Rapid assessment of severity is crucial for timely intervention and improved patient outcomes in heatstroke (HS). However, existing biomarkers are limited in their accuracy and accessibility in prehospital / ER settings. A prospective study was conducted to assess urinary liver fatty acid-binding protein (L-FABP) levels using a point-of-care testing upon HS. Pathophysiological severity was estimated using initial Sequential Organ Failure Assessment (SOFA) scores, and outcomes were measured using modified Rankin Scale (mRS) scores. In 78 severe HS patients, urinary L-FABP concentrations were measured on admission, with a median concentration of 48.3 ng/mL. Positive correlation was observed between urinary L-FABP concentration and pulse rate (r = 0.300), lactate (r = 0.259), and initial SOFA score (r = 0.211). The POCT of L-FABP showed promise in predicting pathophysiological severity, as indicated by higher concentrations in patients with higher initial SOFA scores. Additionally, patients in the POCT positive group had significantly worse outcomes at discharge compared to the negative group, although this difference diminished over time. The study demonstrates the feasibility and potential utility of POCT for initial L-FABP in estimating pathophysiological severity in HS patients. This rapid and accessible testing method may aid in early field triage and intervention, ultimately improving patient outcomes in the management of HS.