High-throughput interspecies profiling of acidic plant hormones using miniaturised sample processing.

Area of Science:

• Plant Science
• Biochemistry
• Analytical Chemistry

Background:

• Acidic phytohormones regulate crucial plant physiological functions.
• Understanding phytohormone profiles (active forms, precursors, metabolites) is vital for plant biology research.

Purpose of the Study:

• To develop and optimize a high-throughput sample preparation method for analyzing acidic phytohormones.
• To enable the analysis of phytohormone profiles from minute amounts of plant material.

Main Methods:

• Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed.
• A method using minimal plant tissue (<10 mg fresh weight) with acidic extraction and miniaturized purification was optimized.
• A 3D-printed 96-place interface facilitated processing of 192 samples per run.

Main Results:

• The optimized method accurately determines 25 acidic phytohormones (auxins, jasmonates, abscisates, salicylic acid).
• The method demonstrated high process efficiency, recovery, and accuracy with minimal matrix effects.
• Validation parameters confirmed the method's reliability for plant phytohormone profiling.

Conclusions:

• The developed method enables high-throughput profiling of acidic phytohormones from minute plant samples.
• This technique is suitable for large-scale, interspecies comparative studies in plant science.