Metabolite Profiling of Clinical Cancer Biofluid Samples by NMR Spectroscopy
Metabolomics is a comprehensive characterization of the small polar molecules (metabolites) in different biological systems. One of the analytical platforms commonly used to study metabolic alterations in biofluid samples is proton nuclear magnetic resonance (1H NMR) spectroscopy. NMR spectroscopy is very specific, quantitative, and highly reproducible. Moreover, sample preparation for NMR experiments is very simple and straightforward, and this gives NMR spectroscopy a distinct advantage over other metabolic profiling methods. It has already been shown that 1H NMR-based profiling of biological fluids can be effective in differentiating benign from malignant lesions and in investigating the efficacy of specific cancer treatments. Therefore, 1H NMR spectroscopy may become a promising tool for early noninvasive diagnosis and rapid assessment of treatment effects in cancer patients. Here, we describe a detailed protocol for 1H NMR metabolite profiling in serum, plasma, and urine samples, including sample collection procedures, sample preparation for 1H NMR experiments, spectral acquisition and processing, and quantitative profiling of 1H NMR spectra. We also discuss several aspects of appropriate study design and some multivariate statistical methods that are commonly used to analyze metabolomics datasets.
Key wordsMetabolomics 1H NMR Quantitative profiling Cancer Serum Plasma Urine
This work was funded by a CRIO-Cancer Grant from Alberta Innovates Health Solutions. We are indebted to all our colleagues that provided clinical samples, and especially to Drs. Aalim Weljie, Farshad Farshidfar, and Karen Kopciuk for discussions about methodology.
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