Determination of huperzine A in human plasma by hydrophilic interaction chromatography coupled to tandem mass spectrometry

Abstract

A simple hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry (HILIC-MS/MS) method was developed and validated for the quantitative analysis of huperzine A in human plasma. The huperzine A and codeine phosphate (internal standard) were separated on an Alltima HP HILIC column using the mobile phase of acetonitrile–ammonium formate (5 mmol L⁻¹; pH 3.0) (13 : 87, v/v). The analyte was quantified using multiple reaction monitoring (MRM) in positive ion electrospray mode and no matrix effect was observed during the analysis. Linearity was established for concentrations in the range of 0.05–5 ng mL⁻¹ with a correlation coefficient (r) of 0.9992 using weighted 1/x²) linear least-squares regression. The lower limit of quantification (LLOQ) was identifiable and reproducible at 0.05 ng mL⁻¹. The present method was successfully applied to the pharmacokinetic study of huperzine A in 24 healthy male volunteers.