The evaporation-induced hydrovoltaic effect based on ion-selective nanochannels can theoretically be employed for high-performance ion sensing; yet, the indeterminate ion-sensing properties and the acquisition of high sensing performance are rarely explored. Herein, a controllable nanochannel regulation strategy for flexible hydrovoltaic devices with highly sensitive ion-sensing abilities is presented across a wide concentration range. By multiple dip-coating of silk fibroin (SF) on an electrospinning nylon-66 nanofiber (NNF) film, the surface polarity enhancement, the fibers size regulation with a precision of ≈25 nm, and the nanostructure firm binding are achieved simultaneously. The resultant flexible freestanding hydrovoltaic device exhibits an open circuit voltage up to 4.82 V in deionized water, a wide ion sensing range of 10−7 to 100 m, and ultrahigh sensitivity as high as 1.37 V dec−1, which is significantly higher than the sensitivity of the traditional solid-contact ion-selective electrodes (SC-ISEs). The fabricated flexible ion-sensitive hydrovoltaic device is successfully applied for wearable human sweat electrolyte sensing and for environmental trace-ion monitoring, thereby confirming the potential application of the hydrovoltaic effect for ion sensing.