Abstract

HPLC-MS METHOD DEVELOPMENT AND VALIDATION OF RASAGILINE

*Km. Pooja, Shilpa Pahwa, Perwaiz Alam and Lalit Kumar Tyagi

ABSTRACT

The rapid advancement in the area of pharmaceutical analysis where sensitive chromatographic and spectral techniques have been evoked for the determination of drugs in pharmaceutical dosage forms and in biological fluids. The most suitable analytical method based on liquid–liquid extraction has been developed and validated for quantification of Rasagiline in human plasma. Rasagiline-13C3 mesylate was used as an internal standard for Rasagiline. Zorbax Eclipse Plus C18 (2.1 mm50 mm, 3.5 mm) column provided chromatographic separation of analyte followed by detection with mass spectrometry. The method involved simple isocratic chromatographic condition and massspectrometric detection in the positive ionization mode using an API-4000 system. The total runtime was 3.0 min. The proposed method has been validated with the linear range of 5–12000 pg/mL for Rasagiline. The intra-run and inter-run precision values were within 1.3%–2.9% and 1.6%–2.2% respectively for Rasagiline. The overall recovery for Rasagiline and Rasagiline-13C3 mesylate analog was 96.9% and 96.7% respectively. This validated method was successfully applied to the bioequivalence and pharmacokinetic study of human volunteers under fasting condition.

Keywords: Spectral techniques; High performance liquid chromatography; Mass spectrometry; Rasagiline; Liquid–liquid extraction.