Development of facile, on-spot, robust, and efficient optical nanosensors for drugs and pesticides is one of the active research area. In this study, a nanosensor based on fairly stable silver nanoparticles (AgNPs) conjugated with specially designed thiourea (TU)-based molecule, N-((4'-methoxy-2'-nitrophenyl) carbamothioyl) dodecanamide (TU3), was prepared using one pot protocol and characterized by Ultraviolet (UV)-Visible spectroscopy, dynamic light scattering (DLS), fourier transform infrared (FTIR) spectroscopy and atomic force microscopy (AFM). The TU3 conjugated AgNPs (TU3-AgNPs) were found to be selective spectrophotometric sensor for an antimicrobial drug, amlodipine (Ad), among the pool of different drugs. TU3-AgNPs based nanosensor allows for the rapid quantitative assay having a detection limit of 0.3 μM in an analytical range of 0.1-100 μM. The proposed sensor is specific in presence of number of other interfering drugs and other matrix. Upto the best of our literature survey, this is first ever colorimetric sensor for quantification of amlodipine in complex matrix. The specificity of the sensor allows for quantitative detection of amlodipine in environmental, biological and pharmaceutical samples.