Abstract

RP-HPLC METHOD DEVELOPMENT AND ITS VALIDATION FOR SIMULTANEOUS ESTIMATION OF AMPICILLIN AND CLOXACILLIN IN ITS PHARMACEUTICAL CAPSULE DOSAGE FORM

M. Reena Sharma*, Smita Khare, Sulakshana Pawar and Sonal Dixit

ABSTRACT

A simple, rapid, accurate and precise reverse phase high performance liquid chromatography (RP-HPLC) method for simultaneous estimation of Ampicillin and Cloxacillin capsule (oral dosage form) was developed. Different analytical parameters, such as linearity, accuracy, precision and robustness, were determined according to the ICH guidelines. The chromatographic separation was carried out on Inertsil C18 analytical column (250×4.6mm;5μm) with acetonitrile: phosphate buffer (potassium dihydrogen ortho phosphate and pH 5 adjusted with sodium hydroxide) 25:75 v/v as mobile phase isocratic flow; at flow rate of 1.0ml/min. UV detector set at 220 nm. The proposed method showed a retention time (Rt) of 1.7 min for Ampicillin and 5.4 min for Cloxacillin. The linearity of the calibration curves for each analyte, in the desired concentration range, was found to be good (r2 > 0.9997). Good recovery was obtained by this developed method for each drug with a relative standard deviation (%RSD) of <2%. Peaks corresponding to each of the drugs exhibited positive values for the minimum peak purity index over the entire range of integrated chromatographic peak indicating high purity of the peaks. Stability analysis revealed that the drugs remained stable for sufficient time. Thus, the developed method was found to be robust and it can be employed to quantify Ampicillin and Cloxacillin in commercial sample.

Keywords: RPHPLC, Inertsil C18, Ampicillin, Cloxacillin.