Validated UV Spectroscopic Method for Estimation of Montelukast sodium from Bulk and Tablet Formulations

 

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

 

Analytical method development and validation play an important role in the discovery, development and manufacture of pharmaceuticals. This research paper described a simple, selective, linear, precise and accurate UV Spectroscopic method was developed and validation for the estimation of Montelukast sodium from bulk and tablet formulations.

 

 

 

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 has clearly demonstrated the ability to ameliorate brochocostriction and indices of airway edema and abnormal mucus production as observed in clinical trials. Montelukast inhibits physiologic actions of LTD4 at the CysLT1 receptor without any agonist activity. The drug commercially available in various forms of once daily oral dosage formulations including oral granules. In oral dosage form, each packet contains Montelukast sodium equivalent to 10 mg of Montelukast^(1,2,3).

 

Fig. 1: Structure of Montelukast sodium^(4,5).

 

Instrumentation:

Double beam UV- visible spectrophotometer, Make: JASCO spectrophotometer, model JASCO V630 connected to a computer loaded with spectra manager software was used for all the spectrophotometric measurements in all proposed spectrophotometric methods. It consists of pair of 10 mm matched quartz cells was used for experiments. The absorption spectra of reference and test solution were carried out in a 1 cm quartz cell over the range 200- 400 nm.

 

MATERIAL AND METHODS:

Reagents and chemicals

All the reagents used were of analytical reagent grade. Montelukast sodium was a gifted sample obtained from Glenmark Pharmaceutical Ltd., Mumbai, India. RomilastTM [Ranbaxy Pharma] is a marketed formulation of Montelukast sodium procured from local pharmacy.

 

Preparation of standard drug solution

Standard stock solution containing Montelukast sodium was prepared by dissolving 10 mg of Montelukast in 15 ml of methanol. It was then sonicated for 10 minutes and the final volume of the solution was made up to 100 ml with distilled water to get stock solutions containing 100 ug/ml.

 

Preparation of Montelukast sodium sample solution:

Randomly selected Romilast tablets were weighed initially and crushed to powder. Powder quantity equivalent to 10 mg was weighed accurately and add in 15 ml of methanol. It was then sonicated for 10 minutes and the final volume of the solution was made up to 100 ml with distilled water to get stock solutions containing 100 ug/ml.

 

Determination of absorption maxima:

By appropriate dilution of standard drug solution with distilled water, a solution containing 10 ug/ml of Montelukast sodium solution was scanned in the range of 200-400 nm to determine the wavelength of maximum absorption for the drugs. Montelukast showed absorbance maxima at 283 nm.

 

Procedure for construction of calibration curve:

To a series of 10 ml volumetric flasks, carefully transferred aliquots of standard drug solution [0.5 to 3.0, 10 ug/ml] and the volume was made with the diluents. The instrument was for photometric mode and the absorbance of each solution was recorded at 283 nm against the blank diluents. Calibration curve was constructed by taking absorbance on ordinates and concentration of the standard Montelukast sodium on abscissa. Mean absorbance values are shown in table 1.

 

Validation parameters for Montelukast sodium:

Linearity, range and calibration curve:

The method was validated according to ICH guidelines. The linearity of analytical method is its ability to elicit test results that are directly proportional to the concentration of analyte in the sample within the range. The range of the analytical method is the interval between the upper and lower levels that have been demonstrated to be determined within a suitable level of precision, accuracy and linearity. The method was found to be linear in the concentration range of 0.3 to 3.0 ug/ ml for both methods. Absorbance spectra of each solution against distilled water as blank were measured at 283 nm and the graph of absorbance against concentration were plotted and the regression equation and correlation coefficient were determined for both the methods.

 

Precision:

The precision of an analytical method is the degree of agreement among individual test results, when the method is applied repeatedly to multiple sampling of homogeneous samples. It provides an indication of random error results and is expressed as relative standard deviation [%RSD).

 

Repeatability:

Repeatability expresses the analytical variability under the same operating conditions over a short interval of time. The repeatability studies were carried out by selecting 10 ug/ml as the standard concentration and repeating it for six times for both methods.

 

Intermediate precision:

Variation of results within the same day [ intraday] and variation of results between days [ interday] were analyzed for both methods. Intraday precision was determined by analyzing Montelukast for 6 times on the preceding day at 283 nm and % RSD was calculated.

 

Accuracy:

Accuracy is the closeness of the results obtained by the method to the true value. This study was carried out 80%, 100% and 120%. The % recovery was calculated.

 

Ruggedness:

The solutions were prepared and analyzed with change in the analytical conditions like different instrument and the different analyst.

 

Limit of detection [LOD] and Limit of quantification [LOQ]:

The sensitivity of the proposed method for the measurement of Montelukast sodium was estimated in terms of Limit of detection [LOD] and Limit of quantification [LOQ] were calculated by using the slope and SD of response. The mean slope value and the SD response were obtained from the calibration curve. The LOD and LOQ calculations were done and reported.

 

Application of the proposed method for pharmaceutical dosage form:

The solution was filtered through whatmann filter paper 0.5 ml of this solution was transferred to 10 ml volumetric flask and final volume made with the help of solvent. It was scanned on a spectrophotometer in the UV range 200-400 nm. The spectrum was recorded at 283 nm against blank solution. Determine the amount of % Montelukast sodium in tablet according to the following formula.

                       

 

                  AT× WS× Sample D.F. × Average weight × PR

% Assay =------------------------------------------------------------

                    AR× Standard D.F. × WT×LA

Where,

WS= weight of standard; WT= weight of sample, AT= absorbance of Montelukast sodium in the test solution, AR= absorbance of Montelukast sodium in the standard solution, Standard D. F. = Standard dilution factor, Sample D. F. = Sample dilution factor, PR= Purity of working standard [%], LA= Labeled amount of Montelukast sodium⁽²⁾.

 

RESULT AND DISCUSSION:

The method was validated according to ICH guidelines in order to determine the linearity, precision, accuracy and ruggedness of the method.

 

The standard stock solution of Montelukast sodium 10 ug/ml concentration was scanned from 200-400 nm and the absorption spectra’s were recorded at 283 nm wavelength in UV spectrophotometer.

 

Linearity:

The absorbance of the solution of Montelukast sodium was determined at a wavelength of 283 nm. The correlation coefficient was found to be 0.9967 and the regression equation was found to be Y= 0.0252x+ 0.0718. The calibration curves and overlay spectra’s of Montelukast sodium was shown in fig.

 

Intermediate precision [reproducibility]:

The precision of the developed method was expressed in terms of percent relative standard deviation [%RSD]. These results show reproducibility of the assay. The % RSD values were found to be less than 2 that indicate this method precise for the determination of the pure form. The interday and intraday precision results were mentioned in table3.

 

Accuracy:

Accuracy is determined by performing recovery studies at 3 levels in which known amount of analyte shall be added and recovery shall be carried out in three replicates of each concentration level and the % recovery was calculated. The accuracy results are shown in table.5.

 

 

Ruggedness:

Ruggedness studies was performed by two different analysts and two different analyst and the results of the study and % RSD obtained was less than 2 which is within the acceptance limits. And % RSD were found to be 0.34was reported in table.6.

 

Limit of detection and limit of quantification:

The parameters LOD and LOQ were determined on the basis of response and slope the regression equation LOD and LOQ values are 0.0868 and 0.2632 respectively.

 

Application of the proposed method for pharmaceutical formulation:

The result of % purity was found to be 98.70 which were shown in table.7.

 

Fig. no.2: Plot of linearity of Montelukast sodium.

 

 

Table no.1: Data for Calibration curve.

 

Table no.2: Repeatability studies of Montelukast sodium.

 

Table no. 3: Intermediate precision.

Concentration taken [ug/ml]	Intraday precision			Interday precision
	Absorbance	SD	% RSD	Absorbance	Mean	% RSD
10	0.3238 0.3231 0.3234	0.0139	0.1396	0.3238 0.3231 0.3234	0.0139	0.1396
15	0.4501 0.4481 0.4485	0.0488	0.3258	0.4504 0.4481 0.4485	0.0488	0.3258
20	0.5689 0.5698 0.5711	0.0439	0.2220	0.5684 0.5698 0.5711	0.0439	0.2220

 

Table no.5: Determination of accuracy results of Montelukast sodium.

Spiked level [%]	Formulation concentration [ug/ml]	Absorbance	Amount recovery	% Amount recovery	SD	RSD
80	10 10 10	0.5233 0.5229 0.5227	17.9167 17.7900 17.8929	98.95 98.75 98.66	0.0121	0.0677
100	10 10 10	0.5233 0.5229 0.5227	19.8452 19.8571 19.8730	98.45 98.57 98.73	0.0139	0.0702
120	10 10 10	0.6208 0.6211 0.6216	21.797 21.809 21.829	98.31 98.41 98.43	0.0160	0.0735

 

Table no.6: Ruggedness of Montelukast sodium for different analyst.

Concentration [ug/ml]	Analyst 1	% mean recovery± S.D.	% R.S.D.	Analyst 2	% mean recovery± S.D.	% R.S.D.
	Absorbance			Absorbance
10	0.3211	99.11±0.34	0.34	0.3201	98.96±0.34	0.35
10	0.3219			0.3215
10	0.3208			0.3208
10	0.3206			0.3227
10	0.3222			0.3209
10	0.3228			0.3211

 

 

Table no.7: Assay studies of Montelukast sodium.

Formulation	% purity	Mean ± SD	% RSD
RomilastTM Tablets	98.7037	98.70± 0.21	0.2168

 

CONCLUSION:

Regression equation like slope [b], intercept [a] and correlation coefficient [R²] using the method of least squares were calculated and are presented in table. 8. The results show that the methods are reasonably precise. The developed UV Spectrophotometric method was found to be simple, economic, easy, accurate, precise, reproducible and highly sensitive and can be used for routine estimation of Montelukast sodium in bulk and formulations.

 

Table no.8: Summary of UV Spectrophotometric method for estimation of Montelukast sodium.

Parameters	Bulk formulation	Tablet formulation
λmax [nm]	283	283
Correlation coefficient	0.9978	0.9966
Regression equation	0.0251x+0.0718	0.0249x+0.0775
Slope	0.0251	0.0249
Intercept	0.0718	0.0775

 

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 29.07.2016        © AJRC All right reserved