Optimization of Human Plasma ¹H NMR Spectroscopic Data Processing for High-Throughput Metabolic Phenotyping Studies and Detection of Insulin Resistance Related to Type 2 Diabetes

Abstract

Optimizing NMR experimental parameters for high-throughput metabolic phenotyping requires careful examination of the total biochemical information obtainable from (1)H NMR data, which includes concentration and molecular dynamics information. Here we have applied two different types of mathematical transformation (calculation of the first derivative of the NMR spectrum and Gaussian shaping of the free-induction decay) to attenuate broad spectral features from macromolecules and enhance the signals of small molecules. By application of chemometric methods such as principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (O-PLS-DA) and statistical spectroscopic tools such as statistical total correlation spectroscopy (STOCSY), we show that these methods successfully identify the same potential biomarkers as spin-echo (1)H NMR spectra in which broad lines are suppressed via T2 relaxation editing. Finally, we applied these methods for identification of the metabolic phenotype of patients with type 2 diabetes. This "virtual" relaxation-edited spectroscopy (RESY) approach can be particularly useful for high-throughput screening of complex mixtures such as human plasma and may be useful for extraction of latent biochemical information from legacy or archived NMR data sets for which only standard 1D data sets exist.

Two-dimensional DOSY experiment with Excitation Sculpting water suppression for the analysis of natural and biological media

“…3). The plasma metabolite assignment generally needs at least two 1D NMR experiments [18]: a pulse-and-acquire sequence, in which large signals of macromolecules preclude small molecules analysis, and a transverse relaxation-edited sequence (also called a Carr-Purcell-Meiboom-Gill (CPMG)) in which small molecules are observed due to the suppression of signals from protons with short T 2 relaxation times. The signal of both types of components can be detected in only one step with the DOSY-ES experiment.…”

GC/TOFMS analysis of metabolites in serum and urine reveals metabolic perturbation of TCA cycle in db/db mice involved in diabetic nephropathy

“…Recently, metabonomics has been applied to the evaluate DM-and DN-related diseases (9,17,18,32,33). These studies suggested potential markers of DM-and DN-related diseases, such as the nonesterified fatty acids and esterified fatty acids, lipids, acyl-carnitines, acyl-glycines, and metabolites related to tryptophan metabolism.…”

Two-dimensional correlation spectroscopy — Biannual survey 2007–2009

“…A number of papers dealing with statistical 2D covariance analysis have recently appeared in the field of NMR spectroscopy, notably by Brüschweiler [102][103][104][105][106], Nicholson [108][109][110][111][112][113][114][115][116] and others [117][118][119][120][121]. For example, Chen et al [102] reported that 2D NMR spectra can be generated by the covariance approach by using a relatively few t 1 increments.…”

“…Johnson et al [112] used the kinetic variant of STOCSY called J-resolved STOCSY to study acyl glucuronide intra-molecular transacylation reaction. Maher et al [113] applied STOCSY for high-throughput metabolic phenotyping for diabetes detection. Lindon et al [114] claimed STOCSY is related to samplesample correlation in the review of NMR application on metabonomics.…”