Pollutant conversions at the air−water interface of a single droplet are crucial for obtaining accurate predictions on environment quality. Herein, droplet ESI MS was carried out to understand the photochemical conversions of pollutants across the water−air interface of a single droplet. A single droplet was kept on a Y-shaped Cu electrode, realizing the in situ electrospray ionization for the examinations. By applying high voltage, in situ MS detection of a droplet was carried out layer-by-layer, from the water−air interface to the inner droplet. By droplet ESI MS, secondary pollutants in a single droplet were observed under visible-light irradiation, while no obvious conversion was recorded in the bulk solution, indicating the complicated and active conversions in droplets. More significantly, different pollutant conversions at the water−air interface and inner droplet were recorded by the dynamic monitoring via droplet ESI MS detections layer-by-layer. By examining the observed dimer and radical intermediates, photochemical mechanisms across the water−air interface were proposed. By providing an effective pathway for single-droplet examinations, this work enlarges applications of ambient MS into environment studies.