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International Journal of ChemTech Research

CODEN( USA): IJCRGG ISSN : 0974-4290
Vol.1, No.4, pp 1105-1108, Oct-Dec 2009

REVERSE PHASE HIGH PERFORMANCE LIQUID
CHROMATOGRAPHIC METHOD FOR SIMULTANEOUS
DETERMINATION OF CEFUROXIME AXETIL AND POTASSIUM
CLAVULANATE IN TABLET DOSAGE FORM

Mahima R. Sengar, Santosh V. Gandhi *, Upasana P. Patil, Vivek S. Rajmane

Department of Pharmaceutical Analysis,
A.I.S.S.M.S. College of Pharmacy, Kennedy Road, Near R.T.O.,
Pune - 411 001.India.

*Corres.author:santoshvgandhi@rediffmail.com

ABSTRACT: A simple, specific, accurate, and precise reverse phase high-performance liquid chromatographic method
for analysis of Cefuroxime axetil and Potassium clavulanate has been developed. Separation of drugs was carried out on
Jasco HPLC system with Hypersil Gold C-18 column (250 mm × 4.6 mm i.d.), using 0.01M Potassium dihydrogen
phosphate: methanol 60: 40 (v/v) as mobile phase. Quantitation of drugs was carried out at wavelength 225 nm. Results
were found to be linear in the concentration range of 5-50 µg/ml for CA and 5-30 µg/ml for PC. Mean retention times
for Potassium clavulanate (PC) and the two isomers of Cefuroxime axetil (CA1 and CA2) were found to be 2.573, 8.293
and 9.987 min, respectively. Intra-day variation, as RSD (%), was 0.328 for Cefuroxime axetil and 0.382 for Potassium
clavulanate. Interday variation, as RSD (%) was 0.545 for Cefuroxime axetil and 0.552 for Potassium clavulanate. The
% assay was found to be 100.976 ± 0.439 for Cefuroxime axetil and 101.053 ± 0.423 for Potassium clavulanate (Mean ±
S.D., n = 6).
KEYWORDS: RP-High performance liquid chromatography, Cefuroxime axetil, Potassium clavulanate

INTRODUCTION
Cefuroxime Axetil (CA), (RS)-1 hydroxyethyl Literature survey reveals spectrophotometric3 and
(6R,7R)-7-[2- (2-furyl) glyoxyl-amido] -3- (hydroxyl HPTLC methods4-6 for CA determination in combination
methyl -8-oxo-5- thia-1- azabicyclo[4.2.0]-oct-2-ene-2- with other drugs. Stability indicating7 and bioanalytical
carboxylate,72-(Z)-(O-methyl-oxime),1-acetate3- chromatographic methods 8, 9 for quantification of CA are
carbamate ) is second generation cephalosporin used to also reported. CA is official in USP which dictates RP-
treat or prevent infections that are proven or strongly HPLC method for CA determination as single drug10. RP-
suspected to be caused by bacteria1. Clavulanic acid HPLC11, 12 determination of PC with other drugs and
administered as potassium salt, is a powerful inhibitor of bioanalytical methods 13, 14 for determination of PC as
β-lactamase enzyme and is most often formulated in single drug are reported. No reports were found for
combination with antibiotics for treatment of infection simultaneous determination of CA and PC by RP-HPLC
caused by lactamase producing bacteria2. method. Aim of present work was to develop simple,
economical, rapid, precise and accurate RP-HPLC
*Corres.author: method for simultaneous determination of binary drug
Gandhi Santosh V. formulation.
AISSMS College of Pharmacy, Kennedy road, Near
R.T.O., EXPERIMENTAL
Pune. 411 001 Maharashtra, India. CHEMICALS AND REAGENTS
Potassium dihydrogen orthophosphate ( AR
Ph. No. +91-20-26058204, +91-9422349792
grade, S.d. fine - Chem Laboratories Pvt. Ltd., Mumbai,
E-mail: santoshvgandhi@rediffmail.com
India), Methanol (HPLC grade, Merck Specialities Pvt.

The injections were repeated six times and the Pharmaceuticals (Bangalore. 1. The peak areas of CA (CA1 + CA2) and PC reproducibility of the method. It was observed that there were no dihydrogen orthophosphate: methanol in ratio 60: 40 as marked changes in the chromatograms. and mobile phase (250 x 4. 0. which mobile phase at a flow rate of 1.538 and 1.1 ml was taken and transferred to 10 ml volumetric Analytically pure sample of CA was kindly flask.778 mg/ml. MD 2010 PDA detector and determined by small but deliberate changes in BORWIN-PDA (Version 1.418 for PC (Mean ± % RSD. Standard stock solution of PC was plots were found to be linear over the ranges 5–50 μg/ml prepared by dissolving 10 mg of drug in 10 ml methanol and 5–30 μg/ml for CA and PC. The pharmaceutical dosage form chromatogram has been given in Fig. Powder equivalent to 10 mg of PC was weighed. µg/ml.741 ± 0. with to get concentration of 1 mg/ml. PROCEDURE FOR ANALYSIS OF TABLET FORMULATION CONCLUSION Twenty tablets were weighed accurately.0 ml/min. The calibration standard solution. Samples were demonstrated that the RP-HPLC method developed is injected using Rheodyne injector with 20 µl loop.01 M Potassium composition (± 2 %). INSTRUMENTS AND CHROMATOGRAPHIC CONDITIONS ROBUSTNESS Jasco HPLC system consisting of Jasco PU-2080 Robustness of the developed method was plus HPLC pump. 41. India) were used in analysis. System suitability parameters for RP- working standard stock solution of concentration 50 HPLC method are listed in Table 1.2009.From this solution Chemie.439 for Cefuroxime axetil and 101.D.056 min for CA2 (mean ± S.D. addition of standard drug solution to pre-analyzed sample solution at three different levels 50 %.50) software was used for chromatographic conditions such as flow rate (± 0. Loba through Whatman filter paper no. RESULTS AND DISCUSSION For RP-HPLC method different mobile phases PROCEDURE were tried and the mobile phase containing 0.d) column using 0. filtered .598 ± 0. From this solution 0.976 ± 0.01 M PREPARATION OF STANDARD STOCK Potassium dihydrogen orthophosphate and methanol in SOLUTION ratio 60: 40 (% v/v) was found to be optimal for obtaining Standard stock solution of CA was prepared by well defined and resolved peaks with retention time dissolving 10 mg of drug in 10 ml methanol to get 2.053 ± plotted of response factor against concentration of drug.987 ± 0. n = 6). respectively. Then volume was made up to the mark with the mobile phase. (Pune. Mumbai.409 and 7. 100 % and 150 %. the This work describes a simple.573 ± 0.). India) used as such without peak areas were recorded. Ltd. n = 6). 0. robust. Results and volume was made up to mark with mobile phase.047 min for PC. recovery studies were carried out by to Clavulanic acid 125 mg per tablet.951 mg/ml.055 min for CA1 and concentration of 1 mg/ml. The LOD and the LOQ for the CA were found to be 2.. wavelength (± 1 nm). 8. Gandhi et al /Int.332 and 0.Santosh V. The % assay was found to were calculated and respective calibration curves were be 100.. respectively. μg/ml of PC was injected and chromatogram was India) and that of PC was supplied by Medriech obtained. Mumbai.02 analysis. respectively. which was used as a working 9. used in this study was Covatil CV 250 film coated tablets (Macleods Pharmceuticals.995 ± 0.J.991 ± ml was further diluted to 10 ml with methanol to get 0. A representative further purification. determination of both the drugs and method can be used transferred to a 10 ml volumetric flask containing about 5 conveniently for quality control purposes.6 mm i.273 ± 0. ml of mobile phase and sonicated for 5 min. All weighing were done on Shimadzu balance (Model AY-120). India) and Water (HPLC grade.423 for Potassium clavulanate (Mean ± S. Detection wavelength selected was 225 nm. Separation was carried out on Hypersil Gold C18 ml/min). accurate and average weight was determined and then ground to a fine sensitive validated RP-HPLC method for simultaneous powder. The % Aliquots of working standard solution of CA and recovery was found to be 100.371 for CA and PC were transferred to separate 10 ml volumetric flasks 100. The % Each solution was injected and chromatogram was RSD values were satisfactorily low indicating recorded. and for PC were PREPARATION OF CALIBRATION CURVE found to be 0. Mumbai India) labeled to RECOVERY STUDIES contain Cefuroxime axetil USP equivalent to Cefuroxime To study the accuracy and precision of the 250 mg and diluted Potassium clavulanate BP equivalent proposed method.095 respectively. of recovery studies are represented in Table 2. This sample solution having concentration 10 supplied by Maxim Pharmaceuticals Pvt. ChemTech Res.1(4) 1106 Ltd.5 correlation coefficients of 0.

1(4) 1107 Table 1: System suitability parameters for RP-HPLC method Parameter PC CA1 CA2 Theoretical 4020 4071 4502 plates Asymmetry 1.070 100.312 CA 20 20 40.2009.15 1.(μg/ml) added.Santosh V. Gandhi et al /Int.J.35 0.(μg/ml) found (μg/ml) 20 10 30.408 20 30 50.205 100.26 1.068 100.41 0.396 * Average of three determinations Figure 1: Chromatogram of standard mixture contaning PC 10 μg/ml (2. .07 2.0062 0.394 10 05 15.987 min).474 10 15 25.41 0.46 0. CA2 9.11 Factor HETP (cm) 0.364 101.23 0.16 0.062 100.128 * With respect to previous peak Table 2: Recovery studies of CA and PC Total Amount Amount amount %Recovery* % RSD* Drug taken.0055 Resolution* _ 2.464 101.386 PC 10 10 20.0061 0.573 min) and CA 20 μg/ml (CA1 8.293 min. ChemTech Res.

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