DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-UPLC METHOD FOR THE DETERMINATION OF RELATED SUBSTANCES IN LEVETIRACETAM DRUG SUBSTANCE

Stability-indicating RP-UPLC methods have been developed for determination and validation of related substances in Levetiracetam bulk drug samples. The method shows adequate separation of Levetiracetam from and their associated related substances and degradation products. An efficient Separation of Levetiracetam related substances and degraded products were achieved by using New Waters HSS T3 column with (100 mm x 2.1 mm, 1.8 μm) dimensions. The buffer Ortho phosphoric Acid at pH 2.0 and Acetonitrile initial gradient was taken in (95.0:5.0 v/v) with a flow rate of 0.5 ml/min and Wavelength at 210 nm. Run time of the method was found to be 6.0 min. The linear regression analysis data for calibration plots show good linear relationship with 0.999. The limit of detection and limit of quantification are determined for Levetiracetam chloramide, Levetiracetam acid and Levetiracetam 0.01%, 0.002%, 0.002% and 0.03%, 0.005%, 0.004% respectively. The signal to noise ratios was found more than 3.4 and 10.5 for all components determined in the method. Intra and Inter-day precision is less than 2.5%. The % recovery of Levetiracetam impurities are between 90.2 – 114.5 %. The drug is subjected to different stress conditions and the resulting degradation products obtained did not interfere with the detection of Levetiracetam. Keywords: Levetiracetam, Levetiracetam Chloramide, Levetiracetam Acid, RP-UPLC, Validation