Hanimi Reddy Bapatu et al.

, IJSID, 2012, 2 (3), 377-385

ISSN:2249-5347

IJSID

International Journal of Science Innovations and Discoveries

Research Article

An International peer Review Journal for Science

Available online through www.ijsidonline.info

LEVOCETIRIZINE AND MONTELUKAST IN PHARMACEUTICAL DOSAGE FORMS

A SIMPLE RP-HPLC METHOD FOR SIMULTANEOUS ANALYSIS OF PSEUDOEPHEDRINE, BAMBUTEROL,

1Department

Hanimi Reddy Bapatu*1, Maram Ravi Kumar2, Useni Reddy Mallu3, R.S. Murthy1 and Harikishan Reddy Ganthi3 AP, India

Reddys Laboratories Ltd, Bachupally, Hyd-72, India; 3Department of Chemistry, Sri Krishnadevaraya University, Anantapur, (RP-HPLC) method has been developed for the estimation of Pseudoephedrine, Bambuterol, Levocetirizine and Montelukast in bulk and tablet dosage forms. Separation A simple and economic reverse phase high performance liquid chromatography ABSTRACT

of Chemistry, JNT University, Kukatpally, Hyderabad, AP, India; 2AR&D, Custom Pharmaceutical Services, Dr.

Received: 13.02.2012 Accepted: 19.06.2012

*Corresponding Author

was achieved on C18 column (150 x 4.6mm i.d., 5m) using gradient mobile phase Sol-A: buffer (weighed accurately 1gm of potassium di-hydrogen phosphate in to 1000mL of 20; 20-25min- sol-A: 20-98 and 25-30min- sol-A: 98-98), pumped in to the column at flow rate of 1.0 ml/min and the detection of eluent from the column was carried out 15.4 and 21.1 min, respectively. The standard curves were linear over the concentration range of 10-60 g/ml. The method was validated as per ICH guidelines. Validation control analysis in bulk and tablet dosage forms. using variable wavelength detector at 210nm. The total run time was 30 min and the column oven temperature was maintained at 35C. The retention times of Pseudoephedrine, Bambuterol, Levocetirizine and Montelukast were 4.1 min, 10.9 min, studies demonstrated that the proposed RP-HPLC method is simple, specific, rapid, INTRODUCTION reliable and reproducible. The suitability of the proposed method for the routine quality HPLC grade water) and Sol-B: acetonitrile gradient program (0-4min, sol-A:98-98; 48min- sol-A: 98-75; 8-12min- sol-A: 75-70; 12-15min- sol-A: 70-45; 15-20min- sol-A: 45-

Address: Name: Hanimi Reddy Bapatu Place: JNT University, Hyderabad, India E-mail: hanimi.b@gmail.com

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Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385 Montelukast sodium is a leukotriene receptor antagonist and used in the treatment of asthma. it is a hygroscopic,

INTRODUCTION

optically active, white to off-white powder and freely soluble in ethanol, methanol, and water and practically insoluble in behavior changes, anxiety, depression, tremors or shaking, severe sinus pain, swelling or irritation or worsening asthma. effectiveness in pediatric patients less than 12 months of age with asthma have not been established. Studies are in progress asthma when montelukast was administered in the evening without regard to time of food ingestion. body to the form.

acetonitrile. Side effects of montelukast are skin rash, bruising, severe tingling, numbness, pain, muscle weakness, mood or

General dose of montelukast is for adults and adolescents 15 years of age and older: one 10-mg tablet, for pediatric patients 6 to 14 years of age: one 5-mg chewable tablet and for pediatric patients 2 to 5 years of age: one 4-mg chewable tablet or one on clinical trials in patients with asthma to evaluate the relative efficacy of morning versus evening dosing. The pharmacokinetics of montelukast are similar whether dosed in the morning or evening. Efficacy has been demonstrated for packet of 4-mg oral granules. For pediatric patients 12 to 23 months of age: one packet of 4-mg oral granules. Safety and

and an ester prod rug of 2 adrenergic agonist and it is known to a producing of terbutaline but has better therapeutic to toxicity ratio and acts as an endogenous reservoir. Bambuterol is a pro-drug of terbutaline that is slowly converted into the younger than 4 years old it can give adverse effects of death.

Bambuterol hydrochloride is a direct acting sympathomimetic with predominantly -adrenergic activity (2-agonist) Pseudoephedrine is a decongestant that shrinks blood vessels in the nasal passages. It is used in the treatment of nasal

and sinus congestion, congestion of the tubes that drain fluid from you inner ears. Do not give pseudoephedrine to a child

Pseudoephedrine

Bambuterol

Levocetirizine

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Montelukast Figure-1: Chemical structures of active ingredients

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cetirizine. It works by blocking histamine receptors and it may not prevent the actual release of histamine from mast cells, but the area and provides relief from the typical symptoms of hay fever. Dosage forms list Tablets Dosage form

prevents in binding to its receptors. This in turn prevents the release of other allergy chemicals and increased blood supply to Montelukast 4 mg chewable tablets Montelukast 5 mg chewable tablets Montelukast 10 mg tablets Psedoepidrine HCl-15mg/60mg Bambuterol Tablets 10 mg Bambuterol Tablets 20 mg Composition

Levocetirizine is a third generation non-sedative antihistamine cetirizine. Chemically it is the active enantiomer of

Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385

Montelukast oral granule packets -- containing 4 mg per packet pseudoephedrine hydrochloride USP 120mg/tablet pseudoephedrine hydrochloride USP 240mg/tablet Levocetirizine dihydrochloride 5mg tablet.

Syrup

Bambuterol hydrochloride 5mg/5ml syrup

pseudoephedrine hydrochloride 30mg/5mL

bambuterol, Pseudoephedrine and Bambuterol for individual and combination drug products but not method available for the determination of these four drugs by HPLC method. Instruments: MATERIALS AND METHODS

determination of four drugs in single time. The present research work is to develop a single and simple method for the

Literature survey reveals that numerous methods (1-27) have been reported for the analysis of Montelukast Sodium,

systems were operated by Empower software. Waters make X-Terra C18 1504.6mm, 5m column, Mettler Toledo made analytical balance were used for this study Materials: Potassium Dr-Hydrogen orthophosphate were used. Mobile phase: Pure (not less than 98.5%) standards of all active ingredients, HPLC grade acetonitrile and water; AR grade of

A waters HPLC system consisting of alliance 2695, agilent 1200 series HPLC instrument with UV-Visible detector, two

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Filtered the final solution through a 0.4m membrane filter; Sol-B: HPLC grade acetonitrile. Diluent: Standard solution: Mixed the HPLC water and acetonitrile in the ratio of 1:1 (v/v).

Sol-A: weighed accurately 1g of potassium hydrogen phosphate, transferred in to 1000ml of HPLC water and mixed.

Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385

well then added 40mg of each Montelukast sodium, Bambuterol and Pseudoephedrine standards in to the same volumetric solution into 50ml with diluent (each standard 40ppm). Test solution: Chromatograph Mobile phase Column Flow rate Run time Chromatographic conditions Gradient program Detection Prepared the all dosage forms to get each active ingredient equal to 40microgram per mL with diluent and analyzed. : Waters/ Agilent HPLC system with Empower software. : Waters make X-Terra C18 1504.6mm, 5m. : 1.0 mL per min : 210nm : 10 L : 35C : 30 min. : Solution-A and solution-B with gradient elution. : (0-4min, sol-A: 98-98; 4-8min-sol-A: 98-75; 8-12min-sol-A: 75-70; 12-15min-sol-A: 70-45; 1520min-sol-A: 45-20; 20-25min-sol-A: 20-98 and 25-30min-sol-A: 98-98;)

flask, 50ml of diluent added and sonicated to dissolve and made up to volume with diluent. Further diluted the 5ml of resulting

Transferred the 40mg of Levocetirizine standard in to 100mL volumetric flask and added 10ml of methanol and shake

Injection volume Retention time Calculation

Column oven temp

: Pseudoephedrine-4.1, Bambuterol-10.9, Levocetirizine-15.4 and Montelukast-21.1. : All active ingredients were quantified with the following calculation.

Sample area x standard concentration x Potency of standard Standard area x sample concentration x 100

-------------------------------------------------------------------------- X 100 RESULTS AND DISCUSSION

Method Development

proportions and at deferent pH values. Flow rates between 0.5 and 1.5/min were studied. A flow rate of 1.0 ml/min gave an

optimal signal to noise ratio with a reasonable separation time. Using a reversed-phase C18 column, the retention times of Pseudoephedrine, Bambuterol, Levocetirizine and Montelukast were 4.1 min, 10.9 min, 15.4 and 21.1 min respectively. Total time of analysis was less than 30 min. The maximum absorption of Pseudoephedrine, Bambuterol, Levocetirizine and showed a complete resolution of all peaks (Figure-3). Montelukast together as detected at 210nm and this wavelength was chosen for the analysis. The chromatogram at 210nm International Journal of Science Innovations and Discoveries, Volume 2, Issue 3, May-June 2012

The mobile phase was chosen after several trials with methanol, acetonitrile, water and buffer solutions in various

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Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385 System suitability 40microgram per mL/five replicate injections) in to the chromatographic system. The percent relative standard deviation for and tabulated the results in table-1 and 2. Precision: The precision of the method was demonstrated by inter day and intraday variation studies. In the intraday studies, six System suitability parameters were established by injecting the freshly prepared standard solution (each active

peak area and retention time results found to be satisfactory. System suitability chromatograms were represented in figure-4 repeated injections of standard and sample solutions were made and the area of drug peaks and percentage RSD were calculated. In the inter day variation studies, six repeated injections of standard and sample solutions were made for three consecutive days and area of drug peaks and percentage RSD were calculated. From the data obtained, the developed HPLC method was found to be precise. Linearity: analyte concentration (10microgram per mL to 60microgram per mL for each ingredient) of the standard solution. Linearity represented in graph-1. Linearity is determined by calculating the regression line using a mathematical treatment of the linearity results vs

graph was plotted against peak area and concentration of solution. The correlation coefficient value found to be within the Robustness and Ruggedness:

limit 0.999. The linearity chromatograms shown in figure-5 and linearity results tabulated in table-4 and linearity plots were factor at the time was changed to estimate the effect. Thus, replicate injections (n = 6) of mixed standard solution at three concentration levels were performed under small changes of three chromatographic parameters (factors). Insignificant differences in peak areas and less variability in retention time were observed

Each factor selected (except columns from different manufacturers) was changed at three levels (0.1, 0 and 0.1). One

Figure-2: Diluent chromatogram International Journal of Science Innovations and Discoveries, Volume 2, Issue 3, May-June 2012

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Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385 Table-1: System suitability (Area %RSD) Inj-2

Active Ingredient Name Pseudoephedrine Bambuterol Levocetirizine Montelukast

Inj-1

Standard solution Area Inj-3 Inj-4 Inj-5

Average

%RSD

16524569 1868913 2043705 2900859

16522905 1789289 2038930 2902302

16477184 1849078 2033310 2901514

16701716 1841880 2027799 2899794

16634850 1852652 2036356 2916066

16572245 1840362 2036020 2904107

0.56 1.64 0.29 0.23

Active Ingredient Name

Pseudoephedrine Bambuterol Levocetirizine Montelukast

Table-2: System suitability (Retention time %RSD)

Figure-3: Standard chromatogram

4.11 10.89 15.38 21.09

Inj-1

Active Ingredient Name

Pseudoephedrine Bambuterol Levocetirizine Montelukast

Table-3: System suitability (USP Tailing factor) Inj-1

4.12 10.9 15.39 21.1

Inj-2

Standard solution Retention time (min) Inj-3 Inj-4 Inj-5

4.12 10.9 15.39 21.11

4.12 10.91 15.39 21.1

4.12 10.91 15.4 21.11

Average

4.118 10.902 15.39 21.102

%RSD

0.11 0.08 0.05 0.04

Resolution between Pseudoephedrine and Bambuterol 11.18 11.54 11.69 11.91 Resolution between Bambuterol and Levocetirizine 21.69 2186 21.6 21.48 Resolution between Levocetirizine and Montelukast 26.37 26.66 26.4 26.69 International Journal of Science Innovations and Discoveries, Volume 2, Issue 3, May-June 2012 Active Ingredient Name Inj-1

Table-4: System suitability (USP Resolution)

1.0 1.2 0.9 0.8

Standard solution Tailing factor Inj-2 Inj-3 Inj-4

1.0 1.1 0.9 0.8

1.0 1.1 0.9 0.8

1.1 1.1 0.9 0.8

Inj-5

1.1 1.1 0.9 0.8

Standard solution Resolution Inj-2 Inj-3 Inj-4

12.11 21.74 26.37 382

Inj-5

Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385

Figure-4: Standard solution system suitability chromatograms Active Ingredient Name Linearity level-2 (50%) Table-5: Linearity Results. Linearity solutions area Linearity Linearity Linearity level-3 level-4 level-5 (75%) (100%) (125%) Linearity level-6 (150%)

Pseudoephedrine Bambuterol Levocetirizine Montelukast

Linearity level1 (25%)

3929981 410793 481666 680955

8152866 870869 1001290 1425061

12291125 1322585 1513533 2167601

16516368 1795780 2028715 2915401

20666550 2232422 2535195 3657614

24676668 2769734 3048322 4401669

Co-relation Coefficient

0.999970 0.999639 0.999994 1.000000

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Hanimi Reddy Bapatu et al., IJSID, 2012, 2 (3), 377-385

Bambuterol, Levocetirizine and Montelukast in both bulk and tablet dosage form. The validation results reveals that, method have good precision and accuracy, which proves the reliability of the proposed method. The retention time of Pseudoephedrine is 4.2min, Bambuterol is 10.9min, Levocetirizine is 15.3min and Montelukast is 21.1min. linearity results 1.000. The short runtime and low solvent consumption are advantageous for applying in quality control analysis. REFERENCES 1. 2. 3. 4. Budavari S, editor. The Merck Index. 12th ed. Whitehouse Station, NJ: Merck & Co Inc; 1996. p. 1070. Limbird LE, Gilman AG, editors. New York: McGraw-Hill; 2001. p. 669. 2001;754:52731 Sweetman SC, editor. Martindale: The Complete Drug Reference. 33rd ed. London: Pharmaceutical Press; 2002. p. 768.

A gradient RP-HPLC method developed and validated for the simultaneous determination of Pseudoephedrine, CONCLUSION

Figure-5: Overlaid Linearity chromatograms

were found to be linear for Pseudoephedrine is 0.9999, Bambuterol is 0.999, Levocetirizine is 0.9999 and Montelukast is

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