International Journal of Pharmaceutics 489 (2015) 5872

Contents lists available at ScienceDirect

International Journal of Pharmaceutics

journal homepage: www.elsevier.com/locate/ijpharm

Implementation of quality by design principles in the development of

microsponges as drug delivery carriers: Identication and optimization
of critical factors using multivariate statistical analyses and design of
experiments studies
Maja Simonoska Crcarevska a,b, * , Aneta Dimitrovska c , Nadica Sibinovska a,b ,
Kristina Mladenovska a,b , Renata Slavevska Raicki a,b , Marija Glavas Dodov a,b
a

Institute of Pharmaceutical Technology, Faculty of Pharmacy, University of Ss. Cyril & Methodius, Majka Tereza 47, 1000 Skopje, Macedonia
Center of Pharmaceutical Nanotechnology, Faculty of Pharmacy, University of Ss. Cyril & Methodius, Majka Tereza 47, 1000 Skopje, Macedonia
c
Institute of Applied Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy, University of Ss. Cyril & Methodius, Majka Tereza 47, 1000 Skopje,
Macedonia
b

A R T I C L E I N F O

A B S T R A C T

Article history:
Received 4 March 2015
Received in revised form 14 April 2015
Accepted 15 April 2015
Available online 17 April 2015

Microsponges drug delivery system (MDDC) was prepared by double emulsion-solvent-diffusion

technique using rotorstator homogenization. Quality by design (QbD) concept was implemented for the
development of MDDC with potential to be incorporated into semisolid dosage form (gel). Quality target
product prole (QTPP) and critical quality attributes (CQA) were dened and identied, accordingly.
Critical material attributes (CMA) and Critical process parameters (CPP) were identied using quality risk
management (QRM) tool, failure mode, effects and criticality analysis (FMECA). CMA and CPP were
identied based on results obtained from principal component analysis (PCA-X&Y) and partial least
squares (PLS) statistical analysis along with literature data, product and process knowledge and
understanding. FMECA identied amount of ethylcellulose, chitosan, acetone, dichloromethane, span 80,
tween 80 and water ratio in primary/multiple emulsions as CMA and rotation speed and stirrer type used
for organic solvent removal as CPP.
The relationship between identied CPP and particle size as CQA was described in the design space
using design of experiments one-factor response surface method. Obtained results from statistically
designed experiments enabled establishment of mathematical models and equations that were used for
detailed characterization of inuence of identied CPP upon MDDC particle size and particle size
distribution and their subsequent optimization.
2015 Elsevier B.V. All rights reserved.

Keywords:
Microsponges
Semisolid dosage form
Quality by design
Principal component analysis
Partial least squares
Response surface method

1. Introduction
The development of pharmaceutical formulation is a complex
and multistage process. In this process many aspect should be
considered in order to achieve the optimal combination of
operational factors that will provide the best characteristic of the
pharmaceutical product and process response. However, a central
challenge in drug product development is the difculty to evolve an
ideal formulation using conventional approach. For this reason, the

* Corresponding author at: Institute of Pharmaceutical Technology, Faculty of

Pharmacy, Ss. Cyril & Methodius University, Majka Tereza 47, 1000 Skopje,
Macedonia. Tel.: +389 23126032; fax: +389 23123054.
E-mail addresses: msimonoska@ff.ukim.edu.mk, maja.simonoska@gmail.com
(M. Simonoska Crcarevska).
http://dx.doi.org/10.1016/j.ijpharm.2015.04.038
0378-5173/ 2015 Elsevier B.V. All rights reserved.

regulatory strategy for pharmaceutical product development is to

encourage industry to innovatively apply the quality by design (QbD)
approach (ICHQ8(R2), 2009). This concept has provided an
opportunity for the industry to evaluate and identify best practices
for key QbD elements of quality target product proles (QTPP),
critical quality attributes (CQA), risk assessment, process characterization for design-space denition, CQA focused control strategies
and extended change protocols. Having in mind, the vast number of
possible formulation and process factors, as well as their combined
effects that might inuence drug product quality, its therapeutic
efcacy and safety, it is more than important to use quality risk
management (QRM) as integral part of QbD (ICHQ8(R2), 2009;
ICHQ9, 2005) to make rationale balance between experiments,
resources and time required for development of pharmaceutical
formulations (Fahmy et al., 2012). In this study we used QbD concept

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

for development of microsponges as drug delivery carriers (MDDC)

with potential to be incorporated into gel intended for topical
application. MDDC are polymeric porous microparticles and like a
true sponge, each microparticle (size ranging from 5 to 300 mm)
consists of a myriad of interconnecting voids loaded with active
agent within a non-collapsible structure, with a large porous surface
(Kaity et al., 2010). The pores in the microparticle form a continuous
arrangement open to the exterior surface of particles which permits
the outward diffusion of the entrapped drug molecule at a controlled
rate depending on the pore size (Srivastava and Pathak, 2012). These
porous, polymeric microparticles are used mostly for topical and
recently for oral administration and may be integrated or formulated
into pharmaceutical dosage forms, such as creams, gels, lotions,
powders, soaps, capsules and tablets (Jadhav et al., 2013; Orlu et al.,
2006). MDDC can entrap a wide range of active ingredients such as
emollients, fragrances, essential oils, sunscreens, anti-infective,
antifungal or anti-inammatory agents (Kaity et al., 2010). High
quantities of active ingredients can be load into the bulk of the MDDC
and/or adsorbed on their surface (Srivastava and Pathak, 2012). In
addition to efcient entrapment of active ingredients, MDDC
technology is believed to contribute toward efcient delivery of
an active ingredient at the minimum dose, reduced side effects,
improved stability, increased elegance, enhanced formulation
exibility and modication of the drug release (Orlu et al., 2006;
Srivastava and Pathak, 2012). Compared to microcapsules and
liposomes, the microsponges have the advantage of higher payload
and cost effectiveness (Aloorkar et al., 2012).
MDDC technology is being used in cosmetics, over-the-counter
skin care and prescription products (Bamane et al., 2014). The
fundamental appeal of the microsponge technology in topical drug
delivery systems stems from the difculty experienced with
conventional formulations in releasing active ingredients over an
extended period of time. Namely, the conventional topical dosage
forms like ointments, lotions, creams, gels or pastes release their
active ingredient upon application, producing a highly concentrated layer of the active ingredient that is rapidly absorbed, but this
may lead to a cycle of short-term overmedication followed by longterm undermedication (Maiti et al., 2011). The microsponges
system can prevent excessive accumulation of ingredients within
the epidermis and the dermis (Kaity et al., 2010). When the
microsponges are used as topical drug delivery carriers, they allow
an even and sustained rate of release and therefore reduce
irritation while maintaining efcacy (Maiti et al., 2011). In addition,
the microsponges particles themselves do not pass through the
skin, but rather, they gather in the tiny nooks and crannies of the
skin and slowly release the entrapped drug as the skin needs it. The
microsponges are also capable of absorbing skin secretions,
therefore reducing oiliness and shine from the skin (Kaity et al.,
2010). When applied to the skin, the microsponges release the
active ingredient according to a determined time mode but it may
also be released in response to other stimuli (rubbing, temperature, pH, etc.) (Castro and Ferreira, 2008).
Microsponges can be customized in term of their physicochemical and biopharmaceutical properties making them suitable
for a specic therapeutic application. It is known that both the
formulation variables (such as the composition of the internal and
external phases, drug to polymer ratio etc.) and process variables
(type of stirrer, stirring rate, temperature, etc.) inuence the MDDC
properties thus impacting its therapeutic efcacy and safety. In
order to simultaneously study factors that inuence drug product
quality and to thoroughly investigate their relationship in
experimental design space, a combined approach of multivariate
analysis (MVA) and design of experiments (DoE) can be used.
Actually, in this study MVA and DoE were used side by side with
denition of QTPP, identication and determination of potential
CQAs, as well as QRM as integral part of QbD concept (ICHQ8(R2),

59

2009). CQA were identied and prioritized using QRM tool failure
mode, effects and criticality analysis (FMECA). Having in mind that
particle size is one of MDDC properties that is critical for their
encapsulation efciency, drug loading capacity and drug release
prole (properties related to drug product efcacy and safety), we
used it as a stand for identication of critical material attributes
(CMA) and critical process parameters (CPP). CMA and CPP were
identied using MVA principal component analysis (PCA) and
partial least squares (PLS) applied on data from experiments
previously conducted in our laboratory, along with available
literature data, knowledge related to potential drug product (gel
for topical application with MDDC) and manufacturing process
understanding. The relationship between the identied CPP and
particle size as CQA was described in the design space using onefactor response surface method (RSM) DoE.
2. Materials and methods
2.1. Materials
Ethylcellulose (EC) was purchased from Sigma, USA. Chitosan
(CTS) (high viscous) was supplied from Aldrich, Island. Tween 80
(T80), span 80 (S80), dichloromethane (DHM) and hydrochloric
acid were obtained from Merck, Germany; while acetone (ACT)
was purchased from Alkaloid, Macedonia. All other used chemicals
and reagents were of analytical grade and were used without any
modications.
2.2. Methods
2.2.1. QbD approach for development of formulation of microsponges
as drug delivery carriers for incorporation into semisolid dosage form,
gel for topical application
QTPP include patient relevant product performance elements.
QTPP describes prospectively the quality characteristics of a drug
product that should be achieved to ensure the desired quality,
taking into account safety and efcacy of the drug product. CQA for
nished product are subsets of QTPP that has a potential to be
altered by the change in formulation or process variables (Chang
et al., 2013; Jain, 2014).
CQA list should also include the acceptance limits for each CQA,
and a rationale for designating these properties as CQA. QTPP and
CQA are dened according to guidelines and prior knowledge and
understanding of the drug product and manufacturing process
(EMA/430501/2013, 2013; ICHQ8(R2), 2009).
QRM is based on FMECA approach (ICHQ9, 2005). The goal is to
quantify the potential risk of formulation and process variables and
assess its impact on the drug product manufacturing. Risk
quantication was performed according to Zhang et al. (2013).
Briey, ranking system covers severity (S), probability (P), and
detectability (D) of each parameter. For each factor, scores for S, P,
and D are multiplied, yielding a risk priority number (RPN) on a
scale from 1 to 36. Any factor with the value of RPN above 10 should
be taken into account in further studies. Ranking scale was as
follows:
- Severity (S)

4 - High, a minor change in the parameter could cause

signicant impact on product quality
3 - Moderate, only if parameter uctuates in a wide range may
cause signicant impact on product quality
2 - Low impact, always not signicant
1 - No impact
- Probability (P)

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M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

3 - Highest probability that the parameter may fail

2 - Failure may occur sometimes
1 - Almost not fails
- Detectability (D)
3 - Failure may be probably not detected
2 - Occasionally not detected
1 - Detected every time
CMA and CPP were identied using available literature data,
knowledge related to drug product and manufacturing process
understanding, as well as PCA and PLS multivariate analysis on
historical data (experiments previously conducted in our laboratory).
The relationship between the selected variables (process
parameters and/or material attributes) and the CQA will be
described in the design space using one-factor RSM DoE.
2.2.2. Method of preparation of MDDC
MDDC were prepared by double emulsion solvent diffusion
technique using rotorstator homogenization (Ultra-Turrax1 T-25,

Dispersing element S25N-10G, IKA, Germany). Primary W/O

emulsion was prepared by addition of internal water phase
(CTS, water (W1) and ACT) to oil phase (EC, S80 and DHM). Multiple
W/O/W emulsion was prepared by addition of primary W/O
emulsion to outer water phase (water (W2), T80) and then, organic
solvents were removed. After complete evaporation of organic
solvents and solidication of MDDC, the resultant dispersion was
cooled to room temperature by magnetic stirring (200 rpm,
Variomag, Multipoint HP 15, Germany). Formulation and process
parameters used for MDDC preparation are presented in Table 1.
2.2.3. Morphological properties
Morphological properties were examined using transmission
emission microscopy (TEM) (JEM-1400, Jeol, Japan) attached to a
digital camera (Veleta TEM Camera, Olympus, Germany) and
controlled by the iTem software v.5.2.
2.2.4. Particle size analysis
MDDC were characterized in terms of particle size and particle
size distribution by laser diffractometry using Mastersizer 2000

Table 1
Formulation composition, manufacturing conditions and MDDC particle size (D50).
Sample Formulation parameters
Internal water
phase

Process parameters

Oil phase

Outer water
phase

Temperature for organic Stirrer type for

solvents removal (
C)
organic solvent
removal

Particles or
aggregates

Particle size 
SD
(D50, mm)

Aggregates
Particles
Particles
Aggregates
Aggregates
Particles
Particles
Aggregates
Aggregates
Particles
Aggregates
Aggregates
Particles
Particles

/
8.631  1.0646
103.179  5.6604
/
/
130.153  7.1242
134.179  7.4531
/
/
17.133  0.8433
/
/
59.289  0.8889
30.928  2.1147

Particles

48.114  0.2011

Particles

42.388  2.1082

Particles

22.189  0.4252

particles

16.402  1.0353

Particles

5.198  0.1847

Particles

10.414  0.2791

Particles

11.641  0.3084

Particles

34.468  3.2502

Particles

27.482  0.0557

Particles

26.889  0.3016

Particles

27.004  0.5189

Particles

21.356  0.0366

Particles

19.551  0.1635

Particles

22.998  0.2638

CTS
08%
(ml)

W1
(ml)

ACT
(ml)

S80 (g)

EC (g)

DHM
(ml)

W2 T80
(ml) (g)

2
3
4
5
6
7
8
9
10
11
12
13
14

1
1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.25

7
7
3.5
3.5
3.5
3.5
3.5
3.5
3.5
3.5
3.5
3.5
3.5
1.75

2
2
1
1
1
1
1
1
1
1
1
1
1
0.5

0.125
0.125
0.0634
0.0634
0.127
0.127
0.127
0.128
0.128
0.128
0.1291
0.1291
0.1258
0.0618

0.25
0.25
0.1261
0.1261
0.1283
0.1283
0.1283
0.125
0.125
0.125
0.1281
0.1281
0.1295
0.0679

25
25
12.5
12.5
12.5
12.5
12.5
12.5
12.5
12.5
12.5
12.5
12.5
6.25

60
60
30
30
30
30
30
30
30
30
30
30
60
30

0.36
0.36
0.1837
0.1837
0.1818
0.1818
0.1818
0.3665
0.3665
0.3665
0.3684
0.3597
0.37
0.1895

6500
6500
6500
6500
6500
6500
6500
6500
6500
6500
24000
24000
24000
24000

25
25
25
25
25
40
25
25
40
25
25
25
40
40

15

0.25

1.75

0.5

0.0678

0.0672

6.25

30

0.18

24000

40

16

0.25

1.75

0.5

0.063

0.0617

6.25

30

0.1866

24000

40

17

0.25

1.75

0.5

0.0625

0.0689

6.25

30

0.1929

24000

40

18

0.25

1.75

0.5

0.0693

0.0625

6.25

30

0.1796

24000

40

19

0.125

1.75

0.25

0.09375 0.03125

6.25

30

0.45

24000

40

20

0.125

1.75

0.75

0.03125 0.03125

6.25

30

0.45

24000

40

21

0.375

1.75

0.75

0.09375 0.03125

6.25

30

0.15

24000

40

22

0.125

1.75

0.25

0.09375 0.09375

6.25

30

0.15

24000

40

23

0.375

1.75

0.25

0.03125 0.09375

6.25

30

0.45

24000

40

24

0.125

1.75

0.75

0.03125 0.09375

6.25

30

0.15

24000

40

25

0.375

1.75

0.75

0.09375 0.09375

6.25

30

0.45

24000

40

26

0.25

1.75

0.5

0.0625

0.0625

6.25

30

0.3

24000

40

27

0.25

1.75

0.5

0.0625

0.0625

6.25

30

0.3

24000

40

28

0.25

1.75

0.5

0.0625

0.0625

6.25

30

0.3

24000

40

Results

Rotation
speed
(rpm)

Magnetic
Rotovapor
Magnetic
Rotovapor
Rotovapor
Magnetic
Magnetic
Magnetic
Magnetic
Rotovapor
Rotovapor
Magnetic
Magnetic
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer
Rotorstator
homogenizer

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

61

Table 2
Coded and actual values of independent experimental variables.
Factor

Level
Low

A: rotation speed (rpm)

B: stirrer type used for organic solvent removal

Actual values

Coded values

Actual values

1
1

6000
Magnetic stirrer

+1
+1

24000
Rotorstator homogenizer

equipped with Hydro 2000S, Malvern Instruments Ltd., UK, for wet
dispersions. An aliquot of prepared MDDC dispersion was
transferred to the optical measurement cell containing the blank
dispersing medium, distilled water to reach the obscuration of 10
12%. Measurements were performed while the sample was in the
cell under stirring (2520 rpm/stirrer rate). At least ve measurements for each sample were performed. Particle size distribution
was expressed in term of SPAN factor (Simonoska Crcarevska et al.,
2008).
2.2.5. PCA and PLS multivariate regression analysis
PCA and PLS multivariate analysis were applied on historical
data for determination and identication of formulation and
process variables inuencing MDDC particle size (Table 1). This
historical data are actually experimental data obtained from MDDC
formulations of previously performed experiments in our laboratory. Some of the experiments resulted with particles formation
while in others aggregates were formed.
PCA exploratory analysis (SIMCA 13, Umetrics AB, Sweden) was
applied for identication of critical parameters inuencing MDDC
particle formation.
PLS multivariate regression method (SIMCA 13, Umetrics AB,
Sweden) was used for identication of statistically signicant
parameters inuencing MDDC particle size (D50) and denition of
design space for subsequent optimization studies.
For multivariate analysis, ratio of the used materials to outer
water phase was used and hence each variable variance was
expressed in scale 1/SD (UV scaled). The number of components
was determined by cross validation.
2.2.6. Experimental design studies and process optimization
In order to nd optimal process conditions for preparation of
the MDDC with desired particle size and distribution, CPP
identied with QRM, were varied in the relevant design space.
Optimization of process condition was performed by one-factor
RSM design, with one numerical (continuous) and one nominal
(categorical) independent factor, using Design-Expert1V8 trial
(Stat-Ease, Inc., Minneapolis, USA) software. Rotation speed (rpm)
Table 3
Experimental design points for studied independent experimental variables in
coded values.
Std

Run Rotation speed

(rpm)

Stirrer type used for organic solvent removal

14
13
8
7
9
11
6
12
3
4
10
5
1
2

1
2
3
4
5
6
7
8
9
10
11
12
13
14

1
1
1
-1
1
1
1
1
1
1
1
1
1
1

0.000
1.000
-1.000
0.000
1.000
0.500
1.000
1.000
0.500
0.500
0.500
1.000
1.000
1.000

High

Coded values

was used as independent continuous variable and was varied on 5

levels in a range of 650024000 rpm. Stirrer type used for organic
solvent removal was set as categorical independent variable and
was varied on 2 levels (magnetic stirrer and rotorstator
homogenizer). The totals of 14 experiments were carried out.
Coded and actual level values of varied variables are presented
in Table 2, while designed experiments are presented in Table 3.
Experiments were carried out at random.
Composition of formulation used for optimization of process
parameters is presented in Table 4.
3. Results and discussion
3.1. QbD approach for development of formulation of microsponges
MDDC were prepared by double emulsion solvent diffusion
technique using rotorstator homogenization technique. TEM
micrographs indicated that prepared MDDC were characterized
with spherical shape and porous structure with open pores
distributed from the core to the surface of MDDC particle (Fig. 1).
The aim of this work was using QbD approach to develop MDDC
with potential to be incorporated into semisolid dosage form, gel
for topical application. Having in mind that the target (nal)
product would be gel with MDDC for topical application
pharmaceutical development should begin by dening its QTPP.
In an hypothetical situation of generic drug product development,
QTPP could be established based on physico-chemical characterization of the reference listed drug product, its packaging insert and
labeling (Chang et al., 2013). Dened and justied QTPP (Table 5)
will be used as guide during pharmaceutical development of
MDDC for incorporation into gel for topical application that should
be pharmaceutically equivalent with reference listed drug product.
Quality attributes for target drug product are presented in
Table 6, and CQA are appropriately marked and justied.
Usually QRM analysis is done on the basis of available literature
and/or historical data. Considering that literature data in form of
original scientic research publications related to microsponges
are limited (Abdelmalak and El-Menshawe, 2012; Arya and Pathak,
2014; Bamane et al., 2014; Comoglu et al., 2003; Dsouza and More,
2008; Deshmukh and Poddar, 2012; Jain and Singh, 2010;
Jelvehgari et al., 2006; Karthika et al., 2013; Mahajan et al.,
2011; Maiti et al., 2011; Manjula, 2011; Nokhodchi et al., 2007; Orlu
et al., 2006; Ravi and Senthil, 2013; Swetha et al., 2011; Veena and

Table 4
Composition of formulation used for optimization of process parameters.
Internal water phase

CTS 0.8% (ml)

W1 (ml)
ACT (ml)

0.375
1.875
3.25

Oil phase

S80 (g)
EC (g)
DHM (ml)

0.03125
0.09375
6.25

Outer water phase

W2 (ml)
T80 (g)

30
0.45

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M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

Fig. 1. TEM micrographs of prepared MDDC (sample 23).

Manjula, 2013; Vyas et al., 2010), our approach to QRM with FMECA
approach combines literature reference data and results derived
from multivariate statistical analysis (PCA and PLS) performed on
experimental data from previously conducted experiments (in our
laboratory) associated with the MDDC prepared by double
emulsion solvent diffusion technique using rotorstator homogenization.
PCA-X&Y analysis decompose X variables matrix in number of
principal components (PC). PCA analysis applied on experimental
data presented in Table 1 resulted with one PC that maximizes the
explained variance in the data set (Fig. 2). The PCA model with one
PC explains (R2X) 63.3% of the variation in the data set (X). The
parameter Q2 obtained with cross-validation of the model has a
value of 53.9% and it refers to the percentage of variation of all X
variables that can be estimated with the model. Values of Q2 > 50%
indicate good predictability. So, model is acceptable and it was
used for identication of formulation and process parameters
inuencing MDDC particles formation.

Score scatter plot might indicate the presence of patterns in the

results, e.g., clusters, trends or results that do not t the model
(outliers). Score scatter plot of performed PCA XY analyses (Fig. 3)
clearly points to existence of 2 clusters: group 1 and group 2. Group
1 is closely related with Y variable particles, while group 2 was
correlated with Y variable aggregates. In depth analysis pointed
that group 2 was consisted of samples 1, 4, 5, 8, 9, 11, 13 which
clearly resulted with formation of aggregates, but also samples 2, 3,
6, 7 and 10 were included in this cluster indicating that, although
this samples were characterized with formation of particles,
somehow they are more related to this group than to samples in
group 1 (all other samples). The variables which in turn affect the
results can be displayed in loading plot (Fig. 4), thus each variable
gets a loading value in each PC. Loading plot was used for
interpretation of the clusters seen on score scatter plot as it
resembles the covariance between X variables and t score vectors.
This plot pointed that group 1 is closely related to rotation speed
(rpm), temperature and stirrer type used for organic solvent
removal (rotorstator homogenizer), while group 2 is correlated

Table 5
QTPP of target drug product, gel with MDDC, for topical application.
QTPP elements

Target

Justication

Dosage form

Hydrogel

Route of
administration
Dosage strength

Topical
% of drug substance (m/m)

Dosage form design

Pharmacokinetics
Stability

Microsponges as drug delivery carriers incorporated/ dispersed into hydrogel

Not applicable, because the drug product is intended for local treatment
Shelf-life not less than 24 months at room temperature

Pharmaceutical equivalence requirement:

same dosage form
Pharmaceutical equivalence requirement:
same route of administration
Pharmaceutical equivalence requirement:
same dosage strength
Match reference listed drug product
Match reference listed drug product
Equivalent or longer shelf-life compared to
reference listed drug product
Pharmaceutical equivalence requirement: fulll
the same quality standards as reference listed
drug product

Drug product quality Physical attributes

attributes
Identication
Assay
Uniformity of content
Degradation products
Residual solvents
Dissolution
Microbiological quality
pH
Rheological behavior
Container closure
Suitable container closure system that will support estimated shelf-life and drug product
system
integrity during the transport, Identical primary packaging as reference listed drug product.
Alternative methods No
of administration

Tube, similar with reference listed drug product,

acceptable for the patient
None are listed on reference drug product
labeling

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

63

Table 6
Quality attributes for target drug product, gel with MDDC.
Drug product quality attributes

Target

Is it a Justication
CQA?

Physical
attributes

Color and shape acceptable

for the patient
No unpleasant odor

No

Appearance
Odor
Microsponges particle size
and particle size
distribution
Rheological behavior
pH

Similar to reference listed

drug

Color and shape are not directly related to the safety and efcacy. Required for
patient acceptability
Odor is not directly related to the safety and efcacy. Required for patient
acceptability and unpleasant odor might be a reason for patient complaints
It might inuence encapsulation efcacy and drug content, as well as drug release. It
is critical for drug product efcacy and safety

No
Yes

Conform to Ph. Eur. 7.02.2.10 Yes

a

Required to demonstrate Q3 (same components in same concentrations with same

microstructure)
Might inuence drug product stability and hence its efcacy and safety. This CQA
can be effectively controlled by quality management system
Identication is critical for safety and efcacy, but this CQA can be efciently
controlled by quality management system and will be followed during dissolution
studies. Formulation and process variables cant inuence identity
Assay variability will have inuence upon safety and efcacy
Variability in tube homogeneity and uniformity of content will affect safety and
efcacy

Conform to Ph. Eur. 7.02.2.3

Yes

Identication

Positive for drug substance

Yesa

Assay
Tube homogeneity and
Tube
(Content)
uniformity
uniformity
Residual solvents

90110% of label claim

Conform to USP <3>

Yes
Yes

Conform to ICH Q3C(R5)

Yes

Degradation products

Conform to ICH Q3B (R2)

Yesa

In vitro dissolution prole

Match reference listed drug

product

Yes

Microbiological quality

Conform to Ph. Eur. 7.05.1.4

Yesa

Preservative content
Efcacy of antimicrobial preservation

80110 % from label claim

Conform to Ph. Eur. 7.05.1.3

No
Yesa

Container closure system

Identical primary packaging

to reference listed drug
product
No failure

No

Microsponges manufacturing procedure include use of organic solvents: acetone

(class 3) and dichlormetane (class 2). Their residual quantity is critical for safety
Degradation products limit is critical for safety. Limit of individual unknown
degradation products must comply with ICH Q3B (R2). Limit for total degradation
products is based on reference listed drug product analysis near the expiry date. If
the active pharmaceutical ingredient do not contain functional groups with
apparent sensitivity to oxidation, hydrolysis, light, heat, acid and base, and stability
is conrmed with forced degradation studies.The possibility that formulation and
process parameters will affect degradation products is very low if during
preformulation compatibility studies performed as part of development no
chemical interaction were observed
Having in mind that in vitro drug release is surrogate for in vivo performance the
target is to ensure similar dissolution prole compared to reference listed drug
product (f2 > 50)
Non compliance of microbiological quality will have impact upon safety. The
formulation and process variables are unlikely to affect it as long as raw materials
comply to microbiological quality requirements
Required to ensure antimicrobial effectiveness
Inefciency of the preservative at declared storage conditions, might result in
microbiological contamination and affect safety, but is unlikely that formulation and
process variables will affect it
Match reference listed drug product. Required for patient acceptability

No

Required for stability, efcacy and safety

Packaging integrity
a

It is unlikely that formulation and process variables will have impact upon this CQA, but it is still part of QTPP.

with the rest of the X variables, except T80 which has the least
inuence on clusters formation.
Similar conclusion can be drawn from the group 2 to group
1 comparison plot (Fig. 5). Variables are represented as bars that
are indicating which variables deviate between groups and the
sign of the bar (minus or plus) point to the direction of deviation.

Contribution of each variable in means of R2 and Q2 in PC score

is presented on Fig 6.

Fig. 2. PCA Summary of t.

Fig. 3. PCA Score scatter plot.

64

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

Fig. 4. PCA loading scatter plot.

Stirrer type used for organic solvent removal (magnetic stirrer

and rotovapor) and S80 had a moderate values for R2, while all
other X-variables except T80 are characterized with high R2 values
pointing to their large loading values for the selected component.
At the same time high Q2 values for the variables are in the favor of
reliability of these R2 and loading values. Low R2 and Q2 for
T80 point to fact that its inuence on micropsonges formation
could not be well explained by this PC.
The PCA analysis can help in revealing similarities and
differences between the samples of interest and analysis, as well
as in discovering the origin of these differences, i.e., to point to the
variables in which samples differ. However, it should be noted that
PCA analysis do not possess power to detect whether the observed
differences will affect the manufacturing process and product
quality (Ferreira and Tobyn, 2014), therefore subsequent analyses
are required, as well as sufcient process and product knowledge
and understanding, and in some cases it can be done only with
additional experiments.

After identication of the variables associated with two groups

of results (particles and aggregates) the next step was to determine
how X-variables inuence the size (D50) of MDDC particles. For
this reason PLS analysis was applied on the experimental results
associated with particles formation (total of 21 samples, Table 5)
PLS analysis resulted with model with two components, with
the rst component explaining 56.1% (R2Y) of variations with
predictive ability (Q2) of 40.6%, while the inclusion of the second
component explains total of 91.9% (R2Y) of variations with the total
predictive ability (Q2) of 84.3% (Fig. 7).
Considering that goodness of t (R2Y) and goodness of
prediction (Q2) values should be as close to 100% as possible,
and both values should be in a reasonable agreement, preferably
not separated by more than 2030% (Malzert-Fron et al., 2010),
the obtained values for R2Y and Q2 indicated that model is
interpretable.
Score scatter plot (t1 vs t2) is presented on Fig. 8. Actually these
scores represent new variables summarizing X-variables. These
scores are completely independent of each other and number of
score vectors depends from components in PLS model. Score
marked as t1 (rst component) explains the largest variation in Xvariables, than t2 score (second component) follows. Similar to
PCA, score scatter plot shows presence of patterns, groups, clusters,
similarities or outliers. Actually from Fig. 8 it can be seen that
samples 2 and 10 are outside of the window of modeled X space.
These samples are outliers and in order to get more plausible
model they can be excluded from modeling. Therefore, remodeling
is conducted on remaining 19 samples. PLS analysis resulted in a
model with a component pointing that 68.1% (R2X) of X variables
explains 91.6% (R2Y) of variations in the data set. PLS model was
characterized with predictability (Q2) of 88.9% (Fig. 9). Score
scatter plot (Fig. 10) points that all 19 samples are in the window of
modeled X-space.
Loading scatter plot from PLS analysis (Fig. 11) shows the
relationship between X and Y variables. X variables with large
values of w*c (positive or negative) are highly correlated with Y and
hence they are situated far away from origin on the plot. Variables
that are opposing each other are negatively correlated, and those
that are close to each other on this graph are positively correlated.

Fig. 5. PCA Score contribution plot, Group 2 to Group 1 comparison.

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

65

Fig. 6. PCA-X&Y component contribution plot.

Fig. 11 points that MDDC particle size (D50) is inversely

correlated with T80 amount, temperature and stirrer type (rotor
stator homogenizer) used for organic solvent removal, as well as
rotation speed (rpm), while all other X-variables (amount of W1,
ACT, CTS, EC, DHM and S80) are positively correlated, except stirrer
type (rotovapor) used for organic solvent removal.
From graphic and tabular representation of the coefcients
(Fig. 12,Table 7) model equation describing inuence of X variables
on MDDC particle size (D50) can be established. Coefcients
magnitude and sign (positive or negative) point to the degree and
manner of inuence of each X-variable upon MDDC particle size
(D50).
Considering coefcients values it can be noticed that positively
correlated W1 and DCM amount as well as inversely correlated
rotation speed (rpm) (coefcients 0.124592) have highest

inuence on MDDC particle size (D50), than stirrer type used

for organic solvent removal, rotorstator homogenizer (inversely
correlated) and magnetic stirrer (positively correlated) with
coefcients 0.119495 follow etc. Performed ANOVA in the cross
validated residuals of a Y variable returned p value indicative of the
statistical signicance of the investigated model (p < 0.05). The VIP
(Variable Importance for the Projection) values summarizing the
importance of the variables both to explain X and to correlate to Y,
are presented on Fig 13.
VIP values are calculated for each X-variable summing the
squares of the PLS loading weights, wak, weighted by the amount of
sum of squares explained in each model component. VIP values
greater than 1 indicate signicant X-variables (amount of W1,
DHM, EC and rotation speed as well as stirrer type used for organic
solvent removal (rotorstator homogenizer and magnetic stirrer)).

Fig. 7. PLS analysis summary of t (21 samples).

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M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

Fig. 8. PLS analysis score scatter plot (21 samples).

VIP values below 0.5 indicate insignicant variables X (stirrer

type used for organic solvent removal (rotovapor)), while the
interval between 1 and 0.5 (the amount of CTS, temperature for
organic solvent removal, amount of ACT, S80 and T80) is called
gray area and in this case the level of signicance has to be
determined by additional experiments.
Literature data are in favor of obtained results regarding
inuence of formulation and process variables on MDDC particle
size. Results from performed PLS analysis indicated that particle
size is increasing with increase of water amount in primary
emulsion, amount of EC (polymer), DHM (organic phase), S80
(lipophilic surfactant) and temperature. In general it can be
anticipated that microsponges particle size is dependent from
emulsion droplet size related to viscosity of phases of multiple
emulsion. Abdelmalak & co. related increased particle size of
microsponges with increased polymer concentration as a result of

increment of polymer amount per formed particle and increased

viscosity of polymer solution hindering breaking of emulsion into
smaller droplets hence resulting in microsponges with larger
particle size (Abdelmalak and El-Menshawe, 2012). Studies of
Nokhodchi et al. resulted in a similar conclusion about the
inuence of organic phase viscosity (polymer is dissolved in
organic phase) on the microsponges particle size (Nokhodchi et al.,
2007). Similar results about inuence of polymer amount on
microsponges particle size can be found in the literature (Arya and
Pathak, 2014; Comoglu et al., 2003; Ravi and Senthil, 2013). This
can be related with the work of Badawi and El-Khordagui which
conrm that viscosity of prepared emulsions is increased with
increment of lipophilic surfactant, organic phase and polymer
amount (Badawi and El-Khordagui, 2014). In addition, the similar
are ndings of Pillay et al. (2013) and Sy et al. (2009) which shows
that viscosity of W/O emulsions increased with polymer

Fig. 9. PLS analysis summary of t (19 samples)

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

67

Fig. 10. PLS analysis score scatter plot (19 samples).

concentration increment, concentration of lipophilic surfactant

(Jiao and Burgess, 2003) and volume fraction of water (Sy et al.,
2009). Jelvehgari & co. related microsponges particle size to the
differences in viscosity between internal and outer phase of
emulsion system thus implying that greater difference resulted
with larger particle size. Due to the higher viscosity of internal
phase the globules of the formed emulsion can hardly be divided
into smaller particles and bigger droplets were found and mean
particle sizes increased (Jelvehgari et al., 2006). Tiranksiz & co.
indicated that globule size in multiple emulsions depend from
concentration of lipophilic surfactant, hence its higher concentration resulted with formation of bigger droplets. This is explained
with probability the excess of lipophilic surfactant to be found in
the organic phase between the internal and external aqueous
phase, and this can lead to the so-called phenomenon of globule
swelling i.e., to increment of globule diameter (Tirnaksiz and
Kalsin, 2005).

PLS analysis points to inverse inuence of T80 to MDDC particle

size (D50). This is probably related to decrease of size of multiple
emulsion droplets by increasing the hydrophilic surfactant
amount. Similar are the observations of Abdelmalak and ElMenshawe (2012) which suggest that most likely a layer is formed
by surfactant molecules i.e., they are adsorbed on the primary
emulsion droplet thus resulting with decrease of surface tension
between W/O and W phases during formation of multiple W/O/W
emulsion, hence as lower is the surface tension the smaller are the
droplets of W/O/W emulsion. On the other hand smaller surfactant
amount would result with incomplete coverage of interphase more
likely leading to droplet coalescence and occurrence (growth) of
droplets/particles with larger diameter (Hecht et al., 2011; Ngan
et al., 2014; Qian and McClements, 2011).
Inuence of temperature on MDDC particle size can be related
to decrease of viscosity by temperature increment and/or
enhanced diffusion of surfactant to the new interface, thus

Fig. 11. PLS loading scatter plot.

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M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

Fig. 12. Coefcients plot for X-variables inuencing MDDC article size (D50).

resulting with temperature direct inuence on the coalescence

(Lindenstruth and Mller, 2004).
Inverse correlation of MDDC particle size with rotation speed
(rpm) can be also supported by literature data. Namely, the higher
rotation speed is the lower the particle size is. Jelvehgari et al.
observed that increase of the mixing speed from 1000 to 4000 rpm
resulted with smaller particle size (603.9  58.4 to 310.3  34.3 mm
appropriately) (Jelvehgari et al., 2006). Similar are the results of
Nokhodchi et al., that explain the increment of MDDC particle size
by globules coalescence tendency and aggregation at lower mixing
(rotation) speed. On the other hand higher mixing speed would
result with faster breaking of formed droplets and lower
probability for coalescence into larger droplets (Nokhodchi
et al., 2007). Similar are the ndings of Jain and Singh as well as
those of Ravi and Senthil (Jain and Singh, 2010; Ravi and Senthil,
2013).
QRM analysis using FMECA approach is based on the results of
multivariate statistical analysis on historical data, available
literature data for microsponges and formulation and process
parameters inuence on particle size, as well as knowledge and
understanding of pharmaceutical-technological process used for
microsponges manufacturing (Table 8).

Based on QRM analysis all process and formulation variables

with RPN above 10 (Zhang et al., 2013) were identied as potential
variables with critical inuence on efcacy of topical dosage form,
gel with MDDC. In addition, a total of 10 variables were identied
as factors that need to be further examined in terms of their
inuence on the properties of the topical dosage form. The
examination should encompass a detailed (qualitative and
quantitative) characterization of the inuence of these factors
on the properties of the dosage form.

3.1.1. Experimental design and process optimization

Characterization of inuence of variables identied by QRM
analysis upon properties of drug dosage form should be conducted
using experimental design studies. Having in mind relatively high
number of identied variables (10) that might have critical
inuence upon drug product quality, use of experimental design
studies that will include all variables at the same time (formulation
and process) will result in a considerable number of experiments.
Having in mind the previous, it is more appropriate to apply
rational approach for the further experiments, and they should be
conducted in two phases, i.e., formulation and process variables

Table 7
Regression coefcients for scaled and centered X-variables related to MDDC particle size (D50).
Variables

M3.CoeffCS[1](D50)

1.89456  M3.CoeffCS[1](D50) cvSE

Constant
W1
ACT
DHM
CTS
S80
EC
T80
Rotation speed (rpm)
Temperature used for organic solvent removal (
E)
Stirrer type used for organic solvent removal

1.08273
0.124592
0.0910824
0.124592
0.100199
0.0749492
0.109179
0.0533568
0.124592
0.0940158
0.119495
0.119495
0

0
0.0323482
0.0372271
0.0323482
0.0247113
0.0602417
0.0367501
0.0418254
0.0323482
0.0829384
0.0313605
0
0.0313605

Rotorstator homogenizer
Magnetic stirrer
Rotovapor

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

69

Fig. 13. VIP values for X variables related to MDDC particle size (D50).

should be examined separately. When it comes to QRM identied

formulation variables (amount of ethylcellulose, chitosan, acetone,
dichloromethane, span 80, tween 80 and water ratio in primary/
multiple emulsions) at rst screening experimental design should
be used, and subsequently optimization design studies should be
carried out. QRM identied only 2 process variables, one numerical
(rotational speed (rpm)) and one categorical (stirrer type used for
organic solvent removal (rotorstator homogenizer and magnetic
stirrer)), so optimization design studies using one-factor RSM
design were carried out. They enabled acquiring of detailed
knowledge and understanding of complex inuence of examined
process variables upon microsponges particle size (D10, D50, D90)

and distribution (Span factor). Results from conducted experiments are presented in Table 9.
Experiments resulted with micropsonges particles with D10 in
range from 1.721  0.0487 to 14.18  0.0857 mm. Inuence of
examined variables on D10 can be described as inverse function
of square root of D10 by 2FI model. One way ANOVA indicated that
variables A, B and AB are signicant model terms. Equation
describing inuence of examined variables on D10 (Eq. (1)) in
terms of coded factors is:
1
0:49 0:20  A  0:036  B  0:056  A  B
SqrtD50

(1)

Table 8
QRM of typical manufacturing process using FMECA approach
Risk area

Risk analysis

Failure mode

Failure effect

Change in particle size

or shape, as well as
surface energy
Inadequate product
homogeneity
Inadequate size of
emulsion droplets
Inadequate size of
MDDC particles
Inadequate porosity of
MDDC
Collapsibility of MDDC
structure
Physical instability,
phase separation,
coalescence
Chemical insatiability
of API and excipients
Inadequate rheological
properties
Inadequate drug
release
Microbiological
contamination

Change in dissolution and impact on clinical efcacy

S P D RPN
CMA
Active pharmaceutical substance

2 1 3

3
4
3
3
4
3
4
2
1

2
2
2
2
2
2
2
2
3

12
16
12
12
16
12
16
8
3

4 3 2

24

4 2 2

16

 Magnetic stirrer

4 2 2

16

 Rotovapor

2 3 1

Temperature for organic solvent removal

2 2 2

Excipients
CTS
EC
S80
T80
W1
ACT
DHM
Gelling agent
Preservative
CPP
Rotation speed (rpm)
Stirrer type used for organic solvent removal
 Rotorstator homogenizer

2
2
2
2
2
2
2
2
1

Impossibility to form primary emulsion MDDC would

not be formed
Impossibility to form multiple emulsion MDDC would
not be formed
Separation of water and organic phase during emulsion
production MDDC particles would not be formed or
aggregates will be formed
Inadequate MDDC particles size, low porosity or
absence of porosity
Lower or higher viscosity of nal drug product (gel) and
change in consistency
All of the above will inuence:
MDDC particle formation
Drug incorporation into MDDC
Drug dissolution prole and drug release kinetics
Efcacy of therapeutic treatment

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M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

Table 9
Coded values of examined varaiables and obtained responses from desgined experiments.

Std

Run

Factor 1
A: rotation speed (rpm)

Factor 2
B: stirrer type used for organic solvent removal

Response 1
D10  SD

10
12
3
14
9
5
6
4
7
11
2
1
8
13

1
2
3
4
5
6
7
8
9
10
11
12
13
14

0.5
1
0.5
0
1
1
1
0.5
0
0.5
1
1
1
1

1
1
1
1
1
1
1
1
1
1
1
1
1
1

6.286
2.378
6.994
4.506
12.510
1.834
1.821
1.721
3.574
5.345
14.032
14.180
8.528
2.810

From the Eq. (1) it can be seen that 1.0/Sqrt(D10) is positively

correlated with variable A (rotation speed (rpm)), and inversely
correlated with variable B (stirrer type used for organic solvent
removal) and their interaction AB.
D50 of microsponges particles is in the range from
3.465  0.0466 to 34.6 1 0.5623 mm. Inuence of examined
variables can be described as inverse function of square root of
D50, by quadratic model. One way ANOVA for this model indicated
A, B and AB as signicant model terms. Equation describing
inuence of examined variables on D50 (Eq. (2)) in terms of coded
factors is:
















0.1133
0.1937
0.0145
0.0340
0.0606
0.0156
0.0091
0.0487
0.0806
0.02180
0.1708
0.0857
0.0509
0.3164

Response 2
D50  SD
26.649
10.595
20.591
14.644
31.023
3.531
3.465
5.681
11.510
33.865
33.681
34.610
19.665
11.432
















0.6640
0.4387
0.3267
0.1075
0.2457
0.0830
0.0466
0.9934
0.1427
0.0830
1.0345
0.5623
0.0682
0.4587

Response 3
D90  SD

Response 4
Span  SD

178.331  7.0423
53.134  4.1807
230.293  3.2591
132.190  7.0541
163.589  14.7843
283.06735.7549
262.312  14.6760
536.571  15.0734
243.538  25.8347
96.270  0.6358
520.818  64.746
561.538  38.4715
58.224  3.1573
61.939  2.2940

6.456  0.1177
4.791  0.3240
10.844  0.0502
8.719  0.5861
4.870  0.4365
79.656  8.1181
75.173  3.1699
94.141  4.1008
20.848  2.1137
2.685  0.0127
15.047  1.4422
15.815  0.9489
2.527  0.1535
5.172  0.0590

1
0:26 0:11  A  0:051  B  0:071  A  B
SqrtD50
0:045  A2

(2)

Eq. (2) points that 1.0/Sqrt(D50) is positively related to variable A

(rotation speed (rpm)) and its quadartic function (A2), and
inversely related to variable B (stirrer type used for organic
solvent removal) and their interaction AB.
D90 of microsponges particles is in the range from
53.134  4.1807 to 561.538  38.4715 mm. Inuence of examined
variables can be described as logarithmic function of D50, by linear

Fig. 14. Plots ilustrating the dependance of the responces from selected model terms.

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

71

Table 10
ANOVA statistical analysis for responses models.
Response

1.0/Sqrt(D10)

1.0/Sqrt(D50)

Log10(D90)

Log10(Span)

Suggested model
Sum of square (SS)
Degrees of freedom
Mean squares (MS)
F-value
Prob > F
SD
Mean
CV%
PRESS
R2
Adj-R2
Pred-R2
Adequate precision

2FI
0.4
3
0.13
45.2
<0.0001
0.054
0.49
11.06
0.053
0.9312
0.9106
0.8761
17.491

Quadratic
0.2
4
0.05
49.56
<0.0001
0.032
0.28
11.19
0.019
0.9566
0.9373
0.9085
19.315

Linear
1.23
2
0.62
18.34
0.0003
0.18
2.26
8.11
0.59
0.7693
0.7273
0.6331
9.302

2FI
3.20
3
1.07
25.7
<0.0001
0.2
1.08
18.83
0.7
0.8852
0.8507
0.8056
11.684

Table 11
Optimized samples with estimated responses and cross validation of the model.
Run

A: rotation speed (rpm)

B: stirrer type used for organic solvent removal

D10

D50

D90

Span

Desirability

24000
1-Bias(%)

Rotorstator homogenizer

2.82723/2.968
4.98

11.7848/11.632
1.3

73.6166/62.967
14.47

4.91402/5.158
4.96

0.626

model. One way ANOVA for this model indicated variable B as

signicant model term. Equation describing inuence of examined
variables on D90 (Eq. (3)) in terms of coded factors is:
log10 D90 2:26  0:11  A  0:28  B

(3)

Eq. (3) points that log10(D90) is inversely related to variable A

(rotation speed (rpm)) and variable B (stirrer type used for organic
solvent removal).
Response particle size distribution of micropsonges (expressed
as Span factor) is in range from 2.527  0.1535 to 94.141  4.1008.
Inuence of examined variables on particle size distribution can be
described as Span logarithmic function by 2FI model. One way
ANOVA for this model indicated variables A, B and AB as signicant
model terms. Equation describing inuence of examined variables
on Span (Eq. (4)) in terms of coded factors is:
log10 Span 1:08 0:22  A  0:42  B  0:20  A  B

(4)

Eq. (4) points that log10(Span) is positively related to variable A

(rotation speed (rpm)) and inversely related to variable B (stirrer
type used for organic solvent removal) and their interaction AB.
Plots illustrating the dependance of the responses from selected
model terms are presented on Fig. 14.
ANOVA statistics for the tted model for all followed responses
is presented in Table 10. The predicted R2 (Pred-R2) and the Adj-R2
should be within 0.20 of each other. Otherwise there may be a
problem with either the data or the model. Adj-R2 and Pred-R2
values were in reasonable agreement, signifying satisfactory
model t. R squared values are higher than 0.76 and adequate
precison values are greater than 4 indicating that all suggested
models can be used to navigate the design space (Makraduli et al.,
2013; Simonoska-Crcarevska et al., 2013).
As targets for optimization the requirements for microsponges
minimal particle size (D10, D50, D90) and minimal distribution
(Span factor) were set. Optimized samples and conrmation report
from the cross-validation of the model and the per cent of relative
error of the predicted and experimental values are presented in
Table 11.
4. Conclusion
QbD tool was used for the development of MDDC with potential
to be incorporated into gel as semi-solid dosage form for topical

application. QTPP was dened and used as a guide for pharmaceutical development. CQA were identied. QRM as a systematic
process of assessment of risks that might inuence pharmaceutical
product quality throughout its lifecycle was applied. Using FMECA
approach CMA and CPP were identied and CQA were prioritized.
CMA (amount of CTS, EC, S80, T80, ACT, DHM, W1) and CPP
(rotation speed (rpm) and stirrer type used for organic solvent
removal (rotorstator homogenizer and magnetic stirrer)) were
identied. Identication was based on available literature data for
microsponges and formulation and process parameters inuencing particle size, as well as knowledge and understanding of
pharmaceutical-technological process used for MDDC manufacturing and per se on the results of multivariate statistical analysis on
historical data for MDDC formulations prepared in our laboratory.
PCA and PLS analysis were performed using statistical software
SIMCA 13 (Umetrics AB, Sweden).
Identied CPP were subsequently optimized by one-factor RSM
design using Design-Expert1 V8 trial (Stat-Ease, Inc., Minneapolis,
USA). Obtained results from statistically designed experiments
enabled establishment of mathematical models and equations that
were used for detailed characterization of inuence of identied
CPP upon MDDC particle size (D10, D50, D90) and particle size
distribution (Span).
Having in mind the number of identied CMA (7) further
research should be conducted using sequential experimental
strategy, where screening DoE for identication and rationalization of signicant formulation variables, will be followed by
characterization and optimization DoE (preferentially RSM).
Overall results will determine the control strategy for
manufacturing process and pharmaceutical product quality
assurance.
Declaration of interest
The authors report no declarations of interest.

Acknowledgments
The work was supported by Faculty of pharmacy, University of
Ss. Cyril & Methodius, Skopje Macedonia.

72

M. Simonoska Crcarevska et al. / International Journal of Pharmaceutics 489 (2015) 5872

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