In this work we demonstrate 'singlet-contrast magnetic resonance imaging' using fumarate, a representative biomolecule. Unlike previous work on this chemical system which used dissolution dynamic nuclear polarization (D-DNP) to generate singlet order , we produce fumarate by chemically reacting para-enriched hydrogen gas with an acetylene precursor in D2O, using a ruthenium trans-hydrogenation catalyst . Fumarate is a metabolite in the citric acid (Krebs) cycle, and is converted into malate by addition of a water molecule; a reaction catalysed by the enzyme fumarase, and of great importance for hyperpolarization-enhanced MRI . The enzymatic conversion to malate renders the fumarate protons chemically inequivalent. Since the fumarate protons originate from parahydrogen, the malate becomes hyperpolarized, and the resulting enhanced NMR signals are antiphase. These PASADENA (Parahydrogen And Synthesis Allow Dramatically Enhanced Nuclear Alignment) signals can be observed by applying a 45° rf pulse . The formation and metabolism of fumarate are shown in Fig. , alongside a comparison between PASADENA and thermal equilibrium 1 H NMR spectra of malate.