A novel, direct and simple colorimetric method employing μicroanalytical paper-based devices (μ-PADs) for the selective determination of bismuth is described. The suggested method exploits the colorimetric variation of bismuth after its rapid reaction with methyl thymol blue (MTB) in an acidic medium (pH ranging between 0.7 and 3.0), modified with nitric acid, on the surface of a paper device at room temperature. The devices are low cost, composed of chromatographic paper and wax barriers and the analytical protocol is easily applicable with minimal technical expertise and without the need for experimental apparatus. The user must add a test sample and read the color intensity of the colored Bi(III)–MTB complex formed at the sensing area using a simple imaging device such as a flatbed scanner. Various chemical variables, such as HNO3 and MTB concentration, reaction time, ionic strength, detection zone size and photo-capture detector are optimized. A study of interfering ions such as K+, Na+, Ca2+, Mg2+, Cl−, SO42− and HCO3− was also conducted. The stability of the paper devices is also studied in different maintenance conditions with particularly satisfactory results, rendering the method suitable for field analysis. The detection limits are as low as 3.0 mg L−1 with very satisfactory precision, ranging from 4.0% (intra-day) to 5.5% (inter-day). Natural water samples are successfully analyzed, and bismuth percentage recoveries were calculated in the range of 82.8 to 115.4%.