Research article

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

  1. Daniel Cuthbertson1,†,
  2. Jasenka Piljac-Žegarac1,2,†,
  3. Bernd Markus Lange1,*

Article first published online: 24 JAN 2012

DOI: 10.1002/bmc.2677

Additional Information

How to Cite

Cuthbertson, D., Piljac-Žegarac, J. and Lange, B. M. (2012), Validation of a microscale extraction and high-throughput UHPLC-QTOF-MS analysis method for huperzine A in Huperzia. Biomed. Chromatogr.. doi: 10.1002/bmc.2677

Author Information

  1. 1

    Institute of Biological Chemistry and M.J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA, USA

  2. 2

    Department of Molecular Biology, Institute Ruđer Bošković, Zagreb, Croatia

  1. †

    Daniel Cuthbertson and Jasenka Piljac-Žegarac contributed equally to this work.

*B. Markus Lange, Institute of Biological Chemistry and M.J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA 99164–6340, USA. E-mail: lange-m@wsu.edu

Publication History

  1. Article first published online: 24 JAN 2012
  2. Manuscript Accepted: 14 NOV 2011
  3. Manuscript Revised: 11 NOV 2011
  4. Manuscript Received: 25 JUL 2011

Funded by

  • National Institutes of Health. Grant Number: 5RC2GM092561-02

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Keywords:

  • alkaloid;
  • Huperzia;
  • huperzine A;
  • mass spectrometry;
  • microscale extraction

ABSTRACT

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.

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