A Comprehensive Targeted Metabolomics Assay for Crop Plant Sample Analysis.

Area of Science:

• Plant biology
• Biochemistry
• Analytical chemistry

Background:

• Metabolomics is crucial in agriculture and health, but comprehensive quantitative plant metabolite analysis is limited.
• Existing methods lack the sensitivity and breadth for detailed plant phenotyping and disease diagnosis.

Purpose of the Study:

• To develop and validate a high-coverage, quantitative mass spectrometry (MS)-based assay for plant metabolite analysis.
• To enable precise quantification of primary and secondary plant metabolites, including hormones.

Main Methods:

• Utilized liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) in both positive and negative modes.
• Employed multiple reaction monitoring (MRM) for targeted quantification of 206 metabolites.
• Developed assay in a 96-well plate format for high-throughput analysis.

Main Results:

• Quantified 206 plant metabolites: 28 amino acids, 27 organic acids, 20 biogenic amines, 40 acylcarnitines, 90 phospholipids, and C-6 sugars.
• Achieved recovery rates of 80-120% and precision below 20% for spiked samples.
• Successfully applied the assay to diverse plant samples including needles, roots, and cannabis.

Conclusions:

• The developed LC-MS/MS assay provides a robust, high-throughput method for quantitative plant metabolomics.
• This assay facilitates detailed plant phenotyping, disease diagnosis, and analysis of large sample cohorts.