Reversible Parahydrogen Induced Hyperpolarization of ¹⁵N in Unmodified Amino Acids Unraveled at High Magnetic Field

Abstract

Amino acids (AAs) and ammonia are metabolic markers essential for nitrogen metabolism and cell regulation in both plants and humans. NMR provides interesting opportunities to investigate these metabolic pathways, yet lacks sensitivity, especially in case of 15N. In this study, spin order embedded in p‐H2 is used to produce on‐demand reversible hyperpolarization in 15N of pristine alanine and ammonia under ambient protic conditions directly in the NMR spectrometer. This is made possible by designing a mixed‐ligand Ir‐catalyst, selectively ligating the amino group of AA by exploiting ammonia as a strongly competitive co‐ligand and preventing deactivation of Ir by bidentate ligation of AA. The stereoisomerism of the catalyst complexes is determined by hydride fingerprinting using 1H/D scrambling of the associated N‐functional groups on the catalyst (i.e., isotopological fingerprinting), and unravelled by 2D‐ZQ‐NMR. Monitoring the transfer of spin order from p‐H2 to 15N nuclei of ligated and free alanine and ammonia targets using SABRE‐INEPT with variable exchange delays pinpoints the monodentate elucidated catalyst complexes to be most SABRE active. Also RF‐spin locking (SABRE‐SLIC) enables transfer of hyperpolarization to 15N. The presented high‐field approach can be a valuable alternative to SABRE‐SHEATH techniques since the obtained catalytic insights (stereochemistry and kinetics) will remain valid at ultra‐low magnetic fields.

J Coupling Constants of <1 Hz Enable 13C Hyperpolarization of Pyruvate via Reversible Exchange of Parahydrogen

“…15 We started with the pH 2 and insensitive nuclei polarization transfer sequence (phINEPT), 44 which has already been used for SABRE to enhance 15 N signals (and will be used for 13 C in this work) and to measure lifetimes for Ir complexes. 43,45 This approach theoretically allows one to probe the J couplings. However, when the simple phINEPT approach is used, it becomes apparent that it is virtually impossible to simultaneously measure two weak 1 H− 13 C J coupling constants (Figure S5).…”