Determination of huperzine A in human plasma by HPLC-MS-MS and its application to a relative bioavailability of huperzine A tablets in healthy volunteers

Objective To establish an HPLC-MS-MS method for the determination of huperzine A in the plasma and to study the relative bioavailability and the pharmacokinetics of huperzine A tablets.Methods With pseudoephedrine hydrochloride as the internal standard,huperzine A was extracted from the plasma sample with acetic ether and analyzed by a Han Bang C18 column(4.6 mm×150 mm,5 μm).HPLC-MS-MS was carried out with a mobile phase consisting of methanol-0.2% methanoic acid(85∶15,v/v) at a flow rate of 0.7 mL·min-1.A single oral dose of 0.2 mg huperzine A(T or R) was administered to 18 healthy volunteers in a randomized crossover study.Huperzine A concentration in the plasma was determined by the newly developed HPLC-MS-MS method,and the pharmacokinetics and bioavailability were studied.Results The calibration curve was linear within 0.050 8~5.080 μg·L-1(r=0.999 8).The method recovery of huperzine A was 101.8%~109.8%(n=5),and the intra-day and inter-day RSD 1.8%~Conclusion The established HPLC-MS-MS method is accurate and sensible.It can be used to study the pharmacokinetics of huperzine A.There is no significant difference between the two tablets,and they are bioequivalent.