Determination and pharmacokinetic studies of arecoline in dog plasma by liquid chromatography–tandem mass spectrometry

• A LC–MS/MS method for the determination of arecoline was developed in dog plasma.
• The method is simple, sensitive and rapid.
• The pharmacokinetics of arecoline hydrobromide in dogs was studied after oral administration.
• The synthesis of arecoline hydrobromide can achieve a yield of more than 78.11%.

A rapid and sensitive high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of arecoline concentration in dog plasma. Plasma sample was prepared by protein precipitation using n-hexane (containing 1% isoamyl alcohol) with β-pinene as an internal standard. Chromatographic separation was achieved on an Agilent C18 column (4.6 × 75 mm, 3.5 μm) using methanol: 5 mM ammonium acetate as the mobile phase with isocratic elution. Mass detection was carried out using positive electrospray ionization in multiple reaction monitoring mode. The calibration curve for arecoline was linear over a concentration range of 2–500 ng/mL. The intra-day and inter-day accuracy and precision were within the acceptable limits of ±10% at all concentrations. In summary, the LC–MS/MS method described herein was fully validated and successfully applied to the pharmacokinetic study of arecoline hydrobromide tablets in dogs after oral administration.