Immunoaffinity chromatography of abscisic acid combined with electrospray liquid chromatography–mass spectrometry

Hradecká, Veronika; Novák, Ondřej; Havlíček, Libor; Strnad, Miroslav
JOURNAL OF CHROMATOGRAPHY Analytical Technologies in the Biomedical and Life Sciences 847 [2]: 162-173, 2007

Keywords: abscisic acid; immunoaffinity chromatography; liquid chromatography-mass spectrometry
Abstract: Polyclonal antibodies with high specificity for C1-immobilised (+)-cis,trans-abscisic acid (ABA) were raised, characterised by enzyme-linked immunosorbent assay (ELISA) and used for preparation of an inummoaffinity chromatography (IAQ gel. The detection limit of the ELISA was approximately 4.6 x 10(-10) mol/L. Sensitive electrospray liquid chromatography-mass; spectrometry (LC-ESI-MS) methods, were also developed with detection limits below 0.1 x 10(-12) mol. The IAC allowed quick, single-step processing of samples prior to the analyses. The LC-ESI-MS and LC-ELISA techniques were used for comparative estimation of endogenous ABA levels in immumoaffinity purified extracts of normal and water-stressed Nicotiana tabacum L. leaves. The analytical approaches were validated using deuterium- and tritium-labelled internal standards, respectively. The LAC method was found to be highly effective, sensitive and convenient for isolating the target analyte from plant material.
IEB authors: Libor Havlíček, Ondřej Novák, Miroslav Strnad