World Journal of Pharmaceutical Research

Karpagavalli et al. SJIF Impact

World Journal of Pharmaceutical Factor 7.523
Research
Volume 6, Issue 2, 940-951. Research Article ISSN 2277 7105

FORMULATION AND EVALUATION OF ZOLPIDEM NASAL IN SITU

GEL

L. Karpagavalli1*, K. Gopalasrsatheeskumar1, N. Narayanan1, A. Maheswaran1, A.

Isakki Raj1 and J. Hari Priya1

Jaya College of Paramedical Sciences, College of Pharmacy, Thiruninravur, Tamilnadu,

India.

ABSTRACT
Article Received on
09 Dec. 2016, In this work, Zolpidem nasal in situ gel was prepared and evaluated.
Revised on 29 Dec. 2016, TheZolpidem nasal in situgel were prepared by Temperature induced
Accepted on 19 Jan. 2017
DOI: 10.20959/wjpr20172-7768 InSitu gelling system using various concentrations of carbopol934p
and poloxamer(F1-F6). The prepared nasal in situgel were

*Corresponding Author characterized for its Appearance,Viscosity, pH, Drug content, Gelation
L. Karpagavalli temperature, Gel strength, Mucoadhesive strength, and invitro drug
Jaya College of Paramedical release profile. The preformulation study results confirmed the
Sciences, College of
compatibility between the drug and other excipients used in the
Pharmacy, Thiruninravur,
formulation. The optimized formulation was selected based on itsgood
Tamilnadu, India.
gelation temperature and gel strength and invitro drug release profile.
Thus it can be concluded that nasal in situ gel containing (F4) 0.5%carbopol&18%poloxamer
concentration is an ideal gel for controlled and sustained nasal drug delivery system.

KEY WORDS: Zolpidem, Carbopol934p, Poloxamer, insitu, Gel strength, Hypnotic.

INTRODUCTION
The generation of a new drug molecule is an expensive and time consuming process. Hence
the safety and efficacy ratio of old drugs can be improved by delivering these drugs at
controlled and slow delivery or targeted delivery.

The human nose has the potential to be an alternative route for the systemic delivery which
has wide range of therapeutic agents. They have been richly supplied with vascular nature of
the nasal mucosa and its high drug permeation makes it a potential route for the
administration of many drugs including proteins, peptides and vaccines.[1] Nasal route is most

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suitable for those drugs which cannot be administered orally due to gastric degradation or
hepatic first pass metabolism.[2]Insitu forming polymeric systems are formulations that are in
solution form before administration into the body but once administered, undergo gelation in
situ, to form a gel.[3] The present work was aimed to formulate and evaluateZolpidem nasal
insitu gel. It is prepared by using polymers to sustain the release of Zolpidem for the
treatment of insomnia.

To achieve the objectives the plan is executed to drugs are orally not absorbed can be
delivered to this route, hepatic first pass metabolism is avoided, enhance the bioavailability,
reduced the dose and dosing frequency, improved patient compliance and improvement in
time residence so it gives the sustained drug release.

MATERIALS AND METHODS

Materials Used
Zolpidem, Carbopol 934p and,Poloxamer407 were purchased from Sigma Aldrich, Propylene
glycol, sodium chloride, propyl paraben was purchased from Loba Chemicals, Mumbai,All
other chemicals and reagents used were of analytical grade.

Methods
Preparation of nasal insitugel formulation by (Temperature induced InSitu gelling
system)
It is prepared by dispersing polymers in distilled water with continuous stirring (Thermostatic
hot plate with magnetic stirrer, Remi) until completely dissolved and allowed to hydrate
overnight. The preparation of solution, first poloxamer was added in distilled water and
allowed to hydrate. Then carbopol was sprinkled over the solution and allowed to hydrate
overnight. After complete hydration of polymers a separate solution of Zolpidem tartrate and
sodium chloride was added to the polymeric solution.[4,9] The resultant solution was
thoroughly mixed,Propylparaben was added and mixed until a uniform and clear solutions
were formed. Finally volume was makeup to required volume of distilled water. All the
formulations were adjusted to pH 4.5 to 5.5 by using freshly prepared 0.5 M sodium
hydroxide solution.

Preparation of Zolpidem tartrate

Accurately weighed amount of polymers mixed with 10ml of distilled water, preheated to
600c followed by addition of propylene glycol (5ml). To this 0.15gm of Zolpidem was

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added and mixed thoroughly to get an homogeneous solution 500c and then add drop wise
Propyl paraben (0.02%w/v) then pre heated at 600c with magnetic stirrer until to get an
homogeneous solution. Then sodium chloride was added drop wise for an 1hour with
continuous stirring at room temperature and it is kept stabilize to get in situ gel. Then the
mixture was left to cool at 5 to 100c for 30mins to enhance the settling of insitu gel.

Several batches namely (F1, F2, F3,and F4, F5, F6) were formulated by changing the drug
and polymer (carbopol934p and poloxamer) ratio.

Table 1: Composition of nasal in situ gel formulation of Zolpidem

INGREDIENTS(mg) F1 F2 F3 F4 F5 F6
Zolpidem 150 150 150 150 150 150
Carbopol934p 0.2 0.3 0.4 0.2 0.3 0.4
Poloxamer 0.3 0.3 0.3 0.4 0.4 0.4
Nacl 180 180 180 180 180 180
Propyl paraben 0.02 0.02 0.02 0.02 0.02 0.02
Propylene glycol(ml) 5 5 5 5 5 5
Distilled water q.s q.s q.s q.s q.s q.s

Characterization of Zolpidem nasal insitu gel

Appearance
The developed formulations were inspected visually for clarity in sol and gel form.

Viscosity and rheological behavior studies

Viscosity of formulations before and after gelation are measured by Brookfield R/S CPS +
Rheometer with software Rheo 3000 and using spindle CP75 at 100 rpm shear rate.[5]

pH of the formulation
The pH of the each formulation was determined by using pH meter.

Drug content
To 1 ml formulation taken in a 50 ml volumetric flask phosphate buffer pH 6.4 was added for
dilution and shaken to dissolve the drug. The solution was filtered through 0.45 PVDF
syringe filter,1ml of above filtrate was pipette out and diluted to 10 ml with phosphate buffer
pH 6.4. The drug content was estimated spectrophotometrically by using standard curve
plotted at 294.2nm.

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Gelation temperature
Gelation Temperature, defined as the temperature at which the liquid phase makes the
transition to a gel, determined by using method described by Miller and Donovan technique.
A 2ml aliquot of gel was transferred to a test tube, immersed in a water bath. [6] The
temperature of water bath was increased slowly and left to equilibrate for 5 min at each new
setting. The sample was then examined for gelation, which was said to have occurred when
the meniscus would no longer move upon tilting the test tube to 900.

Gel strength determination

It is expressed in time (in seconds) required by a 35 g piston for penetration of 5 cm distance,
through the 50g gel formulation. Test was performed using Gel strength apparatus
modified in laboratory.[7]Formulation(50g) was placed in a 100 ml measuring cylinderand
gelation was induced by Simulated Nasal Fluid. Theapparatus i.e. piston for measuring gel
strength (35g) was thenplaced onto the gel. The gel strength was measured as the time(in
seconds) required for moving the apparatus 5 cm downthrough the gel. In cases, that take
more than 5 min to drop theapparatus into the gel, suitable weights were placed on top ofthe
apparatus and gel strength was described by the minimalweights that pushed the apparatus
5cm down through the gel.

Figure 1: Gel strength measuring device

(A) Weights (B) Device (C) Graduated cylinder (D) gel

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Mucoadhesive strength study

Mucoadhesive force of nasal phase transition system was determined using sheep nasal
mucosa and phosphate buffer pH 6.4 as the moistening fluid at the time of testing, a section
of tissue was secured, keeping the mucosal side out, onto each glass vial using a rubber band
and aluminium cap. The diameter of each exposed mucosal membrane was 1.1 cm. On glass
vials, tissues were fixed in a manner that the mucosal side became outer part and properly
fixed. A vial with a section of tissue was connected to the modified balance and suitable
height was maintained. The gel was applied to the exposed tissue of lower vial. The height of
the vial was adjusted so that the gel could adhere to the mucosal tissues of upper vial. After
applying constant weight for several minutes, suitable weights were added to the modified
balance. Minimum amount of weight that detached two vials expressed as mucoadhesive
force (dyne /cm2).[8]
Detachment stress (dynes /cm2) =M+G/A
Where,
Mis the weight added to balance in grams;
G is the acceleration due to gravity taken as 980 cm/sec2;
A is the area of the tissue exposed and is equal to r2 (r, the radius of the circular hole in the
aluminum cap).

Figure 2: Modified balance for bio-adhesive study

A: Modified balance, B: Weighing pan, C: Weight D: Gel, E: Nasal mucosa F:
Polypropylene cylinder.

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In vitro release studies

Invitrodiffusion study of formulated in situ gels was carried out on Franz diffusion cell
having 2.4 cm diameter and 13 ml capacity. Dialysis membrane having cut off molecular
weight1200014000 kDa was used as diffusion membrane. Pieces of dialysis membrane were
soaked in phosphate buffer pH 6.4 for 24 hrs prior to experiment.[10,11] Diffusion cell was
filled with phosphate buffer pH 6.4; dialysis membrane was mounted on cell. The
temperature was maintained at 37 0.5C. The donor compartment contained 3 ml of
artificial nasal fluid. After an equilibration of membrane, formulation equivalent to 1 mg of
Zolpidem tartrate was placed in the donor compartment. At predetermined time points (30,
60, 90, 120, 150,and 180 min), 1 ml samples were withdrawn from the acceptor
compartment, replacing the sampled volume with phosphate buffer pH 6.4 after each
sampling to maintain a constant volume, for a period of 5hr The samples withdrawn were
filtered and used for analysis. Blank samples (without Zolpidem tartrate were run
simultaneously throughout the experiment to check for any interference. The amount of
diffused drug was determined using UV visible spectroscopic method.

Figure 3: Laboratory designed diffusion cell

A- Test tube containing formulation. B- Egg membrane. C-beaker containing simulated nasal
solution. D- Magnetic stirrer.

RESULTS AND DISCUSSION

Characterization of Zolpidem
Description
The physical appearance of Zolpidemtatarate was found to be white crystalline powder form.

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The solubility results of Zolpidemtatarate in buffer were given below,

Viscosity measurements and rheological studies

Viscosities of all the formulation was determined at pH 4.4, the formulations exhibited low
viscosity and were in solution form. An increase in pH to 6.5 (pH of nasal fluid) using 0.5 M
NaOH transformed the solution into gel and showed increasein viscosity.

Table 2: Viscosities of prepared formulation

VISCOSITY OF VISCOSITY
FORMULATION CODE
SOLUTION(cps) OF GEL(cps)
F1 15.06 1138
F2 26.75 1369
F3 74.13 1474
F4 196.33 1512
F5 228.66 1586
F6 285.69 1701

Gelation temperature
Gelation temperature range suitable for nasal gel is 32-35C. As the concentration of the
poloxamer -407 [PF-127] was increased gelation temperature was decreased. Increasing the
concentration of PF-127 to 20% [w/w] causes gelation in the temperature range of 28-34C.
The temperature dependent gelation of PF-127 molecules exhibit a well arranged zigzag
configuration of poloxamer, may be transformed into a close packed meander configuration,
forming a more close pack and more viscous gel. As the concentration of PF-127 increases,
the gel structure becomes more closely packed with the arrangement in the lattice pattern.
Gelation temperature lowering effect of mucoadhesive polymer could be explained by its
ability to bind polyethylene oxide[PEO]. Chains present in the PF-127 molecules promote
dehydration and causes increase in entanglement of adjacent molecules with more extensive
intermolecular hydrogen bonding.

Table 3: Gelation temperature

GELATION
FORMULATION CODE
TEMPERATURE(0C)
F1 29.5 40.2
F2 32.4 37.4
F3 30.5 35.3
F4 32.1 33.2
F5 35.3 37.4
F6 37.4 29.4

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Drug content
The percent drug content of all formulations (F1 to F6) was found to be in range of 92.08
100.50%.

Table 4: Drug content of nasal insitu gel

DRUG
S.NO FORMULATION CODE
CONTENT %
1 F1 93.08
2 F2 99.17
3 F3 96.38
4 F4 95.05
5 F5 98.06
6 F6 99.25.

Gelation studies
The gelling capacity of prepared formulations was observed by visual examination and it is
graded based on the nature of gel formed.
-No gelation occurred.
+ Gelation occurred in few min and it is remained for few hrs.
++ Gelation immediate remained for few hrs.
+++ Gelation Immediate, and for extended period.
++++ Very stiff gel

Table 5: Gelling capacity of insitu gel

FORMULATION
S.NO GELLING CAPACITY
CODE
1. F1 -
2. F2 ++
3. F3 ++
4. F4 +++
5. F5 +++
6. F6 ++++

From the gelation studies, it was observed that all the formulation showed gelation.
Formulation F1 gel does not occur, F2, F3showed weakest gelation Formulations F5-F6
showed good gelation.

Measurement of gel strength

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

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with strength greater than 50 seconds is too stiff and may cause discomfort. Formulation F1,
F2, F3 showed gel strength value below 25 sec. From the results it was found that
formulationsF5 F6 showed suitable gel value.

Table 6: Measurement of gel strength of prepared formulation

FORMULATION CODE GEL STRENGTH(sec)
F1 12.50
F2 15.40
F3 19.30
F4 28.20
F5 29.30
F6 35.40

GELATINE STRENGTH
GELATINE TEMPERATURE

40
35.4
30 28.2 29.3
19.3
(0 C)

20 12.5 15.4

10

0
F1 F2 F3
F4
F5
F6

BATCH CODE

Gel strength

Figure 4: Gelatine strength of nasal insitu gel

Mucoadhesive strength
Mucoadhesive strength was determined in term of detachment stress i.e. force required to
detach the formulation from mucosal surface. All formulations were subjected to in vitro
mucoadhesion studies. Results indicated that the variation in concentration of Poloxamer 407
and carbapol 934P showed changes in mucoadhesive strength. The gradual increase was
observed in mucoadhesive strength with the Poloxamer 407 and carbapol934P level.

Table 7: Measurement of mucoadhesive strength of prepared formulation

MUCOADHESIVE
FORMULATION CODE
STRENGTH(dyne/cm2)
F1 69.30
F2 72.29
F3 75.37
F4 82.13
F5 83.36
F6 87.84

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In vitro diffusion study

From the results it was concluded that the initial release rate was 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 indicates 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 Zolpidem tartrate for the duration of 8
hours. In vitro release study indicated that the release of drug varied according to
concentration of polymers. In situ gels were prepared using carbopol 934 and Poloxamer 407.
In vitro release of drug from formulations F6 indicated that a combination of carbopol- 934 in
poloxamer 407 formulations is highly effective in sustaining the drug release up to 8 hours
due to increased concentration of polymers.

Table 8: Invitro studies of Zolpidem nasal insitu gel

TIME % CUMULATIVE DRUG RELEASE
SOLUTION
(mins) F1 F2 F3 F4 F5 F6
0 0 0 0 0 0 0 0
30 36.06 13.88 9.7 15.45 25.28 11.6 16.60
60 48`04 18`14 14.2 23.38 41.25 23.7 22.18
90 60.50 33.34 18.70 35.34 42.76 25.9 34.86
120 69.68 36.35 27.62 38.98 45.47 39.8 38.99
150 80.69 45.30 39.29 45.67 50.64 55.5 45.28
180 89.26 100.9 67.60 58.56 63.07 65.1 55.68

Figure 5: invitro studies of nasal insitu gel

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SUMMARY CONCLUSION
Zolpidem tartrate for nasal administration could have potential to avoid first-pass effect then
oral route, thus improve bioavailability of drug and as a safe, sustained release nasal delivery
system to control insomnia.

Zolpidem tartrate was successfully in Temperature induced in situ gel system using carbopol
934 as a temperature induced in situ gelling agent in combination with poloxamer 407 as a
viscosity enhancing agent.

The Zolpidem nasal insitu gel were formulated and evaluated for, appearance, viscosity,
gelation temperature and drug content, gelling capacity, gel strength, mucoadhesive strength
and drug release.

The result showed that in vitro drug release for F4 is 96.54, respectively.

The in vitro % drug release of F6 formulation was 96.54 and it was found to be suitable
formulation for the treatment of insomnia patients. Hence it can be concluded that the newly
formulated controlled and sustained release nasal in situ drug delivery system of Zolpidem
may be ideal effective to control the insomnia attacks by allowing the drug to release
continuously for 8 hr.

The temperature induced system is found to best suitable method for preparation of Zolpidem
tartrate nasal insitu gel. Prepared nasal insitu gel showed good gelation temperature and gel
strength. Thus it can be concluded that nasal insitu gel containing (F4)
0.5%carbopol&18%poloxamer concentration is an ideal gel for controlled and sustained
nasal drug delivery system with respect to its evaluation parameters like gelation temperature
and drug release.

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