This procedure includes adding a small drop of concentrated glucose solution to the sample in the NMR tube, mixing, waiting for an equilibration time, and acquisition of a second spectrum. We propose a simple “Add to Subtract” background subtraction method, and show that it can reduce the glucose signals by 98% to allow retrieval of the hidden information. Signals from most metabolites in this region are overwhelmed by the glucose background signals and become invisible. In particular, blood serum/plasma and diabetic urine samples contain high concentrations of glucose, which produce strong peaks between 3.2 ppm – 4.0 ppm. However 1H NMR spectra contain many overlapped peaks. Due to its highly reproducible and quantitative nature, and minimal requirements for sample preparation or separation, 1H nuclear magnetic resonance (NMR) spectroscopy is widely used for profiling small-molecule metabolites in biofluids.
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