Porphyra sensu lato, a highly valuable edible seaweed renowned for its distinctive umami taste, undergoes significant taste variations during the harvest cycle, affecting product quality and pricing. In this study, umami-related metabolites in Pyropia haitanensis were investigated at different harvesting times using GC–MS metabolomic, targeted LC-MS analysis, and an electronic tongue taste evaluation. High concentrations of compounds, including glutamic acid, aspartic acid, and inosine 5′-monophosphate, were identified as the main contributors to the overall umami profile. The concentrations of the compounds and umami-enhancing substances, such as sugars, were negatively correlated as the harvesting period extended. The early harvested P. haitanensis exhibited a superior umami taste, which gradually decreased with subsequent harvest time. Proline, a known cold-resistance metabolite, accumulated as the seawater temperature decreased and the harvest period progressed. These findings provide insights into the optimal cultivation and harvesting practices for maintaining umami quality in P. haitanensis products.