One of the fundamental limitations of hyperpolarized 13C technology is the effective lifetime of the signal, which decays in keeping with the spin-lattice relaxation constant T1. We have developed a robust late stage deuteration methodology which is broadly applicable to amino and alpha hydroxyl acids, including commonly used probes such as alanine and lactate. This methodology enables significant T1 prolongation, yielding an effective doubling of in vivo signal to noise ratio at relevant imaging time-points. We tested [1-13C, 2-2H]alanine prepared via this method both in vitro and in vivo. This broadly applicable methodology may facilitate implementation and translation of hyperpolarized 13C MRI probes.