Hyperpolarization of stable isotope-labeled substrates and subsequent NMR measurement of the metabolic reactions allow for direct tracking of cellular reactions in vitro and in vivo. Here, we report the hyperpolarization of ¹³C₆-glucose- d ₇ and evaluate its use as probes to observe glucose flux in cells. We measured the lifetime of the polarized signal governed by the spin–lattice relaxation time T ₁. ¹³C₆-Glucose- d ₇ exhibited a T ₁ that was over ten times as long as that of ¹³C₆-glucose, and metabolic NMR studies of hyperpolarized ¹³C₆-glucose- d ₇ using tumor cell lysate led to observation of the resonances due to phosphorylated fluctofuranoses generated through aerobic glycolysis.