Determination of Huperzine A Concentration in Human Plasma by LC-MS/MS / 中国药房

ABSTRACT

OBJECTIVE:To develop a high performance liquid chromatography-mass spectrography method (LC-MS/MS) for determination of huperzine A in human serum. METHODS:The clonidine was used as the internal standard,and the test serum samples after being liquid-liquid extracted by ethyl acetate,dried in organic phase and reconstituted in mobile phase were analyzed on a Phenomenex Luna CN column interfaced with LC-MS with Methanol-0.08% formic acid (46∶54 containing 0.2% ammonium acetate). Positive ion SRM detection was performed for ESI iron source. Target ions were at m/z 243. 2→210. 9 for Huperzine A with an impact energy of 30V and m/z 229. 9→212. 9 for the internal standard with an impact energy of 32V during 0.5 s (scan time). RESULTS:The assay was proved to be free from the interference from endogenous substance,and the linear concentration range of Huperzine A was 0.08～8 ng?mL-1(r=0.993 0～0.996 0). The intra-and inter-day precision over the entire concentration range were less than 15%. At low,middle and high concentrations,the mean extraction recoveries of Huperzine A were 69.9%,63.7% and 63.7%,respectively. CONCLUSION:The LC-MS/MS method is rapid,sensitive,simple,accurate and in which little amount of mediator is needed,thus meeting the requirement for the detection of biological specimen and applicable for the treatment of large batch of serum samples in clinical pharmacokinetic study.