Simultaneous UV Spectrophotometric Method for Estimation of Gemifloxacin mesylate and Ambroxol HCl in Combined Dosage form

 

Dharam M. Barad*, Badmanaban R. and Chaganbhai N. Patel

Shri Sarvajanik Pharmacy College, Nr. Arvind Baug, Mehsana (Gujarat), India 384001.

*Corresponding Author E-mail: dharambarad@yahoo.com

 

ABSTRACT:

Two simple, sensitive, rapid, precise UV spectrophotometric methods have been developed and validated for simultaneous estimation of Gemifloxacin mesylate and Ambroxol hydrochloride. Recently this combination of two drugs is approved for the treatment of lower respiratory tract infection mainly pneumonia and bronchitis in adult which is available in tablet dosage form. First method is simultaneous equations method, wavelength selected for estimation of Gemifloxacin mesylate and Ambroxol hydrochloride are 271 nm and 209 nm. The second method is first order derivative method based on measurement at zero crossover point of another drug, measurement of Gemifloxacin mesylate and Ambroxol hydrochloride were carried out at 262 and 218 nm respectively. The linearity was obtained in the concentration range of 2-14 µg/ml for Gemifloxacin mesylate and 1.5-10.5µg/ml for Ambroxol hydrochloride. The proposed methods are successfully applied for the simultaneous determination of both drugs in marketed tablet preparation. The method was validated with respect to linearity, accuracy, precision as per the International Conference on Harmonization (ICH) guidelines.

 

KEYWORDS: Gemifloxacin Mesylate,  Ambroxol Hydrochloride,  UV Spectrophotometry,  Simultaneous equations,  First order derivative.

 

 


INTRODUCTION:

Gemifloxacin, (R,S)-7-(3-aminomethyl-4-syn-methoxyimino-1-pyrrolidinyl)-1-cyclopropyl- 6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylicid methanesulfonate, is a new fluoroquinolone antibacterial compound and structure of gemifloxacin is shown in Fig. 1(a). Gemifloxacin which is widely used in chronic bronchitis, pneumonia, and urinary tract infections.1-3 Ambroxol hydrochloride, chemically 4([(2-amino, 3, 5 dibromophenyl)-methyl] amino)-cyclohexyl or N- [(trans-p-hydroxy cyclo hexyl)-(2-amino 3, 5-dibromo benzyl)-amino] cyclohexanol hydrochloride is a mucolytic expectorant and structure of ambroxol hydrochloride is shown in Fig. 2(a) and used to reduce the viscosity of mucous secretions.4 A fixed dose combination of Gemifloxacin Mesylate (GEM) and Ambroxol Hydrochloride (AMB) is available for the treatment of upper and lower respiratory tract infections. Tablet is available commercially as G-CIN A.

 

On detailed literature survey, it was found that these drugs have been estimated  individually and in combinations by various methods.5-17 To the best of our knowledge, no study has been described for the simultaneous determination of both drugs in combined dosage form by UV spectrophotometric method. The present paper described two simple, rapid, economic methods for simultaneous estimation of Gemi and Ambro by simultaneous equation method and first order derivative method. The proposed methods were validated as per International Conference on Harmonization (ICH) guidelines Q2 (R1). 18-19

 

MATERIALS AND METHOD:

Equipment:

The Shimadzu UV-Visible Spectrophotometer model 1700 was employed. The sample solution was recorded in 1 cm quartz cell against solvent blank over the range 200-400 nm. The optimal condition for recording spectra to achieve good reproducibility include medium scan speed, and slit width at 2 nm.

 

Material:

Pure gift sample of GEM and AMB from Zydus Cadila were used. All analytical grade chemicals and solvent were supplied by Loba Chemi Pvt. Ltd. The pharmaceutical dosage form G-CIN A was used.

 

Figure 1(a):  Chemical structure of Gemifloxacin mesylate

 

Figure 1(b): Chemical structure of Ambroxol  Hydrochloride

 

Procedure:

Standard solutions and calibration curve:

Stock solutions for spectrophotometric measurements were prepared by dissolving GEM and AMB in methanol to obtain concentration of 1 mg/ml.  Aliquots of the standard stock solution were transferred to a series of 10 ml volumetric flask and suitability diluted with methanol to give varying concentrations ranging from 2-14 µg/ml for GEM and 1.5-10.5 µg/ml for AMB. The solutions were scanned in the wavelength range 200-400 nm.

 

Sample solution:

Twenty tablets were weighed, powdered and weighed accurately equivalent to 64 mg of GEM and 15mg AMB were transferred to a 100 ml volumetric flask and dissolved in 50 ml of methanol by ultra-sonication for 20 min. The solution was diluted with same solvent, filtered through Whatmann No.41 filter paper. The filtrate was diluted with methanol to get final dilution 6.4µg/ml of GEM and 1.5µg/ml.

 

Method I: Simultaneous equation method:

From Overlain spectrum, two wavelengths selected were 271 nm for GEM and 209 nm for AMB (figure 1). (A1%, 1 cm) was calculated for each drug from absorbance values for the drug at selected wavelengths. Simultaneous equations were constructed from calculated absorptivity value.

 

Absorbance of sample solution was measured at selected wavelengths as A1 and A2 and concentration of the two drugs in sample was calculated by using following equation.

𝐶𝑥 =               …………… (1)

 

𝐶𝑦 =               …………..… (2)

Where,

A1 and A2 are the absorbances of mixture at 209.0 nm and 271.0 nm respectively,

ax1 and ax2 are absorptivities of AMB at 209.0 nm and 271.0 nm respectively,

ay1 and ay2 are absorptivities of GEM at 209.0 nm and 271.0 nm respectively,

 

Figure 2: Overlain Spectra of Gemifloxacin and Ambroxol

 

Method II: First-Order Derivative Spectroscopy

The spectrums obtained in method I was derivatised to obtain first derivative spectrum. The two spectra were overlain. It appeared that AMB showed zero crossing at 262.0 nm while GEM showed zero crossing at 218.0 nm. At the zero crossing point of AMB (262 nm), GEMI showed a substantial dA/dλ, whereas at the zero crossing point of GEM (218 nm), AMB showed a substantial dA/dλ. Hence the wavelengths 218 nm and 262 nm were selected as analytical wavelengths for determination of AMB and GEM, respectively. These two wavelengths can be employed for the estimation of AMB and GEM without any interference from the other drugs in their combined formulation. Calibration curves were plotted by taking dA/dλ on Y-axis and concentrations on X-axis. Spectrum of sample also derivatised and absorbance of both drug were measured at selected wavelengths and concentration of both drug calculated from calibration curve. Overlain first order derivative spectra of GEM and AMB are shown in Fig. 3.

 

Figure 3: Overlain first order derivative spectra of GEM and AMB.

 


 

Table 1: Statistical parameter

Parameter

Method I

Method II

GEM

AMB

GEM

AMB

Regression Equation

Y = 0.0851x -0.0206

Y = 0.0834x +  0.0079

Y = 0.002x-0.00

Y = 0.005x+0.00

Correlation  Co-efficient

0.9997

0.9993

0.997

0.999

Precision (%RSD)

Intraday

0.18 - 0.92 %

0.68 - 1.02 %

0.12 - 0.65 %

0.34 - 0.66 %

Interday

0.23 - 1.11 %

1.41 - 1.57 %

0.34 - 0.86 %

0.69 - 1.24 %

Accuracy

98.15-101.14 %

98.77-101.93 %.

99.06 -100.88 %

100.14-101.24 %

 

Table 2: Analysis of Tablet formulation

Method

DRUG

Lable claim

% Lable claim estimated (mean   ±  S.D.)*

Simultaneous equation method

(Method I)

GEM

320 mg

98.80±0.69%

AMB

75 mg

103.02±1.04 %

First order derivative

(Method II)

GEM

320 mg

99.80 ±0.60 %

AMB

75 mg

101.05±0.74 %

*Average of five determination

 


RESULT AND DISCUSSION:

GEM and AMB both drugs are soluble in methanol; hence methanol was selected as solvent. zero-order overlain spectra of GEM and AMB at 5 μg/ml and the spectra  showed λmax of 271 nm and 209 nm for GEM and AMB, respectively. Also, both absorbs at the λmax of the each other hence, simultaneous equation method was used to estimate GEM and AMB in presence of each other. For first order derivative method measurement zero crossing were used. Both the methods were validated statistically as per ICH guideline for parameters like linearity, precision and accuracy. The values of coefficient of variation were satisfactorily low. Accuracy was performed by recovery study at three level 80%, 100%, and 120% of the test concentration. The RSD values for intraday and interday precision are less than 2%. The recovery was between 98-102%.so the developed method is accurate and precise. All data are as shown in Table 1. The amounts in terms of % label claim obtained by proposed methods are presented in Table 2.

 

CONCLUSION:

The proposed method was successfully applied to the simultaneous determination of GEM and AMB from bulk and pharmaceutical tablet formulation. The presented method was found to be simple, accurate, precise and rugged. It can be directly and easily applied to the analysis of the combined pharmaceutical tablet formulation of GEM and AMB. Moreover, the present method is quick and cost effective as compared to chromatographic techniques. Therefore, it can be concluded that the proposed method provides an alternative procedure for the quality control of GEM and AMB in pharmaceutical formulations.

 

ACKNOWLEDGEMENT:

We are grateful to Zydus Cadila, Ahemdabad for providing gift samples of drug for my work. We are thankful to the management of Shri Sarvajanik College of Pharmacy, Mehsana for providing laboratory facilities and constant encouragement.

 

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Received on 09.04.2011       Modified on 14.04.2011

Accepted on 30.04.2011      © RJPT All right reserved

Research J. Pharm. and Tech. 4(7): July 2011; Page 1129-1131