In this study, a surface plasmon resonance (SPR) sensor based on modified plastic optical fibers (POFs) was combined with a specific molecularly imprinted polymer (MIP), used as a synthetic receptor, for glyphosate (GLY) determination in aqueous solutions. Since GLY is a non-selective herbicide associated with severe environmental and health problems, detecting glyphosate in environmental and biological samples remains challenging. The selective interaction between the MIP layer and GLY is monitored by exploiting the SPR phenomenon at the POF’s gold surface. Experimental results show that in about ten minutes and by dropping microliter volume samples, the presented optical–chemical sensor can quantify up to three orders of magnitude of GLY concentrations, from nanomolar to micromolar, due to a thin MIP layer over the SPR surface. The developed optical–chemical sensor presents a detection limit of about 1 nM and can be used for onsite GLY measurements. Moreover, the experimental analysis demonstrated the high selectivity of the proposed POF-based chemical sensor.