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

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Mar 1;847(2):162-73. doi: 10.1016/j.jchromb.2006.09.034. Epub 2006 Oct 24.

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 immunoaffinity chromatography (IAC) gel. The detection limit of the ELISA was approximately 4.6x10(-10)mol/L. Sensitive electrospray liquid chromatography-mass spectrometry (LC-ESI-MS) methods were also developed with detection limits below 0.1x10(-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 immunoaffinity 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 IAC method was found to be highly effective, sensitive and convenient for isolating the target analyte from plant material.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abscisic Acid / analysis*
  • Abscisic Acid / chemistry
  • Abscisic Acid / immunology
  • Chromatography, Affinity / methods*
  • Chromatography, Liquid / methods*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Nicotiana / chemistry
  • Nicotiana / drug effects
  • Nicotiana / metabolism
  • Plant Leaves / chemistry
  • Plant Leaves / drug effects
  • Plant Leaves / metabolism
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Water / metabolism
  • Water / pharmacology

Substances

  • Water
  • Abscisic Acid