Estimation of Levocetirizine in Bulk and Formulation by Second Order Derivative Area under Curve UV-Spectrophotometric Methods

 

Audumbar Digambar Mali*, Manojkumar Patil

Department of Pharmaceutics, Sahyadri College of Pharmacy, Methwade, Sangola-413307, Solapur MS India.

*Corresponding Author E-mail: maliaudu442@gmail.com

 

ABSTRACT:

Simple, fast and reliable spectrophotometric methods were developed for determination of Levocetirizine in bulk and pharmaceutical dosage forms. The solutions of standard and the sample were prepared in Methanol. The quantitative determination of the drug was carried out using the second order Derivative Area under Curve method values measured at 247-255nm. Calibration graphs constructed at their wavelengths of determination were linear in the concentration range of Levocetirizine using 5-25μg/ml (r=0.999) for second order Derivative Area under Curve spectrophotometric method. The proposed methods have been extensively validated as per ICH guidelines. There was no significant difference between the performance of the proposed methods regarding the mean values and standard deviations. The developed methods were successfully applied to estimate the amount of Levocetirizine in pharmaceutical formulations.

 

KEY WORDS: Levocetirizine, Second order Derivative, Area under Curve (AUC), Precision, Accuracy.

 

 


INTRODUCTION:

Chemically Levocetirizine chemically is [2-[4- [(r)-(4-chlorophenyl) phenylmethyl]-1- piperazinyl] ethoxy] acetic acid is a third generation non-sedative antihistamine, developed from the second generation antihistamine cetirizine. [1, 2] It is the L enantiomer of the cetirizine racemate. Levocetirizine works by blocking histamine receptors. It does not prevent the actual release of histamine from mast cells, but prevents it binding to its receptors. [3, 4] This in turn prevents the release of other allergy chemicals and increased blood supply to the area, and provides relief from the typical symptoms of hay fever. In our Literature survey reveals that for Levocetirizine Spectrophotometric [5, 6] methods and HPLC [7, 8, 9] methods have been reported for its determination in commercial formulation.

 

To our notice, no UV- spectrophotometric method using Second Order Derivative Area under Curve has been reported for the determination of Levocetirizine in bulk and tablets. Hence an attempt has been made to develop new Second Order Derivative Area under Curve spectrophotometric method for estimation of Levocetirizine in bulk and pharmaceutical formulations with good accuracy simplicity, precision and economy.

 

 

Fig. 1 Structure of Levocetirizine

 

MATERIALS AND METHODS:

Derivative Spectrophotometric Methods:-

The second derivative spectrophotometry was used in the wavelength ranges from 247 and 255 nm.

 

[d2A/dλ2]= f(λ): second order

 

The second derivative spectrum of an absorption band is characterized by a maximum, a minimum, and a cross-over point at the λ max of the absorption band. [10, 11]

 

Area under curve (Area calculation):

In this study area was integrated between wavelength ranges from 247 and 255 nm.

Area calculation: (α+β) =

 

Where, α is area of portion bounded by curve data and a straight line connecting the start and end point, β is the area of portion bounded by a straight line connecting the start and end point on curve data and horizontal axis, λ1 and λ2 are wavelength range start and end point of curve region. [12, 13]

 

Apparatus and instrumentation:

A Shimadzu 1800 UV/VIS double beam spectrophotometer with 1cm matched quartz cells was used for all spectral measurements. Single Pan Electronic balance (Contech, CA 223, India) was used for weighing purpose. Sonication of the solutions was carried out using an Ultrasonic Cleaning Bath (Spectra lab UCB 40, India). Calibrated volumetric glassware (Borosil) was used for the validation study.

 

Materials:

Reference standard of Levocetirizine API was supplied as gift sample by Lupin Laboratory Park, Aurangabad. Methanol was obtained from Research-Lab Fine HYPERLINK "http://www.indiamart.com/company/3928956/"ChemHYPERLINK "http://www.indiamart.com/company/3928956/" Industries, HYPERLINK "http://www.indiamart.com/company/3928956/"Islampur, Mumbai, and Maharashtra. Capsule sample with label claim 5 mg per Tablet were purchased from local market Mangalwedha, Solapur, Maharashtra, India.

 

Method development:

Preparation of Standard and Sample Solutions:-

Stock solution of 10μg/ml of Levocetirizine was prepared in Methanol, for Second Order Derivative Area under Curve spectrophotometric analysis. The standard solutions were prepared by dilution of the stock solution with Methanol in a concentration range of 5, 10, 15, 20 and 25μg/ml with Methanol for Second Order Derivative Area under Curve spectrophotometric methods. Methanol was used as a blank solution.


 

Fig. 2 Second order derivative Area under Curve spectrum of Levocetirizine in Methanol (25g/ml).

 

Fig. 3 Second order derivative spectrum of Levocetirizine in Methanol (25g/ml).


Calibration curve for Levocetirizine:

The dilutions were made from Standard Stock solution to get concentration of 5, 10, 15, 20, and 25g/ml respectively. These solutions were scanned from 400 to 200 nm and First Order Derivative Area under Curve values was integrated in the range of 224-231 nm. The calibration curve was plotted between areas under curve values against concentration.

CONC

 
 


Fig. 4 Linearity of Levocetirizine.

Assay of tablet formulation:-

Twenty tablets each containing 5 mg of Levocetirizine were weighed crushed to powder and average weight was calculated. Powder equivalent to 10 mg of Levocetirizine was transferred in 100 ml of volumetric flask. A 50 ml of Methanol was added and sonicated for 15 minutes. Then solution was further diluted up to the mark with Methanol. The solution was filtered using Whatman filter paper no. 41, first 5 ml of filtrate was discarded. This solution was further diluted to obtain 10g/mL solution with water, subjected for UV analysis using Methanol as blank. This procedure was repeated three times.


 

Fig. 5 Second order derivative Area under Curve spectrum of Levocetirizine of dosage form in Methanol (25g/ml).

Fig. 6 Second order derivative spectrum of Levocetirizine of dosage form in Methanol (25g/ml).

Fig. 7 Second order derivative overlay of Levocetirizine at diff. Concentration.

 

 


Table 1: Assay of tablet dosage form:-

Sr.No.

Sample Solution Concentration (g/ml)

Amount found (%)

Mean % found*

%RSD*

1

25

98.59

 

 

2

25

99.52

99.00

0.2961

3

25

98.90

 

 

*n=3, % RSD = % Relative Standard Deviation.

 

 

Method Validation:[14]

The above method was validated for various parameters such as Accuracy, Linearity, Precision, Limit of detection (LOD) and Limit of Quantitation (LOQ) according to ICH guideline.

 

Accuracy:-

The accuracy for the analytical method was evaluated at 80%, 100% and 120% levels of 25g/ml Sample solution. Second Order Derivative Area under curve (AUC) was measured in wavelength range 247-255 nm and results were obtained in terms of percent recovery. Three determinations at each level were performed and % RSD was calculated for each level.

 

 


Table 2: Accuracy results for Levocetirizine:-

Accuracy level

Sample Conc. (g/ml)

Std. Conc.

Total amount. Added (g/ml)

% Recovery

Mean % Recovery

% RSD

80

25

12

22

100.12

 

 

100

25

15

25

100.29

100.49

0.2397

120

25

18

28

101.08

 

 

 


 

Precision:-

The precision of an analytical procedure expresses the closeness of an agreement (degree of scatter) between a series of measurements obtained from multiple sampling of the same homogeneous sample under the prescribed conditions intraday precision was studied by integrating area of standard solution of 25g/ml concentration at six independent series in the same day. Interday precision studies were performed by integrating area of standard solution of 25g/mlconcentration on three consequent days. The %RSD Was calculated.

 

Table 3: Precision Study:-

Parameter

Intra day

Inter-day

Sample sol conc.g/ml

25

25

AUC (mean)

0.5326

0.5237

%RSD

0.8369

0.8620

 

 

Limit of Detection and Limit of Quantification:-

The Limit of Detection (LOD) is the smallest concentration of the analyte that gives the measurable response. LOD was calculated using the following formula

LOD = 3.3 σ /S

 

The Limit of Quantification (LOQ) is the smallest concentration of the analyte, which gives response that can be accurately quantified. LOQ was calculated using the following formula

= 10 σ/S

Where, σ is standard deviation of the response and

S is the slope of the calibration curve.

LOD and LOQ of Levocetirizine was found to be 0.53g/ml and1.59g/ml respectively.

 

Table 4: Summary of validation parameters:-

Parameter

Result

λ range

247-255

Regression Equation (y=mx+c)

Y=0.023x + 0.003

Linearity range

5-25g/ml

Slope

0.023

Intercept

0.003

Correlation coefficient (R2)

0.999

Limit of Detection (LOD) g/ml

0.53

Limit of Quantitation (LOQ) g/ml

1.59

Accuracy (Mean % Recovery)

100.49

Precision (%RSD)

0.2397

 

RESULTS AND DISCUSSION:

The UV visible spectroscopic method for the Levocetirizine by Second order derivative Area under Curve was found to be simple, accurate, economical and reproducible. The drug concentrations were found to be linear in the range of 05-25 g/ml and the correlation coefficient value of 0.999 indicates that developed method was linear. For Precision the percent relative standard deviation (% RSD) was found to be 0.2397 while, intra-day and inter-day precision results in terms of percent relative standard deviation values were found to be 0.5326 and 0.5237 respectively thus the method is observed as precise. The accuracy of the method was assessed by recovery studies at three different levels i.e. 80%, 100%, 120%. The values of standard deviation were satisfactory and the recovery studies were close to 100%. The % RSD value is ≤ 2 indicates the accuracy of the method. The Limit of Detection and Limit of Quantitation values were found to be 0.53g/ml and 1.59g/ml respectively. The result of the analysis for pharmaceutical formulation by the developed method was consistent with the label claim, highly reproducible and reliable. The method can be used for routine quality control analysis of Levocetirizine in bulk and pharmaceutical formulations.

 

CONCLUSION:

The UV spectroscopic AUC method for the analysis of Levocetirizine by Second order derivative Area under Curvewas found to be simple, precise, and accurate; can be used for assay of bulk drug and pharmaceutical dosage formulations.

 

ACKNOWLEDGEMENT:

The authors are highly thankful to the Sahyadri College of Pharmacy, Methwade, Sangola, Solapur, Maharashtra, India for proving all the facilities to carry out the research work.

 

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Received on 05.07.2015 Accepted on 22.09.2015

Asian Pharma Press All Right Reserved

Asian J. Pharm. Res. 5(3): July- Sept., 2015; Page 145-150

DOI: 10.5958/2231-5691.2015.00022.2