Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Formulation and Evaluation of Ezetimibe Nanoparticles
Yasser A.Ali

*
and Shaimaa N. Abd-Alhammid *

, 1

*
Department of Pharmaceutics, College of Pharmacy, University of Baghdad, Baghdad, Iraq,

Abstract
The aim of this study is to formulate and evaluate ezetimibe nanoparticles using solvent

antisolvent technology. Ezetimibe is a practically water-insoluble drug which acts as a lipid lowering

drug that selectively inhibits the intestinal absorption of cholesterol and related phytosterols. Ezetimibe

prepared as nano particles in order to improve its solubility and dissolution rate.
Thirty formulas were prepared and different stabilizing agents were used with different concentrations

such as poly vinyl pyrrolidone (PVPK-30), poly vinyl alcohol (PVA), hydroxy propyl methyl cellulose

E5 (HPMC), and poloxamer. The ratios of drug to stabilizers used to prepare the nanoparticles were 1:

2, 1:3 and 1:4.

The prepared nanoparticles were evaluated for particle size, entrapment efficiency, dissolution

study, Fourier transform infrared spectroscopy, differential scanning calorimetry, and atomic force

microscopy. The percentage of drug entrapment efficiency of F1-F30 was ranged from 85% ± 1 to 98

% ± 1. On the other hand dissolution rate increasing as the particle surface area is increase due to

reduction of particle size to the nano range.

The results showed that poly vinyl pyrrolidone (PVPK-30) was found to be the best stabilizer.
Keywords: Ezetimibe, Nanoparticles, Particle Size, poly vinyl alcohol.

تيميب بواسطة انجسيمات اننانويةياالز حبيباتوتقييم تصييغ

ياسر عبد انصاحب عهي
*
عبد انحميدنسارشيماء و

*،1

*
.العراق ،بغداد ،جبمعة بغداد ،كلية الصيدلة ،فرع الصيدالنيبت

انخالصة
الحرسدددي مددد جكنىلىجيدددب ببسدددحمدا لعقدددبر االميح بيددد نبنىيدددة جسدددي بت وجقيدددي لصددديب ة هدددى الدراسدددة هددد مددد الهدددد أن

انحقددددبي بشددددك جثدددد الحدددد علددددً الدددد الدددددهىن وهددددى دواء يع دددد ال ددددبء يددددر ايدددد فدددد دواء هددددى اميح بيدددد . مضددددبد ال دددد ي

القببليددددة جحسددددي بغيددددة نبنىيددددة كجسددددي بت أعددددد اميح بيدددد . الصددددلة ات و ال ددددىاد الدهنيددددة األمعددددبء امحصددددبك الكىلسددددحرو مدددد

. االمحصبك ومعد لل وببن

اللبينيددد ببيروليددددون مثددد ممحللدددة بحراكيددد اسدددحمدمث ممحللدددة اسدددحقرار ببسدددحمدا بدددىلي رات صددديغة ثالثدددي إعدددداد جددد

. وبىلىكسددددبمير ،HPMC E5) (السددددليلىم ال يثيدددد بروبيدددد هيدروكسدددد ،(PVA) الكحددددى اللينيدددد وبددددىل ،(PVP) ال حعدددددد

. 2:1 و 2:1 و 2:1 النبنىية ه الجسي بت إعداد ف ال سحمدمة ال ث حبت إلً الدواء وكبنث نس

الشددددك ال يددددبن للححددددرر ودراسددددة انح ددددبد الدددددواء، وكلددددبء مدددد ايددددا الحجدددد الح ي دددد للجسددددي بت " نبنىيددددة جسددددي بت" وقي ددددث

النسدددد ة .ال ريددددة القددددى ومجهددددر الحلبضددددل ( ال سدددد وقيددددبد ، الح ددددراء جحددددث األشددددعة مطيبفيددددةوكدددد لا دراسددددة الحىافدددد ) الدددددواي ،

%.مدددد نبايددددة 85% الددددً 58هدددد مدددد 13الدددددواء للصدددديي الدواييددددة مدددد الصدددديغة االولددددً الددددً الصدددديغة ال ئىيددددة لكلددددبء انح ددددبد

بدددىل أن النحدددبي وأظهدددرت اادددري يددد داد جحدددرر الددددواء كل دددب صدددغر اجددد الجسدددي بت النبنىيدددة ل يدددبد ال سدددباة السدددطحية للجسدددي .

نىية.بىلي ر اسحقرار للجسي بت النب هى أفض ( -13PVP K(الليني ببيروليدون
.انكحول انفينيم بوني انحبيبي، انحجم انجسيمات اننانوية، إزتيميب، -انمفتاحية : كهماتان

Introduction
Solubility is of the most important

parameters to achieve the desired

concentration of a drug in the systemic

circulation for pharmacological response

to be shown. A number of methodologies

can be adapted to improve solubilization

of poor water soluble drug and further to

improve its bioavailability include

chemical modification , pH adjustment,

solid dispersion, complexation, co‐
solvency, and micronization

(1)
.

One of these methods is the

nanosuspension which is colloidal

dispersions of nano-sized drug particles

that are produced by a proper method and

stabilized by a suitable stabilizer
(19)

. The

particle size distribution of the solid

particles in nano suspensions is usually

less than one micron with an average

particle size ranging from 200 and 1000

nm
(2)

. Ezetimibe is a member of new

class of lipid - lowering compounds that

selectively inhibit the intestinal absorption

of cholesterol and decrease cholesterol

absorption
(3)

Ezetimibe is categorized as a class

II agent (poorly water soluble and highly

permeable with a relative

1
Corresponding author E-mail: shaimaa-alsamariai@yahoo.com.

Received: 21/4/ 2015

Accepted: 29/6/2015

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Bioavailability range from 35-65 %
(4)

The aim of this study is to formulate

and evaluate ezetimibe nanoparticles using

solvent antisolvent technology.

Materials and Methods
Materials

Ezetimibe powder, was purchased

from (Provizer Pharma, Gujarat, India).

Poly vinyl pyrrolidone PVP K-30 (BDH

chemicals LTD, Liverpool, England).

Poly vinyl alcohol (Riedal De Haen Ag

Seelze, Hannover, Germany), HPMC

(Provizer Pharma, Gujarat, India).

Poloxamer 188 (BDH chemicals LTD,

Liverpool, England). Methanol (GCC

Analytical reagent, UK). brij35 (Riedal

De Haen Ag Seelze, Hannover,

Germany). All other chemicals were of

analytical grade.

Methods

Preparation of ezetimibe nanosuspension

Nanosuspensions were prepared

by the solvent evaporation technique

which is also called solvent antisolvet

technique
(5)

, as shown in table (1), (2), (3)

that the ezetimibe was dissolved in 10 ml

methanol and poured into 100 ml water

containing different types of stabilizers

(alone and in combination) maintained at

a temperature of 50°C and subsequently

stirred at agitation speed of 500 revolution

per minute (rpm) on magnetic stirrer for 1

hour to allow the volatile solvent to

evaporate. The organic solvents which

contain 10 mg of ezetimibe were added by

means of a syringe drop by drop

positioned with the needle directly into

stabilizers containing water. The ratios of

drug to stabilizers used to prepare the

nanosuspension were 1: 2, 1:3 and 1:4.

Then centrifuge to obtain the

nanoparticles.

Table (1): Composition of ezetimibe nanosuspension using different types of stabilizers at drug:

stabilizer ratio 1:2.

Formula no.

Materials

F1 F2 F3 F4 F5 F6 F7 F8 F9 F10

Ezetimibe(mg) 10 10 10 10 10 10 10 10 10 10

PVP(mg) 20 10 10 10

PVA(mg) 20 10 10 10

HPMC(mg) 20 10 10 10

Poloxamer188

(mg)

20 10 10 10

Methanol (ml) 10 10 10 10 10 10 10 10 10 10

Water (ml) QS 100 100 100 100 100 100 100 100 100 100

Table (2): Composition of ezetimibe nanosuspension using different types of stabilizers at drug:

stabilizer ratio 1:3.

Formula no.

Materials

F11 F12 F13 F14 F15 F16 F17 F18 F19 F20

Ezetimibe(mg) 10 10 10 10 10 10 10 10 10 10

PVP(mg) 30 15 15 15

PVA(mg) 30 15 15 15

HPMC(mg) 30 15 15 15

Poloxamer188 30 15 15 15

Methanol

(ml)

10 10 10 10 10 10 10 10 10 10

Water (ml)

100 100 100 100 100 100 100 100 100 100

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Table (3): Composition of ezetimibe nanosuspension using different types of stabilizers at drug:

stabilizer ratio 1:4

Formula no.

Materials

F21 F22 F23 F24 F25 F26 F27 F28 F29 F30

Ezetimibe(mg) 10 10 10 10 10 10 10 10 10 10

PVP(mg) 40 20 20 20

PVA(mg) 40 20 20 20

HPMC(mg) 40 20 20 20

Poloxamer 188 40 20 20 20

Methanol (ml) 10 10 10 10 10 10 10 10 10 10

Water (ml) QS 100 100 100 100 100 100 100 100 100 100

Evaluation of the prepared

nanosuspension

Particle size determination was

done using ABT-9000 nano laser particle

size analyzer at 25ºC without dilution of

the samples. The average particle size (D)

was measured for all the prepared

formulas at ratios of 1: 2 (F1-F10), 1: 3

(F11-F20) and 1: 4(F21- F30). Each

sample was sonicated for 20 minute

before measuring and each sample was

measured in triplicate.

Determination of drug entrapment efficiency

(EE) of nanosuspension

The freshly prepared

nanosuspension of ezitimibe: stabilizer

ratio 1:2, 1:3 and 1:4 was centrifuged at

20,000 rpm for 20 minutes using

ultracentrifuge. The amount of non

incorporated drug was measured by taking

the absorbance of the appropriately

diluted 25 ml of supernatant solution at

232 nm using UV-visible

spectrophotometer. The entrapment

efficiency (EE %) was calculated by

subtracting the amount of the free drug in

the supernatant from the initial amount of

drug taken. For each formulation the

experiment was repeated in triplicate and

the average was calculated
(6, 7)

The Percentage of drug entrapment

efficiency (% EE) could be achieved by

the following equation :

entrapment efficiency = (Weight initial

drug- Weight free drug) / Weight initial

drug

Freeze drying of nanosuspension

In order to retrieve nanoparticles in

dried-powder state from the

nanosuspensions, water-removal was

conducted through freeze-drying, so that

each formula was lyophilized using

vacuum freeze dryer at a controlled

temperature of (- 45) ˚C and the pump

operating at a pressure of 2.5 × 10

pascal over a period of 48–72 hour.

The yielded powders were used for

further studies
(8)

.
In-vitro dissolution profile of

nanosuspension
In vitro dissolution study was

performed using USP dissolution test

apparatus-II (paddle assembly). The

dissolution was performed using

lyophilized powder in 500 ml of 0.1N

HCL (pH 1.2) and phosphate buffer

solution (pH 6.8) as dissolution mediums

containing 2% brij 35 and maintained at

37 °C and 50 rpm for ezetimibe

lyophilized powder formulas. The freshly

prepared formula F1-F4, F11-F14 and

F20-F24 where freeze dried to get the

nanoparticles then immersed in

dissolution medium. Samples (5ml) were

withdrawn at regular intervals of 5

minutes for 120 minutes and replaced

with fresh dissolution medium to keep
sink conditions. Samples were filtered

through filter paper and assayed spectro

photometrically on UV-Visible

spectrophotometer at 232 nm wave length
(9)

.
Fourier transform infrared spectroscopy

(FTIR)

The fourier transform infrared

spectroscopy (FTIR) spectra were obtained

using FTIR spectroscope. Samples which

studied are: Pure ezetimibe powder, PVP K-

30, Physical mixture of ezetimibe, and PVP K-

30 at ratio (1:4); respectively. Lyophilized

powder (nanoparticles) of the selected formula

(F21), Microcrystalline cellulose PH 102

(Avicel) ® , lactose ,Magnesium Sulphate.

All these samples were grounded and mixed

thoroughly with potassium bromide. The

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

spectrum obtained was in between the wave

number of 4000-400 cm-
1 (10).

Differential scanning calorimetry (DSC)
DSC can be used to determine the

compatibility between the drug and

excipients and also used to evaluate the

crystalline state of drug especially when

converted to nanoparticles.

Thermal characteristics of the same

samples that are studied by FTIR were

determined also by an automatic thermal

analyzer system. Therefore accurately

weighed samples (5mg) were placed in

non hermetically aluminum pans and

heated at the rate of 10 ºC/minute against

an empty aluminum pan as a reference

covering a temperature range of 40 ºC to

300 ºC
(11)

Atomic force microscopy
Atomic force microscopy (AFM) is

capable of scanning the surfaces in

controlled environmental conditions, also

can measure the particle size of the

nanoparticles accurately. The size and

surface morphology of ezetimibe

nanoparticles in F21 were confirmed by

atomic force microscopy after drying of

the formula. The optimized formula F21

were lyophilized and dried 15 minutes in

desiccators. Particle size, 3D-dimension

graph and histogram of particle size

distribution were obtained
(12, 13)

Statistical analysis

The results of the experiments were

given as a mean of triplicate samples ±

standard deviation and were analyzed

according to the paired T test and one way

analysis of variance (Single Factor

ANOVA) at the level of (P < 0.05).

Results and Discussion
Evaluation of nanosuspension

Particle size analysis

The particle size of formulas F1-F4

at drug: polymer ratio 1:2 was ranged

from 95.4 -956.5 nm measured by particle

size analyzer, while for F11-F14 at drug:

polymer ratio 1:3 the particle size ranged

from 66.52-899.1 nm. On the other hand

F21-F24 at drug :polymer ratio the

particle size of these formula range from

35.3- 901.2 using PVP K-30 , PVA,

HPMC and poloxamer 188 as primary

stabilizers .

The formulations containing PVP K-30,

PVA , and HPMC as stabilizers had small

significant particle size in comparison

with the formulation containing

poloxamer 188 that gave larger particle

size (p<0.05). Poloxamer188 (pluronic

F68)® is a block co-polymer, responsible

for the hydrophobic interaction with the

drug molecule ,the crystal growth

inhibition is mainly due to the

hydrophobic polypropylene oxide group

(PPO) in the pluronic polymer , while the

hydrophilic polyethylene oxide (PEO)

chains provide steric hindrance upon

aggregation
(14)

. Poloxamer 188 can form a

valuable mechanical and thermodynamic

barrier at the interface that hinders the

approach and coalescence of individual

emulsion droplets at their optimum level.

Although this mechanism of poloxamer

188, but it gave larger particle size in all

three ratios 1:2, 1:3 and 1:4 drug: polymer

in formulas F4, F14 and F24; respectively.

High particle size of F4, F14 and F24 that

contain poloxamer188 as a stabilizer may

be attributed to the insufficient affinity, of

poloxamer188 to ezetimibe, and possess a

slow diffusion rate and ineffective

adsorption onto the drug particle surface

in the water–methanol mixture. However,

if there is no affinity between the particle

surface and the polymer, the attractive

forces between two particles become

dominant due to depletion of polymer

from the gap of two particles (depletion

force)
(15)

Poly dispersity index values of F1, F2 and

F3 ranged from 0.002 -0.005 indicate that

these formulas are mono disperse

standard. While for F4 that contain

poloxamer 188 PDI value was 0.439

which indicate mid range poly dispersity

system. The surface area values of the

particles in F1, F2 and F3 ranged from

39.2 - 57.53 (m
2
/g) while for F4 that

contain poloxamer 188 particle surface

area value was 3.89 (m
2
/g) which has the

smallest particle surface area (p<0.05) in

comparison with other formulas because it

has largest particle size
(16)

For drug :stabilizer ratio 1:3,

polydispersity index values of F11, F12

and F13 ranged from 0.002 - 0.033

indicate that these formulas are

monodisperse standard, while for F14 that

contain poloxamer 188 PDI value was

0.557 which indicates mid range

polydispersity system .

Specific surface area values of the

particles in F11, F12 and F13 ranged from

66.21 – 99.23 (m
2
/g) while for F14 that

contain poloxamer 188 the SSA value was

4.25 (m
2
/g) which has the smallest SSA

(p<0.05) in comparison with other

formulas because it has largest particle

size.

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

On other hand for drug : polymer ratio 1:4

the poly dispersity index values of

formulas F21, F22 and F23 range from

0.002-0.031 which indicate that these

formulas are mono disperse system ,

while for F24 that contain poloxamer188

PDI value was 0.811 which indicates mid

range poly dispersity system Specific

surface area values of the particles in F23,

F22 and F21 ranged from 122.21 – 557.23

(m
2
/g) while for F24 that contain

poloxamer188 the SSA value was 4.25

(m
2
/g) which has the smallest SSA

(p<0.05) in comparison with other

formulas because it has largest particle

size.

This difference in values of PDI could be

attributed to the efficiency of stabilizers,

which cover the organic/aqueous interface

of the nano droplets and prevent them

from coalescing to each other. From the

obtained results, one can conclude that the

poloxamer188 is not suitable as a primary

stabilizer for nanoparticles because of

poor adsorption and poor affinity of

poloxamer188 to the ezetimibe molecules.

Particle size ranged from 35.57 nm for

PVPK30 to 999 nm for poloxamer188,

which appeared to be affected by relative

viscosity of the polymeric dispersion in

the presence of stabilizers and followed

the trend: PVP ˃ PVA ˃ HPMC >

poloxamer188. Nanosuspension with PVP

K-30 as stabilizers possessed the smallest

particle size while that containing

poloxamer188 had the largest particle size
(17)

Effect of polymer concentration on the

size of ezetimibe nanoparticles

The effect of the polymer

concentration on the particle size of

ezetimibe nanosuspention have been

investigated by depending on three ratios

of drug : polymer concentration (1:2) in

the preparation of F1-F4 and in 1:3 of

drug : polymer ratio in the formulation

of F11-F14. And in the ratio of 1:4 in the

formulas F21- F24. Polymer

concentration affecting on the adsorption

affinity of the stabilizers to the particle

surface.

In general as the concentration of

polymers increase the particle size

decrease at fixed drug concentration

except for poloxamer188, which indicated

that the drug particle surface has been

sufficiently covered well by the stabilizer

molecules
(18)

It has been noticed that the particle sizes

of F1, F2 and F3 using PVP K-30, PVA

and HPMC; respectively as stabilizer

were decreased when the concentration of

polymer increased in the formulas F11,

F12 and F13 and they were further

decreased in particle size when the

concentration of the polymer increased as

shown in the formulas F21, F22 and F23

using the same stabilizers, while F4 , F14

and F24 containing poloxamer 188, as the

only stabilizer, have maintained within the

same value of particle size and not

increased sufficiently even when the

concentration was increased. It can be

interpreted as the fact that poloxamer 188,

by itself have poor adsorption properties

and affinity to the molecules of ezetimibe

that prevent particle agglomeration and

this finding did not agree with the

reported one
(19)

Polymer concentration plays a great role

in the stabilization of nanoparticles

because of too little stabilizer induces

agglomeration or aggregation and too

much stabilizer promotes Ostwald’s

ripening
(20)

.The decrease in the particle

size is accompanied by a rapid, highly

increase in the surface area. Thus, the

process of primary coating of the newer

surfaces competes with the agglomeration

of the uncoated surfaces. Hence, an

increase in the surfactant concentration in

the primary dispersion results in rapid

coverage of the newly formed particle

surfaces
(21)

Effect of combination of two polymers on

the size of ezetimibe nanoparticles

The particle size of F5, containing

PVA and PVP K-30 as stabilizers

combination at drug: stabilizer ratio 1:2

was decreased significantly (p<0.05) from

140 nm to 40.23 nm in F25 at drug:

stabilizer ratio 1:4 ratio when the

stabilizer concentration increased.

Poloxamer 188, when used singly was

not so effective in reducing the particle

size as a stabilizer, rather flocculation was

observed. This could be due to its high

hydrophilicity (HLB = 29) due to which it

may not be undergoing preferential

adsorption on the nanoparticle surface.

However, poloxamer188 in combination

with PVP k-30, PVA and HPMC , it

worked synergistically therefore the

particle size of ezetimibe nanosuspension

was drastically reduced.

It has been found that the combination of

PVPK-30 and poloxamer188 have

reduced particle size this mainly due to

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

that PVP K-30 is reported to be a

protective colloid which is indicative of

its greater adsorption potential for the

nanoparticles
(22)

. This is expected as the

stabilizers used for preparing the

ezetimibe nanosuspensions are either

hydrophilic polymers or non-ionic

surfactants which stabilize the particles by

steric stabilization
(22, 23)

Determination of drug entrapment

efficiency of nanosuspension

The percentage of drug entrapment

efficiency of F1-F30 was ranged from

85% ± 1 to 98 % ± 1 as shown in figure

(11). It is clear that the increase in

stabilizer concentration increased the drug

entrapment efficiency, but the study

revealed that the concentration of

stabilizers at ratio 1:2 drug: stabilizer was

sufficient to give the optimized

entrapment efficiency. F4 containing

poloxamer188 as stabilizer had the lowest

entrapment efficiency, while F21

containing PVP-k-30 as the only stabilizer

had the higher entrapment efficiency. This

may be due to the presence of optimum

stabilizer and optimum stabilizer

concentration
(24)

In vitro dissolution study
The dissolution profile was done

for F1-F4, F11-F14 and F21-24 of drug:

stabilizer ratio 1:2, 1:3 and 1:4

respectively. The dissolution of the

prepared formulas was carried in 0.1N

HCL solution (pH1.2) and phosphate

buffer solution (pH6.8) in the presence of

2% brij-35 to get the selected formula that

can increase the dissolution rate in these

buffers which are simulated to gastric and

intestinal fluids. In the dissolution study

one notice enhancement of dissolution

rate, according to Noyes-Whitney

equation the dissolution rate increasing as

the particle surface area is increase due to

reduction of particle size to the nano

range.

Superior dissolution of ezetimibe

nanoparticles may potentially improve

bioavailability and other drug

performances. PVPk-30 containing

ezetimibe nanoparticles at drug - polymer

ratio 1:4 with particle size of 35.57 nm

showed the highest drug release rate as

100% of drug dissolved in 10 minutes

whereas, poloxamer 188 containing

ezetimibe nanoparticles at drug - polymer

ratio 1:2 with particle size of 999 nm

showed about 33.9% of drug dissolved

within 10 minute of dissolution test in

both 0.1 N HCL (pH 1.2) and phosphate

buffer (pH 6.8)
(25, 26)

Figure (1): Effect of polymer type on the

dissolution profile of ezetimibe

nanoparticles from F1, F2, F3, and F4 in 0.1

N HCl solution (pH 1.2) containing 2% brij-

35 w/v at 50 r.p.m and 37˚C temperature .

Figure (2): Effect of polymer type on

the dissolution profile of ezetimibe

nanoparticles from F11, F12, F13, and

F14 in 0.1 N HCL solutions (pH 1.2)

containing 2% brij-35 w/v at 50 r.p.m

and 37˚C temperature.

Figure (3): Effect of polymer type on the

dissolution profile of ezetimibe

nanoparticles from F21, F22, F23, and F24

in 0.1 N HCL solutions (pH 1.2) containing

2% brij-35 w/v at 50 r.p.m and 37˚C

temperature.

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Figure (4): Effect of polymer type on the

dissolution profile of ezetimibe

nanoparticles from F1, F2, F3, and F4 in

phosphate buffer (pH 6.8) containing 2%

brij-35 w/v at 50 r.p.m and 37˚C

temperature.

Figure (5): Effect of polymer type on the

dissolution profile of ezetimibe

nanoparticles from F11, F12, F13, and F14

in phosphate buffer (pH 6.8) containing 2%

brij-35 w/v at 50 r.p.m and 37˚C

temperature.

Figure (6): Effect of polymer type on the

dissolution profile of ezetimibe

nanoparticles from F21, F22, F23, and F24

in phosphate buffer (pH 6.8) containing 2%

brij-35 w/v at 50 r.p.m and 37˚C

temperature.

Drug content in lyophilized powder
The drug content result showed that 25 mg of

lyophilized powder of the selected formula (F

21) contain 5 mg ±0.1 of ezetimibe when

determined by UV-visible spectrophotometer

at λmax 232 nm.

Fourier transforms infrared spectroscopy

FTIR is one of the most widely reported

spectroscopic techniques for solid-state

characterization. The characteristics absorption

bands of ezetimibe are:

1. O–H stretching band at 3650—2700
cm

-1

2. C-O stretching bands of the lactam
ring at 1725—1714 cm

-1

3. C=C stretching band of the aromatic
ring at 1600—1500 cm

-1

4. C–F stretching band at 1000—1200
cm

-1

5. C–O stretching band at 1300—1000
cm

-1

FTIR spectra of ezetimibe

nanosuspention and tablet show no change in

shifting of the position of the major functional

groups indicating no major interaction between

the drug and the stabilizer PVP K-30 and other

excipients. FTIR spectra of physical mixture

also showed peaks at similar position. Hence,

it can be conclude that there was no possible

interaction between the drug, the stabilizer and

the used excipients
(27, 28)

Figure (9) demonstrate the DSC

thermogram of ezetimibe that showed

sharp characteristic endothermic peak at

165.30ºC and this agrees with the

references. This gives an indication that

the drug has crystalline nature with high

purity. The DSC thermograms of the

ezetimibe of tablet of the selected formula

F21, lyophilized powder and physical

mixture are shown in figure (10) that

show that the drug in the crystal

structure have a melting endotherm while

ezetimibe molecules in the amorphous

state do not exhibit a melting endotherm.

As seen in Figures (9), and (10) the sharp

melting peak of ezetimibe (165.30 °C) is

completely disappeared in these figures

that’s mean the stabilizer PVP K-30

completely converted ezetimibe particles

into amorphous state
(29)

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Figure (7): FTIR spectrum of eztimibe

Figure (8): FTIR spectrum of ezetimibe nanosuspension Differential Scanning Calorimetry

0.00 100.00 200.00 300.00
Temp [C]

-10.00

-5.00

0.00

mW
DSC

165.30x100C

Figure (9): DSC thermogram of ezetimibe powder

0.00 100.00 200.00 300.00
Temp [C]

-2.00

0.00

2.00

4.00

mW
DSC

64.82x100C

147.90x100C

214.60x100C

Figure (10): DSC thermogram of ezetimibe tablet of formula F21

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Evaluation of surface morphology

Atomic force microscopy study

The morphological analysis and particle

size of F21 performed by AFM showing

irregular to spherical shaped nanoparticles

with a size of 31 nm as seen in figure (11) as it

approved by the histogram of particle size

distribution in figure (12) also figure (13)

shows histogram of particle size distribution of

F21 by atomic force microscopy.

The formulation was found to be stable

and no aggregation of particles could be

observed. The particle size of F21 obtained by

AFM was comparable to or equal to that

measured by ABT-9000 nano laser particle

size analyzer (35.57 nm) and this in agreement

with particle size measurements provide the

good size distribution and the stability of

ezetimibe nanparticles
(31)

Figure (11): Atomic force microscopy of formula F21 showing cross section and long tudinal

section of the nanoparticles surface.

Figure (12): Particle size distribution of F21 by particle size analyzer ABT-9000

Iraqi J Pharm Sci, Vol.24(2) 2015 Ezetimibe nanoparticles

Figure (13): Histogram of particle size distribution of F21 by AFM

Conclusion
Nano particulate systems have

great potentials, being able to convert

poorly soluble, poorly absorbed and labile

biologically active substance into

promising deliverable drugs.

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