polymer in the preparation of mucoadhesive
microspheresbecauseofitsgoodmucoadhesiveandbiodegradable
properties. Hence, there is a need to develop an oral drug delivery
systemthatisconvenientforpatients.Varioussynthetic andnatural
polymers like alginate, chitosan and polyesters have been used to
develop drug delivery systems for entrapping and delivering drugs
orally18.Theobjectiveofthepresentinvestigationwastodevelopan
extended and controlled release composition and formulation of
trimetazidine using chitosan polymer along with sodium
tripolyphosphate to reduce dose/dosing frequency in the angina
pectoris,whichotherwisedemandsprolongedchemotherapyandto
identifythemodulationofdrugreleasefromtheformulatedmatrix
devices and demonstrate its utility in pharmaceutical drug carrier
systems.
MATERIALS
Chitosan was obtained from shreeji laboratories, Mumbai.
Trimetazidine was obtained from Nivedita Chemicals (Mumbai,
India).Sodiumtripolyphosphate(TPP)and allotherreagents were
ofanalyticalgrade.
METHODS
PREPARATIONOFMICROSPHERES
Thepreparationofthemicrospheresfollowedthemethoddescribed
by Ko et al with some modifications. Chitosan solutions of varying
concentrations were prepared by dissolving them in dilute acetic
acid(1%v/v).Tween80wasaddedintothesolutionasasurfactant.
The core material, trimetazidine hydrochloride, dissolved in CH2Cl2
(2:10), was mixed with the aqueous phase (chitosan solution) in a
homogenizer at 5000 rpm for 20min. The volume ratio of CH2Cl2:
aqueousphasewas1:10.Theemulsionwascrosslinkedbydropping
throughaspraygunintotheTPPsolution(10%).Aftercrosslinking
was allowed for varying time, microspheres were washed with
distilled water repeatedly and vacuum dried for 12 h. Three
different formulations with drug: polymer ratios (1:1, 1:2, 1:3) are
preparedandcodedasF1,F2andF3.
EVALUATIONPARAMETERS
DRUGPOLYMERINTERACTION(FTIR)STUDY19
IR spectroscopy was performed on Fourier transformed infrared
spectrophotometer(840,Shimadzu,Japan).Thepelletsofdrugand
potassium bromide were prepared by compressing the powders at
20psifor10minonKBrpressandthespectrawerescannedinthe
wave numberrange of 4000 600cm1. FTIRstudy was carriedon
puredrug,physicalmixture,formulationsandemptymicrospheres.
SCANNINGELECTRONMICROSCOPY(SEM)
Scanning electron photomicrographs of drugloaded chitosan
microspheres were taken. A small amount of microspheres was
spreadongoldstub.Afterwards,thestubcontainingthesamplewas
placed in the scanning electron microscopy (SEM) chamber. A
scanning electron photomicrograph was taken at the acceleration
voltageof20KV.
PARTICLESIZEMEASUREMENT20
Thesizeofthepreparedmicrosphereswasmeasuredbytheoptical
microscopy method using a calibrated stage micrometer for
randomlyselectedsamplesofalltheformulations.
PERCENTAGEYIELD20
Percentage practical yield is calculated to know about percentage
yield or efficiency of any method, thus it helps in selection of
appropriatemethodofproduction.Practicalyieldwascalculatedas
theweightofmicrospheresrecoveredfromeachbatchinrelationto
the sum of starting material. The percentage yield of prepared
microsphereswasdeterminedbyusingtheformula.

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 RESULTSANDDISCUSSION

In the present work controlled release microspheres of

Trimetazidine hydrochloride were formulated using chitosan
The prepared microspheres were then characterized for their polymer by ionic crosslinking emulsion technique. Three batches
variousproperties preparedwithdifferentpolymerratios wereevaluated forphysical
DETERMINATIONOFDRUGCONTENT propertieslikeFTIR,SEM,particlesize,Percentageyield,percentage
drug content, encapsulation efficiency, in vitro dissolution, release
Practicaldrugcontentwasdeterminedbytakingaweighedquantity kineticsandXRDofTrimetazidinehydrochloridemicrospheres.
of chitosan microspheres (approximately 100 mg) in a 100mL
Table1:Percentageyield,drugcontent,encapsulation
volumetricflask.Sufficientquantityofwaterwasaddedtomakethe
efficiencyandaverageparticleofTrimetazidinemicrospheres
volume100ml.Thesuspensionwasshakenvigorouslyandthenleft
andDiffusionexponent(n)ofPeppasmodelandRegressionco
for 24 hours at room temperature with intermittent shaking.
Supernatant was collected by centrifugation and drug content in efficient(r2)ofTrimetazidineHydrochloridereleasedatafrom
supernatant was determined by UV spectrophotometry at suitable microspheresaccordingtodifferentkineticmodels.
wavelength (270 nm) using a shimadzu UV visible Parameters F1 F2 F3
spectrophotometer(SHIMADZU,Spectrascan2200,Japan).
%Yield 50 67 76
DETERMINATION OF PERCENTAGE DRUG ENTRAPMENT (PDE) Drugcontent 33.80 21.30 13.70

20
(%)
Efficiencyofdrugentrapmentforeachbatchwascalculatedinterms Encapsulation 77.60 83.90 84.80
ofpercentagedrugentrapment(PDE)asperthefollowingformula: Efficiency(%)
Average 915.24 1105.84 2178.49
particlesize
ZeroOrder 0.9910.0008 0.9760.0006 0.9940.0005
FirstOrder 0.9490.05 0.9650.06 0.9530.09
Higuchi 0.9560.0005 0.9640.0002 0.9520.0006
Theoretical drug content was determined by calculation assuming PeppasModel 0.4860.08 0.3990.05 0.4230.06
that the entire drug present in the chitosan solution used gets (n)
entrapped in microspheres and no loss occurs at any stage of
preparationofmicrospheres.
INVITRORELEASE
Dissolution studies of trimetazidine from microspheres was
performedaccordingtoUSPXXIItypeIdissolutionapparatusinpH
1.2 for first 2 h and subsequent rest of the release study was
performed in phosphate buffer of pH 7.4. The temperature was
maintainedat370.5Candtherotationspeedwas100rpm.The 5
ml of sample was withdrawn at various time intervals and
replenished with an equal volume of fresh dissolution media. The
drugcontentinthesamplewasanalyzedspectrophotometrically at
270 nm. A study was performed concurrently with placebo
microspheres to record for any interference by the microsphere
components.
XRAYPOWERDIFFTACTOMETRY(XRD)STUDY21
Xray diffractometry of the Trimetazidine hydrochloride, physical
mixtureofTrimetazidinehydrochlorideandpolymer,Trimetazidine
hydrochloride microspheres and blank microspheres were Fig.1.FourierTransformInfrared(FTIR)Spectrum.(a)
performed by a diffractometer using model (Joel JDX8030, Japan) Trimetazidinehydrochloride,(b)Chitosanpolymer,(c)
equippedwithagraphitecrystalmonochromator(CuK)radiations PhysicalmixtureofTrimetazidinehydrochlorideandChitosan,
toobservethephysicalstateofTMHinthemicrospheres. (d)Trimetazidinehydrochloridemicrospheres,(e)Blank
microspheres

(a) (b) (c)
Fig.2:ScanningElectronMicrographs(SEM)ofTrimetazidinehydrochloridemicrospheres.(a)Microspherespreparedwith1:1
drug/polymerratio,(b)Microspherespreparedwith1:2drug/polymerratio,(c)Microspherespreparedwith1:3drug/polymerratio

12

 250 100
Average partical size(m)

%cum drug release

200 80

150 60

40 F1
100
F2
20
50 F3
0
0 0 1 2 3 4 5 6
F1 F2 F3
Time (hrs)
Formulations
(c)
Fig.3:AveragediameterofTrimetazidinehydrochloride
microspheres Fig.5:InvitroreleaseofTrimetazidinehydrochloride
microspheres.(a)Invitrodrugreleasewastestedforfirsttwo
hoursinpH1.2andchangetopH7.4,(b)Invitrodrugrelease
100 profileinpH1.2for4hrs,(c)InvitrodrugreleaseprofileinpH
Drug entrapment efficiency

7.4for6hrs
80

60

40

20

0
F1 F2 F3
Formulations
Fig.4:DrugentrapmentefficiencyofTrimetazidine
hydrochloridemicrospheres

100 Fig.6:XrayThermograms(XRD).(a)Trimetazidine
hydrochloride,(b)PhysicalmixtureofTrimetazidine
%cum drug release

80 hydrochlorideandChitosan,(c)Trimetazidinehydrochloride
microspheres,(d)Blankmicrospheres
60 TheFTIRSpectraofTrimetazidinehydrochloride,Chitosan,physical
mixture of Trimetazidine hydrochloride and Chitosan,formulations
40 F1 andblankmicrospheresareshownintheFig1.Fromthisitisclear
F2 thatthepeaksatAlkaneCHstretch(2920.0),secondaryamineNH
20 stretch(3446.6),aromaticC=Cstretch(1602.7),tertiaryamineCN
F3
stretch(1363.6),etherOstretch(1288.4)cm1arepresentinboth
the pure and formulations without any change in their positions
0
indicating no chemical interaction between Trimetazidine
0 1 2 3 4 5 6 7 8 9 10 11 12
hydrochlorideandpolymers.
Time (hrs)
The Controlled release microspheres of Trimetazidine
(a) hydrochloride prepared by ionic crosslinking were found to be
almost spherical and freeflowing. SEM was performed on the
prepared microspheres of 1:1, 1:2 and 1:3 to access their surface
150
andmorphologicalcharacteristicsasshowninFig2.
F1
%cum drug release

F2 It was observed that as the polymer ratio in the formulation

100 F3 increases,theproductyieldalsoincreases.Thelowpercentageyield
in some formulation may be due to microspheres lost during the
washing process. Percentage yield, drug content, encapsulation
efficiencyshowninFig3andaverageparticleshowninFig4were
50 giveninTable1.
Keepingdrugratioconstantandvariedpolymerratioasthepolymer
0
concentration increases viscosity, which influences the interaction
0 1 2 3 4 betweendispersephaseanddispersionmediumthataffectsthesize
Ti me (hrs) distribution of particle. If there was increase in the amount of
polymerconcentration,therewasincreaseinrelativeviscositysoas
(b)

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a result increases in mean particle size. The maximum particles
rangebetween100150.
Trimetazidine release from the microsphere was studied for 12 h
the drug released at constant rate in all these preparation and
showed controlled release. The release of trimetazidine
hydrochloride in two different media (pH 1.2 and 7.4) is shown in
Fig5(b)and5(c).ThepercentofdrugreleaseatpH1.2washigher
in comparison to that at pH 7.4 exhibiting pHsensitivity of the
microspheres.Attheendof4h,97.02%ofdrugloadwasdepleted
from formulation in dissolution media (pH 1.2), in comparison to
68.88%inmedia(pH7.4).theresultswassimilarformicrospheres
prepared with high concentration of chitosan with a release of
85.92%forformulationatpH1.2andsustainedreleaseatpH7.4.An
initial burstreleaseofdrug was observedfromall thebatches that
can be attributed to two reasons, the leaching of drug on the
microspheresoutersurfaceandfasteringressofdissolutionmedium
andsubsequentdiffusionofdrug.However,onchangingthepHfrom
lowertohigherlevel,thedrugreleasedslowedFig5(a).Attheend
of 12 h, 95.82% of drug was released from formulation. Similar
pattern was observed in the case of other formulations. Data
obtainedforinvitroreleasestudieswasutilizedforreleasekinetics.
Thecoefficientofdeterminationindicatedthatthereleasedatawas
best fitted with zero order kinetics. Higuchi equation explains the
diffusion controlled release mechanism. The diffusion exponent n
valuesofKorsemeyerPeppasmodelwasfoundtobeintherangeof
below 0.5 indicating Fickian of drug through Trimetazidine
hydrochloridemicrospheresweregiveninTable1.
Wide angle Xray diffraction patterns of the Trimetazidine
hydrochloride,physicalmixtureofTrimetazidinehydrochlorideand
polymer,formulationandblankmicrosphereswereshowninFig6.
TheXRDdataindicatesthattheTrimetazidinehydrochlorideis still
presentinitslatticestructureinthephysicalmixturewhere asitis
completely amorphous inside the Trimetazidine hydrochloride
microspheres.Thismaybeduetotheconditionsusedtopreparethe
Trimetazidine hydrochloride microspheres lead to cause complete
TMHamorphization
ACKNOWLEDGEMENT
Authors are thankful to Bharathi College of Pharmacy for their
supportencouragementincarryingoutthiswork.
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