GC-MS Analysis of Indole-3-Acetic Acid, Abscisic Acid and Gibberellins in Garlic(Allium sativum L.) Purified by Two HPLC Columns

本研究は、HPLCのみで植物ホルモンを精製するために、容量の多いカラム(粗精製カラム)の次に容量の少ないカラム(本精製カラム)で溶離させる精製方法(2段階精製法)を検討した。両カラムにおいて5つの標準物質は、GA_1、IAA、t-ABA、ABA、GA_4の順に溶出した。本精製カラムの分離能は粗精製カラムより高く、 2段階精製法によって精製能が向上した。さらに、GC-MSにおける標準物質(IAA、ABA、GA_1、GA_4)および内部標準物質(C^^<13>_6-IAA、d_6-ABA、[H^^2_2]-GA_1、[H^^2_2]-GA_4)のマススペクトルを示した。2段階精製法によって精製した休眠期のニンニクの内生ABA(52.5 pmol/g 生重量)をGC-MSによって定量した。In this study, the refinement method (two stage refinement method) that is eluted by a column on a few of capacity (purification column) through a column on a lot of capacity (coarse column) was examined to refine plant hormones only with a high performance liquid chromatography (HPLC). Five authentic standards in both columns were eluted in order of Gibberellin A_1 (GA_1), Indole-3-acetic acid (IAA), trans-Abscisic acid (t-ABA), Abscisic acid (ABA) and Gibberellin A_4 (GA_4). The separation capacity in the purification column was higher than the coarse column, and the refining capacity was progressed by the two stage refinement method. In addition, the mass spectrum of four authentic standards (IAA, ABA, GA_1 and GA_4) and four internal standard substances (C^^<13>_6-IAA,[H^^2_6]-ABA,[H^^2_2]-GA_1,[H^^2_2]-GA_4) in a gas chromatography-mass spectrometry (GC-MS) were shown. The internal ABA level in the dormant garlic (Allium sativum L.), that had refined by two stage refinement method, was determined by GC-MS (52.5 pmol/g FW)