A nonenzymatic electrochemical lactic acid sensor was fabricated by electrodepositing gold microflowers on a screen‐printed carbon electrode and used to determine lactic acid in human serum and sweat quantitatively. Extensive voltammetric studies with lactic acid in phosphate‐buffer saline (PBS) and the analysis of the reduction product proved beyond doubt that lactic acid undergoes two‐electron reduction to form lactaldehyde. The reduction current increased quantitively with lactic acid concentration from 0.5 to 8.5 mM in 0.1 M PBS. The sensor showed excellent sensitivities of 1243 µA mM‐1 cm‐2 and 2752 µA mM‐1 cm‐2 in two linear ranges, 0.5 to 2.5 mM and 2.5 to 8.5 mM, respectively with a detection limit of 0.012 ± 0.0014 mM. The performance of the sensor was validated by estimating lactic acid in human serum and the sweat generated through physical exercise. The estimated lactic acid concentrations are in good agreement with clinical analysis data.This indicates the potential applicability of the sensor in the point‐of‐care detection of lactic acid.