Molecular Structure of Gardenia Blue Pigments by Reaction of Genipin with Benzylamine and Amino Acids

Genipin was reacted with benzylamine and several amino acids to prepare gardenia blue (GB). The time-course of GB formation with benzylamine was monitored by high-performance liquid chromatography (HPLC), liquid chromatography time-of-flight mass spectrometry (LC-TOFMS), and H-1 and C-13 NMR measurements. In this experiment, we determined the molecular structures of some intermediates using accurate masses and additional NMR techniques such as heteronuclear multiple bond correlation (HMBC). GBs with amino acids (GB-AAs) were characterized by both liquid and solid-state NMR measurements. Interestingly, many significant peaks appeared in the solid-state NMR spectra, although the 13C NMR spectra from solution samples did not show any distinct peaks. Therefore, we determined that GB-AAs had an alternating copolymer structure composed of methyne and 5H-2-pyrindine, which was substituted by amino acids at N atom and linked with methyne at 5 and 7 positions. To confirm this molecular structure, the pyrolysis gas chromatography-mass spectrometry (GC-MS) measurement of GB-AAs was carried out, and 5H-2-pyrindine and its methyl derivatives were formed as main pyrolysis products from the polymer chains.

Automated summary

Genipin reacted with benzylamine and amino acids to prepare gardenia blue (GB). The time-course of GB formation was monitored by HPLC, LC-TOFMS, and NMR, and the molecular structures of intermediates were determined using accurate masses and additional NMR techniques. GBs with amino acids were characterized by both liquid and solid-state NMR measurements, revealing an alternating copolymer structure composed of methyne and 5H-2-pyrindine substituted by amino acids and linked with methyne at specific positions.