International Journal of Drug Development and Research

Key words

Cefuroxime axetil, UV, validation, quantitative determination.

Introduction

Cefuroxime is chemically (6R, 7R) – 3 – carbamoyloxymethyl – 7 - [(Z)-2-(2-furyl) – 2 - (methoxyimino) acetamido] – ceph – 3 - em-4- carboxylic acid. Cefuroxime is official in Indian pharmacopoeia and United States of Pharmacopoeia.

It is the first of the series of alpha methoxyiminoacyl substituted cephalosporins that constitute most of the third generation agents available for clinical use. It is active against some beta lactamase strains that are resistant to cefamandole[1,2]. The literature survey revealed that various methods of analysis for cefuroxime alone or in combination with other drugs have been reported, which included, HPLC[3–6], electrokinetic[7], HPTLC[8] and spectrophotometric methods.[9]

Accordingly, the objective of this study was to develop and validate the first order derivative method for the estimation of Cefuroxime axetil in bulk and pharmaceutical formulation as per ICH guidelines [10].

Experimental

Materials

Cefuroxime axetil was a gift sample from Torrent Pharmaceutical Limited, Ahmedabad. All chemicals and reagents used were of analytical grade and purchased from Qualigens Fine Chemicals, Mumbai, India.

Preparation of standard stock solution

Accurately weighed 10 mg of Cefuroxime axetil was transferred to 100 ml volumetric flask, dissolved in 20 ml 0.1N HCL by shaking manually for 10 min. The volume was adjusted with the same up to the mark to give final strength i.e. 100 μg/ml.

Selection of wavelength for analysis of Cefuroxime axetil

Appropriate volume 0.2 ml of standard stock solution of Cefuroxime axetil was transferred into 10 ml volumetric flask, diluted to mark with 0.1N HCL to give concentration of 2μg/ml. The resulting solution was scanned in UV range (200 nm – 400 nm). In spectrum Cefuroxime axetil showed absorbance maximum at 281 nm (Fig. 2).

Validation of the method

The method was validated in terms of linearity, accuracy, precision, and ruggedness.

Linearity study

Different aliquots of Cefuroxime axetil in range 0.04- 0.2 ml were transferred into series of 10 ml volumetric flasks and the volume was made up to the mark with 0.1N HCL to get concentrations 0.4, 0.8, 1.2, 1.6 and 2 mg/ml, respectively. The solutions were scanned on spectrophotometer in the UV range 200 - 400 nm. The spectrum was recorded at 281 nm. The calibration plot was constructed as Absorbance vs concentration (Fig. 3).

Accuracy

To the preanalysed sample solutions, a known amount of standard stock solution was added at different levels i.e. 80%, 100% and 120 %. The solutions were reanalyzed by proposed method.

Precision

Precision of the method was studied as intra-day and inter-day variations. Intra-day precision was determined by analyzing the 0.8, 1.2 and 1.6 μg/ml of Cefuroxime axetil solutions for three times in the same day. Inter-day precision was determined by analyzing the 0.8, 1.2 and 1.6 μg/ml of Cefuroxime axetil solutions daily for three days over the period of week.

Sensitivity

The sensitivity of measurements of Cefuroxime axeti lby the use of the proposed method was estimated in terms of the Limit of Quanfication (LOQ) and Limit of Detection (LOD). The LOQ and LOD were calculated using equation LOD = 3.3 x N/B and LOQ = 10 x N/B, where, ‘N’ is standard deviation of the peak areas of the drugs (n = 3), taken as a measure of noise, and ‘B’ is the slope of the corresponding calibration curve.

Repeatability

Repeatability was determined by analyzing 1.2 μg/ml concentration of Cefuroxime axetil solution for six times.

Ruggedness

Ruggedness of the proposed method is determined for 1.2 μg/ml concentration of Cefuroxime axetil by analysis of aliquots from homogenous slot by two analysts using same operational and environmental conditions.

Determination of Cefuroxime axetil in bulk

Accurately weighed 10 mg of Cefuroxime axetil was transferred into 100 ml volumetric flask containing 20 ml 0.1N HCL and volume was made up to the mark using same. Appropriate volume 0.3 ml of this solution was transferred to 10 ml volumetric flask and volume was adjusted to mark using distilled water. The resulting solution was scanned on spectrophotometer in the UV range 200 - 400 nm. The concentrations of the drug were calculated from linear regression equations.

Application of proposed method for pharmaceutical formulation

For analysis of commercial formulation 5 ml of Cefuroxime axetil eye drop solution was taken in 100 ml volumetric flask and the volume was made up to the mark with 0.1N HCL to give 100μg/ml concentration. From this 1 ml was taken and transferred to 10 ml volumetric flask and volume was made up to the mark with distilled water to give 10 μg/ml concentration. It was scanned on spectrophotometer in the UV range 200 - 400 nm. The spectrum was recorded at 281 nm. The concentrations of the drug were calculated from linear regression equation.

Results and Discussion

Method Validation

The proposed method was validated as per ICH guidelines. The solutions of the drugs were prepared as per the earlier adopted procedure given in the experiment.

Linearity studies

The linear regression data for the calibration curves showed good linear relationship over the concentration range 0.4-2.0 μg/ml for Cefuroxime axetil. Linear regression equation was found to be Y = 0.453 X + 0.0285 (r2 = 0.998). The result is expressed in Table 1.

Accuracy

The solutions were reanalyzed by proposed method; results of recovery studies are reported in Table 2 which showed that the % amount found was between 98.54% to 99.98% with %R.S.D. >2.

Precision

The precision of the developed method was expressed in terms of % relative standard deviation (% RSD). These result shows reproducibility of the assay. The % R.S.D. values found to be less than 2, so that indicate this method precise for the determination of both the drugs in formulation (Table 3).

Sensitivity

The linearity equation was found to be Y = 0.0317 X + 0.0047. The LOQ and LOD for Cefuroxime axetil were found to be 0.038 μg and 0.32 μg, respectively.

Repeatability

Repeatability was determined by analyzing 1.2 μg/ml concentration of Cefuroxime axetil solution for six times and the % amount found was between 98% to 102% with % R.S.D. less than 2 (Table 4).

Ruggedness

Peak area was measured for same concentration solutions, six times. The results are in the acceptable range for both the drugs. The results are given in Table 5. The result showed that the % R.S.D. was less than 2%

Determination of Cefuroxime axetil in bulk

The concentrations of the drug were calculated from linear regression equations. The % amount found was between 99.17% to 100.43% (Table 6).

Application of proposed method for pharmaceutical formulation

The spectrum was recorded at 281 nm. The concentrations of the drug were calculated from linear regression equation. The % amount found was between 98.36% to 101.31% (Table 7).

Conclusion

This UV-spectrophotometric technique is quite simple, accurate, precise, reproducible and sensitive. The UV method has been developed for quantification of Cefuroxime axetil in tablet formulation. The validation procedure confirms that this is an appropriate method for their quantification in the plant material and formulation. It is also used in routine quality control of the raw materials as well as formulations containing this entire compound.

Acknowledgement

The authors are thankful to the Principal and management, R.C. Patel Institute of Pharmaceutical Education and Research, Shirpur (M.S.), India for providing the required facilities to carry out this research work

Tables at a glance

Table 1 Table 2 Table 3 Table 4 Table 5 Table 6 Table 7

Figures at a glance

Figure 1 Figure 2 Figure 3

References

1. United states of Pharmacopoeia and National Formulary, 27th Edn, US Pharmacopieial Convention Inc., Rockville, MD, 2000, pp. 335.
2. Insel PA, Hardman JG eds, Goodman and Gilman’s The Pharmacological basis of Therapeutics 9th Edn, New York, Mc-Graw Hill, 1996, pp. 617.
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5. Peleman R, Hoorekebe H, Pauwels R. Measurement of cefuroxime in human alveolar lavage fluid by high performance liquid chromatography after solid phase extraction. J Chromatogr B Biomed SciAppl 1997; 689: 438–441.
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