INTRODUCTION:
Immediate availability of drug is required for
patients with conditions such as sudden episode of allergic attacks or angina pectoris or
local action desired such as local anaesthetic for toothaches, oral ulcers,
cold sores or teething. This can be
achieved by developing quick disintegrating/ dissolving oral films/tablets.The concept of fast dissolving drug delivery systems
has started because of its rapidly gaining interest in the pharmaceutical
industry for the reason of rapid disintegration
or dissolution, self-administration even without water or chewing.
These systems either dissolve or disintegrate
generally within a few seconds, without the need of water, making them
particularly suitable for paediatric and geriatric patients.The advantages of quick disintegrating oral films are
convenient dosing, taste masking, enhanced stability, improved patient
compliance, the drug enters the systemic circulation with reduced hepatic first
pass effect, dose accuracy in comparison to syrup and the potential advantage
is taste masking is always necessary for development of oral
pharmaceuticals.Fast dissolving oral films are useful in patients such as
paediatric, geriatrics, bedridden. The fast dissolving oral films place as an
alternative in the market due to the consumer’s preference for a
fast-dissolving product over conventional tablets/capsules. The oral thin film
technology is still in the beginning stages and has bright future ahead because
it fulfils all the need of patients. The criteria for selection of the drug are
it should have relatively low dose, bitter less in taste, drug should have
smaller and moderate molecular weight and have good stability and
solubility in water as well as saliva1-2.
For the present investigation, montelukast sodium and
levocetirizine dihydrochloride were selected as a model drugs. montelukast
sodiumis a leukotriene receptor antagonist (LTRA) used for the maintenance
treatment of asthma and to relieve symptoms of seasonal allergies, that is it
blocks the action of leukotriene D4 (and secondary ligands LTC4 and LTE4) on
the cysteinyl leukotriene receptor CysLT1 in the lungs and bronchial
tubes by binding to it. This reduces the bronchoconstriction otherwise caused
by the leukotriene and results in less inflammation3-4.
Levocetirizine
dihydrochloride is a third-generation non-sedative antihistamine, developed from the second-generation antihistamine cetrizine. Chemically, levocetirizine dihydrochloride is the
active enantiomer of cetrizine. Allergic rhinitis is a disorder of the
nose caused by inflammation mediated by immunoglobulin E(IGE) that lines the
membranes of the nose after exposure. It will prevent the release of other
allergy chemicals and increase blood supply to the areas. Combination of
montelukast sodium and levocetirizine
dihydrochloride provides additional benefits in comparison with each agent and
could be considered for patients whose quality of life is impaired by
persistent asthma and allergic rhinitis5-6.
In the present investigation
an attempt was made to develop simultaneous estimation of montelukastsodium and
levocetirizine dihydrochloride in the formulation which is a basic parameter to
quantify the amount of drugs present in a combination. Attempts were also made
to design and characterize fast dissolving film of montelukast sodium and
levocetirizine dihydrochloride to achieve increased dissolution rate, hence the
patient’s compliance and also to develop means of administration to patients
who are unable to swallow ordinary tablets and incase of motion sickness. The
main idea behind the fabrication of such dosage form is use of disintegrating
agent like sodium starch glycolate in different concentrations, which burst the
films immediately after placing over tongue, and release the medicaments in
saliva. In the formulation, sucrose is
included as sweetener. These systems may provide better qualities with enhanced
drug bioavailability.
MATERIALS AND METHODS:
Materials:
Montelukast sodium and levocetirizine dihydrochloride
were received as gift samples from Morpen lab(HP – Solon) and Micro lab
(Bangalore -India) respectively.Sodium starch glycolate obtained as a gift
samples from Wallace Pharma, Goa. Hydroxy propyl methyl cellulose (HPMC) E 15
LV and HPMC E50 LV, Loba chemie, Mumbai. Other chemicals and reagents used in
the study were of AR grade.
METHODS:
I. Preformulation studies:
Preformulation studies such as a melting point, IR
spectral analysis, compatibility studies andλmax, were determined by
appropriate methods to identify and to know the purity of the samples7.
II. Analytical method development for simultaneous
estimation of drugs:
Simultaneous equation method:
The method involves the measurement of absorptivity at
its λmax. Two wavelengths selected for the development of
simultaneous estimation of montelukast sodium (245nm) and levocetirizine
dihydrochloride(231nm) in the formulations. Two equations are constructed based
upon the fact that at λ1 and λ2, the absorbance of mixture is the some
of the individual absorbance of both the drugs.
Where,A1= absorption of sample at 245nm.
A2 = absorption
of sample at 231 nm.
ax1 = absorptivity
of montelukast sodium at 245 nm.
ax2 = absorptivity
of montelukast sodium at 231 nm.
ay1 = absorptivity
of levocetirizine dihydrochloride at 245 nm.
ay2 = absorptivity
of levocetirizine dihydrochloride at 231 nm.
Cx and Cy are concentration of montelukast sodium and
levocetirizine dihydrochloride in sample solution8.
Validation of Analytical method:
Linearity:
For each drug, appropriate dilutions of
standard stock solutions were assayed as per the developed methods. The
Beer-Lambert’s concentration range is 1-15μg/ml for montelukast sodium and
2-15μg/ml for levocetirizine dihydrochloride.
Accuracy and Precision:
Accuracy of the method was confirmed by
intraday and interday analysis. Intra-day
and Inter-day absorbance, standard deviation and relative standard deviation
(RSD) values were calculated. Low RSD value obtained from the analysis of drug
solutions indicated good precision of the method.
Recovery Studies:
The recovery of both the drug from
standard mixture was found to be in the range of 98%-101%.This result
indicates that the method was accurate, reproducible and the concerned drugs
could be quantified by this analytical procedure.
Robustness:
When small changes in the wavelength (± 5
nm) were introduced, it is observed that there was no change in the absorbance
values indicates that method was not influenced by the small errors.
III.Preparation
of drug loaded films:
The fast dissolving films of montelukast sodium andlevocetirizine dihydrochloride were prepared by solvent casting technique. The
quantities drug and non-drug components used in the formulation are as per
Table 4. Film forming polymers such asHPMC E15LV, HPMC
E50LV and HPMC K15M
were used. The calculated amount of polymer was soaked in three forth volume of
phosphate buffer of pH 6.8 solution with continuous stirring for about 8hours
to get uniform dispersion. The required amount of montelukast sodium and levocetirizine dihydrochloride was incorporated in the polymeric solutions. Sodium
starch glycolate, vaniline and sucrose were added to the polymeric solution. To
the above mixture citric acid, PEG400 were added and the final volume was adjusted
up to 12ml usingphosphate buffer of pH 6.8 solution. Now, this solution was set aside for the expulsion of
air bubbles for 3 hrs. The films were casted by pouring the above solution in
the glass moulds of 4×4 cm2 and were allowed to dry at the room temperature.
The dried films were peeled of from the mould, wrapped in aluminium foil and
were kept in the desiccator for further studies.
VI.
Evaluation of mouth dissolving films:
Physical
appearance and surface texture:
Physical
appearance was checked by visual inspection and surface texture was evaluated
by touch or feel of the film.
Weight
uniformity:
3
films of 1 cm2 from each formulation were taken, weighed
individually and average weight was calculated.
Thickness
uniformity:
The
thickness of the films was determined using screw-gauge micrometer. The
thickness of the each film was determined at three different places and the
mean was calculated.
Folding
endurance:
The
folding endurance of the films was determined by repeatedly folding and
unfolding of film at sameplace, till it broke.The number of times the film
could be folded at the same place without breaking gives the value of the
folding endurance. The procedure was repeated three times for each film and the
mean was calculated.
Surface
pH:
The
film to be tested was placed in petridish and moistened with 1ml of distilled
water and kept for 1 hr. The calibrated glass electrode was kept in contact
with the surface of the moistened film and allowed equilibrate for 1 min. The
pH was recorded. The procedure was followed three times for each film and the
mean was calculated.
Tensile
strength measurement:
The
tensile strength of the film was determined using the film of known dimension.
The forcerequired to break the film of 4 cm2 was recorded and the
tensile strength was calculated using the following formula:
Force at break
Tensile
strength =----------------------------
Cross sectional area of film
The
procedure was followed three times for each film and the mean was calculated.
Moisture loss study:
Desiccator containing anhydrous calcium chloride was
used to estimate moisture loss of the film.Initial weight of the film was
recorded before keeping the film in the desiccator. After 3 days final weight
of the film was recorded. The % moisture loss was calculated using following
formula.
Initial weight- final weight
Initial w % moisture loss =--------------------------------
× 100
Initial weight -
Final weight
The procedure was repeated three times for each films
and the mean was calculated.
In- vitro disintegration time:
Disintegration test was performed using USP
disintegration test apparatus. 4cm2 film was placed in the basket.
The assembly was suspended in 0.6% sodium lauryl sulphate and time required by
the film to disintegrate completely was noted.
Drug content determination:
Film of 4 cm2 was taken and transferred to
a 100ml of volumetric flask. The flask was filled with methanol, mixed
thoroughly. Dilute 1ml of the above solution to 100ml withphosphate buffer of pH 6.8 solution. The absorbance of the resulting solution was
measured at 245nm and 231nm using a Shimadzu UV-visible spectrophotometer.
In-vitro
drug release studies:
The dissolution studies were conducted using
dissolution test type – I apparatus (basket apparatus). Dissolution medium used for the study was 300 ml of
phosphate buffer of pH 6.8 solutionand 0.6%
sodium lauryl sulphate using. The rotation speed was adjusted to 50 rpm. The
temperature of dissolution medium was maintained at 37.00 ±0.5oC
throughout the experiment. Periodically 5 ml of aliquots was withdrawn and was
replaced with an equal volume of fresh dissolution medium. The samples were
filtered and suitably diluted with phosphate
buffer of pH 6.8 solution. The absorbance
of the solution was measured at 245 nm and 231nm using UV-Visible
spectrophotometer (Shimadzu UV -1800). Three films from each batch (formulation)
were used for the in-vitro dissolution test and mean values was
calculated.9-12.
Stability Studies:
Stability testing was performed by placing selected films at
40.00 ±0.5’C and 75.00 ±5%RH in a stability chamber for 6 months. During
stability testing drug content uniformity, drug release studies and
disintegration tests were performed13-14.
RESULTS:
The Beer-Lambert’s concentration range is 1-15
μg/mL, and 2-15 μg/mL for montelukast sodium and levocetirizine
dihydrochloride at 245 nm, and 231 nm wavelengths with coefficient of
correlation 0.999, and 0.999 respectively and results are shown in Table 1.
Precision of the method was determined by repeating assay 3 times. To study
intraday precision, method was repeated 3 times in a day and the average % RSD
was calculated. Similarly the method was repeated on three different days and
average % RSD was calculated. the values are reported in Table 2(a) and (b).
Accuracy was confirmed by recovery study at three different concentration
levels 50%, 100%, 150% by replicate analysis (n = 3). From the recovery study
it is clear that the method is accurate for quantitative estimation of
montelukast sodium and levocetrizine dihydrochloride, as the statistical
parameters are within the acceptance range. The results of are shown in Table
3. Mouth dissolving films comprising montelukast sodium and levocetirizine
dihydrochloride were prepared by solvent casting technique using disintegrating
agent, polymers, plasticizers, saliva stimulating agent, and sweetening agent.
Altogether 12 formulations were prepared as per the Table 4. The results of
evaluation parameters such as physical
appearance, surface texture, weight
uniformity, thickness uniformity, folding endurance, surface pH, tensile
strength measurement, moisture loss study, andin-vitro disintegration
time are given in Table 5. Drug content determination andin-vitro drug
release studies was conducted for all the formulations and the results are
shown in Table 6.Among all the formulations, F8 was chosen as a better
formulation and used for stability studies, and the results shown inTable 7. In-vitro
dissolution study was conducted for all formulations and the results are shown
in Figure 1-6.
Table
1: Spectral and Linearity Characteristic Data:
|
Equation method
|
|
PARAMETERS
|
montelukast sodium
|
levocetirizine
dihydrochloride
|
|
λmax (nm)
|
241
|
231
|
|
Linearity range(µg/ml)
|
1-15 µg/ml
|
2-15 µg/ml
|
|
Molar absorptivity(L/mol/cm)
|
0.0338
|
0.0314
|
|
Correlation coefficient (r2)
|
0.999
|
0.999
|
|
Slope (m)
|
0.0346
|
0.0315
|
|
Intercept (C)
|
-0.0015
|
0.0009
|
|
Regression equation
|
y=0.0346x-0.0015
|
y=0.0315x+0.0009
|
Figure 1: Comparative in-vitro release profile
ofmontelukast sodium (formulation
F1–F4)in phosphate buffer (pH 6.8)
Figure 2: Comparative in-vitro release profile
of montelukast sodium(formulation F5–F8) in phosphate buffer (pH 6.8)
Figure 3: Comparative in-vitro release profile
of montelukast sodium (formulation
F9–F12) in phosphate buffer (pH 6.8)
Figure 4: Comparative in-vitro release profile
of levocetrizine dihydrochloride (formulation
F1–F4) in phosphate buffer (pH 6.8)
Figure 5: Comparative in-vitro release profile
of levocetrizine dihydrochloride (formulation
F6–F8) in phosphate buffer (pH 6.8)
Figure 6: Comparative in-vitro release profile of
levocetrizine dihydrochloride (formulation
F9–F12) in phosphate buffer (pH 6.8)
DISCUSSION:
Based on the results of IR and melting point studies,
it was confirmed that the obtained drug samples meet the requirements for
purity. The compatibility study results confirm that there was no interaction
between drugs and excipients. Drugs show good regression values at their
respective wavelengths. Further, the precision of the method was confirmed by
Intraday and Interday analysis.The % RSD values for Intraday and Interday
analysis were found to be 0.0042 to 0.045 for montelukast sodium, 0.009 to
0.031 for levocetirizine dihydrochloride respectively. The % recovery was found
to be in the range of 98.33 to 101.20 for montelukast sodium, 99.66to 101.30
for levocetrizine dihydrochloride. This ensures the accuracy of the method.All
the batches were uniform in weight with no significant difference in the weight
of the individual formulation from the average value. The weights ranged
between 24.01±0.02 to 25.77±0.38 mg. All the films
have uniform thickness and ranged between 0.133±0.001 to 0.213±0.005 mm. The standard deviation values indicate that all the
formulations were having less variation. Folding
endurance gives an indication of brittleness of the film and did not show any
cracks even after folding for number of times. These results indicated that
films are flexible in nature. The surface pH was found to be in the range of
6.35±0.017 to 6.70±0.036 which was close to the salivary pH, which indicated
that films may have less potential to irritate the mucosal lining of the oral
cavity and hence more acceptable by the patients. Tensile strength of all
formulations was found to be in range of 1.15±0.07 to 2.77±0.14 the results of
tensile strength shows that all the batches of films are having sufficient
strength. All the formulations have showed very less quantity of moisture was
loss during and after study, and indicates that films are found to be flexible.
The in-vitro disintegration time was found to be in the range from 110.67±1.66
to 34.01±0.73 sec. the results indicated that the disintegration time is
directly proportional to the concentration of super disintegrating agents. i.e.
as the concentration of superdisintegrats increases the films disintegrates at
faster rate. The drug was content was found to be 91.78±2.32 to 95.54±0.87 and
91.87±4.38 to 95.74±0.43 of montelukast
sodium andlevocetirizine dihydrochloride
and result suggested that the drug content uniformity was maintained throughout
the formulation. All the formulations are subjected for in-vitro
dissolution study using USP type – I apparatus (basket type) using a phosphate buffer of pH 6.8 solution. The prepared films released the 94.40% ±0.48 to 95.66%
±0.24 of drug at the end of 225sec. Based on the above results, formulation F8
was selected as a best formulation because disintegration time (34.25sec), drug
content (95.54% and 95.74% for montelukast sodium and levocetirizine
dihydrochloride) and drug release ( 94.40% and 95.66% montelukast sodium and
levocetirizine dihydrochloride respectively). The promising formulation F8 was
subjected for short term stability studies. The results shows that there is no
change in the physical appearance, a negligible change in the drug content and
release studies were observed.
CONCLUSION:
In
present research work, an attempt has been made to prepared fast dissolving
films containing montelukast sodium and levocetirizine dihydrochloride was repared
by solvent casting method using various polymers like HPMC E15LV, HPMC E50LV,
HPMC K15M and disintegrating agent like sodium starch glycolate with different
concentrations (1-5% w/w). From the study it was observed that that the film
prepared using 5% w/w sodium starch glycolate showed disintegration time, drug
content and in-vitro drug release, and hence F8 was selected as best
formulation.
ACKNOWLEDGEMENTS:
The
authors are thankful to Principal and Management, Maratha Mandal’s College of
Pharmacy, Belgaum for providing necessary facilities to conduct this research
work. The authors are grateful to Morphen lab Ltd. (Solon) and Micro labs
(Bangalore) for providing gift samples of montelukast sodium and levocetirizine
dihydrochloride, and to Wallace pharma (Goa) for providing super-disintegrating
agents.
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