Pure Shift NMR with Solvent Suppression: A Robust and General Method for Determining Quantitative Metabolic Profiles...

Area of Science:

• Analytical Chemistry
• Metabolomics
• Biochemistry

Background:

• Ultrahigh-resolution pure shift Nuclear Magnetic Resonance (NMR) offers quantitative metabolic profiling for studying cancer cell metabolic footprints.
• Previous work demonstrated its application on cancer cell growth medium, necessitating validation for broader biological sample types.

Purpose of the Study:

• To generalize a quantification method using ultrahigh-resolution NMR data to various biological samples.
• To evaluate the robustness and reproducibility of the analytical workflow for metabolite quantification.

Main Methods:

• Implementation of a reference ¹H pure shift spectra library with water suppression for metabolite concentration determination.
• Analysis of the NOESY-presat-PSYCHE-SAPPHIRE spectrum from extracellular medium.
• Assessment of sample preparation's influence on metabolite quantification accuracy using a model mixture.

Main Results:

• Quantification of a model metabolite mixture demonstrated good linearity, trueness, and precision across different sample preparation conditions.
• The analytical protocol showed high reproducibility, independent of sample physicochemical conditions.
• Successful implementation of the quantification protocol for determining metabolite levels in real urine and plasma samples.

Conclusions:

• The developed ultrahigh-resolution pure shift NMR method is robust and reproducible for quantitative metabolite analysis.
• This approach facilitates accurate metabolic profiling of diverse aqueous biological samples, including urine and plasma.
• The study paves the way for utilizing pure shift reference spectra libraries for broad quantitative metabolomics applications.