Here we present a novel Smartphone-based colorimeter and demonstrate its application to the measurements of glucose and protein concentrations in biological samples. The key innovation of our approach was to combine powerful image processing encoded into a mobile phone application with a low cost 3D printed sample holder that allowed to control lighting conditions and significantly improved sensitivity. Different solutions with protein and glucose concentrations ranging from 0 to 2000 mg/dL were prepared and analyzed using our system. The Smartphone-based colorimeter always correctly classified the corresponding reagent strip pads, what confirms that it can be used as a low cost alternative for commercial test strip analyzers.