1D-O1-1625 PDF
GC/MSメタボローム解析に資するcalibration-curve-lockingデータベース構築
Calibration-curve-locking databases (CCLD) have been constructed for semi-quantitative screening by gas chromatography/mass spectrometry (GC/MS) in fields of forensic medicine and residual pesticide measuring. These databases contain the retention time, calibration curve and electron impact ionization mass spectrum for automatic compound search and semi-quantification values of target compounds. To expand this method for metabolomics studies, we constructed a novel CCLD targeted the central carbon metabolism. For reproducible quantitative determinations using CCLD, automated just-in-time sample derivatization and sample loading protocol by a robotic platform was employed. Known amounts of standard compounds with 50 ppm internal standards were subjected to GC-MS after the automated sequential methoxime-TMS derivatization step by PAL RTC. One target (quantifier) ion and one or more qualifier ion were selected for each compound and a calibration curve was obtained by plotting the peak area ratio of the target compound to the IS versus the amount of target compound. These data of all standard compounds were registered as the novel CCLD, which enables automatic compound search and quantification by its target deconvolution and quantification algorithm. We examined the applicability of the constructed database to analyzing serum samples.