The development of a portable device created by 3D printing for colorimetric and fluorometric measurements is an efficient tool for analytical applications in situ or in the laboratory presenting a wide field of applications in the environmental and food field. This device uses a light-emitting diode (LED) as radiation source and a webcam as a detector. Digital images obtained by the interaction between the radiation source and the sample were analyzed using a programming language developed in Matlab (Mathworks Inc., Natick, MA, USA), which builds the calibration curves in real-time using the RGB colour model. In addition, the entire system is connected to a notebook which serves as an LED and detector power supply without the need for any additional power source. The proposed device was used for the determination in situ of norfloxacin, allura red, and quinine in water and beverages samples, respectively. For the validation of the developed system, the results obtained were compared with a conventional spectrophotometer and spectrofluorometer respectively with a t-test at a 95% confidence level, which provides satisfactory precision and accuracy values.