Optimization of Diffusion-Ordered NMR Spectroscopy Experiments for High-Throughput Automation in Human Metabolic...

Area of Science:

• Analytical Chemistry
• Metabolomics
• Nuclear Magnetic Resonance Spectroscopy

Background:

• Diffusion-ordered nuclear magnetic resonance spectroscopy (DOSY) can determine metabolite diffusion coefficients in mixtures.
• Widespread adoption in metabolic profiling is limited by the absence of standardized protocols.

Purpose of the Study:

• To develop and validate a standardized pipeline for DOSY experimental setup and data processing.
• To facilitate the use of DOSY for metabolic phenotyping of biological samples.

Main Methods:

• Optimization of three DOSY experiments: standard, relaxation-edited, and diffusion-edited.
• Development of a pipeline for experimental setup and data processing.
• Application to human blood plasma and urine samples.

Main Results:

• Achieved DOSY metabolic profiles with peak-picked diffusion coefficients for over 90% of signals in ¹H NMR spectra.
• Analysis time reduced to as little as 3 min 36 s.
• Demonstrated straightforward implementation for biofluid analysis.

Conclusions:

• The proposed pipeline standardizes DOSY for metabolic phenotyping.
• This approach enhances the utility of DOSY for analyzing complex biological samples.
• Facilitates efficient metabolic profiling of human blood plasma and urine.