Establishment of LC-MS/MS method for determination of aloperine in rat plasma and its application in preclinical pharmacokinetics

Aloperine, a novel natural active alkaloid derived from Sophora alopecuroides L., has attracted much attention for its anti-inflammatory, antiviral, anti-tumor, anti-allergy and other pharmacological activities. In this study, we first established and validated an efficient and sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of aloperine in rat plasma. Cytisine was used as the internal standard (IS). The separation of aloperine and IS was conducted on a Phenomenex Luna Omega Polar C₁₈ column (2.1 × 50 mm, 1.6 μm) with 0.3% (v/v) formic acid aqueous (containing 5 mM ammonium acetate) and 0.3% (v/v) formic acid acetonitrile using isocratic elution condition at a flow rate of 0.20 mL/min. Aloperine and IS were determined under the transitions of m/z 233.2 → 98.1 and m/z 191.2 → 148.2 (positive ionization mode), respectively. The calibration curve of aloperine was established in the range of 5 (LLOQ) to 2000 ng/mL (r² = 0.994). The well validated method was full compliance with the bioanalytical method validation of FDA, and was applied to the pharmacokinetic study of aloperine in Sprague-Dawley rats after 50 mg/kg oral administration and 5 mg/kg intravenous injection. This study provides valuable references for the further study of Sophora alopecuroides L., especially for the drug development and clinical application of aloperine.