Raman microspectroscopic analysis of triterpenoids found in plant cuticles

The above-ground organs of plants are covered by a cuticle, an extracellular membrane performing important physiological and ecological functions, that consists of the fatty acid-derived polymer cutin and waxes. In the cuticular wax of many species, including the leaves of Prunus laurocerasus, triterpenoids are found at high concentrations. This paper investigates the potential of Raman microspectroscopy for the simultaneous detection of structurally similar triterpenoids in plant cuticles. Relative composition analysis was first performed on artificial triterpenoid mixtures consisting of alpha-amyrin and oleanolic acid, as well as oleanolic acid and ursolic acid, the two triterpenoids abundantly found in the cuticles of P. laurocerasus. The different triterpenoids could be distinguished in the mixture spectra and the resulting calculated triterpenoid ratios were consistent with the expected values. Qualitative analysis of the Raman spectra of P. laurocerasus cuticle demonstrated the in situ detectability of the triterpenoids using this approach. It is shown here that Raman microspectroscopy has the potential to provide useful information concerning the spatial distribution of some key chemical components of plant cuticles. This technique thus offers a valuable complement to the current standard analytical methods used for analyzing the bulk composition of plant cuticles.