Derivative and Absorption Factor Spectrophotometric Estimation of Montelukast Sodium and Levocetirizine Dihydrochloride from Pharmaceutical Formulations

 

Vishnu P. Choudhari*, Anamika N. Kale, Satish A.Polshettiwar, Abhijit S. Sutar, Dhaval M. Patel and Bhanudas S. Kuchekar

Pharmaceutical Analysis and Quality Assurance Department, MAEER’s Maharashtra Institute of Pharmacy, MIT Campus, Paud Road, Kothrud,  Pune, 411038, MS, India.

*Corresponding Author E-mail: viraj1404@rediffmail.com

ABSTRACT:

Two simple and sensitive spectrophotometric methods are described for the determination of Montelukast sodium and Levocetirizine Dihydrochloride in combined dosage form. First method was first order derivative spectroscopy where montelukast and levocetirizine were determined at λmax 340nm and 331nm, respectively in methanol. The second method was based on the absorption factor in which montelukast and levocetirizine exhibit λmax at 232 nm and 279 nm respectively in methanol. Montelukast has some interference at 232nm, while levocetirizine do not show any absorption at 279 nm. Quantitative  estimation  of levocetirizine was  carried  out  by subtracting  the  absorption  due  to montelukast at  232nm  using  experimentally calculated absorption factor. Beer’s law was obeyed for montelukast and levocetirizine in the concentration range of 4-12 µg ml^-1 and 2-6 µg ml^-1 , respectively for both methods. The results of analysis have been validated statistically and recovery studies confirmed the accuracy of the proposed method.

 

 

INTRODUCTION:

Montelukast (MON) is an oral selective leukotriene receptor antagonist that inhibits the cysteinyl leukotriene cysLT1 and has been shown to be effective in the treatment of chronic asthma and Chemically, it is 2-[1-[(R)-[3-[2(E)-(7-chloroquinolin-2-yl)vinyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl-sulfanyl methyl] cyclo propyl]acetic acid sodium salt.Levocetirizine (LEV as Levocetirizine Dihydrochloride) is chemically [2-[4-[(R)-(4-Chlorophenyl)phenyl methyl]-1piperazinyl]ethoxy]-acetic acid Dihydrochloride a third-generation non-sedative antihistamine¹. LEV is official in are official in IP 2007². Literature survey reveals that HPLC and UV Spectroscopic methods are reported ^3-25 for the estimation of MON and LEV individually as bulk and in pharmaceutical formulations. The review of the literature revealed that there is no Spectrophotometric method available for this combination. Aim of the study was to develop simple, rapid, accurate, reproducible and economical first order derivative and absorption factor spectroscopic methods for both the drugs in combined dosage forms.

 

EXPERIMENTAL:

Drugs and Chemicals:

Spectroscopic grade Methanol was used throughout the study. All the solvents and reagents used were purchased from LOBA Chemie Pvt. Ltd., Mumbai. Two brands of tablets used for analysis were Montek LC (Tab I) manufactured by Atoz Life Sciences, Pondicherry, India ( Batch no AF60258) and  Montair-LC (Tab II) manufactured by Cipla LTD., Solan, India (Batch no.D84089) containing  Montelukast Sodium equivalent to Montelukast 10mg and Levocetirizine dihydrochloride 5 mg per tablet. Pure drug sample of MON, % purity 98.5% was kindly supplied as a gift sample by Ranbaxy Laboratories Ltd., Dewas and pure drug sample of LEV,  % purity 99.8 was gifted by Mapro Pharmaceuticals, Gujarat. These samples were used without further purification.

 

METHOD A: FIRST ORDER DERIVATIVE SPECTROSCOPY:

Spectral measurements were made on Varian UV- Visible Spectrophotometer CARY100 model with Spectral band width 1 nm. Wavelength accuracy is 0.5 nm and 1 cm matched quartz cells were used. The UV spectra of MON and LEV in various solvents show no spectral isolation. The concentration of LEV is half times lesser than MON and so simultaneous estimation would have much interference. To overcome this, spectra of two drugs were derivatized from first to fourth orders. To determine derivative amplitude for LEV and MON, solution of increasing and decreasing concentrations of LEV and MON were prepared in combination as shown in table no. 1. These spectrums were converted to first order derivative spectra by using instrument mode. After observing the derivative amplitude of first order derivative spectra, it was observed that there is proportionate increase in amplitude of MON with increase in its concentrations at 340 nm and amplitude of LEV decreased with decreased in its concentrations at 331 nm.   Therefore λmax   340 nm and λmax  331nm  were assigned to MON and LEV,  respectively for the study.

 

Table 1: Determination of derivative amplitude for LEV and MON by first order derivative method (n=6)

Sr. No.	Concentration (μg mL-1)		First order derivative amplitude
	LEV	MON	at 331 nm (LEV)	at 340nm (MON)
1	2	12	1.958	12.69
2	3	10	2.836	10.60
3	4	8	3.793	8.546
4	5	6	4.827	6.482
5	6	4	5.684	4.674

 

METHOD B: ABSORPTION FACTOR METHOD:

Preparation of Standard Solutions:

Separate standard stock solutions of MON and of LEV were prepared by dissolving and diluting 100mg of each in to separate 100ml volumetric flask with methanol.

 

Construction of Calibration Curve:

λmax of MON (8 µg ml^-1) and LEV (4 µg ml^-1 )was determined by scanning the drug solution in methanol and was found to be at 279 nm and 232nm respectively. MON also showed absorbance at 232 nm, while LEV did not show any interference at 279nm. To construct Beer’s plot for MON and LEV dilutions were made in methanol using stock solution of 100 µg ml^-1. Also Beer’s plot was constructed for MON and LEV  in  solution  mixture at  different concentration  (4:6,6:5,8:4,10:3 and 12:2 µg ml^-1) levels. Both the drugs followed linearity individually and in mixture within the concentration range 4-12 µg ml^-1  and 2-6 µg ml^-1 for MON and LEV respectively (Fig.no.2).

 

Fig.1: First order derivative spectrum of MON (8µg mL^-1) and LEV (8µg mL^-1) standard mixture in methanol.

 

Fig.2. Overlay spectrum of MON and LEV in methanol. MON (4-12µg mL^-1) and LEV (2-6 µg mL^-1)

 

Determination of Absorption Factor at Selected Wavelengths:

MON and LEV solution in methanol of known concentrations were scanned against blank on spectrophotometer. The value of absorption factor was found to be 2.688. Quantitative estimation of MON and LEV was carried out using following equation:

Corrected Abs of LEV at 232nm

=Abs232 (MON+LEV) – [(abs 232 (MON)/ abs 279 (MON)] *abs279 (MON+LEV)

Where; abs: Absorption value at given wavelengths.

 

Assay of Tablet Formulation:

Accurately weighed 20 tablets were powdered and quantity of tablet powder equivalent to 10 mg MON (5 mg LEV) transferred to 100ml grade A volumetric flask. To it  80 mL of  methanol was added, mixed   and sonicated  for  5min. Volume was adjusted with methanol and filtered  through What man filter paper No. 41 and portion of filtrate was suitably diluted to get final concentration of LEV 5 µg ml^-1 and MON 10 µg ml^-1. Appropriated aliquots were subjected to above methods, absorbances were noted at selected wavelength and concentrations were calculated.  The results of formulations analysis are given in table no.2.

 

RESULTS AND DISCUSSION:

Practically no interference from tablet excipients was observed in these methods. As  their  λmax  differ more  than 20 nm,  absorption  factor  method  was  tried  for  their  simultaneous  estimation  in formulation. Quantitative estimation of LEV was carried out by subtracting interference of MON using experimentally calculated absorption factor. Both the methods are accurate, simple, rapid, precise, reliable, sensitive, reproducible and economical.  The values of  % RSD  and correlation of coefficient were satisfactory ( table no.2). Results of the formulation analysis are in agreement with the label claim and result of the recovery study (table no.3) indicates that there is no interference due to excipients present in the formulation.

 

 

Table 2: Validation parameters

Parameters			Montelukast sodium				Levocetirizine  dihydrochloride
			Method A		Method B		Method A		Method B
Linearity range (µg/mL)			4-12		4-12		2-6		2-6
Slope			0.0865		0.0733		0.0443		0.0634
Intercept			0.588		0.0018		0.0424		-0.0315
Correlation coefficient (r)			0.996		0.998		0.999		0.999
Precision ( %RSD)			0.25		0.48		0.59		0.53
LOD			0.09		0.107		0.178		0.124
LOQ			0.277		0.326		0.591		0.323
Result of Tablet  Analysis
Formulation	Montelukast			Levocetirizine			%Assay		%RSD
	Label claim(mg)	Amt. determined		Label claim(mg)		Amt. determined	MON	LEV	MON	LEV
Tab I	10	10.11		5		4.97	101.10	99.4	0.36	0.34
Tab II	10	10.16		5		4.99	101.64	99.8	0.51	0.47

 

Table3: Result of the recovery analysis

Compound	Recovery Level (%)	aWt. spiked		Wt. recovered		Recovery (%)		R.S.D.(%) n = 3
		MON	LEV	MON	LEV	MON	LEV	MON	LEV
Tab I	50	4.0	2.0	4.06	1.96	101.50	99.33	0.19	0.38
	100	8.0	4.0	7.74	3.88	98.37	98.50	0.12	0.36
	150	12.0	6.0	11.96	6.11	99.80	101.83	0.51	0.48
Tab II	50	4.0	2.0	3.99	1.98	99.16	99.66	0.27	0.28
	100	8.0	4.0	7.79	3.92	98.75	99.00	0.10	0.18
	150	12.0	6.0	12.08	5.95	100.67	99.50	0.33	0.36

^a Wt. = Weight

 

CONCLUSION:

The proposed methods are simple, precise, accurate and rapid for the determination of MON and LEV in combined tablet dosage forms. Analysis of authentic samples containing MON and LEV showed no interference from the common additives and excipients. Hence, recommended procedure is well suited for the assay and evaluation of drugs in pharmaceutical preparations. It can be easily and conveniently adopted for routine quality control analysis.

 

ACKNOWLEDGEMENT:

The authors are thankful to m/s Ranbaxy Laboratories Ltd. Dewas, India and Mapro Pharmaceuticals, Gujarat, India for providing gift samples of Montelukast sodium and Levocetirizine Dihydrochloride, respectively. The authors are thankful to Management of MAEER’s Maharashtra Institute of Pharmacy, Pune for providing necessary facility for the work.

 

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Received on 09.10.2009          Modified on 02.03.2010

Accepted on 12.11.2010         © RJPT All right reserved