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An improved strategy to analyse strigolactones in complex sample matrices using UHPLC-MS/MS.

Kristýna Floková1,2, Mahdere Shimels3,4, Beatriz Andreo Jimenez3,5

  • 1Plant Hormone Biology Group, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.

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This study introduces a novel, sensitive method for extracting and analyzing plant hormones called strigolactones. The technique enables precise detection of seven strigolactones in small plant samples, aiding agricultural research.

Keywords:
Phosphate starvationPhytohormonesQuantitative analysisSolid phase extraction (SPE)StrigolactonesUHPLC–MS/MS

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Area of Science:

  • Plant Biology
  • Hormone Signaling
  • Agricultural Science

Background:

  • Strigolactones are key plant hormones regulating growth and mediating plant-microbe and plant-parasite interactions.
  • Their low concentrations and instability in complex matrices hinder detailed study.
  • Parasitic plant germination, triggered by strigolactones, poses significant agricultural challenges.

Purpose of the Study:

  • To develop a sensitive and efficient method for simultaneous profiling of multiple strigolactones.
  • To overcome challenges associated with low concentrations, instability, and complex sample matrices.
  • To enable analysis from small amounts of plant root tissue and exudates.

Main Methods:

  • Developed tailored extraction strategies with controlled temperature and organic modifiers for improved stability and purity.
  • Utilized rapid, single-step pre-concentration on polymeric reversed-phase solid-phase extraction (RP SPE) sorbent.
  • Designed an efficient ultra-high-performance liquid chromatography (UHPLC) separation coupled with sensitive tandem mass spectrometry (MS/MS) detection.

Main Results:

  • Achieved attomolar detection limits for seven strigolactones and their precursors (carlactone, carlactonoic acid).
  • Successfully profiled strigolactones in small root tissue (150 mg FW) and exudate (20 ml) samples.
  • Validated the method for four agriculturally important species: sorghum, rice, pea, and tomato.

Conclusions:

  • The established method offers efficient strigolactone extraction and pre-concentration from small samples.
  • Improved strigolactone stability and reduced matrix contaminants enhance UHPLC-MS/MS analysis sensitivity.
  • The method is versatile and applicable to diverse plant species for strigolactone profiling.