Development and Validation of UV Spectrophotometric Method for Simultaneous estimation of Terbinafine hydrochloride and Mometasone furoate in Combined Dosage Form

 

Heta D. Patel*, Mehul M. Patel

Department of Quality Assurance, Ramanbhai Patel College of Pharmacy,

Charotar University of Science and Technology, Changa-388 421, Gujarat, India.

*Corresponding Author E-mail: hetapatel32@gmail.com

ABSTRACT:

Terbinafine hydrochloride (TH) is a antifungal drug and Mometasone furoate(MF) is a glucocorticoid. This combination of drug is useful for relief of corticosteroid responsive dermatoses where fungal infections are present, suspected, or likely to occur. Accurate, precise, rapid and economical method was developed for the simultaneous estimation of Terbinafine hydrochloride (TH) and Mometasone furoate(MF) in cream dosage form. Method is based on the absorption correction method and wavelengths selected for analysis were 282.0nm (λmax of Terbinafine hydrochloride) and 248.0 nm (λmax of  Mometasone furoate) respectively, in methanol. The linearity was obtained in the concentration range of 10-70μg/ml for Terbinafine hydrochloride and 1-7μg/ml for Mometasone furoate. The correlation coefficient of Terbinafine hydrochloride and Mometasone furoate were found to be 0.999 and 0.998 respectively. The proposed procedure was successfully applied for the simultaneous determination of both drugs in commercial cream preparation. The results of the analysis have been validated statistically and by recovery studies have confirmed the accuracy of proposed method.

 

 

INTRODUCTION:

Terbinafine hydrochloride (TH) is a antifungal and enzyme inhibitor drug. Chemically, it is (2E)-N,6,6-Trimethyl-N-(naphthalen-1-ylmethyl)hept-2-en-4-yn-1-amine hydrochloride^[1], clinically used in the treatment of dermatophyte infections of the toenail or fingernail caused by susceptible fungi. Also for the treatment of Tinea capitis (scalp ringworm) and Tinea corporis (body ringworm) or Tinea cruris (jock itch). ^[1]. Mometasone furoate(MF) is a glucocorticoid having anti-inflammatory activity and chemically it is 9,21-Dichloro-11β-hydroxy-16α-methyl-3,20-dioxopregna-1,4-dien-17-yl furan-2-carboxylate and  it is also useful treatment of dermatological diseases^[2].

 

Both the drugs are marketed as combined dose cream formulation in the ratio of 10:1 w/w TH: MF. Literature survey reveals that Terbinafine hydrochloride(TH) can be estimated by spectrophotometrically, by HPTLC and by HPLC individually or with other drugs in bulk drugs and in human plasma^[5-23], while Mometasone furoate(MF) can be estimated by spectrophotometrically, HPLC and HPTLC in combination with other drugs^[24-35]. However, there is no analytical method has been reported for the estimation of TH and MF in a combined dosage formulation. Present work describes the absorption correction method^[3] for simultaneous estimation of TH and MF in cream formulation.

 

MATERIALS AND METHODS:

Instrument

A double-beam Shimadzu-1800 UV- Visible spectrophotometer, with spectral bandwidth of 2 nm, wavelength accuracy ± 0.5 nm and a pair of 1-cm matched quartz cells was used to measure absorbance of the resulting solution.

 

 

Figure 1: UV-Spectra of Terbinafine hydrochloride  (10µg/ml)

 

Figure 2: UV-Spectra of Mometasone furoate (1µg/ml)

 

Figure 3: Overlay UV-Spectra of Terbinafine hydrochloride (10µg/ml), Mometasone furoate(1µg/ml) and mixture(10µg/ml of TH and 1µg/ml of MF)

 

 

Materials

Standard gift sample of Terbinafine hydrochloride was provided by Aarti Drugs sales, Mumbai and Mometasone furoate was povided by Glenmark Generics pharmaceutical, Nasik. Combined dose Terbinafine hydrochloride and Mometasone furoate cream sebifin^TM plus was purchased from local market. Methanol (AR Grade) was used as solvent, procured from Loba Chemie pvt ltd., Mumbai.

 

Stock solutions

Stock solution: Standard stock solutions of TH (1000 μg/ml) and MF (1000 μg/ml) were prepared in methanol.

 

Working solutions

Working solution: Standard working solutions of TH (100 μg/ml) and MF (100 μg/ml) were prepared in methanol and used for the analysis.

 

Spectral characteristics of TH and MF

Solutions of TH (10μg/ml) and MF (1μg/ml) were prepared separately by appropriate dilution of standard working solution (according to ratio of both the drugs in combined dosage form). Both the solutions were scanned in the spectrum mode from 200 nm to 400 nm (fig.1, 2). Mixture containing TH (10μg/ml) and MF (1μg/ml) was also scanned in the spectrum mode from 200 nm to 400 nm (fig.3).

 

Preparation of Calibration curves

Appropriate dilutions of the standard working solution were done separately to get 10, 20, 30, 40, 50, 60,70μg/ml of TH and 1, 2, 3, 4, 5, 6,7μg/ml MF, respectively. The absorption spectra of all solutions were recorded between 200-400 nm. The absorbances were measured at 282.0 nm (λmax of TH), 248.0 nm (λmax of MF). Beer’s lamberts range for TH and MF were selected and calibration curves of both the drugs were plotted separately. (fig.4-6)

 

Determination of Absorptivity Value of TH and MF

Appropriate dilutions of the standard working solution were done and the absorbances were measured for TH at 282nm (λmax of TH), and 248nm whereas for MF at 248nm (λmax of MF). The absorptivity values of the drugs were determined at the selected wavelengths. These absorptivity values are the mean of six determinations.

 

Application of the proposed method for the determination of TH and MF in cream

1gm cream was weighed which is equivalent to 10 mg TH and transferred into 10 ml centrifugal tube, dissolved in methanol and centrifuged for 5 minute at 4000rpm.The solution was filtered through Whatman filter paper and 5ml volume was  taken in 50ml volumetric flask and volume adjusted up to the mark with methanol. This solution contains 100μg/ml of TH and 10 μg/ml of MF.

 

From this solution,4 ml was taken into 10 ml volumetric flask and volume was adjusted up to the mark with methanol.

The absorbance of sample solution was measured against blank solution at 282 nm and 248 nm for absorption correction method. Assay of combined dosage form was done 5 times.(tab. 6). The amount of TH and MF was computed using following equations.

 

For Analysis of TH

        C_T=A₂₈₂/a_T282               -------------------(1)

 

Where,

C_T = Concentration of TH gm/100ml,

A₂₈₂ =Absorbance at 282nm

A_T282 = Absorptivity of TH at 282nm

 

For Analysis of MF

        C_M= (A₂₄₈-a_T248C_T)/a_M248   -----------------(2)

Where,

C_M = Concentration of MF gm/100ml,

A₂₄₈ = Total absorbance of Sample at 248 nm ,

a_{T 248} = Absorptivity of TH at 248nm

a_M248 = Absorptivity of MF at 248 nm

 

Validation^[4]

The method was validated with respect to linearity, LOD, LOQ, Accuracy and Precision. (Table 1-5)

 

RESULTS AND DISCUSSION:

The method discussed in the present work provides a convenient and accurate way for simultaneous analysis of TH and MF. In absorption correction method, wavelengths selected for analysis were 282.0 nm (λmax of TH) and 248.0 nm (λmax of MF). In this method linearity for detector response was observed in the concentration range of 10-70μg/ml for TH and 1-7μg/ml for MF. Percent label claim for TH and MF in cream analysis, by this method was found 101.85% for TH and 98.35% for MF. Low values of LOD and LOQ indicated good sensitivity of proposed method. Accuracy of proposed methods was ascertained by recovery studies. The percent recovery for TH and MF, by this method, was found 101.05% for TH and 99.36% for MF respectively. The proposed method could be employed for routine quality control of TH and MF in combined cream dose formulation.

 

 

Figure 4: Calibration Curve of Terbinafine hydrochloride at 282nm

 

Figure 5: Calibration Curve of Terbinafine hydrochloride at 248nm

 

Figure 6: Calibration Curve of  Mometasone furoate at 248nm.

 

Table 1: Summary of Validation Parameters

Sr. No	Parameter	Result
		Terbinafine hydrochloride	Mometasone furoate
1	Absorption maxima (λ max)	282nm	248nm
2	Range (μg/ml)	10-70	1-7
3	Linearity (μg/ml)	10-70	1-7
4	Standard regression equation	y = 0.026x + 0.016	y = 0.090x + 0.002
5	Intercept	0.016	0.002
6	Slope	0.026	0.090
7	Correlation coefficient (r²)	r² = 0.999	r² = 0.998
8	L.O.D (µg/ml)	0.50	0.105
9	L.O.Q (µg/ml)	1.67	0.318
10	Specific absoptivity	λ282nm=255	λ 248nm=901
		λ 248nm=281

 

Table 2: Accuracy data of Terbinafine hydrochloride

Set	Sample conc.	Standard added	Total conc.	Recovered (µg/ml)	% recovery	Mean % recovery
80	20 µg/ml	16 µg/ml	36 µg/ml	36.63	101.75	101.43
			36 µg/ml	36.33	100.92
			36 µg/ml	36.58	101.61
100	20 µg/ml	20 µg/ml	40 µg/ml	40.53	101.33	101.03
			40 µg/ml	40.48	101.20
			40 µg/ml	40.52	100.55
120	20 µg/ml	24 µg/ml	44 µg/ml	44.11	100.25	100.68
			44 µg/ml	44.59	101.34
			44 µg/ml	44.20	100.45

 

 

 

 

 

 

 

 

 

 

 

 

Table 3: Accuracy data of Mometasone furoate

Set	Sample conc.	Standard added	Total conc.	Recovered (µg/ml)	% recovery	Mean % recovery
80	2 µg/ml	1.6 µg/ml	3.6 µg/ml	3.58	99.44	99.35
			3.6 µg/ml	3.56	98.89
			3.6 µg/ml	3.59	99.72
100	2 µg/ml	2 µg/ml	4 µg/ml	4.01	100.25	100.08
			4 µg/ml	3.98	99.50
			4 µg/ml	4.02	100.50
120	2µg/ml	2.4 µg/ml	4.4 µg/ml	4.32	98.18	98.64
			4.4 µg/ml	4.34	98.64
			4.4 µg/ml	4.36	99.09

 

 

 

 

 

 

 

 

 

 

 

 

Table 4: Intraday Precision data of Terbinafine hydrochloride and Mometasone furoate (where, * is the mean of 3 trials)

Conc. (µg/ml)		Mean absorbance ± SD*		%  RSD*
TH	MF	TH 282nm	MF 248nm	TH282nm	MF248nm
10	1	0.243 ± 0.0023	0.088  ±  0.0007	0.95	0.80
30	3	0.774 ± 0.0052	0.269 ± 0.0022	0.67	0.82
60	6	1.571 ± 0.0069	0.534 ± 0.0049	0.44	0.92

 

Table 5: Interday Precision data of Terbinafine hydrochloride and Mometasone furoate (where, * is the mean of 6 trials)

Conc. (µg/ml)		Mean absorbance ± SD*		%  RSD*
TH	MF	TH 282nm	MF 248nm	TH282nm	MF248nm
10	1	0.243 ± 0.0023	0.086 ± 0.0004	0.95	0.47
30	3	0.777 ± 0.0092	0.270 ± 0.0031	1.18	1.15
60	6	1.574 ± 0.0054	0.532 ± 0.0035	0.34	0.66

 

Table 6: Result of Assay (where, * is the mean of 5 trials)

Label claim (µg/ml)		Conc. Found (µg/ml)
Terbinafine hydrochloride	Mometasone furoate	Terbinafine hydrochloride	Mometasone furoate
40	4	40.73	3.93
40	4	40.74	3.92
40	4	40.75	3.92
40	4	40.77	3.93
40	4	40.71	3.97
Average ± SD*		40.74 ± 0.021	3.93 ± 0.019
RSD*		0.052	0.489
% Assay found*		101.85	98.35

 

 

CONCLUSION:

The developed methods were validated as per ICH guidelines and were found to be within the prescribed limit. It concludes that the developed methods are simple, accurate, sensitive and precise and suitable for both authentic and cream dosage form.

 

ACKNOWLEDGEMENTS:

The authors are very thankful to Dr. R.H PARIKH; Principal, Ramanbhai Patel College of Pharmacy, P D Patel Institute of Applied Sciences and Charotar University of Science and Technology, Changa for providing support and research facilities. We are also thankful to Aarti Drugs sales, Mumbai and Glenmark Generics Pharmaceutical, Nasik for providing the gift samples of Terbinafine hydrochloride and Mometasone furoate respectively.

 

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Received on 04.12.2012         Modified on 20.12.2012

Accepted on 22.12.2012         © AJRC All right reserved