nh-PHIP Hyperpolarization for Biological Mixture Analysis

Non-hydrogenative parahydrogen-induced polarization (nh-PHIP) is an NMR sensitivity enhancement technique that is particularly suitable for studying biological samples. It can provide up to three orders of magnitude signal enhancements for a continuously expanding range of valuable analyte classes like drugs, vitamins, nucleobases, nucleosides and nucleotides, amino acids, oligopeptides, etc. The strengths of nh-PHIP lay in its modest instrumental requirements over a standard commercial NMR spectrometer, its simple operating workflow that is similar to traditional NMR spectroscopy, and its proven capability to offer quantitative information from below the sensitivity barrier of regular NMR. This tutorial will give an overview of the background, the principles, and the present application scope of nh-PHIP hyperpolarization. We provide guidelines for setting up an nh-PHIP experiment, from sample preparation to optimizing the hyperpolarization for a particular analyte class. This includes the description of the basic experimental setup and outlining the considerations for method development. An overview of different available nh-PHIP pulse sequences will be given to assist the reader in implementing nh-PHIP.