Although negative air ionizer is commonly used for indoor air cleaning, few studies examine the concentration gradient of negative air ion (NAI) in indoor environments. This study investigated the concentration gradient of NAI at various relative humidities and distances form the source in indoor air. The NAI was generated by single-electrode negative electric discharge; the discharge was kept at dark discharge and 30.0 kV. The NAI concentrations were measured at various distances (10-900 cm) from the discharge electrode in order to identify the distribution of NAI in an indoor environment. The profile of NAI concentration was monitored at different relative humidities (38.1-73.6% RH) and room temperatures (25.2 ± 1.4 °C). Experimental results indicate that the influence of relative humidity on the concentration gradient of NAI was complicated. There were four trends for the relationship between NAI concentration and relative humidity at different distances from the discharge electrode. The changes of NAI concentration with an increase in relative humidity at different distances were quite steady (10-30 cm), strongly declining (70-360 cm), approaching stability (420-450 cm) and moderately increasing (560-900 cm). Additionally, the regression analysis of NAI concentrations and distances from the discharge electrode indicated a logarithmic linear (log-linear) relationship; the distance of log-linear tendency (λ) decreased with an increase in relative humidity such that the log-linear distance of 38.1% RH was 2.9 times that of 73.6% RH. Moreover, an empirical curve fit based on this study for the concentration gradient of NAI generated by negative electric discharge in indoor air was developed for estimating the NAI concentration at different relative humidities and distances from the source of electric discharge.