A Real-Life Reproducibility Assessment for NMR Metabolomics.

Area of Science:

• Analytical Chemistry
• Biochemistry
• Metabolomics

Background:

• Nuclear magnetic resonance (NMR) metabolomics is increasingly adopted by diverse groups, including those with limited analytical expertise.
• Previous studies confirmed high reproducibility with specialized groups using advanced NMR equipment and standard operating procedures (SOPs).
• The impact of mixed operator expertise on NMR metabolomics reproducibility remains unassessed.

Purpose of the Study:

• To evaluate the quantitation reproducibility of NMR metabolomics in a multipurpose environment with operators of varying expertise.
• To identify limiting factors for data reproducibility, including operator skill, pipetting, NMR tubes, and instrumentation.

Main Methods:

• An interlaboratory assessment was conducted using two NMR instruments across different labs.
• Multiple operators with diverse backgrounds and metabolomics experience participated.
• Variability was assessed for operator influence, automatic pipettes, NMR tubes, and NMR instruments.

Main Results:

• The study estimated a maximum variability of 4% for NMR quantitation reproducibility in a typical multipurpose NMR lab.
• Operator-induced variability was found to be significantly compensated by current NMR equipment and SOPs.
• The assessment identified key sources of variability in a real-world NMR setting.

Conclusions:

• Current NMR technology and established SOPs are robust enough to ensure high quantitation reproducibility, even with operators of mixed expertise.
• Multipurpose NMR environments can achieve reliable results by leveraging advanced instrumentation and standardized protocols.
• The findings support the broader application of NMR metabolomics across various research settings.