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Visualizing usnic acid with anisaldehyde reagent

Published online by Cambridge University Press:  12 December 2024

Bruce McCune Open the ORCID record for Bruce McCune [Opens in a new window] ,
Wilmer H. Perera Open the ORCID record for Wilmer H. Perera [Opens in a new window] ,
Xinhui Yu Open the ORCID record for Xinhui Yu [Opens in a new window]  and
Kerry McPhail Open the ORCID record for Kerry McPhail [Opens in a new window]
Bruce McCune*
Affiliation:
Department of Botany and Plant Pathology, Oregon State University, Corvallis, Oregon 97331, USA
Wilmer H. Perera
Affiliation:
CAMAG Scientific, Inc., Wilmington, NC 28401, USA
Xinhui Yu
Affiliation:
Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, Oregon 97331, USA
Kerry McPhail
Affiliation:
Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, Oregon 97331, USA
*
Corresponding author: Bruce McCune; Email: mccuneb@oregonstate.edu
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Abstract

The pale yellowish tint of usnic acid in a lichen thallus itself is a commonly used character in identification keys, particularly in the genus Cladonia. Furthermore, the presence of usnic acid is phylogenetically significant in numerous groups of lichens. While the distinctive colour of usnic acid is readily visible when present in high concentrations, it is commonly problematic to discern when in low to moderate concentrations. We explored the use of an anisaldehyde reagent for visualizing usnic acid. Using both usnic acid-containing Cladonia samples and pure usnic acid, this reaction quickly yields a bright magenta colour on HPTLC and TLC plates after heating and directly with crude acetone extracts on glass slides heated with a lighter. The same magenta product was observed whether or not the usnic acid was accompanied by barbatic, fumarprotocetraric, psoromic, squamatic or thamnolic acids, each of which alone did not produce any colour with anisaldehyde reagent. However, the merochlorophaeic acids in C. albonigra also produced a red reaction. Analysis by high resolution LC-MS of the reaction mixture between anisaldehyde and usnic acid revealed several ions at m/z 477.1586 ([M+H]+, C27H25O8) and 463.1385 ([M+H]+, C26H23O8), respectively, consistent with aldol condensation of usnic acid and p-anisaldehyde.

Keywords

aldol condensationCladoniaHPTLClichenspot testTLCusnic acid
Type
Standard Paper
Information
The Lichenologist , Volume 56 , Special Issue 5: A Special Issue dedicated to Professor Teuvo Ahti , September 2024 , pp. 193 - 200
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Copyright
Copyright © The Author(s), 2024. Published by Cambridge University Press on behalf of the British Lichen Society

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