Characterization and mapping of carotenoids in the algae Dunaliella and Phaeodactylum using Raman and target orthogonal partial least squares

A method was developed for the characterisation of carotenoid pigments in algal species using Raman spectroscopy in combination with multivariate hyperspectral analysis. Target orthogonal partial least squares (T-OPLS) operates by designating one known reference spectrum as the target. The target spectrum is put as the single y column in an OPLS regression model where the X matrix consists of the unfolded image spectra as variables in its columns. The spectral shape of the OPLS first orthogonal target score enabled us to verify the peak positions of the standard, and detect new peaks, not present in the reference standard. It was shown that the mixture of carotenoids present in the algae did not fully match the reference spectrum, however, the method provided enough information to make an analysis possible also in this case. The image results were constructed from the OPLS loading vectors that were showing a correlation map for the reference spectrum from the predictive loadings and maps of the occurrence of deviations from the orthogonal loadings.