Quantitative NMR Spectroscopy

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Description

Analysis of water-soluble metabolite classes including amino acids, sugars, alcohols, organic acids, amines, TCA cycle intermediates, short-chain fatty acids, etc.

Platform Strengths

Diverse substrates and sample types, inherently quantitative, sample integrity

Quantitative

Absolute

Absolute quantification data is reported for all identified metabolites using Chenomx software. The singlet produced by the DSS methyl group is used as an internal standard for chemical shift referencing (set to 0 ppm) and for quantification. All 1H-NMR spectra are processed and analyzed using the Chenomx NMR Suite Professional Software package (Chenomx Inc, Edmonton, AB). The Chenomx NMR Suite software allows for qualitative and quantitative analysis of an NMR spectrum by manually fitting spectral signatures from an internal database to the spectrum. Specifically, the spectral fitting for metabolite is done using the standard Chenomx 700 MHz metabolite library. Typically 95% of visible peaks are assigned to a compound and more than 95% of the spectral area can be routinely fit using the Chenomx spectral analysis software.

We have also developed an in-house program for automated NMR identification & quantification called MagMET. MagMET allows for qualitative and quantitative analysis of NMR spectra by automatically fitting spectral signatures from an internal database to the spectra. MagMET is currently validated for serum/plasma samples, and will be available for other sample types in the future). With MagMET, typically all visible peaks are assigned. Most of the visible peaks are annotated with a compound name. It has been previously shown that this fitting procedure provides absolute concentration accuracy of 90% or better.

Reference: Ravanbakhsh S, Liu P, Bjorndahl TC, Mandal R, Grant JR, Wilson M, Eisner R, Sinelnikov I, Hu X, Luchinat C, Greiner R, Wishart DS. (2015) Accurate, Fully-Automated NMR Spectral Profiling for Metabolomics. PLoS ONE 10(5): e0124219.

Metabolite Identification (Tier 1, Tier 2, Tier 3)

Tier 1

Example Data

Serum/plasma: Tier 1 - 60 metabolites (up to 125 metabolites screened)

Fecal: Tier 1 - 63 metabolites

Fish blood: Tier 1 - 63 metabolites

Statistical Analysis

Basic statistics included.

Platform Available

NMR

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