A liquid chromatography/electrospray ionisation tandem mass spectrometry method for the simultaneous quantification of salicylic, jasmonic and abscisic acids in Coffea arabica leaves

J Sci Food Agric. 2014 Feb;94(3):529-36. doi: 10.1002/jsfa.6288. Epub 2013 Jul 26.

Abstract

Background: Plants have developed an efficient system of recognition that induces a complex network of signalling molecules such as salicylic acid (SA), jasmonic acid (JA) and abscisic acid (ABA) in case of a pathogenic infection. The use of specific and sensitive methods is mandatory for the analysis of compounds in these complex samples.

Results: In this study a liquid chromatography/electrospray ionisation tandem mass spectrometry method was developed and validated for the simultaneous quantification of SA, JA and ABA in Coffea arabica (L.) leaves in order to understand the role of these phytohormones in the signalling network involved in the coffee defence response against Hemileia vastatrix. The results showed that the method was specific, linear (r ≥ 0.99) in the range 0.125-1.00 µg mL⁻¹ for JA and ABA and 0.125-5.00 µg mL⁻¹ for SA, and precise (relative standard deviation ≤11%), and the limit of detection (0.010 µg g⁻¹ fresh weight) was adequate for quantifying these phytohormones in this type of matrix.

Conclusion: In comparison with healthy leaves, those infected with H. vastatrix (resistance reaction) displayed an increase in SA level 24 h after inoculation, suggesting the involvement of an SA-dependent pathway in coffee resistance.

Keywords: Coffea sp; HPLC/ESI-MS/MS; Hemileia vastatrix; coffee rust; phytohormones; signalling.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Abscisic Acid / analysis
  • Chromatography, High Pressure Liquid / methods*
  • Coffea / chemistry*
  • Coffea / microbiology
  • Cyclopentanes / analysis
  • Disease Resistance*
  • Fungi
  • Oxylipins / analysis
  • Plant Diseases / microbiology*
  • Plant Growth Regulators / analysis*
  • Plant Leaves / chemistry*
  • Plant Leaves / microbiology
  • Reproducibility of Results
  • Salicylic Acid / analysis
  • Signal Transduction
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Tandem Mass Spectrometry / methods

Substances

  • Cyclopentanes
  • Oxylipins
  • Plant Growth Regulators
  • jasmonic acid
  • Abscisic Acid
  • Salicylic Acid