VALIDATED UV SPECTROSCOPIC METHOD FOR ESTIMATION OF MONTELUKAST SODIUM

VALIDATED UV SPECTROSCOPIC METHOD FOR ESTIMATION OF MONTELUKAST SODIUM

Kuldeep Singh^*, Paramdeep Bagga, Pragati Shakya, Arun Kumar, M. Khalid, J. Akhtar and M. Arif

Faculty of Pharmacy, Integral University, Kursi road, Lucknow-226026, Uttar Pradesh, India

ABSTRACT: Montelukast sodium is a leukotriene receptor antagonist (LTRA) used for the maintenance treatment of asthma and to relieve symptoms of seasonal allergies^1-2. Montelukast comes as a tablet, a chewable tablet, and granules to take by mouth³. Montelukast is usually taken once a day with or without food³. The present study describes a simple, accurate reproducible and precise uv spectrophotometric method for the estimation of Montelukast Sodium in bulk and in tablet dosage form. The absorbance maxima (λmax) for Montelukast Sodium were found to be 286.5nm. The method was validated for different parameters such as molar absorptivity, accuracy precision, ruggedness, robustness, detection limit, quantification limit etc, (as per ICH guidelines). The relative standard deviation (RSD) in case of, accuracy precision, ruggedness, robustness was less than 2.0% proving that method was highly accurate, precise and robust. This method can be used for determination of Montelukast Sodium in pharmaceutical formulations without interference of the excipients

Montelukast Sodium, Validation, Leukotriene receptor, Ruggedness

INTRODUCTION: Montelukast sodium (MNLT), (S, E) – 2 - (1-((1-(3-(2- (7-chloroquinolin-2-yl) vinyl) phenyl) – 3 - (2-(2- hydroxypropan-2-yl) phenyl) propylthio)methyl)cyclopropyl)acetic acid. It is a leukotriene receptor antagonist (LTRA) used for the maintenance treatment of asthma and to relieve symptoms of seasonal allergies. It is usually administered orally. Montelukast is a CysLT1 antagonist; it blocks the action of leukotriene D4 (and secondary ligands LTC4 and LTE4) on the cysteinyl leukotriene receptor CysLT1 in the lungs and bronchial tubes by binding to it.

This reduces the bronchoconstriction otherwise caused by the leukotriene and results in less inflammation. The chemical structure of MNLT is shown in Fig.1.

FIG. 1

The drug is 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. Several analytical methods have been reported for determination of Montelukast including derivative spectroscopic ⁴, by colorimetry ⁵, by flouorimetry ⁶, by TLC ⁷, by HPTLC ⁸, by simultaneous UV determination in combination drug formulation ⁹, by voltametry ¹⁰, by high performance liquid chromatography (HPLC) ¹¹ and by LCMS ¹². To our knowledge, there is no simple and accurate UV spectrophotometric method for quantitative determination of Montelukast Sodium in its bulk and in its tablet dosage forms.

The objective of study to develop and validate a simple, eco-friendly UV spectrophotometric method for the determination of Montelukast in Montelukast Sodium oral dosage forms (strength is 10 mg as Montelukast). Also method should be capable to apply in routine Quality control analysis. Analytical parameters for the method have also been established and compared with those established and existing HPLC method.

MATERIALS AND METHODS:

 A shimadzu - 1700 double beam uv/vis. spectrophotometer with 1cm matched quartz cells was used for absorbance measurements. Montelukast sodium was a gifted sample obtained from Sanofi India Ltd., Ankleshwar, India.

Double distilled water was used for analysis. Montelast (Cadila pharma) and Montair (Cipla pharma) were marketed formulation of Montelukast sodium procured from local pharmacy. Water, methanol and sodium hydroxide (loba chemical ltd.,) were of analytical grade was used. Drug was dissolved in methanol and further dilutions made with 0.1N NaOH. All the reagents used were of analytical reagent grade.

 Preparation of stock and standard solution:

Stock solution of Montelukast sodium was prepared by dissolving 10 mg in methanol and final dilution was made by 0.1 N NaOH. Suitable aliquots of the stock solution of Montelukast sodium were pipette and transferred separately into 10ml volumetric flasks. The volume was made up with the same solvent.

Determination of Absorption maxima:

Working standard solutions were scanned in UV range of (200-400nm) using a Shimadzu 1700 UV-Visible spectrophotometer with cells of 10mm length against the same solvent used as blank. It was scanned in the range of 200-400 nm and it shows absorbance maxima at 286.5 nm.

Procedure for Analysis of Tablet Formulation:

Twenty tablets were weighed and powdered. An accurately weighed tablet powder equivalent to10 mg of Montelukast sodium transferred in to 25 ml volumetric flask, small amount of methanol was added and sonicated for 15 min. Volume was made up to the mark with 0.1 N NaOH then solution was filtered through whatman filter paper no.41. From this stock solution, necessary dilutions were made with 0.1 N NaOH to get final concentration of 10 μg/ml. The above solution was then analyzed for the content of Montelukast sodium.The amount of Montelukast sodium present in each formulation was calculated from the respective calibration curve.

TABLE 1: OPTICAL CHARACTERISTICS AND PRECISION OF THE PROPOSED METHOD

TABLE 2: MEAN ABSORBANCE VALUE AND STATISTICAL DATA OF THE CALIBRATION CURVE

Analytical method validation: The method was validated for different parameters like linearity, specificity, selectivity, accuracy, precision, robustness, and ruggedness, limit of quantification (LOQ) and limit of detection (LOD) ^13-14

Linearity and Range:

Solutions containing 5, 10, 15, 20, and 25μg / ml of Montelukast sodium were prepared from standard solution to determine the linearity range. The detection was carried out at 286.5 nm. Spectrums were recorded and absorbance was recorded for all the concentrations. A calibration plot of concentration over the absorbance was constructed against the corresponding concentrations to obtain the calibration graphs and was shown in Fig.2 The optical characteristics such as Beer’s law limits, regression equation and correlation coefficient, mean absorbance value, and statistical data of the calibration curve were calculated and results are presented in Table 1 Linearity was checked by calculating regression coefficient ¹⁴.

FIG.2: STANDARD CALIBRATION CURVE OF MONTELUKAST SODIUM SOLUTION IN 0.1 N NAOH

 Specificity and selectivity:

Specificity and selectivity of the selected method was determined by preparing 10μg/ml of Montelukast sodium solution in 0.1 N NaOH along with and without common excipients (Lactose, Hydroxypropyl methylcellulose and starch) separately. All the solutions were scanned from 200-400 nm at fast speed and analyzed for any change in absorbance and percentage drug recovery at respective wavelengths.

Accuracy:

The accuracy of the method was determined by calculating percentage drug recovery of Montelukast at a concentration of 10 μg/ml (n=6) ¹⁴.

Precision:

The precision of proposed method was determined by varying the practical conditions. Inter-day, intra-day and inter-analyst variations were studied to determine the intermediate precision of proposed analytical method. Drug concentrations of Montelukast, mixture was prepared at three different times in a day and studied for intra-day variation. The same procedure was followed for six days in order to study inter-day variations. Developed method was performed by three different analysts for inter analyst studies and by same analyst at six different times for intra analyst studies. The percent relative standard deviation (% RSD) of prepared concentrations was analyzed for precision studies ¹⁴.

Limit of detection and Limit of quantification: Limit of detection (LOD) and Limit of quantification (LOQ) of Montelukast, mixture by proposed method were determined using calibration standards. LOD and LOQ were calculated as 3.3σ/S and 10σ/S respectively. Where, σ is standard deviation of y-intercept of regression equation and S is the slope of the calibration curve ¹⁴.

Ruggedness and Robustness:

Ruggedness and robustness was evaluated by analyzing Montelukast, mixture under variable experimental conditions (a) Sample solution containing 10μg/ml was assayed for six times prepared from different stock solution and the percentage recovery was checked in each case. (b) Sample solution containing 10μg/ml was analyzed by changing the UV apparatus (Apparatus 1: Shimadzu-1800; Apparatus 2: Cintra) to check the inter-instrument variation in the absorbance. (c) Robustness in the form of stability of Montelukast, in prepared solution was also checked at room temperature. The concentration of drug in solvent was checked at time of preparation and at the interval of twenty-four hours¹⁴

Stability:

Stability of the solutions of Montelukast, used for preparing the calibration curves in the method, was ascertained by observing for changes in the absorbance at their analytical wavelengths over a period of 24 h.

RESULTS AND DISCUSSION:

Standard calibration curve The UV scan of standard solution of Montelukast between 200-400 nm gives the absorption maxima at 286.5nm. The regression coefficient for calibration curve of Montelukast in 0.1 N NaOH was found to be 0.999 with regression equation of Y_286.5nm = 0.034x. The optical and regression characteristics are given in Table 1.

Analytical method validation:

Linearity The linearity was found in the range of 5-25μg/ml for Montelukast in 0.1N NaOH supported by high regression coefficient.

Specificity:

The UV-Spectrum of Montelukast showed no change in the presence of common excipients used for the formulation. Absorption spectrum of pure drug sample was matching with the sample mixed with excipients. Table 2 shows the values for specificity studies of Montelukast  in 0.1 N NaOH. The excellent percentage recovery of shows that there is no effect of excipients on the absorbance.

TABLE 2: RESULTS OF SPECIFICITY AND SELECTIVITY STUDIES OF APPLIED METHOD OF UV ESTIMATION OF MONTELUKAST

Accuracy:

The accuracy of the method was checked by determining the percentage recovery values. Percentage drug recovery (± RSD) of above concentrations of Montelukast in 0.1 N NaOH is shown in Table 3. In all the cases RSD was not more than 2% depicting the accuracy of the developed method.

TABLE 3: RESULTS OF ACCURACY STUDIES OF APPLIED METHOD OF UV ESTIMATION OF MONTELUKAST

Precision:

The precision of the developed method was determined by studying the repeatability of the developed method. In this, the percentage recovery of Montelukast in 0.1 N NaOH was calculated by inter-day, intra-day, inter-analyst and intra analyst conditions over a short interval of time. Results of percentage drug recovery obtained from intra-day studies, inter-day studies and inter-analyst studies are shown in Table 4. From obtained data, it was found that in all the precision studies, RSD was not more than 2 indicating that developed method has excellent repeatability.

TABLE 4: RESULTS OF PRECISION STUDIES OF APPLIED METHOD OF UV ESTIMATION OF MONTELUKAST (CONC. OF DRUG TAKEN 10μg/ml)

Limit of Detection and Limit of Quantification:

LOD and LOQ were calculated to be 0.471μg/ml and 1.42 μg/ml respectively.

Robustness:

Robustness of the selected method was checked by inter-stock solution, inter-instrument, and by calculating percentage drug recovery after twenty-four hours of preparation of stock solution (Table 5). Overlaid spectra of stock solution at zero and twenty-four hours (Fig. 7) showed no change in absorbance and λ_max of the solution. Further, data showed that in all the above cases, RSD was found to be less than 2 confirming the robustness of the developed method.

TABLE 5: RESULTS OF ROBUSTNESS STUDIES OF APPLIED METHOD OF UV ESTIMATION OF MONTELUKAST (CONC. OF DRUG TAKEN 10μg/ml)

CONCLUSION: The most striking feature of this method is its simplicity and rapidity, non- requiring- consuming sample preparations such as extraction of solvents, heating, degassing which are needed for HPLC procedure. These are new and novel methods and can be employed for routine analysis in quality control analysis. The described methods give accurate and precise results for determination of Montelukast, sodium in marketed formulation.

REFERENCES:

1. Lipkowitz, Myron A. and Navarra, Tova: The Encyclopedia of Allergies. (2nd ed.) New York, Edition 2: 178.
2. http://www.mascothealth.com
3. Montelukast article on Medline Plus http://www.nlm.nih.gov/medlineplus/druginfo/meds/a600014
4. Patel PG, Vaghela VM, Rathi SG, Rajgor NB and Bhaskar VH: Derivative spectrophotometry method for simultaneous estimation of Rupatadine and Montelukast in their combined dosage form. Journal of Young Pharmacists. 2009; 1:354- 358.
5. Shanmukha kumar JV, Ramachandran D, Settaluri VS and Shechinah Felice C. Spectrophotometric methods for estimation of leukotriene receptor antagonist in bulk dosage forms. RASĀYAN Journal of Chemistry. 2010; 3:166-171.
6. Pattana Sripalakita, Bungon Kongthongc and Aurasorn Saraphanchotiwitthayad. A simple bioanalytical assay for determination of Montelukast in human plasma: Application to a pharmacokinetic study. Journal of Chromatography B. 2008; 869:1-2, 38-44.
7. Smita Sharma MC, Sharma DV, Kohli and Sharma AD. Development and validation of TLC densitometry method for simultaneous quantification of Montelukast sodium and Levocetirizine dihydrochloride pharmaceutical solid dosage form. Der Pharmacia Lettre. 2010; 1:489- 494.
8. Sane RT, Ajay Menezes, Mandar Mote Atul Moghe and Gunesh Gundi. HPTLC Determination of Montelukast Sodium in Bulk Drug and in Pharmaceutical Preparations. Journal of Planar Chromatography. 2004; 17:75-78.
9. Varun Pawar, Sanjay Pai and Roa GK.Development and Validation of UV Spectrophotometric Method for Simultaneous Estimation of Montelukast Sodium and Bambuterol hydrochloride in Bulk and Tablet Dosage Formulation. Journal of Pharmaceutical Sciences. 2008; 2:152-157.
10. Alsarra IM, Al-Omar EA, Gadkariem and Belal F. Voltammetric determination of Montelukast sodium in dosage forms and human plasma, Il-Farmaco. 2005; 60:563–567.
11. Radhakrishnaa T, Narasarajua A, Ramakrishnab M and Satyanarayana A. Simultaneous determination of Montelukast and Loratadine by HPLC and derivative spectrophotometric methods, Journal of Pharmaceutical and Biomedical Analysis. 31:359-368.
12. Vijaya Bharathi D, Kishore Kumar Hotha, Jagadeesh B, Ramesh Mullangi A and Naidu. Quantification of Montelukast, a selective cysteinyl leukotriene receptor (CysLT1) antagonist in human plasma by liquid chromatography–mass spectrometry: validation and its application to a human pharmacokinetic study, Biomedical Chromatography. 2009; 23:804-810.
13. ICH Q2B, 1996. Guidelines on validation of analytical procedure: methodology. Federal register. 60:27424.
14. Bolton S, Linear regression and correlation. In: Bolton S, Pharmaceutical statistics, 3^rd New York (USA), Marcel Dekker Inc; 1997.
    

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    How to cite this article:Singh K, Bagga P, Shakya P, Kumar A, Khalid M, Akhtar J and  Arif M: Validated UV Spectroscopic Method for Estimation of Montelukast Sodium. Int J Pharm Sci Res 2015; 6(11): 4728-32.doi: 10.13040/IJPSR.0975-8232.6(11).4728-32.

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