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International Journal of Analytical and Bioanalytical Chemistry

Universal Research Publications. All rights reserved

ISSN-2231-5012
Original Article
Application of Ratio Derivative Spectrophotometry for Simultaneous
Determination of Mometasone furoate and Salicylic acid in Semisolid dosage form
Dhaval R. Vanani1, Samil D. Desai2, Kalpana G. Patel1, Purvi A. Shah1
1
Department of Quality Assurance, Anand Pharmacy College, Near Townhall, Anand, Gujarat, India.
2
Torrent Research Centre, Village – Bhat, Ahmedabad.
Email: purvi2210@yahoo.co.in
Received 05 June 2013; accepted 01 July 2013
Abstract
Ratio derivative spectrophotometric method has been developed for the simultaneous determination of Mometasone
furoate (MF) and Salicylic acid (SA) in Methanol. In this method, the overlapping spectra of MF and SA were well
resolved by making use of the first-derivative of the ratios of their direct absorption spectra. The derivative ratio
absorbances of MF and SA were measured at λmax 247.60 and λmax 284.80 nm, respectively for their quantification. MF and
SA were determined in the concentration range of 2-12 μg/mL and 5-50 μg/mL respectively. The method was validated as
per the ICH guideline and accuracy, precision are found to be within the acceptable limit. The limits of detection and
quantitation were found to be 0.21 and 0.63 μg/mL, respectively for MF and 0.57 and 1.72 μg/mL, respectively for SA.
The proposed ratio first derivative spectrophotometric method is novel, rapid, simple, sensitive, accurate, precise and does
not require separation of MF and SA hence successfully applied for simultaneous estimation of MF and SA in marketed
semisolid dosage form.
© 2013 Universal Research Publications. All rights reserved
Keywords: Simultaneous determination, Mometasone furoate, Salicylic acid, Ratio derivative spectrophotometric method
diseases, such as neurodermatitis, eczema, atopic dermatitis
1. Introduction and psoriasis of the skin caused by skin inflammation and
Mometasone furoate [MF], 9, 21 – dichloro-11b, 17 – itching. [4, 6]
dihydroxy-16a-methyl-pregnane-1, 4 – diene – 3, 20 – Scientific literature reports that there are many methods
dione 17 – (2 – furoate ester), (Fig. 1) is a synthetic reported for the determination of MF individually and in
glucocorticoid with anti-inflammatory, anti-allergy effect. combination with other drugs like fucidic acid, terbinafine
Mometasone furoate is effective for various skin diseases, hydrochloride, nadifloxacin and formoterol fumarate etc.
such as neurodermatitis, eczema, atopic dermatitis and based on reversed-phase HPLC method [7-12]. For the
psoriasis of the skin caused by skin inflammation and determination of SA either alone or in combination with
itching. [1-5] other drugs several analytical methods were reported
Salicylic acid [SA], is a monohydroxybenzoic acid, a type includes UV spectroscopic method, HPLC, HPTLC and
of phenolic acid and a beta hydroxy acid [Fig. 2]. It has Capillary Electrophoresis [13-17]. MF and SA are official in
bacteriostatic, fungicidal and keratolytic actions. It has IP, BP, and USP individually. [18-20]
been extensively used in dermatologic therapy as a To the best of our knowledge, no spectrophotometric
keratolytic agent, relieves pain and reduces swelling. method has been reported for the estimation of mentioned
Moreover, SA is effective to treat warts, skin ulcer, drugs in formulation. Therefore, the goal of present work is
psoriasis and other skin conditions. [1, 2] to develop a simple procedure that could be applied in
Nowadays, MF has been marketed in combination with SA quality control laboratories for the simultaneous
in semisolid dosage forms, which have lesser side effects determination of both drugs. This work aims to present
and patient specificity. MOMAT-S (Mometasone furoate simple, accurate and precise ratio-derivative
0.1% and 5% Salicylic acid) is used for glucocorticoid with spectrophotometric method for the simultaneous
anti-inflammatory, anti-allergy effect mainly in skin determination of MF and SA in semisolid dosage form.

International Journal of Analytical and Bioanalytical Chemistry 2013; 3(3): 67-71

67
2. MATERIALS AND METHODS
2.1 Instrumentation
An UV-Visible Spectrophotometer (Simadzu-1800, Japan)
with 10 mm matched quartz cells was used for
Spectrophotometric method. All weighing were done on
electronic balance (Model Shimadzu AUW-220D).
Ultrasonicator (Model 5.5 150H) was used for sample
solution preparation.
Fig 1: Structure of Mometasone furoate
Fig 2: Structure of Salicylic acid
2.2 Reagents and chemicals
Analytical pure samples of MF and SA were obtained as a
gift samples from Torrent Pharma, Ahmedabad. These
samples were used without further purification. Semisolid
formulation „MOMAT-S‟ manufactured by Glenmark
Pharmaceutical Industries-Vadodara, was purchased from
the local market containing MF (1 mg) and SA (50 mg) per
ointment (10 g). Analytical grade methanol purchased from
Merck, Mumbai was used throughout the study.
Fig 3: Zero order Overlay spectra of mometasone furoate
and salicylic acid
International Journal of Analytical and Bioanalytical Chemistry 2013; 3(3): 67-71
68
2.3 Preparation of Standard Solutions and Calibration
Curve
Standard stock solutions each containing 1000 μg/mL of
MF and SA were prepared separately in the methanol. The
working standard solutions (100 μg/mL) of mentioned
drugs were obtained by dilution of the respective stock
solution in methanol. For verification of Beer‟s law, a
series of dilutions in the concentration range of 2-12 μg/mL
for MF and 5-50 μg/mL for SA were prepared separately to
establish calibration curve.
2.4 Ratio first derivative Spectrophotometric method
The method involves dividing the spectrum of formulation
by the standardized spectra of each of the analyte and
deriving the ratio to obtain spectrum that is dependent of
concentration of analyte used as a divisor. Using
appropriate dilutions of standard stock solution, the
standard solutions of MF (2 μg/mL) and SA (10 μg/mL)
were prepared and their zero order spectra recorded over
the range 200-400 nm using methanol as blank. The ratio
spectra of different MF standards at increasing
concentrations were obtained by dividing each with the
stored zero order spectrum of standard solution of SA (10
μg/mL) and the first derivative of these spectra traced with
the interval of ∆λ= 8 nm, illustrated in Fig.4. Similarly, the
ratio derivative spectra of the solutions of SA at different
concentrations were obtained by dividing each with the
stored zero order spectrum of standard solution of MF (2
μg/mL) and the first derivative of these spectra traced with
the interval of ∆λ= 8 nm, illustrated in Fig.5 From Fig. 4
and 5, 247.60 nm and 284.80 nm as wavelength maxima
(λmax) was selected for the simultaneous determination of
MF and SA in marketed semisolid formulation,
respectively.
Fig 4: (a) Ratio derivative spectra of mometasone furoate,
10 μg/mL Salicylic acid as divisor and (b) Ratio first order
derivative spectra of mometasone furoate (λmax 247.60, Δλ
= 8)

(a)
(b)
Fig 5: (a) Ratio derivative spectra of salicylic acid, 2
μg/mL mometasone furoate as divisor and (b) Ratio first
order derivative spectra of salicylic acid (λmax 284.80, Δλ =
8)
2.5 Method Validation
The method was validated as per ICH Q2 (R1) guideline.
[21]
Intraday and interday precision was studied by
analyzing three replicates of standard solutions at three
concentrations level. The accuracy studies were carried out
at different concentrations by spiking (50, 100 and 150%) a
known concentration of standard drug to the pre-analyzed
sample and contents were reanalyzed by the developed
method. The limit of detection (LOD = 3.3 σ/s, where σ is
the standard deviation of response and s is slope) and limit
of quantitation (LOQ=10σ/s) of MF and SA was calculated.
2.6 Analysis of marketed semisolid dosage form
For the analysis of marketed semisolid formulation, 10 g
ointment was weighed accurately and a quantity equivalent
to 1 mg of MF and 50 mg of SA was weighed and
dissolved in 50 mL methanol with the aid of ultrasonicator
Table 1: Analytical parameters of proposed method
Parameters
Wavelength (nm)
Linearity range (μg/mL)
Regression equation
Correlation coefficient
SD of intercept
SD of slope
CI of intercepta
CI of slopea
LOD (μg/mL)
LOQ (μg/mL)
n=5 replicates, CI means confidence interval; SD means standard deviation
a
Confidence interval at 95% confidence level and four degree of freedom (t=2.776)
International Journal of Analytical and Bioanalytical Chemistry 2013; 3(3): 67-71
69
for 15 min and solution was filtered through Pre-filter +
PVDF (0.45 μm) into a 100 mL volumetric flask and
volume was made up to mark with methanol as a diluent.
The solution was suitably diluted with methanol to get a
concentration of 5 μg/mL of MF and 50 μg/mL of SA, by
standard addition of standard solution of MF. The prepared
solution were analysed in triplicate as per method given
under 2.4 section and the amount of MF (CMF) and SA
(CSA) in formulation was calculated as per following.
CMF = Derivative amplitude at λmax 247.60
CSA = Derivative amplitude at λmax 284.80
3. RESULTS AND DISCUSSION
3.1 Ratio first derivative Spectrophotometric method
The ratio spectra of different MF standards at increasing
concentrations in methanol obtained by dividing each with
the stored zero order spectrum of standard solution of SA
are shown in Figs.4(a) and the first derivative of these
spectra traced with the interval of ∆λ= 8 nm are illustrated
in Fig.4(b). Similarly, the ratio derivative spectra of the
solutions of SA in different concentrations in methanol
traced with the interval of ∆λ= 8 nm by using the zero order
spectra of MF as divisor by computer aid is demonstrated
in Fig. 5. Here, the standard spectra of 2.0 μg/mL of MF
and 10.0 μg/ml of SA were considered as suitable for the
determination of SA and MF respectively, as divisor. The
∆λ found as optimum for the first derivative of their ratio
spectra was 8 nm. From the Fig. 4(b) and Fig. 5(b),
wavelength maxima 247.60 nm and 284.80 nm were
selected for the determination of the MF and SA
respectively in the assay of pharmaceutical preparation,
semisolid dosage form, due to its lower R.S.D. value and
more suitable mean recovery.
3.2 Method validation
Validation of the methods has been performed according to
ICH recommendations.
3.2.1 Linearity
The calibration range for MF and SA was established
through considerations of the practical range necessary
according to Beer–Lambert‟s law. The linearity response
was determined by analyzing 6 independent levels of
concentrations in the range of 2-12 µg/mL and 5-50 µg/mL
at 247.60 nm for MF and at 284.80 nm for SA respectively.
The values of correlation coefficients of MF and SA were
close to unity indicating good linearity, the characteristic
parameters for the constructed equations are summarized in
Table 1.
MF SA
247.60 284.80
2-12 5-50
y =0.076x ± 0.076 y=0.074x± 0.400
0.9989 0.9989
0.0037 0.00489
0.000503 0.00039
0.0796-0.0718 0.4057-0.3954
0.0766-0.0756 0.0752-0.0743
0.21 0.57
0.63 1.72
3.2.2 Precision at 50, 100 and 150% level and analyzed by the proposed
The intraday precision was carried out through three method, in triplicate.
replicate analysis of 4, 8 and 12 µg/mL of MF and 10, 30 3.2.4 Sensitivity
and 50 µg/mL of SA. The interday precision was also The limit of detection and limit of quantitation were
evaluated through three replicate analysis of the pure drug determined based on the standard deviation of response (y-
samples for three consecutive days at above mentioned intercept) and slope of the calibration curve according to
concentration levels. The developed method is found to be ICH guideline.[21] The limits of detection and quantitation
precise as the % RSD values for intraday and interday were found to be 0.21 and 0.63 μg/mL, respectively for MF
precision were less than 2% (Table 2). and 0.57 and 1.72 μg/mL, respectively for SA.
3.2.3 Accuracy 3.3 Analysis of marketed semisolid dosage form
Accuracy of the methods was assured by applying the The proposed method was applied for the simultaneous
standard addition technique where good percentage determination of MF and SA in commercial semisolid
recoveries were obtained, confirming the accuracy of the formulation and amount of MF and SA were found to be
proposed methods (Table 3). The recovery studies were 98.07% and 100.03% respectively as shown in Table 4.
carried out by adding known amount of standard to samples The percent recoveries of the amount of MF and SA in
Table 2. Precision studies
Intraday precision Interday precision
Amount of drug
Amount of drug found ± SD Amount of drug found ± SD
(μg/mL) %RSD %RSD
(μg/mL) (μg/mL)
MF
4 3.952 ± 0.040 1.02 3.908 ± 0.060 1.54
8 7.816 ± 0.023 0.29 7.807 ± 0.072 0.93
12 11.939 ± 0.042 0.35 11.930 ± 0.064 0.54
SA
10 10.914 ± 0.078 0.71 10.973 ± 0.115 1.05
30 29.712 ± 0.138 0.47 29.838 ± 0.279 0.94
50 50.144 ± 0.109 0.22 51.162 ± 0.446 0.87
n=3 replicate; SD means standard deviation; %RSD means relative standard deviation
Table 3: Recovery studies for determination of MF and SA in semi-solid dosage form
Taken Amount of std Total amount of drug
Drugs % Level % Recovery ± SD % RSD
(μg/mL) added (μg/mL) Found (μg/mL)
50% 0.5 1.45 96.22 ± 0.38 0.39
MF 1 100% 1.0 1.96 97.33 ± 0.57 0.59
150% 1.5 2.46 97.74 ± 1.16 1.19
50% 25 74.32 98.90 ± 0.20 0.20
SA 50 100% 50 99.09 99.05 ± 0.03 0.03
150% 75 124.21 99.39 ± 0.04 0.043
n= 3 replicates; SD means standard deviation; %RSD means relative standard deviation
Table 4: Determination of MF and SA in semi-solid dosage form
Formulation Drug Label Claim % Assay ± SD % RSD
MF 1mg 98.07±1.41 1.44
Momate-S
SA 50mg 100.03±0.25 0.25
n=3 replicates; SD means standard deviation; %RSD means relative standard deviation
semisolid dosage form, expressed as a percentage assay with a relatively inexpensive instrumentation for
were in good agreement with the label claims thereby simultaneous estimation of MF and SA in their binary
suggesting that there is no interference from any of the mixtures.
excipients that normally present in ointment. 5. REFERENCES
4. CONCLUSION 1. The Merck Index, An Encyclopedia of Chemicals,
The proposed ratio first derivative spectrophotometric Drugs, and Biologicals, Merck & Co., Inc.,
method was found to be novel, rapid, simple, sensitive, Whitehouse Station, twelth ed., No:6324, pp1067.
accurate, precise and easy to be understood and applied. 2. “Martindale” The Extra Pharmacopeia” 32 th ed., The
Distinct advantages of the proposed method include the Pharmaceutical Press, London, (1989) 1047.
simplicity and rapidity of sample preparation, good 3. H. P. Rang, J. P. Dale, J. M. Ritter, R. J. Flower, Rang
sensitivity and a cost effective methodology. Hence, the and Dale‟s Pharmacology, six ed., Churchill
proposed method could be regarded as useful alternative to Livingstone Elsevier; 2007.
the chromatographic techniques (HPLC) in the routine 4. K.D Tripathi, Essentials of Medical pharmacology,
quality control of title drugs either alone or in combination six ed., Jaypee Brothers medical publishers (P) LTD;
International Journal of Analytical and Bioanalytical Chemistry 2013; 3(3): 67-71
70
 2008.
5. L. Brunton, K. Parker, D. Blumenthal, L. Buxton,
Pharmacology and Therapeutics, Mc graw hill
Medical; 2007.
6. Formulation approval http://www.freepatents online.
com /EP0735885.pdf
7. Prakash A, Benfield P. Topical Mometasone: A review
of its pharmacological properties and therapeutic use in
the treatement of dermatological disorders. Drugs 55
(1998) 145-63.
8. U. Rekha, B. P. Manjula, Formulation and evaluation
of microsponges for topical drug delivery of
mometasone furoate, IJPP. ISSN-0975-1491,
3, (2011). (http://www.ijppsjournal.com/Vol3Issue4/
2534.pdf)
9. S. Shaikh, M. S. Muneera, O. A.
Thusleem, Muhammad Tahir, V. K. Anand, A simple
13. and benzalkonium chloride in nasal spray solution,
Journal of trace analysis in Food and Drugs. (2013) 14-
21.
14. A. Iqbal, Faiyaz HM Vaid, Determination of benzoic
acid and Salicylic acid in commercial benzoic and
Salicylic acids ointments by spectrophotometric
method, pak. J. pharm. Sci. 22 (2009) 18-22.
15. F. SAnas, J.J. Nevado, A. E. Mansilla, A new
spectrophotometric method for quantitative multi-
-component analysis resolution of mixtures of Salicylic
and Salicyluric acids, Talanta. 37 (1990) 347-51.
16. F. A. El-Yazbi, H. H. Hammud, S. A. Assi, Derivative-
ratio spectrophotometric method for the determination
of ternary mixture of aspirin, paracetamol and Salicylic
acid, PMID: 17350884 Spectrochim Acta A Mol
Biomol Spectrosc. 68 (2007) 275-8.
17. A. N. Jhariya, A. Joshi, A. K. Parashar, R. K. Umar,
Application of sodium citrate as hydrotropic agent in
Source of support: Nil; Conflict of interest: None declared
International Journal of Analytical and Bioanalytical Chemistry 2013; 3(3): 67-71
71
RP-HPLC method for the simultaneous quantitation of
chlorocresol, Mometasone furoate, and fusidic acid in
creams, J Chromatogr Sci. 47 (2009) 178-83.
(http://lib.bioinfo.pl/paper:1298382)
10. K. Srinivasarao, V. Gorule, R. Venkata, K. Venkata,
Validated Method Development for estimation of
formoterol fumarate and mometasone furoate in
metered dose inhalation form by high performance
liquid chromatography, Srinivasarao. J anal Bioanal
techniques 3:7 (2012).
11. A. A. Kulkarni, R. K. Nanda, M. N. Ranjane, P. N.
Ranjane, Simultaneous estimation of nadifloxacin and
mometasone furoate in topical cream by HPTLC
method, Der Pharma Chemica. 2(3) (2010) 25-30.
12. K. A. Shaikh, A. T. Patil, S. Katare, Stability-
Indicating HPLC Method for the determination of
mometasone furoate, oxymetazoline, phenyl ethanol
spectrophotometric analysis of salicylic acid, IJRAPR.
3(2013) 36-38.
18. A. W. Clayton, R. E. Thiers, Direct spectrophotometric
determination of Salicylic acid, acetylsalicylic acid,
Salicylamide, caffeine, and phenacetin in tablets or
powders, Journal of pharmaceutical sciences. 55
(1966) 404–407.
19. Indian Pharmacopoeia 2010. Vol-II. Ministry of Health
& Family Welfare. Published by Indian
Pharmacopoeia Commission.2010, pp1700-03.
20. British Pharmacopoeia 2009. Vol-II. Published by
British Pharmacopoeia Commission. 2009, pp1395-96.
21. United state Pharmacopoeia 2012. USP 35 and NF 30,
Vol-II. Published by United state Pharmacopoeia
Commission.2012, pp 3943-45, 877-881.
22. Validation of analytical procedures: text and
methodology, in: International Conference on
Harmonization (ICH), Q2 (R1), IFPMA, Geneva,
Switzerland, 2005.