Certain dyes are deleterious to the biological system, including animals and plants living in the water sources, soil sources, and so on. Thus, the analysis of these dyes requires a potent, quick, and cost-effective approach to the environmental samples. The present research work shows a modest, low-cost, and eco-friendly electrochemical device based on poly(dl-phenylalanine)-layered carbon nanotube paste electrode (P(PAN)LCNTPE) material for indigo carmine (ICN) detection in the presence of tartrazine. The cyclic voltammetric, field emission scanning electron microscopy, and electrochemical impedance spectroscopic methods were operated for the detection of the redox nature of ICN and electrode material surface activities, respectively. In better operational circumstances, P(PAN)LCNTPE provided better catalytic activity for the redox action of ICN than the bare carbon nanotube paste electrode. The P(PAN)LCNTPE showed good electrochemical activity during the variation of ICN concentrations ranging from 0.2 µM to 10.0 µM with improved peak current, and the limit of detection was about 0.0216 µM. Moreover, the P(PAN)LCNTPE material was performed as a sensor of ICN in a tap water sample and shows adequate stability, repeatability, and reproducibility.