A rapid method for analysis of abscisic acid (ABA) in crude extracts of water stressed Arabidopsis thaliana plants by liquid chromatography-mass spectrometry in tandem mode

We describe a quick, simple method for the extraction and quantification of the phytohormone (+)-abscisic acid (ABA) in samples of plants subjected to different water deficit treatments. The method includes an extraction with acetone/water/acetic acid (80:19:1, v/v), evaporation of the extracts and finally injection into the liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) system in multiple reaction monitoring (MRM) mode. The objective of this work has been to show the applicability of the method to quantify the endogenous content of ABA in Arabidopsis thaliana (Columbia, Col-0) leaves at three different degrees of water stress. Control plants, had almost constant low levels of ABA (2-3 ng g-1, f.w.) throughout the 3 weeks of the experiment. Nevertheless, stressed plants increase the ABA content between the first and the second week (from 10 to 21 ng g-1, f.w.). The results suggest that this method is useful for quantifying ABA from plant material and that it avoids tedious and time-consuming extraction, purification and/or derivatization processes.