Method development and validation of Montelukast sodium in bulk and tablet formulation by HPLC

 

Dipti G. Phadtare2*, Amol R. Pawar1*, Rajashri R. Kulkarni2, Govind K. Patil3

1Department of Pharmaceutical Chemistry, RG Sapkal College of Pharmacy, Anjaneri, Nashik.

2Department of Quality Assurance Techniques, RG Sapkal College of Pharmacy, Anjaneri, Nashik.

3University Institute of Chemical Technology, NMU, Jalgoan.

*Corresponding Author E-mail: amolpharma9@gmail.com

 

 

ABSTRACT:

A high performance liquid chromatography method has been developed and validated method for the simultaneous estimation of Montelukast sodium in bulk and tablet dosage form. The method was carried out using Princeton SPHER ULTIMA C18 100A 5µ 250*4.6 mm using OPA: Methanol (10:90 v/v) as mobile phase at flow rate 1ml/min. HPLC separation was carried out at 284 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.9982 and 0.9979 for Montelukast sodium in bulk and tablet dosage form respectively in the concentration range of 1-6 µl/ml for Montelukast sodium in bulk and tablet dosage form. The method was validated for precision, robustness, specificity and accuracy. The limit of detection and quantitation were 0.0011 and 0.0033 µg/ml. the proposed HPLC method can be applied for identification and quantitative determination of Montelukast sodium in bulk and tablet dosage form.

 

KEYWORDS: Montelukast sodium, tablet, HPLC, validation.

 


 

INTRODUCTION:

Montelukast is a potent, selective and orally active leukotrines receptor antagonist that inhibits the cysteinyl leukotrines CysLT1 receptor used in the treatment of asthma. Montelukast sodium is described chemically as (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 hygroscopic and optically active white to off white powder and it is freely soluble in ethanol, methanol, and water. Montelukast sodium used in the treatment of chronic asthma and allergic rhinitis.

 

Literature survey reveals that Montelukast sodium is estimated individually or in combination with other drugs by UV spectrophotometry, spectrofluorometry, RP- HPLC, HPTLC, plasma HPLC, LC/MS, and stability indicating HPLC methods have been reported.

Above literature suggests that no method has been reported for simultaneous determination of Montelukast sodium in bulk and tablet formulation by HPLC method is sensitive, rapid and less time consuming. In HPLC many samples are simultaneously used and solvent requirement is low. So, the present study is designed for the development and validation of simple, precise and accurate HPLC method for the simultaneous determination of Montelukast sodium in bulk and tablet dosage form. The proposed method is validated as per ICH guidelines.

 

Fig. 1: Structure of Montelukast sodium.

 

EXPERIMENTAL:

Materials:

Working standards of pharmaceutical grade Montelukast sodium was obtained as gift samples from Glenmark Pharmaceuticals, Mumbai and Cipla Pharmaceuticals, Goa, India. Romilast TM [Ranbaxy Pharma] is a marketed formulation of Montelukast sodium procured from local pharmacy. All chemicals and reagents of analytical grade were purchased from Merck Chemicals, Mumbai, India.

 

Selection of analytical wavelength:

Stock solutions of drugs were prepared in methanol separately. UV spectrum of 10 ug/ ml of individual drug were taken. Further, in situ HPLC spectral overlain of Montelukast sodium in bulk and tablet dosage form was taken.

 

Instrumentation and chromatographic conditions:

Instruments and apparatus

The chromatography was performed on a LC 20 Shimadzu, Japan  HPLC instrument equipped with  detector Photon Diode Array [PDA] , C18 column was used as stationary phase.

 

Reagents and materials

Standard samples of Montelukast sodium in bulk and tablet dosage form were obtained from Glenmark [Mumbai] and Cipla [Goa] pharmaceuticals and tablet formulation of Ranbaxy Pharma were procured form local pharmacy. Triple distilled water, methanol, Acetonitrile, tri ethyl amine used were of HPLC grade.

 

Standard solutions and calibration graphs:

Accurately weighed Montelukast sodium in bulk and tablet dosage form were transferred to 50 ml of volumetric flask, dissolved in OPA and diluted up with methanol to obtain stock solution of Montelukast sodium in bulk and in tablet formulation, each. From the above stock solution, an aliquot of the solution was transferred to 50 ml volumetric flask, and diluted up to the mark with mobile phase to obtain a working standard solution of Montelukast sodium in bulk as well as tablet formulation.

 

Preparation of calibration curve

Aliquots of 1, 2, 3, 4, 5, 6 ml solutions of Montelukast sodium were used for mixed working standard solution were transferred in series of 10 ml volumetric flasks, and the volume was made up to the mark with mobile phase. Aliquot of each solution was injected under the operating chromatographic conditions as described above and responses were recorded. Calibration curve were constructed by plotting the peak areas versus the concentrations, and the regression equations were calculated of Montelukast sodium in bulk and tablet dosage form respectively. Each response was average of three determinations.

 

Fig. 2: Calibration curve for Montelukast sodium in bulk.

 

Fig. 3: Calibration curve for Montelukast sodium in tablet formulation.

 

METHOD VALIDATION

The optimized HPLC method was validated with respect to the following parameters as per the ICH guidelines.

 

Precision:

Precision of the method was determined with the standard and the test sample. The precision of the method was verified by repeatability [intraday] and intermediate precision studies. Repeatability studies were performed by analysis of three different concentrations of working standard of 2, 3, 4 µg/ml for Montelukast sodium in bulk and tablet dosage form respectively. Method repeatability was achieved by repeating the same procedure six times on the same day for intra- day precision. the intermediate precision of the method was checked by performing same procedure on different days under the same experimental conditions. The repeatability of sample application and measurement of peak area were expressed in terms of relative standard deviation [% R.S.D.].

 

Robustness:

The robustness was studies by evaluating the effect of small but deliberate variations in the chromatographic conditions. The robustness was studies by analyzing the same samples of Montelukast sodium in bulk and in tablet dosage form by deliberate variation in the method parameters. The changes in the responses were noted. Robustness of the method was studies by changing the extraction time of Montelukast sodium in bulk and in tablet dosage form.  

 

Limit of detection and limit of quantitation:

The detection limit of an individual analytical procedure is the lowest amount of analyte in a sample that can be detected but not necessarily quantitated as an exact value. The quantitation limit of an individual analytical procedure is the lowest amount of analyte in a sample that can be quantitatively determined with suitable precision and accuracy.

LOD = 3.3 × δ/S

LOQ = 10 × δ /S

 

Accuracy:

Accuracy of the proposed method was carried out by applying the method to pharmaceutical dosage form to which known amounts of Montelukast sodium in bulk and in tablet dosage form corresponding to 80, 100 and 120% of label claim had been added. The absolute recovery was calculated by comparing the peak areas obtained from standard solution of Montelukast sodium in bulk and tablet dosage form with the peak areas of samples of different concentration. Six determinations at each level of concentration were performed and the results obtained were compared with expected results.

 

RESULTS AND DISCUSSION:

Selection of analytical wavelength:

UV spectrum of Montelukast sodium showed maximum absorbance at 284 nm.

 

Fig. 4: UV spectrum of Montelukast sodium.

 

Fig. 5: Typical chromatogram of Montelukast sodium.

 

Fig.6: Graph which shows purity.

 

Fig. 7: Peak profile.

 

Table 1: Regression analysis data and summary of validation parameter for the proposed method

Parameters of HPLC method

Montelukast sodium in bulk

Montelukast sodium in tablet dosage form

Concentration range [µg/ml]

1-6

1-6

Slope

26217

26198

Intercept

84362

84530

Correlation coefficient

0.9982

0.9979

% Recovery

99.73± 0.0605

98.71± 0.1131

 

Table 2: Assay results for tablets using the proposed method

Formulation

% Purity

Mean ± S. D.

% R.S.D.

RomilastTM

Tablet [10 mg]

98.7100

98.71 ± 0.1131

0.1146

 

Table 3: Recovery studies of Montelukast sodium in bulk and in tablet dosage form

Drug

Amount  present

Amount added [%]

% Recovery ± SD

 

MLT

[Bulk]

2

80

99.142 ± 0.0780

2

100

99.106 ± 0.0343

2

120

99.799 ± 0.0235

 

MLT

[Tablet]

2

80

99.14 ± 0.07

2

100

99.10 ± 0.034

2

120

99.79 ± 0.023

 

Table 4: System suitability parameters for Montelukast sodium

Parameters

HPLC method

Retention time

7.295

Tailing factor

1.118

Asymmetry factor

1.920

Theoretical plates

5248

 

CONCLUSION:

The developed HPLC method was validated and the system suitability studies were performed and all parameters combined with the simplicity and ease of operation ensures that the validated method can successfully used for routine analysis of Montelukast sodium in bulk and tablet dosage form.

 

REFERENCES:

1.               Hitesh Vekaria, Vipul Limbasiya, Piyush Patel, Development and validation of RP- HPLC method for simultaneous estimation of Montelukast sodium and Fexofenadine hydrochloride in combined dosage form, Journal of Pharmacy Research, Elsevier, [2013]. Page no.:134-139.

2.               K. Pallavi and Srinivasa Babu, Validated UV Spectroscopic method for estimation of Montelukast sodium from bulk and tablet formulations, International Journal of Advances in Pharmacy, Biology and Chemistry, Volume 1[4], Oct- Dec, 2012.

3.               B. V. V. Ravi Kumar, Patnaik A. K., et al., A RP- HPLC method development and validation for the estimation of Montelukast sodium in bulk and pharmaceutical dosage forms, International Research Journal of Pharmacy, Volume 3[11], 2012.

4.               N. S. Dighe, A. S. Balsane, et al., Method development and validation of Rupatadine fumarate and Montelukast sodium by RP- HPLC, International journal of Pharmaceutical Chemistry, Volume 5[2], 2015.

5.               Bonthu M. Gandhi, Atmakuri L. Rao, Jangala V. Rao, Method development and validation for simultaneous estimation of Montelukast sodium and Desloratadine by RP-HPLC, American Journal of Analytical Chemistry, 2015.

6.               Yakkala Manasa, Naidu Rao, Meghana Reddy, Mrthod development and validation for simultaneous estimation of Fexofenadine HCl and Montelukast sodium by RP- HPLC in pure and combined tablet dosage form, World Journal of Pharmacy and Pharmaceutical Sciences, Volume 2[6], 2013.

 

 

 

Received on 22.06.2016         Modified on 26.07.2016

Accepted on 30.07.2015         © AJRC All right reserved

Asian J. Research Chem. 2016; 9(7): 339-342.

DOI: 10.5958/0974-4150.2016.00051.1