Derivatization-Assisted Acoustic Ejection Mass Spectrometry for High-Throughput FAHFA Prescreening

Area of Science:

• Lipidomics
• Biochemistry
• Analytical Chemistry

Background:

• Fatty acid esters of hydroxy fatty acids (FAHFAs) are bioactive lipids involved in metabolic regulation, inflammation, and insulin sensitivity.
• Low abundance and wide polarity range of FAHFAs challenge high-throughput screening.
• Existing methods like LC-MRM MS are time-consuming for comprehensive FAHFA analysis.

Purpose of the Study:

• To develop a novel, high-throughput prescreening strategy for FAHFAs.
• To overcome the limitations of current methods in analyzing diverse FAHFA species.
• To accelerate the discovery of novel FAHFA structures and their biological roles.

Main Methods:

• Development of derivatization-assisted acoustic ejection mass spectrometry (DAEMS).
• Integration of N,N-dimethylethylenediamine (DMED) derivatization with acoustic ejection mass spectrometry (AEMS).
• High-throughput screening of over 2800 potential FAHFA species in under 2 hours.

Main Results:

• DAEMS achieved ultrafast analysis (1-3 samples/sec) with enhanced sensitivity.
• Screened over 2800 FAHFA species, significantly faster than conventional LC-MRM MS.
• DAEMS demonstrated 83% coverage and 79% accuracy compared to LC-MRM MS.
• Identified novel oxidized FAHFA species in edible fungi, suggesting new biosynthetic pathways.

Conclusions:

• DAEMS is a promising tool for rapid FAHFA prescreening.
• The method significantly accelerates the analysis of FAHFA diversity.
• Discovery of oxidized FAHFAs offers new insights into lipid metabolism and biochemistry.