Method Development and Validation for the Simultaneous Estimation of    Levofloxacin and Cefpodoxime Proxetil by Using Rp-Hplc in Combined Tablet Dosage form

Fig3 Accuracy Chromatograms-50% of Levofloxacin and  Cefpodoxime proxetil    Click here to View Figure

inter-day variation studies, six repeated injections of standard and sample solutions were made for three consecutive days and response factor of drugs peaks and percentage RSD were calculated. 

Fig4: Accuracy Chromatograms-100%of Levofloxacin and  Cefpodoxime proxetil   Click here to View Figure

Fig5: Accuracy Chromatograms-150% of Levofloxacin and  Cefpodoxime proxetil  Click here to View Figure

Table 2

Table 3: Accuracy for Levofloxacin

Table 4: Accuracy for Cefpodoxime proxetil

LINEARITY AND RANGE

The linearity of the method was determined at five concentration levels. The calibration curve was constructed by plotting response factor against concentration of drugs. The slope and intercept value for calibration curve was Y=46746X(R²=1) for Levofloxacinand Y=38111X(R²=1) for Cefpodoxime proxetil. The results shows that an excellent correlation exists between areas and concentration of drugs within the concentration range indicated above. The overlay chromatograms of Linearity for Levofloxacin and Cefpodoxime proxetil shows in Fig 6 and the results for calibration curves are given in Fig 7&8.

Fig7: Linearity Curve for Levofloxacin   Click here to View Figure

Fig8: Linearity Curve for  Cefpodoxime proxetil   Click here to View Figure

Fig9: Overlay chromatograms of Linearity for Cefpodoxime proxetil and Levofloxacin   Click here to View Figure

ROBUSTNESS

Robustness of the method was determined by making slight changes in the chromatographic conditions. It was observed that there were no marked changes in the chromatograms, which demonstrated that the RP HPLC method developed, are robust (Table-5&6).

Table5: Robustness for Levofloxacin

Table6:Robustness for Cefpodoxime proxetil

LOD&LOQ

Limit of quantification and detection were predicted by plotting linearity curve for different nominal concentrations of Levofloxacin and  Cefpodoxime proxetil. Relative standard deviation (σ) method was applied, the LOQ and LOD values were predicted using following formulas (a) and (b). Precisionwas established at these predicted levels.

(a) LOQ = 10σ / S

(b) LOD = 3.3 σ / S

 Where

σ = residual standard deviation of response 

S = slope of the calibration 

Figure 10 Click here to View Figure

Table 7: LOD and LOQ For Levofloxacin and  Cefpodoxime proxetil

RESULTS AND DISCUSSION

System suitability results were given by table1 and system suitability parameters are retention time, resolution, tailing and plate count were shown uniformity and %RSD was less than 1. So we can say system is suitable for analysis method specificity was concluded by fig:1 and fig:2 those figures are Levofloxacin and  Cefpodoxime proxetil standard chromatogram and other one is formulation they were not observed placebo and excipients peaks interference with standard and analytic peak so it proves method is selective. The result given in table 2 says that the method precision passed for both Levofloxacin and  Cefpodoxime proxetil studies. The method accuracy was evaluated by recovery studies. Levofloxacin and  Cefpodoxime proxetil recovery was founded 100% as per ICH 97%- 103% and also percentage RSD was very low so method is accurate shown in table 3&4. Linearity calibration curve was given below fig: 7&8 and plot the graph three different concentrations versus areas to construct the linear regression equation and to calculate the value of correlation co-efficient.Linear correlation was found to be Y=46746 for Levofloxacin and y = 38111 for Cefpodoxime proxetil Method robustness results were given by table 5&6, LOQ and LOD Results were given by table 7.

CONCLUSION

The proposed HPLC method was found to be simple, precise, accurate and sensitive for the simultaneous estimation of    Levofloxacin and Cefpodoxime proxetil pharmaceutical dosage forms. Hence, this method can easily and conveniently adopt for rountine quality control analysis of    Levofloxacin and Cefpodoxime proxetil pure and its pharmaceutical dosage forms.

ACKNOWLEDGEMENT

I am thankful to department of Pharmaceutical Analysis and Quality Assurance of Guru Nanak Institute of pharmacy, Jawaharlal Nehru Technological University, Hyderabad, for providing instruments and analytical support.

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