Sample preparation issues in NMR-based plant metabolomics: optimisation for Vitis wood samples.

Area of Science:

• Plant metabolomics
• Analytical chemistry
• Organic chemistry

Background:

• Nuclear magnetic resonance (NMR) is a key technique in plant metabolomics.
• Reproducibility and simplicity of NMR are high, but sample preparation significantly impacts data quality.

Purpose of the Study:

• To investigate and optimize sample preparation protocols for untargeted NMR-based metabolomics.
• Focus on Vitis vinifera L. (grapevine) wood samples.

Main Methods:

• Evaluation of various extraction solvents (e.g., ethyl acetate, methanol, dichloromethane) and deuterated solvents (e.g., methanol-d4, chloroform-d-methanol-d4).
• Assessment of optimal extract concentration, chemical shift stability, and peak area repeatability.
• Utilized multivariate data analysis for cultivar-based variation estimation.

Main Results:

• Ethyl acetate proved to be the most effective extraction solvent for diverse secondary metabolites in grapevine wood.
• Optimal extract concentration determined at 10 mg/mL.
• 7:3 chloroform-d-methanol-d4 (v/v) identified as the superior deuterated solvent system for NMR analysis.

Conclusions:

• Careful selection of extraction and deuterated solvents is essential for comprehensive metabolomic surveys.
• Optimized protocols enhance the quality and reliability of NMR-based metabolomics data for Vitis vinifera wood.