Recent progress in triboelectric nanogenerators (TENGs) has demonstrated their promising potential as a high-efficiency mechanical energy harvesting technology, and plenty of effort has been devoted to improving the power output by maximizing the triboelectric surface charge density. However, due to high-voltage air breakdown, most of the enhanced surface charge density brought by material/surface optimization or external ion injection is not retainable or usable for electricity generation during the operation of contact-separation-triggered TENGs. Here, the existence of the air breakdown effect in a contact-separation mode TENG with a low threshold surface charge density of ≈40–50 µC m−2 is first validated under the high impedance external load, and then followed by the theoretical study of the maximized effective energy output as limited by air breakdown for contact-separation-triggered TENGs. The effects of air pressure and gas composition are also studied and propose promising solutions for reducing the air breakdown effect. This research provides a crucial fundamental study for TENG technology and its further development and applications.