INTRODUCTION:

Cefixime (CEF) is an oral third generation cephalosporin antibiotic. Chemically, it is (6R,7R)-7-{[2-(2-amino-1,3-thiazol-4-yl)-2-(carboxymethoxyimino)acetyl]amino}-3-ethenyl 8-oxo-5-thia-1- azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid, clinically used in the treatment of susceptible infections including gonorrhoea, otitis media, pharyngitis, lower respiratory-tract infections such as bronchitis, and urinary-tract infections^1-5.

Ofloxacin (OF) is chemically 9-fluoro-2, 3-dihydro-3-methyl-10- (4-methyl-1-piperazinyl)-7-Oxo-7H-pyrido (1,2,3-di)- 1,4-benzoxazine carboxylic acid. It is a fluoroquinolone derivative⁶. It is used mainly as an antibacterial. It is official in USP⁷ (United State Pharmacopoeia), BP⁸(British Pharmacopoeia) and EP⁹ (European Pharmacopoeia). More ever, the literature survey revealed that so far, no method has been reported for estimation of CEF and OFL in combined dosage form, hence Multicomponent methods have been developed for simultaneous estimation of CEF and OFL in tablet dosage form^10,11.

Extensive literature survey reveals, none of the method is available that is based on estimation of Cefixime trihydrate and Ofloxacin by simultaneous equation method and Multicomponent methods. Aim of present work was to develop simple, precise, accurate and economical Spectrophotometric methods for simultaneous determination of Multicomponent methods of binary drug formulation.

MATERIALS AND METHODS:

The sampling wavelengths 262,269,278,281,286 nm was selected on trial and error basis. The concentrations of individual drug (i.e. CEF and OFL) in the respective six mixed standard solutions (Table 1) were feed to multicomponent mode of the instrument. All the six mixed standards were scanned in the range of 200 nm to290 nm and graph was shown in (figure no 1).

The basic necessity for the application of the proposed method is that at all the selected sampling wavelengths the mixed standard solutions must follow the Beer-Lambert’s law. The study shows that at all the selected sampling wavelengths, the mixed standard solutions obey the Beer-Lambert’s law.

Analysis of pharmaceutical Formulation:

Twenty tablets ‘Mahacef-Plus’ (containing 200 mg of CEF and 200 mg of OFL) were weighed and ground to fine powder. A quantity of sample equivalent to 10 mg of Cefixime trihydrate and 10 mg of Ofloxacine was transferred into 100 ml volumetric flask containing 0.1 N acidic methanol, sonicated for 10 min; the volume was made up to the mark and filtered through Whatmann filter paper (no. 41). 0.5 ml of this solution was transferred to 10 ml volumetric flask, dissolved and volume was adjusted to mark.

Fig 1. Linearity graph of mixture at five selected wavelength

Table 1:- Range for mixture

Validation of proposed method:

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

Accuracy:

To assess the accuracy of proposed method, recovery experiment was performed. To the preanalyzed sample solution of CEF and OFL, a known amount of standard drug solution was added that is 2 µg/ml and absorbance were recorded. The % recovery was then calculated.

Precision:

Precision of the method was assessed by repeatability; determined by analyzing 10 µg/ml of CEF and 10 µg/ml OFL of drug solutions for five times; results were recorded [Table 2]. Method precision was studied as intra-day and inter-day variations.

Table 2:- Analysis of standard laboratory mixture

S. N	Amount of drugs taken (µg/mL)		Amount of drug estimated (µg/mL )		% of drugs estimated.
S. N	CEF	OFL	CEF	OFL	CEF	OFL
1	2	2	1.99	1.98	99.5	99
2	4	4	3.97	3.99	99.25	99.75
3	6	6	5.96	5.89	99.33	98.16
4	8	8	7.98	7.99	99.75	99.87
5	10	10	9.94	9.96	99.4	99.6

Statistics

Drug	Mean	± SD	%RSD
CEF	99.44	0.193	0.194
OFL	99.27	0.707	0.712

Table 3. Analysis Data of Tablet Formulation

S.N.	Amount of tablet taken (µg/mL)	Amount of drug estimated ( µg/mL)		% of labeled claim
S.N.	Amount of tablet taken (µg/mL)	CEF	OFL	CEF	OFL
1	10	10.02	10.12	100.01	100.23
2	10	9.86	9.84	99.24	99.20
3	10	9.90	10.02	98.67	100.01
4	10	10.15	10.00	100.08	100.00

Statistics

Drug	Mean	± SD	%RSD
CEF	99.49	51.87	1.912
OFL	99.86	52.048	1.913

Ruggedness:

Ruggedness of the method was determined by analysis of aliquots from homogeneous slot by two analyst using same operational and environmental conditions [Table 4].

Table 4- Results of ruggedness studies

Drugs	Parameter	Method precision	Intermediate Precision
Drugs	Parameter		Inter day	Intraday	Different Analysts
CEF	Mean ± SD	99.18 ±1.2945	98.15 ± 1.124	99.91 ± 1.350	100.5 ± 0.912
CEF	% RSD	1.2857	1.145	1.364	0.907
OFL	Mean ± SD	99.91 ±0.5635	99.83 ± 1.004	98.74 1.675	99.78 ± 0.473
OFL	% RSD	0.5639	1.005	1.696	0.474

Recovery Study:

To check the accuracy of the developed methods and to study the interference of formulation additives, analytical recovery experiment was carried out by standard addition method. From the total amount of drug found, the percentage recovery was calculated. The results are reported in [Table 3].

RESULT AND DISCUSSION:

The UV Spectrophotometric methods were found to be simple, accurate, economic and rapid for routine simultaneous estimation of CEF and OFL, in tablet dosage forms. For both the methods, linearity was obtained in concentration range of 2-10 μg/ml and 5-25 μg/ml for CEF and OFL respectively. Both the drug show good regression values at their respective wavelengths. Recovery was in the range of 99-101%; the value of standard deviation and % R. S. D are found to be < 2%; shows the high precession of the method. The method was found to be rugged as the percentage purity of the drugs determined by two different analysts were 99.8± 0.05 for CEF, 99.5± 0.06for OFL and 99.7± 0.01for CEF, 99.8± 0.04for OFL.

CONCLUSION:

The results of validation showed that the proposed method is simple, linear, accurate and precise and it can be employed in routine assay of CEF and OFL in combined dosage form.

ACKNOWLEDGEMENT:

The authors are thankful to head Shri Sureshdada Jain Institute of Pharmaceutical Education and Research Jamner (Jalgaon) for providing laboratory facilities. The authors are also thankful to JCPL, Jalgaon, for providing gift samples of drugs.

REFERENCES:

1. Sweetman S.C., Martindale, The Complete Drug Reference. 32nd Edn. Pharmaceutical Press, London, 1999, 165-166.

2. Budavari S., The Merck Index, 13th Edn., Merck Research Lab, Division of Merck and co. Inc., Whitehouse Station, NJ, 2001, 1213,1229.

3. Maheshwari R.K., Kinariwala M., Saxena M., Gahlot M., Chaki R. and Jagwani Y., Spectrophotometric Analysis of Cefixime Trihydrate Tablets using Metformin Hydrochloride as Hydrotropic Solubilizing Agent, Asian J. Chem., 2008, 20, 6047-6050.

4. Khan I.U., Sharif S., Ashfaq M. and Asghar M.N., Simultaneous Determination of Potassium Clavulanate and Cefixime in Synthetic Mixtures by High-Performance Liquid Chromatography, J. AOAC Int. 2008, 91, 744-749.

5. Jovanovic S.E., Agbaba D., Stakic D.Z. and Vladimirov S., HPTLC determination of ceftriaxone, cefixime and cefotaxime in dosage forms, J. Pharm. Biomed. Ana., 1998, 18, 893-898

6. United State Pharmacopoeia, 24/National formulary 29, Asian Edn., USP Conv.Inc. Rockville, USA; 2006; 1574.

7. Zendelovska D., Stafilov T. and Milosevski P., High-Performance Liquid Chromatographic Method for Determination of Cefixime and Cefotaxime in Human. Plasma. Bull. Chem. Technol., Macedonia, 2003, 22, 39–45.

8. Khan I.U., Sharif S., Ashfaq M. and Asghar M.N., Simultaneous Determination of Potassium Clavulanate and Cefixime in Synthetic Mixtures by High-Performance Liquid Chromatography, J. AOAC Int. 2008, 91, 744-749.

9. Rathinavel G., Mukherjee P.B., Valarmathy J., Samueljoshua L., Ganesh M., Sivakumar T. and Saravanan T., A Validated RP – HPLC Method for Simultaneous Estimation of Cefixime and Cloxacillin in Tablets, E-Journal of Chem., 2008, 5, 648-651.

10. Jovanovic S.E., Agbaba D., Stakic D.Z. and Vladimirov S., HPTLC determination of ceftriaxone, cefixime and cefotaxime in dosage forms, J. Pharm. Biomed. Ana., 1998, 18, 893-898

11. British Pharmacopoeia, published by the London Stationary Office under department of health 4th Edn, 2002; 1, 124

Received on 28.12.2010 Modified on 02.02.2011

Asian J. Research Chem. 4(5): May, 2011; Page 708-710

Name of drug

Concentration in (µg/mL)