A line-broadening free real-time ³¹P pure shift NMR method for phosphometabolomic analysis

Phosphometabolomics by ³¹P NMR can be challenging, since overlapping multiplets of homonuclear coupled phosphorus nuclei complicate spectral analysis. Pure shift NMR allows to simplify such spectra by collapsing multiplets into singlets, but most pure shift methods require substantially elongated measurement times or cause disturbing spectral line broadening. Herein, we combine established pure shift NMR and artefact suppression techniques to record ³¹P pure shift NMR spectra without penalties in measurement time or line width. Examples are demonstrated in resolution of a mixture of nucleotide triphosphates and a biological sample of ¹⁸O labelled ATP isotopomers.