Metabolites in a urine extract are signal-enhanced by SABRE† hyperpolarization and detected using a benchtop NMR spectrometer. Quantification by standard addition is demonstrated for endogenic urinary nicotinamide (vitamin B3). Even higher sensitivity is achieved in an automated setup for multi-scan SABRE experiments. This hyperpolarization scheme is able to expedite biomarker detection and quantification, while maintaining low infrastructure requirements.