Abstract

STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE RELATED SUBSTANCES OF LEVETIRACETAM IN INJECTABLE DOSAGE FORM

C. N. Nalini* and K. M. Mohamed Adhil

Abstract

A straightforward, exact, reliable, robust, and selective stabilityindicating reverse phase high performance liquid chromatographic approach for levetiracetam and its impurities separation and quantification in Bulk and injectable dosage forms. Materials and Procedures: The analysis and description of the enhanced RP-HPLC method for levetiracetam and its impurities' separation and quantification is provided. A YMC- Pack ODS AQ (150 x 4.6 mm, 3μm column is used for reverse phase (RP-HPLC) analysis of samples, and the mobile phase is made up of dipotassium hydrogen phosphate buffer pH adjusted to 4.53 with dilute 85% orthophosphoric acid and Acetonitrile in the ratio of (95:5 v/v). 0.9 ml/min is the flow rate. 25μL of injection volume, a fixed wavelength of 205 nm, and a temperature difference of 25°C and 25°C were maintained for the column and sample, respectively. The limit of detection (LOD) and limit of quantitation (LOQ) for Levetiracetam standard and its impurities 0.01% and 0.03%, all the impurities in the specified concentration range are found satisfactory, with a correlation coefficient greater than 0.99. The accuracy studies were shown as % recovery for Levetiracetam and its impurities at specification level. The limit of % recovered shown is in the range of 80 and 120% and the results obtained were found to be within the limits. The new LC technique was verified in terms of robustness, linearity, specificity. To identify the degradation pathways and demonstrate that the approach indicates stability, a forced degradation research was carried out. As a result, this approach is verified and suitable for both regular and stable sample analysis.

Keywords: Levetiracetam, Levetiracetam RC-A, Levetiracetam RC-B, pyridin-2-ol, Method development, Validation, ICH guidelines.