In the present work, we performed a comparative investigation of spectroelectrochemical and biosensor application of isomeric thienylpyrrole derivatives. For this purpose two thienylpyrrole derivatives were synthesized characterized and electrochemically polymerized. Characterizations of the resulting polymers were performed by cyclic voltammetry (CV), UV-vis spectroscopy. Moreover, the spectroelectrochemical, electrochromic properties and biosensing applications of the polymer films were investigated. The resulting polymer films have distinct electrochromic properties and show five different colors. The 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl) aniline (SNS-NH2) and 3-(2,5- di(thiophen-2-yl)-1H-pyrrol-1-yl) aniline P(SNS-mNH(2)) films show maximum optical contrast (DT%) of 41.5%, 25.4% at 431 nm, 422 nm with a response time of 1.5 s. For biosensing studies, P(SNS-NH2) and P(SNS-mNH(2)) were polymerized on graphite electrodes electrochemically and used as immobilization matrices. After electrochemical deposition, glucose oxidase (GOx) was immobilized on the modified electrodes as the model enzyme. Effects of the position of the amine group on spectroelectrochemical properties and biosensing capability of the polymers were investigated.