Nowadays, gold nanoparticles Au nanoparticles (AuNPs) capture great interest due to their chemical stability, optical properties and biocompatibility. The success of technologies based on the use of AuNPs implies the development of simple synthesis methods allowing, also, the fine control over their properties (shape, sizes, structure). Here, we present the AuNPs fabrication by nanosecond pulsed laser ablation in citrate-solution, that has the advantage of being a simple, economic and eco-sustainable method to fabricate colloidal solutions of NPs. We characterized the stability and the absorbance of the solutions by Ultraviolet-Visible (UV-Vis) spectroscopy and the morphology of the AuNPs by Transmission Electron Microscopy. In addition, we used the AuNPs solutions as colorimetric sensor to detect the amount of glyphosate in liquid. Indeed, glyphosate is one of the most widely used herbicides which intensive use represents a risk to human health. The glyphosate presence in the colloidal AuNPs solutions determines the aggregation of the AuNPs causing the change in the color of the solution. The variation of the optical properties of the colloidal solutions versus the concentration of glyphosate is studied.