In vitro Comparative Forced Degradation Study of Different Brands
and Active form of Montelukast sodium using UV
Spectrophotometer
Md. Jakaria*, Md. Hazrat
Ali, Md. Areeful Haque,
Mohammed Abu Sayeed, Shoayeb
Ahmed
Department of
Pharmacy, International Islamic University Chittagong (IIUC), Chawkbazar, Chittagong-4203, Bangladesh
*Corresponding Author E-mail: pharmajakaria@rocketmail.com
ABSTRACT:
The objective
of this study was to develop the degradation studies of different brands of montelukast sodium in the market and its active form.
Forced degradation is a process that involves degradation of drug products and
drug substances at conditions more severe than accelerated conditions and thus
generates degradation products that can be studied to determine the stability
of the molecule. Montelukast Sodium
(1-[[[(1R)-1-[3-[(1E)-2-(7-chloro-2-quinolinyl) ethenyl]
phenyl]-3-[2-(1-hydroxy-1- methylethyl) phenyl] -propyl] thio] methyl] cyclopropaneacetic acid, monosodium salt is a white colored
powder and it is freely soluble in ethanol, methanol, and water. It is a leukotriene receptor antagonist and, used for the
maintenance treatment of asthma and to relieve symptoms of seasonal allergies
This drug was subjected to different stress conditions as per International
Conference on Harmonization guidelines (ICH). An ultraviolet UV spectroscopic
method was developed for analysis of the drug in the presence of the degradation
products. Methanol was used as a solvent. The amount of degraded drugs was
calculated by taking the absorbance at 285 nm. According to the assay limit of
USP specified that the content should not be less than 95% and not more than
105% of labelled amount. All brands and its active
form were degraded on basic pH and on acidic pH. In
addition to heat exposure all brands and active form were also degraded. It was
concluded that all brands and active form of montelukast
degraded from ranges for all the stresses applied for degradation studies.
KEYWORDS: Degradation
studies, cyclopropaneacetic acid, montelukast
sodium, leukotriene receptor, assay.
1. INTRODUCTION:
Chemical stability of pharmaceutical
molecules is a matter of great concern as it affects the safety and efficacy of
the drug product. The FDA and ICH guidance’s state the requirement of stability
testing data to understand how the quality of a drug substance and drug product
changes with time under the influence of various environmental factors
Knowledge of the stability of molecule helps in selecting proper formulation
and package as well as providing proper storage conditions and shelf life,
which is essential for regulatory documentation. Forced degradation is a
process that involves degradation of drug products and drug substances at
conditions more severe than accelerated conditions and thus generates
degradation products that can be studied to determine the stability of the
molecule.
The ICH guideline states that stress
testing is intended to identify the likely degradation products which further
helps in determination of the intrinsic stability of the molecule and
establishing degradation pathways, and to validate the stability indicating
procedures used.[1-5]
Montelukast Sodium
(1-[[[(1R)-1-[3-[(1E)-2-(7-chloro-2-quinolinyl) ethenyl]
phenyl]-3-[2-(1-hydroxy-1-methylethyl) phenyl]-propyl]
thio] methyl] cyclopropaneacetic
acid, monosodium salt is a white colored powder and it is freely soluble in
ethanol, methanol, and water. Molecular weight of Montelukast
Sodium is 608.2 g/mol and formula is C35H35ClNO3
S.Na., for structure refer figure 1.[6-7] It is a leukotriene receptor antagonist and, used for the
maintenance treatment of asthma and to relieve symptoms of seasonal
allergies.[8-9] The drug is commercially available in various forms of once
daily oral dosage formulations including oral granules. In oral dosage form,
each packet contains Montelukast Sodium equivalent to
10 mg of Montelukast.[4]
Figure 1: Structure of Montelukast sodium
Several analytical methods have been
reported for determination of Montelukast including
derivative spectroscopic [10], by colorimetry [11],
by flouorimetry [12], by TLC [13], by HPTLC [14], by
simultaneous UV determination in combination drug formulation [15], by voltametry [16], by high performance liquid chromatography
(HPLC) [17] and by LCMS.[18] The aim of the present work was to develop and
validate a simple UV spectrophotometric method to be applied for analysis of montelukast sodium degradation in tablets and its active
form as per ICH guidelines, which serves as a tool for the quality control of
pharmaceutical dosage forms. Our research group has done these types of
degradation studies of different brand drugs such as atorvastatin,
rabeprazole, amytriptyline,
domperidone [2-5] and these are very helpful for
health care professionals.[19-21]
2. EXPERIMENTAL:
2.1. Reagents
Analytical grade reagents were used 0.1 N
sodium hydroxide, 0.1 N hydrochloric acid, de-mineralized water and distilled
water.
2.2. Glass wares
Volumetric flask, funnel, beakers,
Measuring cylinder, pipette, and stirrer used were of Pyrex type and were
washed with chromic acid followed by thorough washing with water and finally
rinsed with distilled water which was freshly prepared in the laboratory.
2.3. Instruments
Theses include
Spectrophotometer:
UV-Vis spectrophotometer, UV mini-1240, Shimadzu.
Corvettes
Weighing
Balance: Precision balance, LF224DR, Shinko Denshi Co., ltd.
Water
Bath: Stainless-steel, thermo station, HH-S
2.4. Wavelength Selection
About 100 ppm of montelukast sodium was accurately prepared in methanol. The
wavelength maxima (λmax) was observed at 285 nm
and this wavelength was adopted for absorbance measurement.
2.5. Preparation of 0.1 N Sodium Hydroxide
0.4 grams of sodium hydroxide were taken
and transferred it in a 100ml volumetric flask and dissolved it in small
quantity of water and finally make up the volume up to the mark of the flask
with de-mineralized water.
2.6. Preparation of 0.1 N Hydrochloric Acid
8.36 ml analytical grade hydrochloric acid
(37%, 12N) was taken in a volumetric flask and de-mineralized water was added
to making up to the volume.
2.7. Standard Stock Solution
All tablets of brand were labelled to contain montelukast
10 mg per tablet. Showing manufacturing and expire date of different brands and
active form (Table 1). Weigh and finally crushed tablets accurately for making
primary solutions of Montelukast 10 mg, Aeron (0.2482
gm) Healthcare Pharmaceuticals Limited., Montene
10 (0.1738 gm) Square Pharmaceuticals Ltd., Lumona
10 (0.3042 gm) Eskayef Bangladesh Ltd., Monas 10 (0.2502 gm) ACME Laboratories Ltd. and active form
(0.10 gm) equivalent to 10 mg were weighed accurately and introduced in 100 ml
volumetric flasks. Methanol was added and shaken vigorously and was making up
the volume up to 100 ml to make the strength of the solution 100 ppm in 100 ml.
2.8. Procedure
2.8.1. For Acid
To study the effect of acid, 5 ml of 100 ppm solution of each brand and active form was taken in
five separated test tubes, then 5ml of 0.1N HCl is
added in each test tube. They were then left for a period of 1 hour. Upon
completion of the time period, solutions were transferred to a cuvette separately and then absorbance of the solutions was
recorded at the wavelength of 285 nm.
2.8.2. For Base
To study the effect of base, 5 ml of 100 ppm solution of each brand and active form was taken in
five separated test tubes, then 5 ml of 0.1N NaOH is
added in each test tube. The samples were then left for a period of 1 hour.
Upon completion of time period, solutions were transferred to a cuvette separately and then absorbance of the solutions was
recorded at the wavelength of 285 nm.
2.8.3. For Heat
To study the effect of heat, 5 ml of 100 ppm solution of each brand and active was taken in five separated
test tubes, each containing 5 ml of water, then place these solutions in water
bath for 1 hour and absorbance of the solutions was recorded at the wavelength
of 285 nm.
3. RESULT:
This research was performed with the
purpose to compare the degree of degradation in four different brands and
active form of montelukast sodium 10 mg tablet. Table
2 shows the variation in absorbance after the effect of different degradation
parameters. After acidic pH and basic pH effect, percent of the assay was found
94.85%-43.41% (Table 3, 4 and Figure 2, 3). In addition to heat exposure
percent of the assay was found 69.97%-90.74% (Table 5 and Figure 4).
Figure 2: Showing acidic pH effect on the % of the assay
Figure 3: Showing basic pH effect on the % of the assay
Figure 4: Showing heat effect on the % of the assay
Table 1: Showing
manufacturing and expire date of different brands and active form
|
Sl No. |
Brand name |
Mfg. Date |
Exp. Date |
|
1. |
Aeron |
August,
2014 |
August,
2016 |
|
2. |
Montene |
September,
2014 |
August,
2016 |
|
3. |
Lumona |
July,
2014 |
July,
2017 |
|
4. |
Monas |
December,
2014 |
December,
2016 |
|
5. |
Active
form |
November,
2014 |
November,
2015 |
Table 2: Showing absorbance
of drug in different parameters
|
Sl. No. |
Brand name |
Absorbance of standard |
Absorbance after acidic pH effect |
Absorbance after basic pH effect |
Absorbance after heat effect |
|
1. |
Aeron |
2.971 |
1.533 |
2.735 |
2.494 |
|
2. |
Montene |
2.971 |
1.527 |
2.735 |
2.440 |
|
3. |
Lumona |
3.029 |
1.315 |
2.705 |
2.308 |
|
4. |
Monas |
2.971 |
1.385 |
2.404 |
2.079 |
|
5. |
Active
form |
2.971 |
1.508 |
2.818 |
2.696 |
Table 3: Showing effect of
acidic pH on different brands and active form
|
Sl. No. |
Brands |
% Assay |
|
1. |
Aeron |
51.59% |
|
2. |
Montene |
51.39% |
|
3. |
Lumona |
43.41% |
|
4. |
Monas |
46.61% |
|
5. |
Active
form |
50.75% |
Table 4: Showing effect of
basic pH on different brands and active form
|
Sl. No. |
Brands |
% Assay |
|
1. |
Aeron |
92.05% |
|
2. |
Montene |
92.05% |
|
3. |
Lumona |
89.30% |
|
4. |
Monas |
80.91% |
|
5. |
Active
form |
94.85% |
Table 5: Showing effect of
heat on different brands and active form
|
Sl No. |
Brands |
% Assay |
|
1. |
Aeron |
83.94% |
|
2. |
Montene |
82.12% |
|
3. |
Lumona |
76.19% |
|
4. |
Monas |
69.97% |
|
5. |
Active
form |
90.74% |
4. DISCUSSION:
The purpose of degradation studies was to
investigate those changes, to get a shelf life of the drug product and to
recommend storage conditions, which will be applicable to all future batches of
the tested drug product manufactured and packaged under similar
circumstances.[3] Montelukast was acidic drug, in
acidic medium percent of the assay less than basic medium. Forced degradation
study of montelukast was done by HPLC and result was
found that montelukast was degraded in effect of
acidic pH, basic pH and heat.[22] In our forced degradation study of montelukast was done by UV spectrophotometer. The limit of
the assay by USP specified that the content should not be less than 95% and not
more than 105% of labelled amount. According to this
USP specified limit, all brands were degraded in acidic and basic pH. After the heat exposure all brands were also degraded.
Effect of acidic pH, basic pH and heat exposure no brands of montelukast does comply with this USP specified limit.
5. CONCLUSION:
Montelukast was found to be degraded in
almost all types of stress conditions and was found to be less stable. It was
used to study the stress-degradation studies as per ICH guidelines. The method
was used is accurate and precise as well as reproducible and economical and can
be successfully used degradation studies of different dosage form. It was
concluded that all brands degraded from ranges for all the stresses applied for
degradation studies.
6. ACKNOWLEDGEMENT:
The authors greatly acknowledge to the
management system of Department of Pharmacy, International Islamic University
Chittagong (IIUC), for the supports of valuable instruments and equipments
during period of laboratory works.
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Received on 09.12.2014 Accepted on 11.01.2015
© Asian Pharma
Press All Right Reserved
Asian J. Pharm. Ana. 5(1): Jan.- March 2015; Page 26-30
DOI: 10.5958/2231-5675.2015.00005.8