Novel multiple emitting amphiphilic conjugated polythiophene‐coated CdTe quantum dots for picogram level determination of the 2,4,6‐trinitrophenol (TNP) explosive are developed. Four biocompatible sensors, cationic polythiophene nanohybrids (CPTQDs), nonionic polythiophene nanohybrids (NPTQDs), anionic polythiophene nanohybrids (APTQDs), and thiophene copolymer nanohybrids (TCPQDs), are designed using an in situ polymerization method, which shows highly enhanced fluorescence intensity and quantum yield (up to 78%). All sensors are investigated for nitroexplosive detection to provide a remarkable fluorescence quenching for TNP and the quenching efficiency reached 96% in the case of TCPQDs. The fluorescence of the sensors are quenched by TNP through inner filter effect, electrostatic, π−π, and hydrogen bonding interactions. Under optimal conditions, the detection limits of CPTQDs, NPTQDs, APTQDs, and TCPQDs are 2.56, 7.23, 4.12, and 0.56 × 10−9
m, respectively, within 60 s. More importantly, portable, cost effective, and simple to use paper strips and chitosan film are successfully applied to visually detect as little as 2.29 pg of TNP. The possibility of utilizing a smartphone with a color‐scanning APP in the determination of TNP is also established. Moreover, the practical application of the developed sensors for TNP detection in tap and river water samples is described with satisfactory recoveries of 98.02−107.50%.