Toxicity of the compound on the nervous system has been reported in development of pharmaceuticals such as toxicity of the compound and medicinal efficacy evaluation. In nonclinical study for the purpose of pharmaceuticals development, in in vivo using model animals are being conducted to examine the efficacy of the drug. However, the labor required for the experiment is excessive and it has a challenges low reproducibility and accuracy. Therefore, there is a need to develop a high accuracy and objective method of pain evaluated, but there method are not established. In this study, we aimed to development of pain evaluation system to evaluate of effectiveness and side effect of drugs. Using complementary metal-oxide semiconductor microelectrode array (CMOS-MEA) with nerve cells electrical activity can measure an ultra-high temporal resolution, the response of single sensory neurons to pain was obtained. Furthermore, based on the electrical activity obtained from 236,880 electrodes, we constructed a method to identify single cell, calculation of conduction velocity of axons and profiling of firing pattern each single cells.