Validation of a microscale extraction and high-throughput UHPLC-QTOF-MS analysis method for huperzine A in Huperzia

Herein we report on an improved method for the microscale extraction of huperzine A (HupA), an acetylcholinesterase‐inhibiting alkaloid, from as little as 3 mg of tissue homogenate from the clubmoss Huperzia squarrosa (G. Forst.) Trevis with 99.95% recovery. We also validated a novel UHPLC‐QTOF‐MS method for the high‐throughput analysis of H. squarrosa extracts in only 6 min, which, in combination with the very low limit of detection (20 pg on column) and the wide linear range for quantification (20–10,000 pg on column), allow for a highly efficient screening of extracts containing varying amounts of HupA. Utilization of this methodology has the potential to conserve valuable plant resources. Copyright © 2012 John Wiley & Sons, Ltd.