Rapid UPLC-MS/MS Determination of Amantadine and Rimantadine in Human Blood and Urine for Forensic Toxicology

Abstract

A rapid UPLC-MS/MS method was developed and validated for quantifying amantadine (AMD) and rimantadine (RIM) in human blood and urine. Sample preparation involved acetonitrile protein precipitation with amantadine-D6 as internal standard. Separation used an ACE C₁₈-PFP column and a mobile phase of 0.1% aqueous formic acid/5 mmol/L ammonium formate-acetonitrile (85:15, v/v). ESI⁺-MRM detection was employed. Acetonitrile provided optimal extraction. The method showed excellent linearity (1.0–1000.0 ng/mL; R² > 0.99), LOD/LOQ , mean recoveries (82.16–105.43%), and precision (RSD 3.57–10.39%). This sensitive, reliable method is suitable for forensic and public health applications.