Indo American Journal of Pharmaceutical Research, 2014

ISSN NO: 2231-6876

SPECTROSCOPIC METHODS FOR THE SIMULTANEOUS ESTIMATION

MOMETASONE FUROATE AND FORMOTEROL FUMARATE IN ROTACAPS.

OF

Aarti. S. Zanwar*1, Dhanya B. Sen1, Dipti B Ruikar2, Dr A.K. Seth1.

1

Department of Pharmacy, Sumandeep Vidyapeeth, Piparia,Vadodara-391760,Gujarat.

Pharmacy Department, The Maharaja Sayajirao University Of Baroda, Vadodara, 390 002, Gujarat, India.

ARTICLE INFO
Article history
Received 21/12/2014
Available online
31/12/2014
Keywords
Mometasone Furoate,
Formoterol Fumarate,
Spectrophotometric Method,
Analysis.

ABSTRACT
Two spectrophotometric methods Zero crossing derivative and ratio- spectra derivative
spectrophotometric methods were developed for the determination of Mometasone furoate
(MF) and Formoterol fumarate (FF) in the pharmaceutical dosage form. Zero crossing
dervative spectrophotometry involves amplitudes measurement of the first derivative spectra
of the standard and sample solution at 267.56 nm for MF and 306 nm for FF. Ratio spectra
derivative spectrophotometry involves amplitudes measurement of the ratio first derivative
spectra at 263nm and 267.30 nm for MF and 293.08 and 302.60 nm for FF. The calibration
graph follows beers law in the range of 30-200 g /ml of MF and 2-6 g /ml for FF. The
accuracy and precision of the method were validated statistically. So it can be a preferable
method for routine analysis due to its simplicity and economical advantages.

Copy right 2014 This is an Open Access article distributed under the terms of the Indo American journal of Pharmaceutical
Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Please cite this article in press as Aarti. S. Zanwar et al. Spectroscopic methods for the simultaneous estimation of Mometasone
furoate and Formoterol fumarate in rotacaps. Indo American Journal of Pharm Research.2014:4(12).

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Corresponding author
Aarti. S. Zanwar
Department of Pharmacy,
Sumandeep Vidyapeeth, Piparia,
Vadodara-391760,Gujarat
aarti.zanwar @ gmail.com

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INTRODUCTION
Nowadays combinational therapy of mometasone fumarate and formoterol furoate has been used in the treatment of COPD
and asthma as it produces the additive effect for improving the symptom, lung function and reduces exacerbation in patient.[1]
Mometasone furoate (MF) (Fig.1) is a highly potent synthetic chlorinated glucocorticosteriod. It is used in the treatment of
glucocorticoid responsive disorders of dermatology, seasonal and perennial allergic rhinitis and asthma. [2]. Formoterol fumarate (Fig
2) appear to be more effective than shorter acting 2-agoinst in the treatment of nocturnal and exercise induced asthma. It acts locally
in the lung as a bronchodilator [3]. Literature survey reveals that serval analytical method have been published for the estimation of
MF alone or in combination with other drugs like fucidic acid, terbinafine HCl, miconazole, eberconazole nitrate etc. Some of these
methods include HPLC, GC, supercritical fluid chromatography and UV spectrophometry[4-11]. Various method has also been
reported for the estimation of FF alone and in combination with other drugs including HPLC, GC, and UV spectrophometry [12-14].
However, no UV method has been reported for the simultaneous estimation of MF and FF in rotacaps by zero crossing derivative and
ratio derivative spectrophotometry method. So, the aim of the present work was to develop simple, sensitive and validated two
spectrophotometric methods for the simultaneous determination of MF and FF in their pharmaceutical preparations.

Fig1 Chemical structure of MF

Fig 2 Chemical structure of FF

Experimental work
Instrumentation:
Shimadzu UV-1800 spectrophotometer connected to computer loaded with Shimadzu UV probe 2.10 software was used for
all the spectrophotometeric measurements. All weighing were done on electronic balance( model Ohaus adventurer Pro).
Ultrasonicator was used for sample solution preparation.
Reagents and chemicals:
Analytical pure sample of MF and FF were obtained as a gift sample from Sun Pharma, Vadodara. These sample were used
without further purification. The formulation Evocort manufactured by Cipla was purchased from the local market containing MF
(200g) and FF (6 g) per rotacap. Analytical grade methanol was purchased from Merck Mumbai was used throughout the study.
Preparation of standard solution and calibration curve:
Individual standard stock solution of MF (100 g/ ml) and FF (100 g/ ml) were prepared by dissolving 10 mg of MF and 10
mg of FF in 100 ml methanol in separate volumetric flask. The std stock solution were further diluted separately to obtained working
standard solution of the conc. 30-200 g/ ml of MF and 2-6 g/ ml of FF.

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First derivative zero crossing spectrophometry:

The working standard solutions of both the drug were scanned between the range 200-400 nm in 1 cm cell against blank. The
zero order spectrum of 10 g/ml of both the drug were tried for the first- fourth order derivative absorption spectra. And then first
order derivative method was selected. The recorded spectrums were converted into first order derivative spectra with = 8 and
scaling factor 32. For the estimation of MF amplitude 267.56 nm was selected which was the zero crossing of formoterol fumarate and
for the estimation of FF amplitude 306 nm was selected which was the zero crossing of mometasone furoate as shown in fig 4.

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Fig.3 Overlain zero order spectra of the

standard solution of MF and FF.

Aarti. S. Zanwar et al.

ISSN NO: 2231-6876

Fig.4 Overlain first derivative spectra of MF and FF, for the estimation
of MF when FF spectra is showing zero crossing at 267.56 nm and
for FF when MF is showing zero crossing at 306 nm.

Fig. 5 First order derivative spectra for the estimation

of MF at 267.56 nm for linearity.

Fig. 6 First order derivative spectra for the estimation

of FF at 306 nm for linearity.

Fig.7 Ratio derivative spectra of mometasone furoate.

at different concentration using a standard spectrum
of FF (6 g/ml) as a divisor

Fig. 8 Ratio derivative spectra for formoterol fumarate

at different concentration using a standard spectrum
of MF (30 g/ml) as a divisor

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Ratio spectra derivative method:

According to the theory of the ratio spectra derivative method, for the estimation of MF the stored UV absorption spectra of
the standard solution of the MF were divided wavelength by wavelength by a standard spectrum of FF(6 g/ ml). The first derivative
was calculated for the obtained spectra with = 2 and scaling factor 4. The amplitudes at 263 nm and 267.30 nm were measured. For
the FF, the stored spectrum UV absorption spectra of the std solution of FF were divided wavelength by wavelength by a standard
spectrum of MF( 30 g/ml ). The amplitudes at 293.08 nm and 302.60 nm were measured.

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Analysis of Sample formulation:

The content powder of 20 rotacaps were emptied, accurately weighed and mixed. A portion of powder equivalent to 2 mg of
mometasone furoate which will also contain 0.06 mg of formoterol fumarate was transferred to 25 ml of volumetric flask containing 5
ml of methanol. This solution was sonicated for 30 minutes and diluted up to mark with methanol. The sample was filtered. Further
dilution was carried out with methanol to obtain the conc. of both the drug within the calibration range. The procedures describe above
for zero crossing derivatives and ratio derivative were followed and the concentrations of MF and FF were calculated from the
calibration curved of respective method.
Method Validation :
Both the method was validated as per ICH Q2 (R1) guideline[15]. The standard calibration curve was plotted for MF and FF
in both the method at their respective peak amplitude and correlation coefficient was calculated. The limit of detection (LOD= 3.3 /s,
where is the standard deviation of the response and s is the slope) and limit of quantitation (LOQ=10 /s) of MF and FF was
calculated. Intraday and interday precision was studied by the analyzing three replicates of the standard solution at three concentration
levels. To ascertain the accuracy of the proposed method recovery studies were carried out by standard addition method, adding
know amount of each drug to the preanalysed capsule powder at three levels 80% , 100%, 120% of the label claim.
RESULTS AND DISCUSSION
The zero-crossing first order derivative and ratio- spectra first derivative spectrophotometry method permits a more selective
identification and determination of the two drugs in mixture. The zero crossing method involves measurements of the absolute value
of the total derivative spectrum at an abscissa value corresponding to the zero crossing wavelengths of the derivative spectra of the
individual component. Fig (3-5) shows the D1 spectra of MF and FF. The selection of the optimum wavelength has the best linear
response to the analyte concentration, it is not affected by the conc. of any other component and gives a near zero intercept on the
ordinate axis of the calibration curve. Therefore 267.56 nm (zero crossing wavelength point of FF) and 306 nm (zero crossing
wavelength point of MF) were chosen as optimum working wavelength for the simultaneous determination of MF and FF in a binary
mixture. Measurements of the absolute value of the derivative spectrum taken at these wavelengths gave the best linear response to the
analyte concentration.
Table 1.Regression parameter of the proposed zero-crossing first order derivative (Method D1) and ratio- spectra
first derivative.
(Method RD) spectrophotometry method for simultaneous determination of MF and FF .
Method D1
MF
FF
Wavelength (nm)
267.56
306
Beers law range ( g/ml)
30-200
2-6
Correlation Coefficeint(R2)
0.9992
0.9991
Slope
0.042
-0.031
Intercept
0.009
0.016
SD of slope*
0.00571
0.00570
SD of intercept*
0.004359 0.008025
LOD
0.3424
0.8542
LOQ
1.037
2.654
Intraday precision (% RSD)** 0.87
0.98
Interday precision(% RSD)** 1.15
1.26
* for five replicate and ** for three replicate

MF
263
30-200
0.9991
3.382
-0.004
0.07638
0.09018
0.0879
0.266
0.81
1.56

267.3
30-200
0.9994
-3.020
-0.328
0.0503
0.0851
0.0929
0.2817
0.69
1.06

Method RD1
FF
293.08
302.60
2-6
2-6
0.9982
0.9986
0.238
-0.257
0.251
-1.200
0.02685 0.02525
0.01752 0.02163
0.2429
0.0183
0.7361
0.8416
0.89
0.96
1.38
1.32

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Table 2: Recovery analysis for the proposed spectrometric method D1 and RD1 applied to the Rotacaps.
% Recovery of MF
% Recovery of FF
Spike level(%) Method D1
Method RD1
Method D1
Method RD1
267.56nm
263nm
267.3nm
306nm
293.08nm
302.60nm
80
99.24 0.32 99.730.56
99.380.53
98.440.25
99.260.44 98.670.44
100
100.180.47 99.540.68
99.770.50
97.7830.51 98.780.33 99.160.84
120
99.17 0.52 100.10 0.86 100.350.63 98.100.59
98.250.32 99.060.23

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Parameter assessed

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Table 3: Analysis results of Rotacaps.

Drugs

Labeled claim (g per rotacap)

MF
FF

200
6

Method 1
% S.D*(n=6)
100.400.736
98.910.562

Method 2
% S.D*(n=6)
101.20 0.6377
99.200.812

The main advantage of the ratio derivative method is the chance of easy measurement in corresponding to peak so it permits
the use of the wavelength of the highest value of analytical signal. Moreover, the presence of a lot of maxima and minima is another
advantage by the fact that these wavelengths give an opportunity for the determination of active compounds or excipients which
possibly interfere with the analysis. The influence of the and scaling factor for the obtaining the first derivative of the ratio spectra
as well as the effect of divisor concentration on the calibration graphs for the proposed mixture was studied in order to select the best
factors affecting the determination of the analytes. Results indicated that =2 and scaling factor 4 was the most suitable one.
Determination of both drugs was done by dividing the absorption spectra of MF by that of standard spectra of FF (6 g/ml) while the
absorption spectra of FF were divided by that of the standard spectra of MF (30 g/ml).
The proposed methods for the simultaneous estimation of MF and FF in combined dosage form were found to be accurate,
simple and rapid which can be well understood from validation data as given in table (1) &(2). Linearity was observed by linear
regression equation method for MF and FF in different conc. range. The correlation coefficient of these drugs was found to be close to
1.000 indicating good linearity. Percentage RSD for intraday and inter day was found to be less than 2 and the % recovery for both the
drug was found within the range(98.10%-100.18%)as mention in table (2), which indicates the validity of both method. The analysis
results of the rotacaps are given in the table (3).
CONCLUSION
Zero crossing derivative and ratio- spectra derivative spectrophotometric methods were applied successfully for the
simultaneous analysis of MF and FF in the rotacaps. The proposed methods were simple, cost effective, accurate and precise which
can be used for the routine quality control analysis of MF and FF in combined dosage forms.

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ACKNOWLEDGMENT
The authors are thankful to the Research Director , Sumandeep Vidyapeeth for providing all the facility for doing the research work.

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