High-throughput miniaturised solid phase extraction - supercritical fluid chromatography tandem mass spectrometry...

Area of Science:

• Plant Science
• Analytical Chemistry
• Biochemistry

Background:

• Acidic phytohormones are crucial low-abundance signaling molecules in plants.
• Accurate and efficient profiling of these compounds is essential for understanding plant physiology.

Purpose of the Study:

• To develop a high-throughput workflow for profiling acidic phytohormones.
• To introduce complementary selectivity in sample cleanup and chromatographic separation.

Main Methods:

• Coupling of miniaturized solid-phase extraction (RP-μSPE) with ultra-high performance supercritical fluid chromatography tandem mass spectrometry (SFC-MS/MS).
• Optimization of SFC-MS/MS conditions including stationary phase, mobile phase, and gradient.
• Validation of the workflow for 19 acidic phytohormones.

Main Results:

• The SFC-MS/MS workflow reduced run time from 19 to 11 minutes compared to RP-LC.
• Achieved accuracies within 85-115% and precision of 0.2-14.9%.
• Demonstrated applicability through time-course profiling in wounded Arabidopsis thaliana leaves.

Conclusions:

• The established SFC-MS/MS workflow provides an efficient and selective method for acidic phytohormone profiling.
• This method offers advantages in speed and selectivity over traditional RP-LC workflows.
• The workflow is suitable for plant research, including stress response studies.