Metabolic profiling of flavonoids in Lotus japonicus using liquid chromatography Fourier transform ion cyclotron resonance mass spectrometry

• Published in: Phytochemistry (Oxford) 2008, Vol.69 (1), p.99-111
• Main Authors: Suzuki, Hideyuki, Sasaki, Ryosuke, Ogata, Yoshiyuki, Nakamura, Yukiko, Sakurai, Nozomu, Kitajima, Mariko, Takayama, Hiromitsu, Kanaya, Shigehiko, Aoki, Koh, Shibata, Daisuke, Saito, Kazuki
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Chemical constitution; Chromatography, High Pressure Liquid; Cyclotrons; Flavonoids; Flavonoids - analysis; Flavonoids - chemistry; Flavonoids - metabolism; Flavonols - chemistry; Fourier Analysis; FT-MS; Fundamental and applied biological sciences. Psychology; glycosides; Glycosides - analysis; gossypetin; high performance liquid chromatography; isorhamnetin; LC-FTICR/MS; Leguminoseae; Loteae - chemistry; Loteae - growth & development; Loteae - metabolism; Lotus corniculatus var. japonicus; Lotus japonicus; mass spectrometry; Mass Spectrometry - methods; Metabolite profiling; metabolomics; Nitrogen - chemistry; Plant Components, Aerial - chemistry; Plant Components, Aerial - growth & development; Plant cytology, morphology, systematics, chorology and evolution; Plant physiology and development; Quercetin - analogs & derivatives; Spectrophotometry, Ultraviolet; Spermatophyta; Systematics (diagnosis, chromosome numbers)
• ISSN: 0031-9422; 1873-3700
• DOI: 10.1016/j.phytochem.2007.06.017

A comprehensive flavonoid analysis using LC-FTICR/MS established the presence of 61 flavonoid metabolites including gossypetin and isorhamnetin derivatives in Lotus japonicus accessions MG-20 and B-129. Flavonoid profiling demonstrated that anthocyanins and flavonol glycosides accumulated in a tissue- and development-dependent manner. Flavonoids detected from a model legume plant, Lotus japonicus accessions Miyakojima MG-20 and Gifu B-129, were profiled using liquid chromatography Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR/MS). Five flavonols and two anthocyanidins were detected as aglycones. LC-FTICR/MS facilitated simultaneous detection of 61 flavonoids including compounds that have not been reported previously. Chemical information of the peaks such as retention time, λ max, m/ z value of the quasi-molecular ion, m/ z value of MS/MS fragment ions, and relative intensity of MS/MS fragments was obtained, along with the molecular formulas and conjugate structures. Fourteen were completely identified by comparison with authentic compounds. The high accuracy of m/ z values, being 0.081 ppm between observed and theoretical values, allowed prediction of molecular formulas of unknown compounds with the help of isotope peak information for determination of chemical composition. Based on a predicted elemental composition, the presence of a novel nitrogen-containing flavonoid was proposed. A comparison of flavonoid profiles in flowers, stems, and leaves demonstrated that the flowers yielded the most complex profile, containing 30 flower-specific flavonoids including gossypetin glycosides and isorhamnetin glycosides. A comparison of flavonoid profiles between MG-20 and B-129 grown under the same conditions revealed that the accumulation of anthocyanins was higher in B-129 than MG-20, particularly in the stem. Developmental changes in the flavonoid profiles demonstrated that kaempferol glycosides increased promptly after germination. In contrast, quercetin glycosides, predominant flavonoids in the seeds, were not detectable in growing leaves.