[NMR analysis blog] Binning and NMR Data Analysis

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Binning and NMR Data Analysis

Yesterday I mentioned that many NMR arrayed experiments suffer from unwanted chemical shift variations due to fluctuations in experimental conditions such as sample temperature, pH, ionic strength, etc. This phenomenon is very common in NMR spectra of e.g. biofluids (metabonomics/metabolomics) but also exists in many other experiments such us Relaxation, Kinetics and PFG NMR spectra (diffusion).

This problem negatively affects the reliability of quantitation using, for instance, peak heights, and for this reason integration is, in general, a more robust procedure as these spectral variations are mitigated by averaging data points over the integral segment. In this post, I just want to show you one simple trick which helps to understand, in a pictorial way, why integration is useful to remove the major part of chemical shift scattering.


First, consider the following experiment depicted in the figure below. It shows a triplet and as you can see, some minor peaks shifts are present from spectrum to spectrum



If peak heights are determined at a fixed position, this might introduce appreciable errors in the posteriori quantitative analysis (e.g. exponential fitting). As described in my former post, this could be circumvented in some extent by using parabolic interpolation or peak searching of the maximum in a predefined box.
Nevertheless, integration is a very simple solution as can be appreciated in the figure below. Instead of using the Peak Integrals tool in the Data Analysis module, I will show now a complementary procedure. Basically, what I have applied to all spectra is the well-known binning operation which consists of dividing each spectrum in equally sized (e.g. 0.01 ppm in this case) bins, so that integral (area) of each bin represents a new point in the binned spectrum


As seen in the figure above, binning clearly removes the effect of chemical shift changes but of course, at the cost of a significant reduction in data resolution.






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Source: NMR-analysis blog