{"id":283,"date":"2015-01-20T07:30:37","date_gmt":"2015-01-20T07:30:37","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=283"},"modified":"2020-04-23T11:23:55","modified_gmt":"2020-04-23T11:23:55","slug":"simultaneous-estimation-of-montelukast-and-bambuterol-in-tablet-dosage-forms-by-rp-hplc","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol1no1\/simultaneous-estimation-of-montelukast-and-bambuterol-in-tablet-dosage-forms-by-rp-hplc\/","title":{"rendered":"Simultaneous Estimation of Montelukast and Bambuterol in Tablet Dosage forms by RP-HPLC"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Montelukast 1<\/sup> is a selective leukotriene receptor antagonist used in the management of chronic asthma. Chemically it is 1-[[[(1R)-1-[3-[1E)-2-(7-chloro-2-quinolinyl)ethenyl]phenyl]thio]methyl]cyclopropane acetic acid1<\/sup>. Its molecular weight is 586.18 and molecular formula is C35<\/sub>H36<\/sub>NSClO3.<\/sub> Montelukast is a selective cysteinyl leukotrien type I receptor antagonsit4. <\/sup>Bambuterol3<\/sup> is inactive prodrug of terbutaline, a direct acting sympathomimetic with predominantly beta-adrenergic activity and a selective action on beta2<\/sub>ireceptors. It is used as bronchodilator for persistent asthma. Chemically it is Dimethyl carbamic acid 5-[2-[(1,1-dimethyl ethyl)amino]-1-hydroxy ethyl]-1,3,-phenylene ester2<\/sup> with molecular weight 367.4 and molecular formula C18<\/sub>H29<\/sub>N3<\/sub>O5. <\/sub>Literature survey reveals a few chromatographic methods 6-12<\/sup> to deterimine the montelukast in tablet dosage form and in biological fluids and only one spectrophotometric method5<\/sup> was reported for the estimation of bambuterol in tablet dosage form. So far, no chromatographic methods were reported for the simultaneous estimation of Montelukast & Bambuterol in pharmaceutical dosage forms. The availability of an HPLC method with high sensitivity and selectivity will be very useful for the determination of Montelukast & Bambuterol in pharmaceutical formulations. The aim of the study was to develop a simple, precise and accurate reverse-phase HPLC method for the estimation of Montelukast & Bambuterol in bulk drug samples and in pharmaceutical dosage form.<\/p>\n\n\n\n
\u00a0\"Scheme<\/td>\n\n

Scheme 1<\/strong><\/p>\n

Click here to View\u00a0scheme<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Experimental<\/strong><\/p>\n

Materials and Methods<\/strong><\/p>\n

Montelukast & Bambuterol were obtained as a gift samples from MSN Pharmachem Pvt.Ltd, Hyderabad. Acetonitrile and water used were of HPLC grade (Qualigens). Commercially available tablets (Montair Plus, Cipla) were procured from local market.<\/p>\n

Instrument<\/strong><\/p>\n

Quantitative HPLC was performed on liquid Chromatograph, Waters separation 2996, PDA detector module equipped with automatic injector with injection volume 20 \u00b5l, and 2693 pump. An Inertsil-ODS-3V C18 column (250×4.6 mm i.d; particle size 5 \u03bcm) was used. The HPLC system was equipped with Empower Software.<\/p>\n

HPLC Conditions<\/strong><\/p>\n

The contents of the mobile phase were 0.1% Trifluoro Acetic acid: Methanol in the ratio of 10:90 v\/v. They were filtered before use through a 0.45 \u03bcm membrane filter, and pumped from the respective solvent reservoirs to the column at a flow rate of 0.8 ml\/min. The run time was set at 12.0 min and the column temperature was ambient. Prior to the injection of the drug solution, the column was equilibrated for at least 30 min with the mobile phase flowing through the system. The eluents were monitored at 220 nm.<\/p>\n

Preparation of Standard Stock solution<\/strong><\/p>\n

A standard stock solution of the drug was prepared by dissolving 193.5 mg of Bambuterol hydrochloride (equivalent to 175mg of Bambuterol) and 182 mg of Montelukast sodium (equivalent to 175 mg of montelukast) in 100 ml volumetric flask containing 30 ml of methanol as diluent, sonicated for about 15 min and then made up to 100 ml with methanol to get standard stock solution of 1.75 mg\/ml each of montelukast and Bambuterol.<\/p>\n

Working Standard solution<\/strong><\/p>\n

10ml of the above stock solution was taken in 50 ml volumetric flask and made up to 50 ml with methanol as diluent to get a concentration of each 350\u00b5g\/ml of Montelukast and Bambuterol.<\/p>\n

Preparation of Sample solution<\/strong><\/p>\n

Twenty tablets (Montair Plus, Cipla) were weighed, and then powdered. A sample of the powdered tablets, equivalent to mixture containing 175 mg of montelukast and 175 mg of bambuterol active ingredients, was mixed with 30 ml of methanol as diluent in 50 ml volumetric flask. The mixture was allowed to stand for 1 hr with intermittent sonication to ensure complete solubility of the drug, and then filtered through a 0.45 \u03bcm membrane filter, followed by adding methanol up to100 ml to obtain a stock solution each of 1.75mg\/ml of Montelukast and Bambuterol. 10ml of the above sample stock solution was taken in 50 ml volumetric flask and made up to 50 ml with methanol as diluent to get a concentration of each 350 \u00b5g\/ml of Montelukast and Bambuterol.<\/p>\n

Linearity<\/strong><\/p>\n

Aliquots of standard Montelukast & Bambuterol stock solution were taken in different 10 ml volumetric flasks and diluted up to the mark with the mobile phase such that the final concentrations of Montelukast & Bambuterol are in the range of 17.5-420 \u03bcg\/ml. Each of these drug solutions (20 \u03bcL) was injected three times into the column, and the peak areas and retention times were recorded. Evaluation was performed with PDA detector at 220 nm and a Calibration graphs were obtained by plotting peak area versus concentration of Montelukast & Bambuterol (Fig 2). The plot of peak areas of each sample against respective concentration of Montelukast & Bambuterol was found to be linear in the range of 17.5-420 \u03bcg\/ml.with correlation coefficient of 0.9999. Linear regression least square fit data obtained from the measurements are given in table I. The respective linear regression equation being Y= 26705.167x+67644.9694 for Bambueterol and Y=75702.306x+114141.09 for Montelukast. \u00a0The regression characteristics, such as slope, intercept, and %RSD were calculated for this method and given in Table I.<\/p>\n\n\n\n
\"Figure<\/td>\n\n

Figure 1:\u00a0 Typical Chromatogram of Montelukast & Bambuterol by RP-HPLC.<\/strong><\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table I: Linear Regression Data for Calibration curves.<\/strong><\/p>\n\n\n\n\n
Parameter<\/strong><\/td>\nMontelukast<\/strong><\/td>\nBambuterol<\/strong><\/td>\n<\/tr>\n
Conc.range (\u00b5g\/ml)<\/p>\n

Slope (m)<\/p>\n

Intercept (b)<\/p>\n

Correlation coeff.<\/p>\n

% RSD<\/p>\n

Standard error of estimate<\/td>\n

17.5-420<\/p>\n

75702.306<\/p>\n

114141.0925<\/p>\n

0.9999<\/p>\n

0.93<\/p>\n

118206.6442<\/p>\n

 <\/td>\n

\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 17.5-420<\/p>\n

26705.167<\/p>\n

67644.9694<\/p>\n

0.9999<\/p>\n

0.35<\/p>\n

 <\/p>\n

48619.7117<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\u00a0\"Figure<\/td>\n\n

Figure 2: Caliberation curves of the Montelukast & Bambuterol by RP-HPLC<\/strong>.<\/p>\n

Click here to View figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Assay<\/strong><\/p>\n

20 \u00b5l of sample solution was injected into the injector of liquid chromatograph. The retention times were found to be 2.373min for Bambuterol and 6.899 for Montelukast.. The amount of drug present per tablet was calculated by comparing the peak area of the sample solution with that of the standard solution. The data are presented in Table II.<\/p>\n

Table II: Assay and Recovery of Montelukast & Bambuterol in Tablet dosage form.<\/strong><\/p>\n\n\n\n\n\n\n\n
 <\/p>\n

 <\/p>\n

Tablet formulation<\/strong><\/td>\n

Amount claim<\/strong><\/p>\n

(mg\/tablet)<\/strong><\/td>\n

Amount claim<\/strong><\/p>\n

(mg\/tablet)<\/strong><\/td>\n

 <\/p>\n

Amount Obtained (mg)*<\/strong><\/p>\n

by proposed method<\/strong><\/td>\n

 <\/p>\n

** % Recovery by the<\/strong><\/p>\n

Proposed method<\/strong><\/td>\n<\/tr>\n

Monteleukast<\/strong><\/td>\nBambuterol<\/strong><\/td>\nMontelukast<\/strong><\/td>\nBambuterol<\/strong><\/td>\nMontelukast<\/strong><\/td>\nBambuterol<\/strong><\/td>\n<\/td>\n<\/tr>\n
1<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 10<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0 10<\/td>\n9.84<\/td>\n9.87<\/td>\n99.5<\/td>\n100.3<\/td>\n<\/td>\n<\/tr>\n
2<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 10<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0 10<\/td>\n9.88<\/td>\n9.88<\/td>\n99.0<\/td>\n101.1<\/td>\n<\/td>\n<\/tr>\n
3<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 10<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0 10<\/td>\n9.80<\/td>\n9.86<\/td>\n99.1<\/td>\n99.2<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

*Average of three determinations.<\/p>\n

** After spiking the sample.<\/p>\n

Recovery Studies<\/strong><\/p>\n

Accuracy was determined by recovery studies of Montelukast & Bambuterol; known amount of standard was added to the preanalysed sample and subjected to the proposed HPLC analysis. Results of recovery study are shown in Table II. The study was done at three different concentration levels.<\/p>\n

Results and Discussion<\/strong><\/p>\n

The system suitability tests were carried out on freshly prepared standard stock solutions of Montelukast & Bambuterol. Parameters that were studied to evaluate the suitability of the system are given in Table III.<\/p>\n

Table III:\u00a0 Validation Summary<\/strong><\/p>\n\n\n\n\n\n
Parameter<\/strong><\/td>\nMontelukast<\/strong><\/td>\nBambuterol<\/strong><\/td>\n<\/tr>\n
System Suitability<\/u><\/p>\n

TheoreticalPlates(N)<\/p>\n

Tailing factor<\/p>\n

Retention time(min)<\/p>\n

Resolution<\/p>\n

% Peak Area<\/td>\n

 <\/p>\n

2400.38<\/p>\n

1.14<\/p>\n

6.899<\/p>\n

17.52<\/p>\n

99.98<\/td>\n

 <\/p>\n

2007.45<\/p>\n

1.15<\/p>\n

2.373<\/p>\n

17.36<\/p>\n

99.82<\/td>\n<\/tr>\n

LOD (\u00b5g\/ml)<\/p>\n

LOQ (\u00b5g\/ml)<\/td>\n

0.35<\/p>\n

1.05<\/td>\n

0.175<\/p>\n

0.525<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Limit of Detection (LOD) and Limit of Quantification (LOQ)<\/strong><\/p>\n

The limit of detection (LOD) is 0.175mcg and 0.35 mcg for Bambuterol and Montelukast respectively. The limit of quantification (LOQ) is 0.525mcg for Bambuterol and 1.05mcg for Montelukast.<\/p>\n

From the typical chromatogram of Montelukast & Bambuterol as shown in Fig 1, it was found that the retention times were 2.373min. for bambuterol and 6.899 min. for Montelukast. A mixture of 0.1% Trifluoro Acetic acid: Methanol in the ratio of 10:90 v\/v was found to be the most suitable as mobile phase to obtain the peaks well defined and free from tailing. In the present developed HPLC method, the standard and sample preparation required less time and no tedious extractions were involved. A good linear relationship (r=0.9999) was observed between the concentration range of 17.5-420 \u00b5g\/ml. Low values of standard deviation are indicative of the high precision of the method. The assay of Montelukast & Bambuterol tablets was found to be 98.48% and 98.78% respectively. From the recovery studies it was found that about 99.14% of Montelukast & 100.2% of Bambuterol was recovered which indicates high accuracy of the method. The absence of additional peaks in the chromatogram indicates non-interference of the common excipients used in the tablets. This demonstrates that the developed HPLC method is simple, linear, accurate, sensitive and reproducible. Thus, the developed method can be easily used for the routine quality control of bulk and tablet dosage forms of Montelukast & Bambuterol within a short analysis time.<\/p>\n

Acknowledgements<\/strong><\/p>\n

The authors are grateful to M\/s MSN Pharmachem Pvt.Ltd, Hyderabad for the supply of as a gift sample of Montelukast and\u00a0Bambuterol and to the Management, Sultan-Ul-Uloom college of Pharmacy, Hyderabad, for providing the necessary facilities to carry out the research work.<\/p>\n

References<\/strong><\/p>\n

    \n
  1. The Merck Index (monograph#953), 14, 1080, (2006).<\/li>\n
  2. The Merck Index (monograph#6258), 14, 161, (2006).<\/li>\n
  3. \u00a0Martindale-The Complete Drug Reference, 34, 781, (2005).<\/li>\n
  4. Martindale-The Complete Drug Reference, 34, 788, (2005).<\/li>\n
  5. S. Appala Raju, Shobha Manjunath, Asian J. Chem. 15, 1117-1120, (2003).<\/li>\n
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  7. Chauhan B, Rani\u00a0Shubha ; Nivasarkar M.\u00a0 PadhH. ; Indian journal of pharmaceutical sciences, 68(4), 517-520\u00a0(2006).<\/li>\n
  8. Alsarra I, Khalil NY, Sultan M, Al-Ashban R,\u00a0 Pharmazie, 60(11), 823-6, (2005).<\/li>\n
  9. Smith GA, Rawls CM, Kunka RL. Pharm Res, 21(9):1539-44, (2004).<\/li>\n
  10. Radhakrishna T, Narasaraju A, Ramakrishna M, Satyanarayana A. J Pharm Biomed Anal. 31(2), 359-68, (2003).<\/li>\n
  11. Al-Rawithi S, Al-Gazlan S, Al-Ahmadi W, Alshowaier IA, Yusuf A, Raines DA.\u00a0<\/u>J Chromatogr B Biomed Sci Appl, 754(2), 527-31 (2001).<\/li>\n
  12. Ochiai H, Uchiyama N, Takano T, Hara K, Kamei T. J Chromatogr B Biomed Sci Appl. 713(2), 409-14, (1998).<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

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