Colour pigments of Trichoderma harzianum: Preliminary investigations with thin-layer chromatography–Fourier transform infrared spectroscopy and high-performance liquid chromatography with diode array and mass spectrometric detection

Abstract

The colour pigments of Trichoderma harzianum fermentation broth were separated and the main fractions were tentatively identified by reversed-phase thin-layer chromatography–Fourier transform infrared spectroscopy (RP-TLC–FT-IR), RP-HPLC–diode array detection and RP-HPLC–MS. It was established that the multistep gradient elution developed for RP-TLC separation of pigments can be successfully used as a pilot method for the rational design of gradient elution in RP-HPLC for the separation of the same pigments. FT-IR and MS measurements were unable to identify the exact chemical structures of the main pigment fractions, the presence of OH, =CH and C=O (RP-TLC–FT-IR) and OH and NH 2 substructures (RP-HPLC–MS) was confirmed. It was assumed that the main pigment fractions are oxidation polymers originating from monomer molecules containing polar substructures and double bonds in the alkyl chain which are liable for oxidation during the aerobic fermentation process.