Colorimetric detection of fatty acids during biological interactions is extremely difficult since they are optically silent. Here, fatty acids are found to function as ion-exchangers in ultrasmall polymeric nanospheres to facilitate the protonation of chromoionophores, causing a vivid color change between red and blue. With an excellent detection limit of 1.8 μg mL-1 for oleic acid, colorimetric assays for lipase and albumin are developed with quick response, high sensitivity, and low cost.