Program

Oral Sessions

Day 2, May 16（Wed.）　16:20-16:40　Room B (Seiun 1)

2B-O2-M-1620　　PDF

Highly accurate detection and identification methodology of xenobiotic metabolites based on LC/HRMS/MS

(¹Kyushu Univ., ²Sumitomo Chemical, ³Sojo Univ., ⁴Osaka Univ.)
^oMasatomo Takahashi¹, Yoshihiro Izumi¹, Fukumatsu Iwahashi², Yasumune Nakayama³, Mitsuhiko Iwakoshi², Motonao Nakao¹, Seiji Yamato², Eiichiro Fukusaki⁴, Takeshi Bamba¹

Recently, a generally applicable discovery method of xenobiotic metabolites is becoming important for safety and effective development of xenobiotics. In the present study, we have proposed an advanced methodology for detection and identification of comprehensive xenobiotic metabolites by a combination of stable isotope labeling, liquid chromatography coupled to a benchtop quadrupole Orbitrap high-resolution tandem mass spectrometry (LC/HRMS/MS) analysis, data mining techniques (alignment, peak picking, and paired-peaks filtering), in silico metabolism prediction, and time-dependent profiling. The LC/HRMS analysis was carried out using Arabidopsis T87 cultured cells treated with unlabeled, or ¹³C- or ²H-labeled 2,4-dichlorophenoxyacetic acid (2,4-D). The paired-peaks filtering, which is a filtering technique by searching LC/HRMS paired-peaks generated from unlabeled and labeled 2,4-D, enabled the accurate detection of 83 candidates for 2,4-D metabolites without false positive peaks derived from solvents or biological matrix. In addition to confirming 10 kinds of 2,4-D metabolites reported so far, we successfully identified 16 novel 2,4-D metabolites using in silico metabolism prediction, time-dependent profiling, and HRMS/MS spectra. Our established method has several advantages over conventional techniques, including the accurate detection and identification of comprehensive xenobiotic metabolites, and represents a potentially useful tool to elucidate the xenobiotic metabolism.