DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR DETERMINATION OF LEVAMISOLE IN BULK AND DOSAGE FORM

Abstract

ABSTRACT

Objective: To develop a reverse phase isocratic HPLC was developed and validated for the determination of levamisole in bulk and tablet dosage forms.

Method: Method development was carried out on Welchrom C₁₈ isocratic column, (250 mm × 4.6 mm i.d., particle size 5 μm, maintained at ambient temperature), Shimadzu LC-20AT Prominence Liquid Chromatograph. The mobile phase was a mixture of Methanol: Acetonitrile: Water 50:30:20 v/v, with apparent pH of 4.6 and the flow rate was set at 1.0 ml/min and UV detection at 225 nm. Results: Validation parameters were evaluated for the method according to the ICH (Q2R1) guidelines. In the linearity study, linearity was observed from 2-10 μg/ml with correlation coefficient of 0.9999 and regression coefficient of 0.9998. The limit of detection and limit of quantitation for the method were 0.1219 μg/ml and 0.3695 μg/ml, respectively. The statistical analysis shows that the method was found to be accurate, reliable, simple and reproducible. The intra- and inter-assay precisions were satisfactory; the values of relative standard deviations did not exceed 2%. The accuracy of the method was proved; the mean recovery of levamisole was 99.36% to 100.56%. The chromatographic retention time of proposed method was 2.790 min and the mean assay of content was found to be 102.135 ± 0.933%.

Conclusion: The proposed method was successfully applied for the quantitative determination of levamisole in bulk form and could be used for routine analysis with phenomenal accuracy and precisions.

Keywords: RP-HPLC, Levamisole, Validation, ICH guidelines.