A SENSITIVE AND ACCURATE METHOD FOR SIMULTANEOUS DETERMINATION OF ARTESUNATE AND ITS METABOLITE DIHYDRO- ARTEMISININ IN HUMAN PLASMA BY USING LC-MS/MS METHOD
Pikkili Viswanath, Chamarthi Nagaraju* and Doddipalli Venkata Ramana Reddy
ABSTRACT
A sensitive, accurate, precise and rugged method was developed and validated by using LC-MS/MS for Artesunate (AS) and its metabolite dihydroartemisinin (DHA) with internal standards. The extraction procedure carried out by liquid-liquid extraction using t-BME solvent and separated on a Thermo, Hypersil BDS, C18 (100 mm × 4.6 mm, 5 μm) column. The mobile phase was composed of 2 mM Ammoniumformate (pH 2.5): Acetonitrile in the ratio of 15:85 as a mobile phase at a flow rate of 0.5 mL/min in isocratic with 2.5 mins of run time. Artesunate (AS) and metabolite dihydroartemisinin (DHA) were ionized in positive polarity and detected in the selective reaction monitoring mode (SRM) using m/z 402.124→163.006, m/z 302.244→163.186, and internal standards Artesunate D3 and dihydroartemisinin D4 (AS-D3), (DHA-D4) m/z 406.172→163.078 and m/z 305.189→166.159 respectively. The linearity range used for quantification is 5.000-500.000 ng/mL for Artesunate (AS) and 10.000-1000.000 ng/mL for dihydroartemisinin (DHA) respectively.
Keywords: Artesunate, Dihydroartemisinin, Human plasma, Prodrug.
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