Atmospheric pressure plasma (APP) has intrigued the interest of researchers for various applications such as disinfection, air purification, etc. In this context, a deeper understanding of the correlation between APP’s characteristics like discharge parameters, active species composition, and eradication of airborne microorganisms with varying relative humidity (RH) has been examined using a dielectric barrier discharge based atmospheric pressure plasma (DBD-APP) source. One of the electrodes of the developed DBD-APP source has been coated with TiO2 nanoparticles to enhance the generation of reactive species during the discharge process. The results show that, even with the same peak-to-peak applied voltage, the peak-to-peak current and discharge power decrease with increasing RH. Optical emission spectroscopy (OES) is used to determine the relative emission intensity of the reactive species, whereas spectrophotometry is used to quantify the reactive species produced by the plasma at various parameters. Instead of UV radiation, the plasma-produced highly energetic electrons activates the TiO2 nanoparticles for electron-hole pair generation. The geometry of the discharge has played an important role in generating high energy electrons. From the developed DBD-APP source, the airborne microorganism’s disinfection efficiency of ~ 95.8% and ~ 98.7% has been achieved in the total bacterial counts (TBCs) and total fungal counts (TFCs) at an RH range of 70% – 90%, in just 20 minutes of continuous operation. However, in the RH range of 20% – 40%, the inactivation efficiency dropped to ~ 78.8% and ~ 87.5% for the TBCs and TFCs, respectively. The outcome indicates that higher humidity levels are better for indoor air purification using DBD-APP sources and that plasma with a circulation system can effectively disinfect indoor environments.