Simultaneous UPLC Estimation of Fexofenadine HCl and Montelukast Sodium Tablets

 

Mohamed Mustafa, S. Amuthalakshmi*, C.N. Nalini

Department of pharmaceutical analysis, C.L. Baid Metha College of Pharmacy, Thorapakkam, Chennai-600097.

 

ABSTRACT:

A new simple, rapid and sensitive Gradient Ultra Performance Liquid Chromatography (UPLC) method has been developed for the determination of Fexofenadine HCl and Montelukast Sodium. The method employs Thermo Scientific UPLC system on Waters (symmetry) (c18 1.8 micron 4.6 x 50mm) column. Best chromatographic separation was achieved by using acetonitrile: 20 mM potassium dihydrogen phosphate 80:30 (v/v) adjusted to pH 5.5 using orthophosphoric acid as mobile phase at a flow rate of 1 ml/min and detection at 230 nm. Separation was completed within 10 min. The retention time of Fexofenadine HCl and Montelukast Sodium was found to be 1.022 and 3.281. The proposed method was found to have linearity in concentration range of 80-120 µg/ml and 96-144 µg/m. The developed method has been statistically validated and was found to be simple, precise, reproducible and accurate. The developed and validated method was successfully used for the quantitative analysis of commercially available dosage form.

 

 

INTRODUCTION:

Fexofenadine HCl is an anti histaminic drug, chemically (2-(4-{1-hydroxy-4-[4-(hydroxydiphenylmethyl) piperidin-1-yl] butyl} phenyl)-2-methylpropanoic acid (fig. 1(a)). It interacts with histamine receptor complex [1] and shares some of the pharmacological properties of the histamine [2-4]. Montelukast sodium prevents asthma and is chemically 2-[1-({[(1R)-1-{3-[(E)-2-(7-chloroquinolin-2-yl) ethenyl] phenyl}-3-[2-(2-hydroxypropan-2-yl) phenyl] propyl] sulfanyl} methyl) cyclopropyl] acetic acid [5] (fig. 1(b)).

 

This combination is available in tablet dosage form as fexofenadine HCl 120mg and montelukast sodium 10mg.

 

Literature survey reveals good number of analytical methods is available for the estimation of this drug. Various analytical methods including, HPTLC [6], HPLC methods [7-13], RP-LC-PDA [14-24], Fourier Transform Convolution Emission Data Under Non- Parametric Linear Regression Method [25], are available for the estimation of fexofenadine HCl and montelukast sodium in pharmaceutical dosage forms.

 

Fig. 1: Structure of (a) Fexofenadine HCl and (b) Montelukast Sodium

 

In the present study we developed a simple UPLC method for the simultaneous estimation of fexofenadine HCl and montelukast Sodium by UPLC in pharmaceutical dosage forms. The proposed method was optimized and validated as per the International Conference of Harmonization (ICH) guidelines [26].

 

MATERIALS AND METHODS:

The pharmaceutical grade working standards of fexofenadine HCl and montelukast sodium were obtained as a gift from Skan pharmaceutical (Pondicherry, India). Fixed dosage combination tablet fexofenadine HCl 120 mg and montelukast sodium 10 mg were purchased from local market, Chennai India. All the chemicals were HPLC grade purchased from SD Fine Chem., Mumbai.

 

a)       Chromatographic conditions:

The Waters (symmetry c18 1.8 micron 4.6×50mm) was equilibrated with the mobile phase, acetonitrile:20 mM potassium dihydrogen phosphate 80:30 (v/v); pH 5.5. The flow rate was maintained at 1 ml /min. Eluent were monitored with UV detector at 230 nm, and the injection volume was 10ml. Total run time was kept 10 min.

 

b)       Preparation of Stock Solution:

Montelukast (10 mg) and fexofenadine (120 mg) was weighed accurately, transferred in to a 100 volumetric flask separately and sufficient ethanol was added to dissolve it. Volumes were made the up to the mark with ethanol. Aliquot from the stock solutions of montelukast and fexofenadine of and were appropriately diluted with mobile phase to obtain working standard of 10µg/mL of montelukast and 120µg/ml fexofenadine.

 

RESULTS AND DISCUSSION:

i.         Method development and optimization of chromatographic conditions

To achieve good separation between the two components different buffer pH-conditions and different proportions of solvents like ethanol, acetonitrile and water tested binary and tertiary eluents. However, acetonitrile:20 mM potassium dihydrogen phosphate 80:30 (v/v) adjusted to pH 5.5 using orthophosphoric acid as mobile phase at a flow rate of 1 ml/min and detection at 230 nm achieved good satisfactory results.

 

ii.       Method validation

The method was validated for accuracy, precision, linearity, detection limit, quantitation limit and robustness

 

a)       System Suitability

System Performance parameters of developed UPLC method was determined by injecting standard solutions. Parameters such as number of theoretical plates (N), tailing factor, resolution (R), retention time (RT) and %RSD for replicate injections were determined (table 1). The results were within the limits and were presented in fig. 2.

Table 1:  System Suitability Results

 

 

Fig. 2: System Suitability chromatogram

 

b)       Specificity

Specificity is the ability of a method to discriminate between the analyte(s) of interest and other components that are present in the sample. Studies are designed to evaluate the degree of interference, if any, which can be attributed to other analyte, impurities, degradation products, reagent "blanks" and excipients.  Blank chromatogram does not show any disturbance peak at the retention times of fexofenadine HCl and montelukast sodium (fig.3) and hence the method is specific.

 

Fig. 3: Blank chromatogram of fexofenadine HCl and montelukast sodium

 

c)       Accuracy

To determine the Accuracy of the proposed method, recovery studies were conducted; known amount of pure drug concentrations was spiked in placebo at three different levels, ie, 50%, 100% and 150% and was calculated. Accuracy was calculated as the percentage of recovery. The results were tabulated in table 2.

Table 2: Accuracy data

Parameter	Amount taken(g)	Amount recovered(g)	% Recovery	Mean Recovery	%RSD
Monteleukast sodium
50%	10	9.99	99.9	99.71	0.92
100%	20	19.88	99.4
150%	30	29.95	99.83
Fexofenadine HCl
50%	15	14.98	99.86	99.77	0.95
100%	30	29.91	99.7
150%	45	44.89	99.75

 

d)       Precision:

Precision was evaluated at three levels: repeatability, intermediate precision and reproducibility. Each level of precision was investigated by six replicate injections of concentrations of 96, 108, 120, 132,144 and 156 µg/ml of fexofenadine and 80, 90, 100, 110,120 and 130 µg/ml of montelukast sodium. The result of precision was expressed as % RSD, given in table 3.

 

Table 3: Method precision Results

Parameter	Results
	Monteleukast Sodium	Fexofenadine HCl
Repeatability
Mean %RSD of Retention Time	0.15	0.06
Mean % RSD of Peak Area	0.27	0.88
Mean % Assay	100.23	99.05
Reproducibility
Mean %RSD of Retention Time	0.16	0.07
Mean % RSD of Peak Area	0.26	0.82
Mean % Assay	100.82	99.9
Intermediate Precision
Mean %RSD of Retention Time	0.18	0.09
Mean % RSD of Peak Area	0.29	0.81
Mean % Assay	99.98	99.52

 

e)       Linearity

The linearity was evaluated by measuring different concentrations (80% to 130%) of the standard solutions. The solutions were examined by the assay procedure. The calibration curve was plotted using response factor (peak area ratio of the standard peak area and internal standard peak area) Vs concentration of the standard solution. From the calibration curve, the regression equation was computed. The summary of the parameters are shown in table 4.

 

Table 4: Regression equation parameters

Parameter	Monteleukast Sodium	Fexofenadine HCl
Linearity Range(µg/ml)	80-120	96-144
Correltion coeffecient	0.97	0.99
Slope	86804	71452
Y-intercept	419233	544283

 

f)        Detection limit (DL) and quantitation limit (QL)

Estimation of DL and QL considered the acceptable signal-to-noise ratios 3:1 and 10:1 respectively. The limit of detection and quantitation determined was 0.17 and 0.55 µg/ml for fexofenadine, 0.142 and 0.313 µg/ml for montelukast sodium respectively.

 

g)       Robustness and Ruggedness

The robustness of an analytical method was unaffected by deliberate change in the Flow rate, pH, mobile phase composition and column temperature were performed at 100% concentration.

 

The ruggedness of the proposed analytical method was performed in different condition like different columns, analyst, instrument, laboratories analysis of the same sample.

 

iii.     Mobile phase stability

The stability of the mobile phase used in the current method was observed for 24 and 48 hours at room temperature. There were no significant changes observed in peak areas, theoretical plates, tailing factors, retention time and resolution and it was stable.

 

iv.      Analysis of marketed sample

The proposed method was applied for the analysis of montelukast (10 mg) and fexofenadine (120 mg) in tablet dosage forms, the results were found to be between 98 and 99 (table 5).

 

Table 5: Assay results

Drug	Labelled amount	Amount found	% assay
Montelukast Sodium	10	9.91	99.01
Fexofenadine HCL	120	119.89	98.9

 

v.        Forced degradation studies

a)       Stock solution:

Working standards equivalent to 10 mg of montelukast sodium and 120 mg of fexofenadine weighed accurately and transferred into 50 ml volumetric flask, 30 ml of diluent was added and dissolved; further the volume was made with the diluent. Results of Forced degradation are shown in table 6.

 

 

a)       Acidic degradation:

To 10 ml of stock solution, 10ml of 1N hydrochloric acid was added and kept at 80ºC for about 12 hours in water bath, cooled and the volume was made up to 100ml with mobile phase. The solution was filtered through 0.22 micron membrane filter.

 

b)       Alkali degradation:

To 10 ml of stock solution, 10ml of 0.5N sodium hydroxide was added and kept at 80ºC for about 12 hours in water bath, cooled and the volume was made up to 100ml with mobile phase. The solution was filtered through 0.22 micron membrane filter.

 

c)       Oxidative degradation:

To 10 ml of stock solution, 5ml of 3% hydrogen peroxide was added and kept at 80ºC for about 12 hours in water bath, cooled and the volume was made up to 100ml with mobile phase. The solution was filtered through 0.22 micron membrane filter.

 

d)       Thermal degradation:

The 10 ml of stock solution was kept at 70ºC for about 10 days, cooled and the volume was made up to 100ml with mobile phase. The solution was filtered through 0.22 micron membrane filter.

 

Table 6: Degradation studies of Fexofenadine HCl and Montelukast Sodium

Condition	Montelukast	%Rec	Fexofenadine	%Rec
acid	1502998	95.08	1842471	99.01
base	1396675	89.63	1637266	94.69
perox	1566183	98.65	1794025	99.44
heat	1534041	95.83	1797059	99.57
UV	1494948	95.91	1729483	100.05

 

CONCLUSION:

Based on the results of the above studies, it is concluded that the method for determination of assay of fexofenadine HCl 120mg and montelukast sodium 10mg oral tablets is precise, linear over the concentration range, stability indicating, and rugged. The method is robust with respect to variation in flow rate, column temperature and buffer composition. The method is specific for the quantization of assay of fexofenadine HCl and montelukast sodium Tablets. So the developed method can be easily applied for routine analysis of fexofenadine HCl and montelukast sodium in bulk and pharmaceutical dosage form.

 

The method was found to be simple accurate economical and rapid and it can be applied for routine analysis in laboratories is suitable for the quality control of bulk and pharmaceutical formulations.

 

ACKNOWLEDGEMENT

The authors are thankful to Department of pharmaceutical Analysis, C.L. Baid Metha College of Pharmacy, Thorapakkam, Chennai, and Skan pharmaceuticals India for providing standards, lab facilities and for the encouragement.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest for this article.

 

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Accepted on 16.01.2017           © RJPT All right reserved