Bullet-DNP Enables NMR Spectroscopy of Pyruvate and Amino Acids at Nanomolar to Low Micromolar Concentrations #DNPNMR

Published: Wednesday, 02 October 2024 - 10:00 -0400

Author: Thorsten Maly

Narwal, Pooja, Nils Lorz, Masoud Minaei, Sami Jannin, Karel Kouřil, Alvar Gossert, and Benno Meier. “Bullet-DNP Enables NMR Spectroscopy of Pyruvate and Amino Acids at Nanomolar to Low Micromolar Concentrations.” Analytical Chemistry, September 3, 2024, acs.analchem.4c00618.

https://doi.org/10.1021/acs.analchem.4c00618.

Hyperpolarized pyruvate is a widely used marker to track metabolism in vivo and a benchmark molecule for hyperpolarization methods. Here, we show how a combination of improved bullet-DNP instrumentation, an optimized sample preparation and a further sensitivity increase via a 13C−1H polarization transfer after dissolution enable the observation of pyruvate at a concentration of 250 nM immediately after dissolution. At this concentration, the experiment employs a total mass of pyruvate of only 20 ng or 180 pmol. If the concentration is increased to 45 μM, pyruvate may be detected 1 min after dissolution with a signal-to-noise ratio exceeding 50. The procedure can be extended to observe a mixture of amino acids at low micromolar concentrations.