Development of Microemulsion For Solubility Enhancement of Poorly Water Soluble Drug Valsartan

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Int. J. Pharm. Sci. Rev. Res.

, 22(2), Sep – Oct 2013; nᵒ 45, 246-251 ISSN 0976 – 044X

Research Article

Development of Microemulsion for Solubility Enhancement of


Poorly Water Soluble Drug Valsartan
Shruti G. Shahu*, Rita N. Wadetwar, Gouri R. Dixit
J. L .Chaturvedi College of Pharmacy, Electronic Zone Building, MIDC, Hingna Road, Nagpur, India.
*Corresponding author’s E-mail: shrutishahu1@gmail.com

Accepted on: 05-08-2013; Finalized on: 30-09-2013.


ABSTRACT
Valsartan is orally active, and specific angiotensin II antagonist acting on the AT1 receptor subtype (angiotensin receptor blocker).
Valsartan is poorly water soluble drug. The aim of the investigation was to design and develop micro emulsion of valsartan for
enhancing its solubility hence the oral bioavailability. Solubility of valsartan was determined in various vehicles and maximum
solubility was found in Capmul MCM (oil), Cremophor EL (surfactant) and Transcutol HP (co-surfactant). These components were
used to construct pseudo-ternary phase diagrams to identify the micro emulsion existing zone. The micro emulsion was prepared by
phase titration method. Optimized micro emulsion was characterized for its transparency, particle size, drug content, viscosity, and
stability study etc. Particle size of optimized micro emulsion was found to be 51.32 nm. Drug content of the micro emulsion
formulation was 98.29% ± 0.91%. The viscosity data indicated the micro emulsion to be O/W type. 78.49% and 71.53% of the drug
was found to be released in 4hrs in the in-vitro and in-vivo intestinal permiability studies respectively. Hence, by formulating into
micro emulsion, the solubility of valsartan was significantly enhanced which may increase its bioavailability. Thus, it could be
concluded that micro emulsion formulation could be used as a possible alternative to traditional oral formulations of Valsartan to
improve its bioavailability.
Keywords: Micro emulsion, Pseudo-ternary phase diagrams, Solubility, Valsartan.

INTRODUCTION thermodynamic stability, reversibility, simple


manufacturing, and scale up feasibility, and do not

O ral route still remains the favorite route of drug


administration in many diseases and till today it is
the first way investigated in the development of
new dosage forms. Successful oral delivery of drugs has
require any special equipment. Oil-in-water (o/w) micro
emulsion is the most suitable formulation, which is
expected to increase the solubility by dissolving the
compounds with low water solubility into an oil phase.5
always remained a challenge to the drug delivery field,
Thus, Valsartan is considered to be a good candidate for
since approximately 40% of the new drug candidates have
micro emulsion drug delivery system to enhance its oral
poor water solubility, and thus oral delivery is frequently
bioavailability by reducing the droplet size, hence
associated with implications of low bioavailability.1
increasing the rate of absorption due to surfactant
In recent years, much attention has focused on lipid- induced permeability changes.
based formulations to improve the oral bioavailability of
MATERIALS AND METHODS
poorly water soluble drug compounds. In fact, the most
popular approaches is the incorporation of the active Materials
lipophillic component into inert lipid vehicles such as oils,
Valsartan was a gift sample from Zim Laboratories Limited
surfactant dispersions, nano emulsions, micro emulsions,
(Kalmeshwar, Maharashtra, India). Cremophor EL was a
self-emulsifying formulations, self nano or micro
gift sample from BASF India Ltd (Mumbai, India), Capmul
emulsifying formulations, emulsions and liposomes.2
MCM was a gift sample from Indchem International
Valsartan is a nonpeptide, orally active, and specific (Mumbai, India) and Transcutol HP was a gift sample from
angiotensin II antagonist acting on the AT1 receptor Gattefosse India Pvt. Ltd. All other chemicals used were of
subtype. It is categorized in angiotensin receptor blocker. analytical reagent grade and used as received without
Valsartan is poorly soluble and the aqueous solubility is further purification. Double-distilled water was used
reported to be less than 1 mg/mL. The drug has oral throughout the study. The animal requirement was
bioavailability of about 23% due to its poor aqueous approved by the Institute Animal Ethics Committee, and
solubility.3 all experiments were conducted as per the norms of the
Committee for the Purpose of Supervision of Experiments
Micro emulsions are clear, transparent, optically isotropic
on Animals, India.
and thermodynamically stable systems comprising of oil,
surfactant, co-surfactant and aqueous phase.4 Micro Methodology
emulsion (ME), a novel drug delivery system, has been
Solubility of the oil phase, surfactant and co-surfactant
reported to improve the rate and extent of absorption of
lipophilic drugs.11 Micro emulsions are used as edge as Selection of the oil phase was based upon the maximum
potential drug delivery vehicles because of their solubility of the drug. Different oils including Capmul
International Journal of Pharmaceutical Sciences Review and Research
Available online at www.globalresearchonline.net 246
Int. J. Pharm. Sci. Rev. Res., 22(2), Sep – Oct 2013; nᵒ 45, 246-251 ISSN 0976 – 044X

MCM, castor oil, rice bran oil, oleic acid, labrafil 1944 CS 650 nm with distilled water taken as blank and three
and captex 355 were taken for the studies. Also various replicates were performed for each sample.6,8,11
surfactants like tween 80, span 20, cremophor EL, and
pH determination
cremophor RH 40 were taken and co-surfactants like iso-
propanlol, ethanol, transcutol HP, transcutol P, capryol The apparent pH of all the selected micro emulsions and
90, polyethylene glycol (PEG) 400 and propylene glycol the plain micro emulsion was determined at 25°C by
(PG) were taken for solubility studies. 2 ml of each of the immersing the electrode directly into the micro emulsion
selected vehicle were added to each cap vial containing using a digital pH meter.8,12
an excess of valsartan. After sealing, mixtures were
o Refractive index
shaken with shaker at 25 C for 48 hr. After reaching
equilibrium, each vial was centrifuged at 500 rpm for 15 Refractive indices of the prepared micro emulsions were
min, and excess insoluble valsartan was discarded by determined at 25°C by Abbe’s refractometer by placing
filtration using a membrane filter (0.45 µm, 13 mm, one drop of micro emulsion on the slide.6,9
Whatman). The concentration of valsartan was
Viscosity measurement
determined in each oils, surfactants, and co-surfactants
by UV spectrophotometer at their respective wavelength Micro emulsions are generally low viscosity systems. The
250 nm.6 viscosity of the prepared micro emulsion was measured
at 25°C at 60 rpm by LV spindle no. 63 using a Brookfield
Pseudo-ternary phase diagram
viscometer (model LVDVE230; Brookfield Engineering
Pseudo-ternary phase diagrams were constructed to Laboratories, Inc).6,8,12
obtain the appropriate components, and their
Determination of Drug Content in the micro emulsion
concentration ranges that resulted in a large existence
area of micro emulsion were chosen. In order to optimize The drug content of the micro emulsion formulation was
the concentration of oil phase, surfactant and co- determined by dissolving 1 ml (equivalent to 10 mg drug)
surfactant, different batches of varied concentration were of the formulation in 10 ml of methanol. After suitable
prepared and titrated with distilled water till dilutions with methanol, absorbance was determined
transparency persisted. Ternary phase diagram was using the UV spectrophotometer keeping blank micro
prepared by using a constant ratio of surfactant to co- emulsion as control at wavelength 250 nm and three
surfactant. Four ratios of surfactant (Cremophor EL) and replicates were performed for each sample.5,6,8
co-surfactant (Transcutol HP) were selected (1:1, 2:1, 3:1
Particle size Determination
and 4:1). At each specific Smix weight ratio, the ratio of
oil to the Smix was varied as 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, The particle size of the plain micro emulsion and final
7:3, 8:2 and 9:1. Each mixture of oil, surfactant and co- formulation F4 was determined by using particle size
surfactant, at specific weight ratio were titrated with analyzer Nanophox (NX0088) Sympatec, Germany
water along water dilution lines. If clear and transparent performed at Sinhagad Institute of Pharmacy, Pune.9
mixtures were visualized after stirring, the samples were
Drug release studies
considered to be monophasic and were marked as points
in the phase diagrams and areas covered by these points In-vitro drug release
were considered as the micro emulsion region of The diffusion study was carried out using a modified Franz
existence region. Maximum micro emulsion region was diffusion cell. The receptor compartment was filled with
optimized for further studies and four ratios of the 30 ml of Phosphate buffer (pH 5.5). The donor
optimized formulation were characterized. compartment was fixed with cellophane membrane and it
Preparation of Valsartan micro emulsion system contained micro emulsion (Valsartan) and plain drug
suspension separately. At predetermined time intervals of
Valsartan containing micro emulsion was prepared by
30 minutes samples were withdrawn from receptor
water titration method. Valsartan was first dissolved in
compartment and analyzed for drug content by UV
the pre-measured volume of oil by stirring on a magnetic 10
Spectrophotometer at 250 nm.
stirrer. A mixture of the surfactant and co-surfactant at a
fixed ratio (v/v) was added to the above resulting In-vitro intestinal permeability study
mixture. Finally this mixture was titrated with distilled The methods employed were modified from experimental
water. The optimized micro emulsion drug delivery procedures. Male albino rats (250-300 g) were killed by
8
system was also subjected for accelerated stability study. overdose with pentobarbitone administered by
Characterization of micro emulsion intravenous injection. To check the intra-duodenal
permeability, the duodenal part of the small intestine was
Percentage Transmittance
isolated and taken for the in vitro diffusion study. Then
Transparency of micro emulsion formulation was this tissue was thoroughly washed with cold Ringer’s
determined by measuring percentage transmittance solution to remove the mucous and lumen contents. The
through U.V. Spectrophotometer (UV-1601 SHIMADZU) at equivalent dose of optimized micro emulsions and plain
drug solution were prepared. One side of the intestine
International Journal of Pharmaceutical Sciences Review and Research
Available online at www.globalresearchonline.net 247
Int. J. Pharm. Sci. Rev. Res., 22(2), Sep – Oct 2013; nᵒ 45, 246-251 ISSN 0976 – 044X

was tightly closed, resultant samples (1 mg/mL) were Drug stability


injected into the lumen of the duodenum using a syringe
Stability studies of selected formulation were carried out
and then other side of the intestine was tightly closed.
at ambient temperature (30°C±2°C), under cold condition
Then the tissue was placed in a chamber of organ bath
4 ± 2°C and at elevated temperature 40 ± 5°C. After every
with continuous aeration and a constant temperature of
15 days the micro emulsion was analyzed for phase
37˚C. The receiver compartment was filled with 30 ml of
separation, % transmittance, pH and refractive index.5
phosphate-buffered saline (pH 5.5). After a particular
time interval of 30 minutes the 1ml sample was RESULTS AND DISCUSSION
withdrawn and diluted to 10 ml. The absorbance was
Solubility of the oil phase, surfactant and co-surfactant:
measured using a UV-VIS spectrophotometer at a
wavelength of 250 nm, keeping the respective blank. The Solubility studies were carried out to identify the
percent of cumulative drug diffusion was calculated potential ingredients for the formulation of micro
against time and plotted on a graph. Similarly, suspension emulsion. The solubility studies were performed using
of plain drug was prepared and compared with the different oils, surfactants and co-surfactants to find out
optimized formulation.10 the highest solubility of the drug valsartan in all the
components of micro emulsion. The results are shown in
table 1. The drug shows highest solubility in the oil
Capmul MCM, surfactant Cremophor EL and co-surfactant
Transcutol HP.
Table 1: Solubility of valsartan in various oils, surfactants and co-surfactants
Oils Solubility mg / ml Surfactant Solubility mg/ml Co-surfactant Solubility mg / ml
Castor oil 534 Tween 80 501 Propanolol 1006
Capmul MCM 823 Cremophor EL 615 Ethanol 1025
Captex 355 3 Span 20 166 PEG 400 909
Rice bran oil 4 Propylene glycol 339
Oleic acid 23 Transcutol P 1261
Cremophor RH 40 489
Transcutol HP 1621
Labrafil 1944 19
Capryol 90 1021

F1 F2 F3 F4
Figure 1: Ternary phase diagrams representing the micro emulsion formed with Capmul MCM (oil), Cremophor EL
(surfactant) and Transcutol HP (co-surfactant).
Pseudo-ternary phase diagram formed. Thus, four ratios of the optimized formulation
were selected.
Pseudo-ternary phase diagrams were constructed by sing
Capmul MCM (oil), Cremophor EL (surfactant) and Characterization of the micro emulsion
Transcutol HP (co-surfactant) to identify the micro
The optimized formulations were further studied for
emulsion existing zone from which appropriate
percent transmittance, drug content, pH determination,
concentration ranges of components of micro emulsion
refractive index and viscosity of the different ratios of the
can be obtained. The ternary phase diagrams of all the
selected system:
ratios are shown in figure 1. Formation of micro emulsion
systems (the shaded area) was observed at room Percentage Transmittance
temperature. Phase behavior investigations of this system
The percent transmission carried out on UV
demonstrated the suitable approach to determine the
spectrophotometer at 650 nm was found to be in the
water phase, oil phase, surfactant concentration, and
range of 98.23 % to 99.37 % for formulations F1 to F4
cosurfactant concentration with which the transparent,
along with the plain formulation which confirms good
one-phase, low-viscous microemulsion system was
transparent nature of formulations.

International Journal of Pharmaceutical Sciences Review and Research


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Int. J. Pharm. Sci. Rev. Res., 22(2), Sep – Oct 2013; nᵒ 45, 246-251 ISSN 0976 – 044X

Drug Content Viscosity


The drug content at 250 nm was found to be in the range All formulations F1 to F4 were found to have rather low
of 99.23% to 99.72% in the optimized F1 to F4 viscosities, ranging from 63 to 96 cps. The viscosity of the
formulations. micro emulsion increased with increasing concentration
of the surfactant.
pH determination
Particle size determination
For the formulations F1 to F4, the pH value was found to
be in the range of 3.66 to 4.02. The particle size was found to be 51.32 nm for the drug
formulation F1, which was found to be in the range for
Refractive index
micro emulsions (10-100nm). The result is shown in figure
The refractive index carried out by Abbe refractometer 2.
was found to be in the range of 1.3618 to 1.3646 of F1 to
F4 formulations along with the plain formulation.
Table 2: Results for percent transmittance, drug content, pH determination, refractive index and viscosity of optimized
formulation along with plain system
Formulation code Percent transmittance (%) pH Refractive index Viscosity (cp) Drug content (%)
Without drug 99.2 ± 0.04 3.66 ± 0.07 1.3648±0.0006 63.33±4.1 -
F1 (1:1 ratio) 99.37 ± 0.16 3.96 ± 0.20 1.3620 ± 0.004 66.36±5.7 99.72 ± 0.31
F2 (2:1 ratio) 99.32 ± 1.33 3.92 ± 0.05 1.3618 ± 0.002 76.46±4.77 99.4 ± 1.75
F3 (3:1 ratio) 98.47 ± 1.02 3.94 ± 0.02 1.3620 ± 0.005 83.33±4.34 99.32 ± 1.33
F4 (4:1 ratio) 98.23 ± 1.91 4.02 ± 0.12 1.3618±0.0012 96.66±5.74 98.23 ± 1.91

Figure 2: Shows the particle size determination of the drug loaded formulation.
Drug release studies In-vitro intestinal permeability study
In-vitro drug release study From the figure 4, it was seen that after 4 hours of
diffusion, 99.478% of the drug was released from the
It was seen that after 4 hours of diffusion, the drug
formulation F1 which is higher than that of the other
released from the formulation F1 faster and more than
ratios. Hence, F1 formulation was selected as optimized
that of the other ratios i.e., 93.410%. The results are
formulation and was considered for comparison with the
shown in figure 3.
plain drug suspension. From the comparison study of F1
formulation and plain drug suspension, it can be
concluded that the extent of diffusion of valsartan from
the Capmul micro emulsion is greater than the plain drug
suspension as shown in figure 5.
Stability studies
Stability studies of selected formulation were carried out
at 30°C±2°C, 4°C±2°C and 40°C±5°C. The results of
stability are shown in table 3.

Figure 3: In-vitro cumulative average drug release


formulations F1, F2, F3 and F4
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Int. J. Pharm. Sci. Rev. Res., 22(2), Sep – Oct 2013; nᵒ 45, 246-251 ISSN 0976 – 044X

Table 3: Stability studies of micro emulsion formulation F1


Phase Drug Percent
Temperature Days Viscosity pH Refractive index
separation content transmittance
15 No 60.66 ± 4.12 99.63± 0.04 3.87± 0.04 99.22± 0.07 1.3618 ± 0.0002
30 No 55.36 ± 4.70 99.42± 0.03 3.83± 0.03 99.14± 0.03 1.3601 ± 0.0012
4 ± 2°C
45 No 51.06 ± 5.7 99.22 ± 0.04 3.78± 0.06 98.97± 0.03 1.3585 ± 0.0008
15 No 56.36 ± 4.3 99.46± 0.03 3.78± 0.02 99.17± 0.04 1.3615 ± 0.0006
30 No 47.66 ± 5.71 99.16 ± 0.03 3.73± 0.08 99.02± 0.03 1.3510 ± 0.0006
30°C±2°C
45 No 43.36 ± 4.23 98.82± 0.04 3.66± 0.05 98.92± 0.03 1.3512 ± 0.0007
15 No 44 ± 3.464 99.24± 0.03 3.72± 0.04 99.06± 0.04 1.3550 ± 0.0037
30 No 39.26 ± 4.20 98.84± 0.05 3.68± 0.04 98.91± 0.05 1.3548 ± 0.0055
40 ± 5°C
45 No 35 ± 7.071 98.52± 0.04 3.63± 0.03 98.72± 0.08 1.3482 ± 0.0032

permeability studies, the prepared microemulsion


showed the in vitro intestinal permeability release of
about 99.47% within 4 hours which is more than that of
plain drug suspension 60.33%. The stability studies
confirmed that the optimized formulation is stable for a
period of 45 days. Thus, it can be concluded that valsartan
having poor solubility can be formulated as
microemulsion, which increase the solubility of the drug
and hence, which will increase its oral bioavailability than
the other dosage forms.
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Source of Support: Nil, Conflict of Interest: None.

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