Tanaji Nandgude et al.

: Formulation and Evaluation of pH Induced… 177

Research Paper

Formulation and Evaluation of pH Induced In-situ

Nasal Gel of Salbutamol Sulphate
Tanaji Nandgude*1, Rahul Thube2, Nitin Jaiswal2, Pradip Deshmukh1, Vivek Chatap1, Nitin Hire2
1
B.R. Nahata College of Pharmacy & BRNSS Contract Research Center, Mhow – Neemuch Road,
Mandsaur (M.P.), 458001.
2
M. C. E. Society’s, Allana College of Pharmacy, 2390-k, B. Hidayatullah road, Azam campus, Pune-411 001

ABSTRACT: Nasal solutions of Salbutamol Sulphate were prepared for sustaining its release and improving
its bioavailability. Carbopol was used as a key ingredient to effect pH induced sol to gel conversion of the
formulations. Different formulations were prepared by varying the concentrations of Carbopol 934 and
Hydroxyl Propyl Methyl Cellulose. These formulations were evaluated for parameters like pH, drug content,
viscosity, gel strength and drug release. Release profile of some formulations showed rapid phase while
some showed slow phase. At extreme low concentrations of the polymers, the formulations drained out due
to poor viscosity while at higher concentrations of the same the formulations formed stiff gel and showed slow
release of drug. Finally optimized formulation with specific concentrations of carbopol 934 and Hydroxyl
Propyl Methyl Cellulose showed pH induced sol-gel conversion, sustained release and higher bioavailability.
KEY WORDS: pH induced sol to gel conversion, Carbopol 934, HPMC, sustained release.

Introduction
For systemic therapy, drugs are traditionally administered
by oral and parenteral routes. However in many instances,
oral administration is unsuitable when the drug undergoes
significant degradation in the gastrointestinal tract or is
metabolized to a high degree via the first pass effect in the
liver (Illum et al., 2003). Researchers resorted to the
parenteral route as an easy solution to this problem. But
parenteral route is undesirable or impractical if a drug is
intended for the treatment of chronic diseases. Therefore
an alternative route of administration would be preferred.
For the past few decades, the transdermal route has been
explored for a number of drugs, but its use is limited due to
low permeability of the skin to many drugs (Mao et al.,
2004) Nonparenteral routes for drug delivery include nasal,
buccal, pulmonary and transdermal routes. All these
application routes are suitable for self-administration in an
ambulatory setting. The nasal application is the only route
of administration, which offers rapid onset of action; high
absorption of small molecular weight hydrophobic drugs,
relatively high bioavailability, avoidance of first pass
effect and ease of administration by patients favors this
route of application (Osth K et al., 2002). Traditionally, the
nasal route has been used for delivery of drugs for local
treatment such as nasal congestion, allergy and infections.
More recently, nasal drug delivery has generated
widespread interest among the scientific community as an
alternative route for the administration of drugs and
biomolecules that are susceptible to enzymatic or acidic
degradation and first pass hepatic metabolism or
incompletely absorbed in the gastrointestinal tract or gives
undesirably slow effects when administered orally. Nasal
*Corresponding author: Tanaji Nandgude
Tel: 07422-284224, Mobile: 09926878701
e-mail: tanajinandgude@gmail.com
route may also be preferred when rapid onset of action is
required and when small molecular weight polar drugs,
peptides and proteins are not easily administered via other
routes than by injection. (Wadell C et al., 1999)
Salbutamol Sulphate is a selective β2 - adrenoreceptor
agonist and is readily absorbed from G.I.T. but it is
subjected to considerable first pass metabolism. Thus its
oral bioavailability is only 50%. To increase patient
compliance, bioavailability and to have convenience of
administration nasal gel of Salbutamol Sulphate was
prepared by using pH induced gelling polymer which
increased the residence time. Gel point was adjusted in
such a way that formulation is in the liquid state during the
shelf life and gels upon administration. For improving the
bioavailabilty of Salbutamol Sulphate following objectives
were set. Formulate the nasal in- situ gel that can be easily
administered as drop and attains semisolid properties at
nasal pH, Improve bioavailability by avoiding the first pass
effect, thus reducing the dose and dosing frequency,
Prolong the contact time with nasal mucosa.
Materials and Methods
Materials
Salbutamol Sulphate was obtained as a gift sample from
Litaca Pharmaceuticals Pvt. Ltd., Pune, India. Batch No.
SS/2005/039. Hydroxypropyl Methyl cellulose K4M was
obtained as a gift sample from Colorcon Asia Pvt. Ltd.,
Singapore-056283. Benzalkonium Chloride was obtained
from Research Lab Fine Chem Industries, Mumbai-400
002.Batch No. 6591203. Sodium Metabisulphite, Carbopol
934P & Sodium Chloride were obtained from Loba Chem
Pvt. Ltd., Mumbai-400 002. Batch No. V260905,
V1181043 & V278205.

177
178 International Journal of Pharmaceutical Sciences and Nanotechnology Volume 1 • Issue 2 • July - September 2008

Table 1 Compositions of in situ nasal formulations.

Ingredients Formulations
B1 B2 B3 B4 B5 B6 B7 B8
Salbutamol sulphate (%w/v) 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25
Carbopol 934 (%w/v) 0.1 0.2 0.3 0.4 0.5 0.4 0.4 0.4
HPMC K4M (%w/v) - - - - - 0.5 1.0 1.5
NaCl (%w/v) 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.9
Benzalkonium chloride
0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02
(%v/v)
Sodium metabisulphite
0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1
(%w/v)
Distilled water (ml) q.s q.s q.s q.s q.s q.s q.s q.s

Methods 2. pH of the Gel (Paranjothy KLK 1993)

Preparation of Standard Curve of Salbutamol
Sulphate (Moffat AC et al., 1999)
Standard curve of Salbutamol Sulphate was prepared in
phosphate buffer solution pH 7.4 (PBS 7.4). The
absorbance of the solution was measured against PBS 7.4
as a blank at 276 nm, using double beam UV visible
spectrophotometer (Model No. V-530, Make- Jasco
Corporation Tokyo, Japan). The graph of absorbance v/s
concentration was plotted.
Formulation of in Situ Gel (Charoo A et al., 2003, Keny
RV et al., 2004, Lin HR et al., 2000, Rao KP et al., 2005)
The formulations as given in Table 1 were prepared by
dispersing carbopol in distilled water with continuous
stirring (Thermostatic hot plate with magnetic stirrer,
Model No. MS 5836) until completely dissolved and
allowed to hydrate overnight. For the preparation of
solution, first HPMC K4M was added in distilled water and
allowed to hydrate. Then Carbopol was sprinkled over this
solution and allowed to hydrate overnight. After complete
hydration of polymers a separate solution of Salbutamol
Sulphate and Sodium Chloride was added to the polymeric
solution. The resultant solution was thoroughly mixed,
Benzalkonium Chloride and Sodium Metabisulphite were
then added and mixing was confirmed until a uniform and
clear solutions were formed. Final volume was made by
adding required volume of distilled water. All the
formulations were adjusted to pH 4.5 by 0.5 M Sodium
Hydroxide solution.
Evaluation of in Situ Gels
1. Appearance (Balasubramanium J et al., 2003)
The developed formulations were inspected visually for
clarity in sol and gel form.
pH of the each formulation was determined by using
pH meter (Model No. CL 54, Make- Toshniwal
Instrument Pvt. Ltd., Ajmer, India). The pH meter was
first calibrated using solutions of pH 4 and pH 7.
3. Gelation Studies (Balasubramanium J et al., 2003)
Gelation studies were carried out in gelation cells,
fabricated locally using Teflon. The cells were
cylindrical reservoirs holding 3 ml of gelation solution.
Within the cells at the bottom, a 400 μl transparent
plastic cup was located to hold the gel sample in place
after its formation. The studies were carried out using
0.5 M Sodium hydroxide, which resembles nasal
fluid. The formulation (250μl) was carefully placed
into the cavity of the cup using a micropipette and 2 ml
of gelation solution was added slowly. Gelation was
assessed by visual examination.
4. Drug Content (Charoo A et al., 2003, Shakleya DM
et al., 2000)
Each formulation (0.5 gm) was taken in a 50ml
volumetric flask diluted with PBS pH 7.4 and shaken to
dissolve the drug in PBS pH 7.4. The solution was
filtered through whatman filter paper, 1ml of above
filtrate was pipette out and diluted to 10 ml with PBS
pH 7.4. The content of the drug was estimated
spectrophotometrically by using standard curve plotted
at 276 nm.
5. Viscosity Measurement (Srividya B et al., 2001)
The apparent single viscosity values were measured
with the Brookfield digital viscometer (Model No.
CAP-2000, Make- Brookfield Engineering Lab. Inc.
Middleboro, MA-02346, USA). Viscosity was
measured at 10 rpm after 30 seconds for liquid
formulations and for gels. Formulations B1 and B2 did
not form a gel. So they were not subjected to further
evaluation parameters.
 Tanaji Nandgude et al. : Formulation and Evaluation of pH Induced…
6. Measurement of Gel Strength (Choi HG et al., 1998,
Elhady SS et al., 2003, Kim CG et al., 1998, Yong
CS et al., 2001)
A sample of 50gm of the formulation was placed in a
100 ml graduated cylinder and gelled by neutralizing
with 0.5 M NaOH. A weight of 35 gm was then placed
onto gel. The gel strength, which is an indication for
the viscosity of the nasal in situ gel at physiological
condition, was determined by time in seconds the
weight took to penetrate 5 cm down through the gel.
7. In Vitro Release Study (Maitini Y et al., 1992,
Maitini Y et al., 1991)
In vitro release study of the formulated in situ gel was
carried out in two-chamber diffusion cell through
dialysis membrane-70 (Himedia) with molecular
weight cut off 1200-1400 KDa. Diffusion cell
(Fabricated by Sai Enterprises, Pune) of diameter
1.2 cm and 16 ml capacity consisted of upper
cylindrical compartment open from above and
diffusion membrane at its base. To prepare artificial
membrane, pieces of dialysis membrane were soaked in
PBS pH 7.4 for 24 hrs before mounting on diffusion
cell. Dialysis membranes were mounted in a two
chamber cells. In situ gels of Carbopol loaded with
drug were placed in the donor compartment. 16 ml of
PBS 7.4 was placed in the receptor compartment. The
temperature of receiver compartment was maintained at
the 370C ± 1.00C during experiment and the Table 4
content of the receiver compartment was stirred using
magnetic stirrer. The position of the donor
Table 2 Absorbance values of salbutamol sulphate in PBS 7.4 at 276 nm for preparation of standard curve.
Sr. No. Conc.
1 0
2 10
3 20
4 30
5 40
6 50
7 60
8 70
9 80
10 90
11 100
∗Mean (n=3) ± SD < 0.011
179
compartment was adjusted so that dialysis membrane
just touches the diffusion medium. An aliquot of 1 ml
was withdrawn from receiver compartment initially
after 15 and 30 min and then at 1-hr interval and
replaced with same amount of fresh medium. Aliquot
so withdrawn were suitably diluted and analyzed using
UV spectrophotometer at 276 nm for drug. In vitro
drug release study was carried out for 8 hrs.
Results and Discussion
The absorbances of Salbutamol Sulphate in PBS 7.4 at
276 nm (Table 2). The standard calibration curve of
Salbutamol Sulphate in PBS 7.4 is depicted in Fig. 1. In
situ nasal gel of Salbutamol Sulphate was prepared
successfully by using Carbopol 934 P alone and by using
Carbopol 934 P and HPMC K4M. It was found that all the
formulations had clear appearance. All the formulations
were found to have pH value in between 5.72 to 6.42
(Table 3). Results of the gelation studies are shown in
Table 3. As shown in Table 4 all the formulations were
found to have drug content in between 98.21 – 99.58%
w/w. Viscosities of all formulations in solution and gel
form are given in Table 4. The gel strength values between
25-50 seconds were considered sufficient as gel strength
less than 25 seconds may not preserve its integrity and may
erode rapidly while gels with strength greater than
50 seconds is too stiff and may cause discomfort
(Table 4). Table 5 and Fig. 2 & 3 shows release profile of
drug carbopol alone and carbopol with HPMC K4M. The
formulation B7 showed maximum cumulative percent of
drug release i.e. 86.21 ± 0.38%.
Absorbance∗
0
0.0543
0.1124
0.1710
0.2395
0.2740
0.3380
0.3625
0.4387
0.4960
0.5467
180 International Journal of Pharmaceutical Sciences and Nanotechnology Volume 1 • Issue 2 • July - September 2008

Fig. 1 Standard calibration curve of Salbutamol sulphate.

Equation of regression line: Y= 0.0054 X + 0.0048
R2= 0.9971

Table 3 pH and gelling capacities of prepared formulations

Formulations pH Gelling capacity

B1 6.4 -
B2 6.8 -
B3 6.5 ++
B4 6.4 ++
B5 6.1 ++++
B6 6.2 +++
B7 6.2 +++
B8 6.5 ++++
– No gelation
++ Gelation occured immediately and remained for a few hrs
+++ Gelation occured immediately and remained for extended period
++++ Very stiff gel

Table 4 Drug content, viscosity and Gel strength of prepared formulations

*Drug content Viscosity of liquid Viscosity of *Gel strength

Formulations
(%w/w) formulations (Pa.S) gels (Pa.S) (seconds)
B1 99.41 ±0.38 1.920 - -
B2 98.96 ±0.62 2.625 - -
B3 98.63 ±0.80 2.848 32.581 22.33 ±2.05
B4 99.38 ±1.18 3.285 34.900 27.00 ±0.84
B5 99.05 ±1.08 3.908 38.380 30.66 ±0.47
B6 99.58 ±1.15 3.315 36.920 28.00 ±0.94
B7 99.46 ±0.28 3.812 38.230 39.33 ±1.24
B8 98.21 ±1.12 4.214 40.160 51.66 ±2.05
∗Mean (n=3) ± SD
 Tanaji Nandgude et al. : Formulation and Evaluation of pH Induced… 181

Table 5 In vitro release profile of drug from prepared formulations

Sr. No. Time (hrs) Cumulative percent drug released

B3 B4 B5 B6 B7 B8
1 0 0 0 0 0 0 0
2 0.25 24.96±0.39 21.24±0.05 14.41±0.34 12.98±0.24 13.78±0.63 11.17±0.84
3 0.5 38.24±0.36 30.54±0.09 23.74±0.24 21.41±0.84 25.87±0.28 21.36±0.35
4 1 51.25±0.39 43.58±0.29 31.42±0.64 28.97±1.06 29.59±0.23 25.62±0.83
5 2 63.49±1.09 55.69±0.43 42.31±0.83 37.52±0.27 39.35±0.93 33.24±0.21
6 3 72.95±0.81 66.47±0.41 49.36±1.02 43.78±0.98 49.78±0.18 39.25±0..47
7 4 84.39±0.49 73.34±0.91 54.28±0.92 53.63±0.65 57.49±29 47.21±0.92
8 5 92.46±0.57 81.05±0.91 63.12±0.61 56.36±0.35 64.36±0.74 53.78±0.24
9 6 99.86±0.11 89.64±0.79 67.41±0.21 64.85±0.18 71.35±0.37 59.64±0.74
10 7 - 99.49±0.64 72.21±0.78 74.19±0.14 77.47±0.84 64.52±.0.84
11 8 - - 79.18±0.74 79.84±0.71 86.21±0.38 71.59±0.42

Fig. 2 Release profile of drug from prepared Formulations (B3, B4, B5).

Fig. 3 Release profile of drug from prepared Formulations (B6, B7, B8).
182 International Journal of Pharmaceutical Sciences and Nanotechnology
From the results it was found that standard calibration
curve at 276 nm followed Beer-Lamberts law in the
concentration ranging from 10-100 μg/ml. For the
formulation of in situ gels, carbopol 934 P was selected as
pH inducing gelling agent. HPMC K4M was added as
viscosity builder and mucoadhesive agent in order to
reduce the concentration of carbopol just sufficient to
induce gelation while retaining the same mucoadhesion as
that of higher concentration of carbopol. All the
formulations had clearance appearance. All the
formulations had pH values in the nasal pH range. From
gelation study it was observed that carbopol solutions with
concentration equal to or less than 0.2 %w/v had free
flowing property at non-physiological condition, however
these composition could not form strong gel at
physiological condition. On the other hand, for the
carbopol concentration equal to or greater than 0.5%w/v
very stiff gel was formed at physiological pH. As the
concentration of carbopol was increased solution become
highly acidic and it would not be neutralized by the
buffering action of the nasal pH, which might irritate nasal
mucosa. The carbopol solution of 0.4%w/v retained liquid
state at pH 4.5 and gelled at physiological pH. It was also
found that a reduction in the concentration without
compromising in situ gelling properties can be achieved by
adding a suitable viscosity enhancing polymers such as
HPMC which improved and strengthen the gel integrity.
From the results of gelation study the optimum
concentration of carbopol to be used as in situ gel forming
agent was selected as 0.4%w/v, which was used for further
studies with HPMC. All the formulations were having
drug content within the limit i.e. 97.5%-102.5%. The
formulations should have an optimum viscosity, which will
allow its easy administration into nasal cavity, as a liquid,
which will then undergo rapid sol to gel conversion. From
the results it was found that marked increase in viscosity
values was observed in all formulations at nasal pH due to
sol to gel conversion, except B1 and B2. Hence these two
formulations did not have any importance for further
evaluation and study. Formulation B3 and B8 had gel
strength value beyond the limits of 25-50 seconds. From
the results it was found that formulations B4, B5, B6 and
B7 showed suitable gel strength, did not have postnasal
drip and were easily administered as drops. From the
results it was concluded that the initial release rate were
very rapid due to incomplete gel formation, but the release
became slow after complete gel formation and remained
so. The release profiles exhibited an inflection point, which
indicated gel formation on the diffusion membrane in
donor compartment of diffusion cell. During gel formation,
formulation got converted into the gel phase and thus drug
release became slow. The results showed that the formed
gels had the ability to retain Salbutamol Sulphate for the
duration of 8 hours. In vitro release study indicated that the
Volume 1 • Issue 2 • July - September 2008
release of drug varied according to concentration of
polymers. As gel prepared using carbopol alone cannot
sustain release below 0.5% and concentration above 0.4%
may irritate nasal mucosa, in situ gels were further
prepared using carbopol and HPMC in combination. In
vitro release of drug from these formulations B6, B7 and
B8 indicated that a combination of HPMC in carbopol
formulations is highly effective in sustaining the drug
release. Amongst these formulations formulation B7 was
considered as optimized formulation as it showed the best
in-vitro release profile i.e. 86.21%.
Conclusion
From the results it can be concluded that Salbutamol
sulphate was successfully formulated as a pH induced in
situ nasal gelling system using Carbopol 934. The
optimized formulation B7 provided sustained in vitro
release of drug over an extended period of 8 hrs. The
optimized formulation can be a competent alternative to
conventional nasal drops. As a consequence of its
enhanced bioavailability and longer residence time, it
avoids the first pass effect and reduces the dosing
frequency as well.
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