Drug Delivery

ISSN: 1071-7544 (Print) 1521-0464 (Online) Journal homepage: https://www.tandfonline.com/loi/idrd20

Ketoprofen mesoporous silica nanoparticles

SBA-15 hard gelatin capsules: preparation and in
vitro/in vivo characterization

Ahmed A. Abd-Elrahman, Mohamed A. El Nabarawi, Doaa H. Hassan & Amal

A. Taha

To cite this article: Ahmed A. Abd-Elrahman, Mohamed A. El Nabarawi, Doaa H. Hassan

& Amal A. Taha (2016) Ketoprofen mesoporous silica nanoparticles SBA-15 hard gelatin
capsules: preparation and in�vitro/in�vivo characterization, Drug Delivery, 23:9, 3387-3398, DOI:
10.1080/10717544.2016.1186251

To link to this article: https://doi.org/10.1080/10717544.2016.1186251

Published online: 03 Jun 2016.

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ISSN: 1071-7544 (print), 1521-0464 (electronic)

Drug Deliv, 2016; 23(9): 3387–3398

! 2016 Informa UK Limited, trading as Taylor & Francis Group. DOI: 10.1080/10717544.2016.1186251

RESEARCH ARTICLE

Ketoprofen mesoporous silica nanoparticles SBA-15 hard gelatin

capsules: preparation and in vitro/in vivo characterization
Ahmed A. Abd-Elrahman1, Mohamed A. El Nabarawi1, Doaa H. Hassan2, and Amal A. Taha2
1
Department of Pharmaceutics and Industrial Pharmacy, College of Pharmacy, Cairo University, Cairo, Egypt and 2Department of Pharmaceutics
and Industrial Pharmacy, College of Pharmacy, Misr University for Science and Technology, Sixth of October City, Giza, Egypt

Abstract Keywords
SBA-15 is used to enhance the bioavailability of poorly soluble ketoprofen (KP) through In vitro in vivo study, immersion-rotavapor
stabilization of its amorphous state. Additionally, the current work provides a complete in vitro loading method, loading method meso-
and in vivo study on preformulated KP-SBA-15 sample and formulated KP-SBA-15 in hard porous silica nanoparticle, SBA-15
gelatin capsule. Loading of KP was done by a novel method called immersion-rotavapor
method. KP was quantified by extraction and thermal gravimetric analysis (TGA). History
Characterization of the loaded SBA-15 sample was done by high resolution transmission
electron microscopy (HRTEM), small angle X-ray diffraction (SAXRD), nitrogen adsorption/ Received 22 February 2016
desorption isotherms, differential scanning calorimetry (DSC), Fourier transform infrared (FT-IR) Revised 26 April 2016
spectroscopy and dissolution profiles. The loaded sample was formulated in hard gelatin Accepted 2 May 2016
capsule. The anti-inflammatory and analgesic studies were carried out on 24 adult male albino
rats. TGA and extraction results showed 54.4 wt% of drug incorporated. Characterization of
KP-SBA-15 sample confirmed the successful encapsulation of KP into the carrier pores in a
molecular amorphous state. Additionally, loading of KP did not affect the mesoporous internal
structure. During the first 5 min, the dissolution study showed very high release rates;
nearly 50% of KP was released. These results were reflected on the in vivo study resulting in
82% inhibition in edema after 1 h and maximum analgesia after 30 min from the administration
of the formulated sample. SBA-15 mesoporous silica nanoparticle proved to be a very
promising drug delivery carrier that can be used as a facile way to enhance the bioavailability of
poorly soluble drugs.

Introduction soluble drugs creates one of the most challenging tasks that
the scientists are currently facing (Salazar et al., 2014).
The majority of new innovative drug candidates suffer from
The applications of mesoporous silica nanoparticles
poor pharmacokinetics that reduces their bioavailability,
(MSNs) in the pharmaceutical field are increasing day by
especially in oral drug delivery that is still the most
day and receiving precise attention owing to their highly
common and preferable administration route. The problems
ordered structures, large pore sizes and high surface area.
with poor bioavailability are usually related to the poor
Thanks to its stable mesoporous structure and well-defined
solubility of the drug, and therefore one of the main
surface properties, mesoporous materials seem ideal for the
challenges in pharmaceutical industry is to find ways to
encapsulation of pharmaceutical drugs, proteins and other
enhance the solubility of the drugs without affecting its
biogenic molecules. Currently, employing MSNs for hosting
stability and pharmaceutical activity (Williams et al., 2013).
and further delivering of a variety of molecules of
The number of poorly soluble compounds among drugs under
pharmaceutical interest has appeared (Safari & Zarnegar,
development increased rapidly, straight after high-throughput
2014).
screening (HTS) was introduced in the drug discovery. HTS
MSNs show great promise for the future, as every
greatly strengthened the generation of leads, but the drawback
dissolution-enhancing formulation approach used nowadays
of this new method was that it also caused a shift toward leads
is known to depend on the active compound’s physicochem-
with higher molecular weights and lipophilic properties that
ical profile. The potential of MSNs to yield physically stable,
are typically related to low aqueous solubility (Carnero,
dissolution enhancing formulations, irrespective of a drug’s
2006). Therefore, enhancing the oral bioavailability of poorly
physicochemical profile, therefore seems very promising. In
addition, considering the relatively low cost of their synthesis
Address for correspondence: Mohamed A. El Nabarawi, Department of and the simplicity of the drug loading process, it is clear that
Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo
University, Cairo, Egypt. Tel: +201001424439. E-mail: mesoporous silica drug-carrier constitutes a potentially great
nabrwima@hotmail.com expansion of the formulation scientist’s toolbox to overcome
3388 A. A. Abd-Elrahman et al.
dissolution-limited bioavailability (Van Speybroeck et al.,
2009).
Vallet-Regi et al. (2001) reported the use of mesoporous
silica MCM-41 for the host of ibuprofen. This was the first
attempt to enhance the dissolution by encapsulation of
hydrophobic drug (ibuprofen) in MSN (MCM-41). Since
that time, these inorganic materials are being explored as
carriers for oral drug delivery. In particular, MSNs are widely
used as a delivery reagent because silica possesses favorable
chemical properties, thermal stability and biocompatibility.
Their unique properties render them suitable carriers for
active pharmaceutical ingredients (Van Speybroeck et al.,
2009): pore diameters that can be tuned between 2 and 30 nm,
high specific surface areas (up to 1500 m2/g) creating a high
potential for adsorption, large pore volumes (up to 1.5 cm3/g),
and a silanol containing surface that can be functionalized to
modify drug release. It is interesting to know that MSNs can
be used in both enhancing the drug release and in controlling
the release rate, depending on its type and morphology (Kwon
et al., 2013).
Several studies have shown that different biologically
active agents can be loaded and, subsequently, released from
MSNs (Fu et al., 2012; Lopes et al., 2013). The dissolution of
poorly soluble drugs from the MSNs particles is usually faster
than that of bulk APIs. The fast dissolution from the particles
is related to the confined space inside the pores that prevents
long range ordering, thus preventing the crystallization of the
loaded substances (Mellaerts et al., 2007). The stabilized
amorphous form of the molecules improves the drug dissol-
ution rate. In this context, it must be mentioned that the
crystallization inside the pores is not totally impossible. Even
though the drug typically is in its amorphous form, it may
appear as small, nanosized crystals as well. The solubility of
the nanocrystals is much higher than that of the bulk material.
Therefore, this form is also advantageous considering drug
absorption, and the stability of the product is better than in the
case of amorphous drug. The loaded substances have been
suggested to interact with the surfaces of the carrier via
hydrogen bonding (Vallet-Regi, 2010). This increases the
surface area of the APIs as compared to their crystalline
counterparts, leading to further improvement in the drug
dissolution rate (Bahl & Bogner, 2006).
SBA-15 is a widely researched type of MSNs for drug
delivery (Fu et al., 2012; Wang et al., 2012). SBA-15, with
2D-hexagonal p6mm structure, was firstly synthesized in a
highly acidic media using amphiphilic triblock copolymer of
poly(ethylene oxide)–poly(propylene oxide)–poly(ethylene
oxide) (EO20PO70EO20, P123) as template in 1998 (Zhao
et al., 1998; He et al., 2009). SBA-15 stands for ‘‘Santa
Barbra Amorphous’’. SBA-15 possesses cylindrical ordered
hexagonal mesopores (Zhao et al., 2013).
Most of the previous works concerning loading of APIs in
MSNs focused mainly on the in vitro effect. The current work
does not only focus on the in vitro effect of loading a model
drug (KP) into SBA-15 but also provide a complete in vivo
study assessing the pharmacokinetic effect regarding the anti-
inflammatory effect and central analgesic properties of the
loaded drug compared to a commercial product.
Ketoprofen (KP) is an NSAID with analgesic and
antipyretic properties (Zhu et al., 2011). KP was chosen as
Drug Deliv, 2016; 23(9): 3387–3398
a model drug as it is a BCS Class II drug substance (Shohin
et al., 2012). Consequently, for drugs exhibiting poor
aqueous solubility and high intestinal permeability, it
would be anticipated that dissolution would be the rate-
limiting step to drug absorption. Therefore, it is possible to
study the effect of incorporating poorly aqueous soluble
drugs in SBA-15 on their dissolution properties and hence
their bioavailability.
Materials and methods
Materials
KP was obtained as a gift sample from Egyptian International
Pharmaceutical Industries Company (Cairo, Egypt),
Tetraethyl orthosilicate (TEOS) was obtained from Sigma
Aldrich (St. Lewis), and Pluronic P123 (BASF GmbH,
Ludwigshafen, Germany). Other reagents and solvents were
of HPLC grade.
Methods
SBA-15 synthesis
SBA-15 synthesis was performed according to a standard
method (Zhao et al., 1998). About 6 g of pluronic P123 was
dispersed in 45 g of H2O, then 180 g of HCl (2 M) and
12.8 g of tetraethyl orthosilicate (TEOS) was added to the
mixture. This mixture was then stirred for 24 h at 40  C and
then heated at 100  C for another 24 h under a static
condition. The mesoporous materials were recovered by
filtration, washed with water, air-dried at room temperature
and calcined at 550  C for 6 h to remove the template
forming SBA-15.
Drug loading procedure
Loading was performed by a novel method called immersion-
rotavapor method. KP was dissolved in ethanol (200 mg/ml)
and then adequate amount of the SBA-15 was added, while
protected from light to yield drug:SBA-15 in the ratio 4:1.
The mixture was stirred for 6 h and then the solvent was
evaporated for 15 min using a rotavapor. Subsequently, the
mixture was left to dry under vacuum for 24 h at room
temperature. The resultant sample was denoted as SBA-
15-KP.
Characterization
KP was quantified by extraction and thermal gravimetric
analysis (TGA). Accurately weighed 10 mg of SBA-15-KP
sample was extracted in 20 ml ethanol for 6 h with continu-
ous stirring on magnetic stirrer. Then, the sample was
centrifuged at 5000 rpm for 30 min. The supernatant was
collected and the amount of drug was spectrophotometrically
determined at 259.0 nm. Each analysis was carried out in
triplicate.
TGA test was done by SDT Simultaneous TGA/DSC Q600
USA. The test was carried out as follows: 1 mg of SBA-15-KP
sample was accurately weighed, placed in an aluminum-
sealed cell, and the sample was then scanned by raising the
temperature at a rate of 10  C/min over a 0–600  C range. TG
used nitrogen as purge gas (20 ml/min). TG was calibrated by
DOI: 10.1080/10717544.2016.1186251
the melting point of indium (156.6 ± 0.2  C) and zinc
(419.5 ± 0.3  C).
High resolution transmission electron microscope (HRTEM)
HRTEM imaging of the SBA-15 unloaded and loaded was
recorded using Tecnai G20 Super twin double tilt (Hillsboro,
OR) operating at 200 kV.
Small angle X-ray powder diffraction (SAXRD)
SAXRD patterns were recorded on Scintage XDS 2000
diffractometer (Cupertino, CA). The scan rate was 0.02 deg/
min, step size of 0.01 , and a 2y range of 0.60–10.00 .
Nitrogen adsorption–desorption analysis
The nitrogen adsorption–desorption isotherms and the struc-
tural parameters were recorded using Nitrogen Adsorption/
Desorption analyzer (Nova 3200, West Palm Beach, FL). The
nitrogen adsorption/desorption isotherms were measured at
196.15  C. Prior to analysis, the samples were degassed
under vacuum at 80  C for 24 h. The specific surface areas of
the samples were calculated using the Brunauer–Emmett–
Teller (BET) method (Fagerlund, 1973). The total pore
volume was assessed using the t-plot method illustrated by
Lippens and de Boer (de Boer et al., 1966). The pore size
distribution was calculated from the adsorption branch of
isotherm using the Barrett–Joyner–Halenda (BJH) method
(Barrett et al., 1951).
Differential scanning calorimetry (DSC)
DSC thermograms were recorded by SDT Simultaneous
TGA/DSC Q600 (Delaware). The samples scanned by rising
the temperature at a rate of 10  C/min over a range between 0
and 200  C.
Wide angle X-ray powder diffraction (WAXRD)
WAXRD patterns were recorded on Scintage XDS 2000
diffractometer. The scan rate was 0.03 deg/min, step size of
0.05 and a 2y range of 5.00–35.00 .
Fourier transform infrared spectroscopy (FT-IR)
FT-IR studies were done by spectrometer Jasco FT/IR 460
plus (Oklahoma City, OK) using the KBr disc method. The
disks were made by grinding 2–3 mg of each sample with 25–
50 mg of potassium bromide, and compressed by a hydraulic
press model at 10 tons pressure for 30 s. The scanning range
was 400–4000 cm1 and the resolution was 4 cm1.
Table 1. Composition of the hard gelatin capsule blend.
Formula
F#1 SBA-15-KP Aerosil Mg stearate
91.7 0.18 0.91
*Sodium lauryl sulfate.
SBA-15 hard gelatin capsules 3389
In vitro dissolution study
The dissolution profiles were carried out using USP stand-
ard dissolution apparatus I Dr. Schleuniger Pharmaton
(Switzerland). Accurately weighed 50 mg of KP and equiva-
lent amounts of 50 mg KP from SBA-15-KP sample was used.
The dissolution studies were performed in 900 ml of 0.1 N
HCl. The temperature of the dissolution medium was
maintained at 37  C ± 0.5 and the rotation speed was kept at
50 rpm. Aliquot samples of 5 ml were withdrawn at
predetermined intervals with the replacement by equivalent
volumes of fresh dissolution medium. Samples were filtered
using 0.45 mm Millipore filters and assayed spectrophotomet-
rically at 261 nm.
Formulation in hard gelatin capsule
The loaded sample was formulated in hard gelatin capsule.
The composition of the capsule blend is shown in Table 1.
The prepared formula will be denoted as F#1.
In vivo studies
The in vivo study was approved by the Ethical Committee of
Faculty of Pharmacy, Cairo University
The anti-inflammatory and analgesic studies were carried
out on 24 adult male albino rats weighing 150–180 gm and 24
albino male mice weighing 20–25 gm, respectively. Animals
were randomly allocated into four groups per each experi-
ment, each of six animals. They were kept fasted for 16 h prior
to the experiment, but allowed free access to water.
Tested groups
The tested groups are shown in Table 2. They were divided
into four groups, the first served as the control group, the
second group represented the carrier, while the third and
fourth groups represented the standard reference (commercial
KetofanÕ ) and the prepared formula (F#1), respectively.
Anti-inflammatory study
The anti-inflammatory effect after oral administration of the
tested substances was measured by Carrageenan-induced paw
Table 2. Composition of different tested groups.
Group Treatment Composition
I Control Distilled water
II Carrier 0.1% SBA-15
III Standard reference KetofanÕ capsule content
IV Prepared formula (F#1) SBA-15-KP + Aerosil + Mg
Stearate + Avicel 101+
Ac-Di-Sol + SLS
Ingredients (mg)
Microcrystalline cellulose Crosscarmellose sodium SLS*
27.53 4.58 1.83
3390 A. A. Abd-Elrahman et al.
edema in rats. This effect was determined according to the
method described by Winter et al (Winter et al., 1962).
The exact weight of each rat was accurately recorded
before the start of the experiment. The content of the prepared
capsule formula as well as KetofanÕ capsule were given as
suspensions to the respective animal groups at a dose of 6 mg/
kg. As for the control group, each animal was administered
1 ml distilled water. Finally, regarding group II that received
the carrier was administered 0.1% SBA-15 in a volume of
5 ml/kg (equivalent to the amount of SBA-15 in the tested
formula). A mark was made at the ankle joint (tibio–tarsal
joint) of each rat. Paw edema was induced by subplantar
injection of 0.1 ml of 1% sterile carrageenan in saline into the
right hind paw and 0.1 ml of saline was injected into the left
hind paw. The rats received the control or the tested
substances 30 min before carrageenan injection.
The hind paw volume up to the ankle joint was measured
immediately before carrageenan injection and at selected
times (1/2, 1, 2, 3, 4 and 5 h) by water displacement method
using plethysmometer Ugo Basile 7140 (Comerio, Italy).
Results were expressed as the paw volume in milliliters. The
percentage change in edema volume and the percentage
inhibition was calculated.
% change in oedema volume ¼ ðVa  Vb=VbÞ  100,
where Va and Vb are the edema volume after and before
carrageenan injection, respectively.
% inhibition in oedema volume ¼ 100  ðVt=Vc  100Þ,
where Vt and Vc are the mean percentage change in edema
volume of carrageenan injected in paw of tested and control
group, respectively.
Central analgesic activity study
The central analgesic activity was evaluated using hot plate
method as described by Turner (1965). The hot plate method
is a sensitive method for screening central analgesic effect of
compounds (Wani et al., 2012). Grouping and dosing was
done as per the anti-inflammatory experiment. For three
consecutive days preceding the experiment, mice were
adapted on the hot plate by placing them on a plate (Ugo
Figure 1. TGA charts of ketoprofen, SBA-15
and SBA-15-KP.
Drug Deliv, 2016; 23(9): 3387–3398
Basile 7280, Comerio, Italy) maintained at room temperature
for 15 min each day. Each mouse was then placed gently onto
the hot plate maintained at 50 ± 1  C. Latency in reaction time
to exhibit nociceptive responses, such as licking paws or
jumping off the hot plate, was determined at 0, 30, 60, 90 and
120 min after administration of the test substances or the
control.
Statistical analysis of data
Statistical analysis was carried out using repeated measures
one-way ANOVA followed by the Tukey test for multiple
comparisons at p50.05. Statistics were performed on the
percentage change in edema volume and latency time at each
selected time interval. The statistical study was done by
GraphPad Prism version 5 software (La Jolla, CA).
Result
Characterization
TGA measures the mass loss that is proportional to the total
drug content in the samples. On the other hand, the extraction
depends on extracting the loaded sample in ethanol and then
the amount of loaded KP is spectrophotometrically deter-
mined. The TGA curves for free KP, SBA-15 and SBA-15-KP
are shown in Figure 1. KP showed no weight loss in the range
between 10 to 100  C. Whereas, a sharp decline in the TGA
curve started at 223  C with endset at 290  C pointing to the
decomposition of KP over the range from 220 to 295, causing
99.68% weight loss. Meanwhile, unloaded SBA-15 showed a
weight loss of 3.57% of its total weight in the region below
100  C. This lost weight is due to the evaporation of
physically adsorbed water (Marzouqa et al., 2012). SBA-15
had a total weight loss over the entire temperature range equal
to 3.59%. On the other hand, SBA-15-KP had two distinct
regions of weight losses; in the region below 100  C, and in
the region from 240 to 600  C. Table 3 shows the results from
both the TGA and the extraction method; the average wt% for
the loaded sample was calculated from both the results giving
total amount of 54.4 wt% of drug incorporated.
DOI: 10.1080/10717544.2016.1186251 SBA-15 hard gelatin capsules 3391

Table 3. Quantification of the amount of KP loaded into SBA-15 by TGA and extraction.

TGA Extraction
Weight loss below Total weight Weight loss of Weight loss of Amount of KP Amount of KP Average KP
Sample 100  C (%) loss (%) KP (mg/g) KP (wt%) extracted (mg/g) extracted (wt%) loaded (wt%)
SBA-15 3.59 3.59 - - - -
SBA-15-KP 3.04 57.36 543.2 54.32 544.8 54.48 ± 0.1 54.40 ± 0.1

Figure 2. HRTEM images of SBA-15 (left image) and SBA-15-KP (right image).

Figure 3. SAXRD patterns of Ketoprofen,

SBA-15 and SBA-15-KP.

HRTEM results for free and loaded SBA-15 are shown in
Figure 2. Clearly, both the samples had a characteristic
uniform hexagonal array structure. Figure 3 presents the
SAXRD pattern of KP, SBA-15 and SBA-15 KP. KP diffrac-
tion pattern showed no diffraction peaks at 2 range of 0.60–
10.00 . Free SBA-15 exhibited three well-resolved diffraction
peaks at 2 ¼ 0.84, 1.55 and 1.74 , indexed as the (1 0 0), (1 1
0) and (2 0 0) reflections, typical for SBA-15 materials (Zhao
et al., 1998; Li et al., 2004; Wu et al., 2004). The
corresponding d spacing was 95, 55 and 47.7 Å, respectively
3392 A. A. Abd-Elrahman et al. Drug Deliv, 2016; 23(9): 3387–3398

giving lattice constant of ao ¼ 109 Å as shown in Table 4.
Regarding SBA-15-KP diffraction patterns; showing well-
defined peaks in the 2 range of 0.8–3 . However, the
intensity of diffraction peaks became weaker and shifting to
higher degrees. Meanwhile, the d spacing related to the (1 0 0)
diffraction peak was calculated and it gave a corresponding
lattice constant of 109 Å.
The X-ray diffraction for KP, SBA-15 and SBA-15-KP is
shown in Figure 4. The X-ray diffractogram of KP revealed its
crystalline nature. KP diffractogram showed its characteristic
peaks at 2 diffraction angle located at 14.385, 18.332 and
22.824  . SBA-15 diffraction pattern, showed its characteristic
amorphous peak corresponding to the amorphous silica
(Musić et al., 2011). X-ray diffractograms of SBA-15-KP
exhibited no peaks corresponding to the drug.
DSC curves illustrated in Figure 5 shows that free KP had
an endothermic peak at 96.6  C, while unloaded SBA-15
showed no sharp endothermic peaks, which proves its
amorphous nature. In the meantime, it showed a broad peak
corresponding to physically adsorbed water as observed in the

Table 4. Structural and textural parameters of free and loaded SBA-15.

Specific surface Total pore Pore Cell* Pore diameter distribution

Material area (m2/g) volume (cc/g) diameter (nm) parameter ao (nm) ranges (nm)
SBA-15 780 0.83 6.24 10.96 5.1–7.0
SBA-15-KP 340 0.18 5.45 10.99 4.2–5.8

*The distance from centers of two adjacent pores.

Figure 4. X-ray diffractogram of Ketoprofen,

SBA-15 and SBA-15-KP.

Figure 5. DSC thermograms of Ketoprofen,

SBA-15 and SBA-15-KP.
DOI: 10.1080/10717544.2016.1186251
TGA curve. On the other hand, SBA-15-KP thermogram
revealed complete disappearance of the characteristic endo-
thermic peak of KP.
FT-IR of KP, SBA-15 and SBA-15-KP is shown in
Figure 6. KP spectrum showed a broad band at 3200–
2500 cm1 due to O–H stretching and at 2970, 2930 cm1 due
to C–H stretching of the methyl group. Moreover, there was a
stretching band at 1695 cm1 due to C¼O stretching vibration
of the carboxylic group. Finally, there were different peaks in
the fingerprint region below 1500 cm1 that confirmed the
racemic form of the drug (Liversidge, 1981). As for the
unloaded SBA-15 spectrum, there were broad bands of
absorption at around 3450 cm1 corresponding to molecular
water hydrogen bonded to each other and to Si–OH groups,
they can be assigned to stretching vibrations of O–H and Si–
OH bonds. The bands at around 1620 cm1 related to bending
vibrations of O–H bonds in OH groups, overlapped with C–
O–C stretching vibrations. The bands at around 1080 cm1 is
due to asymmetrical stretching vibrations of Si–O–Si, over-
lapped with Si–O–C, C–O–C and Si–C bond vibrations. The
bands in 980–950 cm1 referred to stretching vibrations of
free silanol (Si–OH) groups on the surface of the amorphous
solid samples also, C–O stretching vibration bonds were
Figure 6. FT-IR Spectra of Ketoprofen, SBA-
15 and SBA-15-KP.
Figure 7. Adsorption/desorption isotherms of
SBA-15 and SBA-15-KP.
SBA-15 hard gelatin capsules 3393
placed in this range. Symmetrical stretching vibrations of Si–
O–Si bonds belonging to ring structures were observed
around 795–790 cm1. The bands in 480–460 cm1 could be
assigned to associate Si–O–Si bond bending vibrations
(Azimov et al., 2012). Regarding the FT-IR spectra of SBA-
15-KP sample, it was clear that the characteristic peaks of
SBA-15 were maintained. In addition, no characteristic peaks
corresponding to KP were observed meanwhile, a new peak
appeared nearly at 1730 cm1.
Nitrogen ads/des results for free and loaded SBA-15 are
shown in Table 4 and Figure 7. SBA-15 isotherm showed the
typical IV isotherms, characteristic for mesoporous molecular
sieves (Zhao et al., 2013) with H1 hysteresis loop.
Subsequently, the capillary condensation step occurred in
the range between 0.6 and 0.8 relative pressures. Similarly,
SBA-15-KP sample exhibited type IV isotherm with H1
hysteresis loop. Meanwhile, the capillary condensation pres-
sures decreased. Hence, the adsorption/desorption isotherm
branches shifted toward lower pressure. Moreover, the results
obtained from calculating specific surface area (m2/g) pore
diameters and total pore volumes (cc/g) for the free and
loaded SBA-15 showed that the value of these parameters
were reduced in the loaded sample.
3394 A. A. Abd-Elrahman et al. Drug Deliv, 2016; 23(9): 3387–3398

Figure 8. Dissolution profile of Ketoprofen and SBA-15-KP.

Table 5. Release mechanism of KP from SBA-15.

In vitro dissolution study
The dissolution profiles of plain KP and SBA-15-KP are Release models Kinetic parameters Free KP Loaded KP
shown in Figure 8. 0.1 N HCl media was selected as the First order R 2
0.995 0.961
dissolution media as it is well-known that comparative AIC 29.53 50.88
dissolution studies at low pH especially 1.2, are more k1(min1) 0.005 0.127
t1/2 (min) 135.22 5.46
discriminating with respect to solubility-enhancing formu- Higuchi R2 0.861 0.231
lation approaches than at higher pH values (Shohin et al., AIC 66.75 100.03
2012). It was clear that the dissolution of the KP that is in kH (min1/2) 3.59 11.9
amorphous state inside the SBA-15 pores was very rapid t1/2 (min) 193.91 17.64
Korsmeyer–Peppas R2 0.994 0.782
compared to the plain crystalline KP. During the first AIC 33.03 70.19
5 min, the release rate of KP from SBA-15 was very fast n value 0.91 0.14
and nearly 50% of KP was dissolved, while for plain KP it KKP (minn) 0.611 53.054
t1/2 (min) 126.31 0.65
took 120 min to reach 45%. The maximum percentage of Weibull R2 0.997 0.994
KP dissolved from free KP was about 45% after 120 min, AIC 27.06 39.83
while maximum percentage of KP released and dissolved  248.26 2.48
from the loaded SBA-15 was 98% after 105 min. It was Td (min) 184.81 7.05
 1.05 0.57
important to estimate the extent of difference or similarity t1/2 (min) 130.86 4.75
between the release profiles under investigation. If both the
test and reference (plain KP) show more than 85%
dissolution within 15 min, the profiles are considered
In vivo studies
similar if not, as in our case, dissolution profiles compari-
son must be done (Sharma & Jain, 2012). Thus, the Table 6 shows the anti-inflammatory results, it is clear that
difference factor f1 and the similarity factor f2 of the loaded the carrier SBA-15 did not have anti-inflammatory effect; the
KP were calculated compared to the free KP. The f1 and f2 percentage inhibition did not exceed 7% when compared to
values were 307.25 and 9.37, respectively. To study the the control. On the other hand, the prepared formula F#1 after
release mechanism of KP from SBA-15 four models were half an hour showed percentage change of 7% in edema and
used, which are first order, the Higuchi, the Korsmeyer– 85% in inhibition. As for the tested reference (KetofanÕ ) after
Peppas and the Weibull. The statistical criteria used for half an hour, it showed percentage change of 27% and 44% in
evaluating the goodness of fit of each model were the edema volume and percentage of inhibition, respectively.
coefficient of determination (R2) and the Akaike Nearly the same results were achieved after 1 h, where the
Information Criterion (AIC). As shown in Table 5, the formula had a percentage inhibition of 82% in edema, while
best fitting model was the Weibull model. the standard reference showed 52%. The maximum anti-
As our goal was to study the effect of incorporating KP inflammatory effect of KetofanÕ was attained after 2 h,
into SBA-15, the loaded sample was formulated in hard whereas F#1 had maximum anti-inflammatory effect after
gelatin capsules. Figure 9 depict the dissolution of F#1 half an hour. At the end of the experiment, both the formula
compared to the dissolution of the SBA-15-KP. It was clear and the reference showed percentage inhibition of 59% and
that the dissolution rate of KP from F#1 showed little 48%, respectively. The statistical results showed that the
improvement when compared to the loaded sample. Clearly, carrier had no significant differences when compared to the
by calculating f1 and f2 values 2.03 and 84.52, respectively it control. On the other hand, on comparing the percentage
was evident that the percentages of KP released from SBA- change in edema volume between F#1 and KetofanÕ after 30
15-KP and the F#1 were pharmaceutically equivalent. and 60 min from carrageenan injection, a very high significant
DOI: 10.1080/10717544.2016.1186251 SBA-15 hard gelatin capsules 3395
Table 6. Ant-inflammatory activity results.

Control Carrier KetofanÕ Formula (F#1)

Mean % change Mean % change Mean % change Mean % Change
in edema in edema in edema in edema
Hours (volume ± S.Da) % Inhibition (volume ± S.D) % Inhibition (volume ± S.D) % Inhibition (volume ± S.D) % Inhibition
1/2 48.9 ± 6.1 – 46.8 ± 5.7 4.3 27.1 ± 4.4 44.5 7.1 ± 2.4b 85.4
1 70.9 ± 8.5 – 70.0 ± 5.2 1.2 32.0 ± 5.4 52 12.4 ± 2.6b 82.5
2 83.2 ± 9.8 – 78.3 ± 5.0 5.8 32.1 ± 5.1 62.4 28.1 ± 1.7 66.2
3 95.4 ± 9.6 – 88.8 ± 8.4 6.9 45.5 ± 7.6 52.3 32.2 ± 3.8 66.2
4 96.7 ± 12.4 – 90.2 ± 7.7 6.7 43.2 ± 10.5 57.5 40.2 ± 7.6 58.4
5 103.9 ± 14.8 – 99.9 ± 3.5 3.8 50.5 ± 7.4 48.4 40.6 ± 6.3 59
a
Standard deviation.
b
Very highly significant.

solution hence, the stirring has to be maintained for 24 h

Table 7. Central analgesic activity results. (Limnell et al., 2011). On the other hand, in rotavapor
method, the diffusion and loading occur mainly during
Control Carrier KetofanÕ Formula F#1 evaporation of the solvent, resulting in a high concentration
Mean Mean Mean Mean gradient of the drug. However, the main drawback is the
Minutes R.Ta ± S.Db R.T ± S.D R.T ± S.D R.T ± S.D uneven distribution of drug inside the pores (Limnell et al.,
0 (Baseline) 4.0 ± 0.6 4.0 ± 0.6 4.0 ± 0.3 3.8 ± 0.4 2011). Therefore, by combining the two methods, we
30 3.4 ± 0.7 3.1 ± 0.5 5.8 ± 0.3 11.9 ± 0.8c managed to cut the loading time to about 2 h with a very
60 3.8 ± 0.5 3.8 ± 1.5 8.3 ± 1.0 11.4 ± 0.7c high loading percentage and uniform distribution of KP
90 3.6 ± 0.5 3.7 ± 0.7 9.2 ± 0.7 11.4 ± 1.0c
120 3.6 ± 0.7 3.9 ± 1.1 9.9 ± 1.1 10.5 ± 0.9 inside SBA-15 pores. The successful loading and the
uniform loading inside the SBA-15 pores with no evidence
a
Reaction time. of surface adsorbed drug particles and no disruption to the
b
Standard deviation.
c
Very highly significant. SBA-15 structure was clearly reflected on the characteriza-
tion results.
In the TGA results, free KP had sharp decline at 223  C
difference was found. Finally, after 2 h till the end of the with endset at 290  C causing 99.68% weight loss. On the
experiment the statistical results showed no significant other hand, the decomposition behavior of the loaded KP
differences between the comparable treatments. sample revealed that the decomposition was nearly by the
The central analgesic activity was evaluated using hot plate same rate over the temperature ranging from 240 to 560  C.
method; the results are shown in Table 7. Regarding the This referred to the uniform distribution of KP inside SBA-15
carrier, it did not show any analgesic activity throughout the pores. Consequently, the decomposition of KP over extended
whole experiment. F#1 caused maximum analgesia (increase temperature range would assert the successful loading of KP
threshold potential of pain) only after 30 min from its into SBA-15. Furthermore, the evaporation temperature for
administration, while the highest analgesic effect caused by the loaded KP was increased and extended compared to the
KetofanÕ appeared after 120 min. free substance. This phenomenon could be explained by
These results reflected on the statistical analysis outcomes; capillary forces in the pores, as has been stated by Peng Zhang
the carrier showed no significant differences when compared et al (Zhang et al., 2014).
to the control. On the other hand, on comparing F#1 with Another important observation was that TGA and extrac-
KetofanÕ after 30, 60 and 90 min from the administration, tion results were in a very good agreement, which strongly
there was a very high significant difference in their mean imply that during the decomposition of a drug no interactions
reaction time. Finally, at 120 min (the end of the experiment) occurred between decomposed drug substances and the
the statistical results showed no significant differences carrier. It is known that if the interactions occurred at TGA
between the formula and KetofanÕ . measurements could give underestimated drug loading values
(Salonen et al., 2005). Therefore, as the TGA and extraction
Discussion
results were very close, the average wt% for the loaded sample
In the current study, we used a novel method in loading KP was calculated from both the results.
into SBA-15 namely immersion rotavapor method. This HRTEM results asserted the successful loading of KP
method is a combination between two well-known methods inside the SBA-15 without affecting its internal structure
for loading drugs into MSNs, which are immersion and where the free and loaded SBA-15 had ordered two-dimen-
rotavapor method. By combining these two methods, we sional channel mesostructure as well as mesoporous highly
beneficiated from the advantages and controlled the draw- ordered honeycomb array structure (Nhu et al., 2010). This
backs of the two methods. The immersion method gives was further confirmed by the SAXRD results, it was clear that
uniform loading inside the MSNs pores meanwhile it needs a the hexagonal pore arrangement was not affected by drug
very high concentration of the drug as well as stirring for loading showing well-defined peaks in the 2 range of 0.8
about 24 h. It is known that drug diffusion as well as loading to 3 . In addition, by calculating the d spacing related to the
occurs during stirring of the particles in the concentrated drug (1 0 0) diffraction peak and its corresponding lattice constant,
3396 A. A. Abd-Elrahman et al.
it was clear that no cell parameter changes occurred after
KP incorporation into the SBA-15 (Zhao et al., 2013).
Consequently, the SAXRD confirmed the successful loading
by showing reduction in the peak intensities of the loaded
sample with shifting to higher degrees. This pointed to the
decrease in pore channels diameter associated with drug
incorporation (Jusoh et al., 2013).
The nitrogen ads/des results showing that both the free and
loaded SBA-15 exhibited typical IV isotherms with H1
hysteresis loop added more conformation for the successful
loading of KP with no evidence of MSNs disruption.
Moreover, SBA-15 isotherm that showed step position at
high relative pressures pointing to the presence of large pore
sizes (above 5 nm) (Cao & Kruk, 2014). In addition, the
sharpness of the adsorption branches (located at a relative
pressure from 0.6 to 0.8) indicates a narrow mesopore sizes
distribution characteristic for a good quality SBA-15 (Wang
Figure 9. Dssolution profile of SBA-15-KP and F#1.
Figure 10. ?Pore size distribution of SBA-15
and SBA-15-KP.
Drug Deliv, 2016; 23(9): 3387–3398
et al., 2013). This was also confirmed by the pore size
distribution curve and pore diameters distribution range
shown in Table 4 and depicted in Figure 10. As for the
loaded sample, a clear decrease in the capillary condensation
pressures was observed. This reduction was due to loading of
KP inside SBA-15 pores (Fu et al., 2012), which was
associated with the reduction in the pore diameter. However,
KP encapsulated inside the pores do not fully occupy the
available space, such that there was still available space for
nitrogen gas to adsorb inside the pores.
The FT-IR results indicated that loading of KP did not
affect the mesoporous chemical structure. However, a new
peak appeared that arises from hydrogen bonding of Si–OH
(silanol group) with the COOH (carboxyl group) of KP
(Song et al., 2005). Consequently, this peak confirmed the
successful loading of KP into SBA-15 pores (Song et al.,
2005).
DOI: 10.1080/10717544.2016.1186251
The X-ray diffraction and DSC results, which showed
complete disappearance of any peaks related to KP,
pointed to two important aspects. First, the successful
encapsulation of KP into the carrier pores in a molecular
amorphous state. Second, no surface-adsorbed drug par-
ticles occurred.
The presence of KP in amorphous state, the large
surface area and the appropriate pore diameters provided by
SBA-15 have reflected clearly on the dissolution profile of
the loaded KP sample, where nearly 90% of the drug
dissolved after 20 min. For further analysis of the dissol-
ution results, the difference factor f1 and the similarity
factor f2 was estimated. The high degree of difference
between the free and loaded KP asserted the prominent
effect of loading KP into SBA-15 on its dissolution rate. In
order to study the release mechanism, the goodness of fit
of each model was determined by the coefficient of
determination (R2) and the Akaike Information Criterion
(AIC). For more precise studying of the dissolution results,
the Weibull model parameters were calculated. The Weibull
model parameters include the scale factor  that relates to
the time scale of the process, parameter  which describes
the shape of the profile either exponential ( ¼ 1; case 1),
sigmoid (41; case 2) or parabolic with a higher initial
slope compared to the exponential (51; case 3) (Adams
et al., 2002) and Td that is defined by  ¼ (Td) . Td
represents the time interval needed to dissolve 63.2% of the
drug (Costa & Sousa Lobo, 2001). Regarding the parameter
, it was found that the loaded sample releasing profiles
was of the parabolic type; case 3 i.e (51) while the plain
drug exhibited sigmoid-shaped curve; case 2 i.e (41). As
for the time scale  and Td parameters, the general
decrease in their values in case of SBA-15-KP would point
to the very fast dissolution rates of the loaded KP. On the
other hand, the results obtained from the dissolution profile
of the F#1 when compared to the loaded sample did not
show a significant improvement in the dissolution rates.
This was very evident when similarity and difference
factors were calculated. This would indicate that the
presence of different excipients did not affect the release
rate of KP. Meanwhile, the rate of drug release was solely
affected by its amorphous state inside the SBA-15 pores as
well as the pore diameter of the SBA-15.
The very fast release rates of KP from SBA-15 had
reflected on its onset of action as clearly observed in the in
vivo study. The F#1 should have maximum anti-inflammatory
and analgesic effect after 30 min from the beginning of the
experiments. This result was in a very good agreement with
the dissolution results, where almost 96% of the formula was
released after 30 min from the beginning of the dissolution
experiment.
Conclusion
SBA-15 was studied as a drug delivery carrier for KP in
order to improve its bioavailability. The results elucidated
the influence of loading KP into SBA-15 on improving its
bioavailability based on both the in vitro and in vivo studies.
Our current work established a new way to improve the
bioavailability of poorly water-soluble drugs, opening a new
SBA-15 hard gelatin capsules 3397
avenue for the utilization of SBA-15 in drug delivery
system.
Acknowledgements
The authors would like to thank Prof. Dr. El-Zeiny M. Ebeid
of Tanta University and MUST University, for helping in
mesoporous silicate synthesis.
Declaration of interest
The authors declare that there is no conflict of interest for this
work. The authors alone are responsible for the content and
writing of this article.
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