Spectrophotometric Estimation of Nadifloxacin in Pharmaceutical Dosage form

 

Amol A Kulkarni*, Rabindra K Nanda, Meenal N Ranjane and Poonam N Ranjane

Padm. Dr. D. Y. Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune-411018, M,S. India.

*Corresponding Author E-mail: amolk301@rediffmail.com

 

ABSTRACT:

Three simple, precise and economical UV methods have been developed for the estimation of Nadifloxacin in pharmaceutical dosage form. Nadifloxacin has the absorbance maxima at 296.5 nm (Method A), and in the first order derivative spectra, showed sharp peak at 278.0 nm (Method B). Method C applied was area under curve (AUC) in the wavelength range of 291-301 nm. Linearity for the detector response was observed in the concentration range of 5-25 μg/ml for all three methods. The proposed methods were successfully applied for the simultaneous determination of Nadifloxacin in commercial pharmaceutical preparation. The results of the analysis were validated statistically and by recovery studies it was found to be satisfactory.

 

KEYWORDS: Nadifloxacin, UV spectrophotometry, derivative spectroscopy, area under curve.

 


INTRODUCTION:

Nadifloxacin (NAD), chemically know as (9-Fluoro-6,7-dihydro-8-(4-hydroxy piperidino)-5-methyl-1-oxo-1H, 5H-benzo [ij] quinolizine-2-carboxylic acid), is a Fluoroquinolone antibiotic used for the treatment of skin infections with susceptible bacteria, Acne vulgaris ( Inflamed lesions) (1 – 3).

 

Literature survey reveals HPLC determination of Nadifloxacin in rat plasma (4)  & UV–visible spectroscopic studies about the interactions between polyamidoamine (PAMAM) dendrimers and quinolones (nadifloxacin and prulifloxacin) (5). No UV method is reported so far for the estimation of drug in pharmaceutical formulations; hence we have developed a UV method for the estimation of nadifloxacin in bulk and pharmaceutical formulation.

 

MATERIALS AND METHODS:

Apparatus:

A double-beam Shimadzu UV- Visible spectrophotometer, 1700 Pharmaspec, with spectral bandwidth of 2 nm, wavelength accuracy ± 0.5 nm and a pair of 1-cm matched quartz cells was used to measure absorbance of the resulting solution.

 

Materials:

Standard gift sample of Nadifloxacin was provided by Cipla Ltd., Goa. Nadifloxacin cream was purchased from local market.

 

Solvent used: Methanol was used as a solvent.

Stock solution: Standard stock solutions of NAD (50μg/ml) was prepared and used for the analysis.

 

PROCEDURE:

Method A -- Absorption Maxima Method:

For the selection of analytical wavelength, 25 μg/ml solution of NAD was prepared by appropriate dilution of standard stock solution and scanned in the spectrum mode from 400 nm to 200 nm. From the spectra of drug (Fig. 1), lmax of NAD, 296.5 nm was selected for the analysis. The calibration curve was prepared in the concentration range of 5-25 μg/ml at 296.5 nm. By using the absoptivity value, the concentration of the sample solution can be determined.

 

Method B-- First Order Derivative Spectroscopy:

In this method, 25 μg/ml solution of NAD was prepared by appropriate dilution of standard stock solution and scanned in the spectrum mode from 400 nm to 200 nm. The absorption spectra thus obtained were derivatized from first to fourth order. First order derivative spectra were selected for analysis of the drug. First order derivative spectra of drug (Fig. 2), showed a sharp peak at 278.0 nm, which was selected for its quantitation. The calibration curve for NAD was plotted in the concentration range of 5-25 μg/ml at wavelength 278.0 nm. The concentration of the drug present in the mixture was determined against the calibration curve in quantitation mode.

 

Method C-- Area under Curve Method:

For the selection of analytical wavelength, 25 μg/ml solution of NAD was prepared by appropriate dilution of standard stock solution and scanned in the spectrum mode from 400 nm to 200 nm. From the spectra of drug, area under the curve in the range of 291.0- 301.0 nm was selected for the analysis. The calibration curve was prepared in the concentration range of 5-25 μg/ml at their respective AUC range. By using the calibration curve, the concentration of the sample solution can be determined.

 

Fig.-1: Zero order spectra of Nadifloxacin

 

Fig.-2: First order derivative Spectra of Nadifloxacin

 

Application of the proposed method for the determination of NAD in cream:

For the estimation of drugs in the commercial formulations, weighed cream equivalent to 10 mg of NAD (i.e. 1g cream) in 100 ml volumetric flask, added 30 ml of methanol and warmed 2-3 mins, sonicated 10 mins with shaking and made the volume up to 100 ml. Filtered the solution with the help of whatman filter paper. Diluted 1 ml of above filtrate up to 10 ml with methanol to obtain concentration of 10 µg/ml. In Method-A, the concentration of NAD was determined by measuring the absorbance of the sample at 296.5 nm in zero order spectrum modes. By using the absorptivity value, the concentration of the sample solution can be determined. Method-B, the concentration of NAD was determined by measuring the absorbance of the sample at 278.0 nm, in first order derivative mode. The results of the cream analysis were calculated against the calibration curve in quantitation mode. For Method-C, the concentration of NAD was determined by measuring area under curve in the range of 291.0-301.0 nm. By using the calibration curve, the concentration of the sample solution can be determined. Results of cream analysis are shown in Table No. 1.

 

Validation:

The proposed method was optimized and validated as per the International Conference on Harmonization (ICH) guidelines (6). The methods were validated with respect to linearity, accuracy, precision and selectivity.

 

Accuracy:

To ascertain the accuracy of proposed methods, recovery studies were carried out by standard addition method at three different levels (80%, 100% and 120%). Percent recovery for NAD, by all the methods, was found in the range of 98.80 % to 101.38 %.

 

Linearity:

The linearity of measurement was evaluated by analyzing different concentration of the standard solution of NAD. Beer-Lambert’s concentration range was found to be 5-25 μg/ml for all three methods.

 

Precision:

The reproducibility of the proposed method was determined by performing cream assay at different time intervals (morning, afternoon and evening) on same day (Intra-day assay precision). For inter-day precision studies was carried out at three concentration levels on three different days. Result of intra-day and inter-day precision is expressed in % RSD. Percent RSD for Intraday assay precision was found to be 0.90, 1.30 and 1.47 for Method A, B and C, respectively. Interday assay precision was found to be 1.13, 1.02 and 1.14 for Method A, B and C respectively.

 

RESULTS AND DISCUSSION:

The methods discussed in the present work provide a convenient and accurate way for simultaneous analysis of Nadifloxacin in its pharmaceutical dosage form. Absorbance maxima of Nadifloxacin at 296.5 nm (Method A) and in the first order derivative spectra, sharp peak at 278.0 nm (Method B) were selected for the analysis. Method C was area under curve (AUC) and the wavelength range for quantitation was 291.0-301.0 nm. Linearity for detector response was observed in the concentration range of 5-25 μg/ml for all three methods. Percent assay for NAD in cream analysis, by all the methods, was found in the range of 99.79 % to 100.56 %. Standard deviation and coefficient of variance for six determinations of cream sample, by all the methods, was found to be less than ± 2.0 indicating the precision of the methods. Accuracy of proposed methods was ascertained by recovery studies and the results are expressed as % recovery. Percent recovery for NAD, by all the methods, was found in the range of 98.80 % to 101.38 % values of standard deviation and coefficient of variation was satisfactorily low indicating the accuracy of all the methods. Based on the results obtained, it is found that the proposed methods are accurate, precise, reproducible and economical and can be employed for routine quality control of Nadifloxacin in its cream formulation.


Table No. 1: Results of Analysis of Cream Formulation

Method

Drug

Label Claim (% w/w)

Amount of drug estimated (mg/g of cream)

% Label Claim* ± S.D.

% Recovery**

A

NAD

1

9.98

99.79 ± 0.90

99.56

B

NAD

1

10.06

100.56 ± 1.30

99.74

C

NAD

1

10.02

100.18 ± 1.46

99.77

*Mean of six determinations, ** Mean of nine determinations

 


CONCLUSION:

The most striking feature of this method is its simplicity and rapidity. This method can be employed for routine analysis in quality control. The described method gives accurate and precise results for determination of Nadifloxacin in cream.

 

ACKNOWLEDGEMENTS:

The authors are very thankful to Dr. Avinash D. Deshpande, Director of Pharmacy, Pad. Dr. D. Y. Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune for providing necessary facilities. The authors are also thankful to Cipla Pvt Ltd, Goa for providing gift samples of Nadifloxacin.

 

REFERENCES:

1.       Sweetman SC. Martindale. The Complete Drug Reference, Pharmaceutical Press, London. 1999; 32: 228.

2.       Merck Index, John Merck research laboratories division of Merck & Co., INC. Whitehouse station, NJ, US. 2006; 14: 1097-1098.

3.       Jacobs MR and Appelbaum PC. Nadifloxacin: a quinolone for topical treatment of skin infections and potential for systemic use of its active isomer, WCK 771. Expert Opin. Pharmacotherapy. 2006; 7: 1957.

4.       Lunn G, Wiley J. HPLC Methods for Recently Approved Pharmaceuticals. J. Wiley & Sons INC publication, New York, NY, USA. 2005; 424.

5.       Cheng Y et. al. Polyamidoamine (PAMAM) dendrimers as biocompatible carriers of quinolone antimicrobials: An in vitro study. European Journal of Medicinal Chemistry. 2007; 42(7): 1032-1038.

6.       ICH Q2 (R1), Harmonised Tripartite Guideline, Validation of Analytical   Procedures: Text and Methodology, 2005.

 

 

 

 

Received on 30.07.2009       Modified on 28.09.2009

Accepted on 21.10.2009      © RJPT All right reserved

Research J. Pharm. and Tech. 3(2): April- June 2010; Page 429-431