Improving Multivariate Data Analysis Results of NMR Data by

Reference Deconvolution
Parvaneh Ebrahimi
1
, Sren Balling Engelsen
1
, Mathias Nilsson
1,2

1
Quality & Technology, Department of Food Sciences, Copenhagen University
2
School of Chemistry, University of Manchester

parvaneh@life.ku.dk

Abstract
NMR data inevitably suffer from hardware inconsistencies, and frequency, phase and line-
shape can differ significantly between samples. Clearly, this is a source of highly undesirable
error in studies that rely on quantitative pattern recognition such as metabolomics. Many of
these inconsistencies, however, can be corrected by reference deconvolution
(1, 2)
, which has
been used to good effect in NMR for many years, but appears neglected by the
metabolomics/chemometrics community. The method exploits the fact that most of these
inconsistencies are the same for all signals in the spectrum, and it works by comparing an
experimental reference signal to that of a perfect synthetic signal. The correction function for
that single signal can then be applied to the whole spectrum. In order to investigate the effect
of reference deconvolution on the results of multivariate analysis of NMR data, a ternary
design was prepared with 136 mixture samples of lactic acid, propionic acid, lactose, and a
constant metabolic background consisting of eight different amino acids and carbohydrates.
The recorded 1D-
1
H NMR data was processed with and without reference deconvolution.
Figure 1 shows the PCA scores plots of mean-centered data, both before and after reference
deconvolution with a 3Hz Gaussian lineshape for the reference TSP signal. It is clear that
reference deconvolution improves the results substantially; the triangular design is recovered
much better in the scores plot from reference deconvoluted data.



Figure1. Scores plot from applying PCA on the data a) before and b) after reference deconvolution

The results strongly suggest that reference deconvolution can be used as a fast and simple
operation to improve results of multivariate analysis of NMR data by cleaning up the NMR
data before multivariate data analysis. It is thus recommended that reference deconvolution
becomes routine in quantitative NMR research including metabolomics.

Main References
1. Morris et al. Reference deconvolution methods Progress in Nuclear Magnetic Resonance
Spectroscopy 31 (1997) 197-257.
2. http://dosytoolbox.chemistry.manchester.ac.uk/dosytoolbox/doku.php