GC-MS analysis of indole-3-acetic acid, abscisic acid and gibberellins in garlic (Allium sativum L.) purified by two HPLC columns

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 A1 (GA1), Indole-3-acetic acid (IAA), trans-Abscisic acid (t-ABA), Abscisic acid (ABA) and Gibberellin A4 (GA4). 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, GA1 and GA4) and four internal standard substances ((sup(13)C6-IAA, [sup(2) H6]-ABA, [sup(2)H2]-GA1 and [sup(2)H2]-GA4 ) 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).