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

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. Desai

2, Kalpana G. Patel

1, Purvi A. Shah

1

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

1. Introduction

Mometasone furoate [MF], 9, 21 – dichloro-11b, 17 –

dihydroxy-16a-methyl-pregnane-1, 4 – diene – 3, 20 –

dione 17 – (2 – furoate ester), (Fig. 1) is a synthetic

glucocorticoid with anti-inflammatory, anti-allergy effect.

Mometasone furoate is effective for various skin diseases,

such as neurodermatitis, eczema, atopic dermatitis and

psoriasis of the skin caused by skin inflammation and

itching. [1-5]

Salicylic acid [SA], is a monohydroxybenzoic acid, a type

of phenolic acid and a beta hydroxy acid [Fig. 2]. It has

bacteriostatic, fungicidal and keratolytic actions. It has

been extensively used in dermatologic therapy as a

keratolytic agent, relieves pain and reduces swelling.

Moreover, SA is effective to treat warts, skin ulcer,

psoriasis and other skin conditions. [1, 2]

Nowadays, MF has been marketed in combination with SA

in semisolid dosage forms, which have lesser side effects

and patient specificity. MOMAT-S (Mometasone furoate

0.1% and 5% Salicylic acid) is used for glucocorticoid with

anti-inflammatory, anti-allergy effect mainly in skin

diseases, such as neurodermatitis, eczema, atopic dermatitis

and psoriasis of the skin caused by skin inflammation and

itching. [4, 6]

Scientific literature reports that there are many methods

reported for the determination of MF individually and in

combination with other drugs like fucidic acid, terbinafine

hydrochloride, nadifloxacin and formoterol fumarate etc.

based on reversed-phase HPLC method [7-12]

. For the

determination of SA either alone or in combination with

other drugs several analytical methods were reported

includes UV spectroscopic method, HPLC, HPTLC and

Capillary Electrophoresis [13-17]

. MF and SA are official in

IP, BP, and USP individually.

[18-20]

To the best of our knowledge, no spectrophotometric

method has been reported for the estimation of mentioned

drugs in formulation. Therefore, the goal of present work is

to develop a simple procedure that could be applied in

quality control laboratories for the simultaneous

determination of both drugs. This work aims to present

simple, accurate and precise ratio-derivative

spectrophotometric method for the simultaneous

determination of MF and SA in semisolid dosage form.

Available online at http://www.urpjournals.com

International Journal of Analytical and Bioanalytical Chemistry

Universal Research Publications. All rights reserved

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

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

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)

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

(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

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.

Table 1: Analytical parameters of proposed method

Parameters MF SA

Wavelength (nm)

Linearity range (μg/mL)

Regression equation

Correlation coefficient

247.60

2-12

y =0.076x ± 0.076

0.9989

284.80

5-50

y=0.074x± 0.400

0.9989

SD of intercept 0.0037 0.00489

SD of slope 0.000503 0.00039

CI of intercepta 0.0796-0.0718 0.4057-0.3954

CI of slopea 0.0766-0.0756 0.0752-0.0743

LOD (μg/mL) 0.21 0.57

LOQ (μg/mL) 0.63 1.72

n=5 replicates, CI means confidence interval; SD means standard deviation

aConfidence interval at 95% confidence level and four degree of freedom (t=2.776)

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

3.2.2 Precision

The intraday precision was carried out through three

replicate analysis of 4, 8 and 12 µg/mL of MF and 10, 30

and 50 µg/mL of SA. The interday precision was also

evaluated through three replicate analysis of the pure drug

samples for three consecutive days at above mentioned

concentration levels. The developed method is found to be

precise as the % RSD values for intraday and interday

precision were less than 2% (Table 2).

3.2.3 Accuracy

Accuracy of the methods was assured by applying the

standard addition technique where good percentage

recoveries were obtained, confirming the accuracy of the

proposed methods (Table 3). The recovery studies were

carried out by adding known amount of standard to samples

at 50, 100 and 150% level and analyzed by the proposed

method, in triplicate.

3.2.4 Sensitivity

The limit of detection and limit of quantitation were

determined based on the standard deviation of response (y-

intercept) and slope of the calibration curve according to

ICH guideline.[21]

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.

3.3 Analysis of marketed semisolid dosage form The proposed method was applied for the simultaneous

determination of MF and SA in commercial semisolid

formulation and amount of MF and SA were found to be

98.07% and 100.03% respectively as shown in Table 4.

The percent recoveries of the amount of MF and SA in

Table 2. Precision studies

Amount of drug

(μg/mL)

Intraday precision Interday precision

Amount of drug found ± SD

(μg/mL) %RSD

Amount of drug found ± SD

(μg/mL) %RSD

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

Drugs Taken

(μg/mL) % Level

Amount of std

added (μg/mL)

Total amount of drug

Found (μg/mL) % Recovery ± SD % RSD

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

Momate-S MF 1mg 98.07±1.41 1.44

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

were in good agreement with the label claims thereby

suggesting that there is no interference from any of the

excipients that normally present in ointment.

4. CONCLUSION The proposed ratio first derivative spectrophotometric

method was found to be novel, rapid, simple, sensitive,

accurate, precise and easy to be understood and applied.

Distinct advantages of the proposed method include the

simplicity and rapidity of sample preparation, good

sensitivity and a cost effective methodology. Hence, the

proposed method could be regarded as useful alternative to

the chromatographic techniques (HPLC) in the routine

quality control of title drugs either alone or in combination

with a relatively inexpensive instrumentation for

simultaneous estimation of MF and SA in their binary

mixtures.

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Source of support: Nil; Conflict of interest: None declared