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

The colour pigments of Trichoderma harzianum fermentation broth were separa ted and the main fractions were tentatively identified by reversed-phase th in-layer chromatography-Fourier transform infrared spectroscopy (RP-TLC-FT- LR), 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 gradi ent elution in RP-HPLC for the separation of the same pigments. FT-IR and M S 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, substructures (RP-HPLC-MS) was confirmed. It was assumed tha t the main pigment fractions are oxidation polymers originating from monome r molecules containing polar substructures and double bonds in the alkyl ch ain which are liable for oxidation during the aerobic fermentation process. (C) 2000 Elsevier Science B.V. All rights reserved.