Calculating analytical uncertainties as a part of method validation is a relevant aspect of field and laboratory practices in instrumental analytical chemistry subjects, which usually require complex algorithms. This work describes the development and didactic use of an automatic and straightforward informatics tool, implemented in an Excel macro, for calculating and interpreting the uncertainty of an analytical method against a reference method on field measurements. The software was initially developed for field testing of low-cost air quality monitoring analytical methods against reference methods, and the present work shows its adaptation to a didactic environment. The uncertainty calculation software was implemented through an Excel macro based on Visual Basic as a graphical user interface. It finds a best-fit line that describes the relation between concentrations determined by the candidate and reference methods. The software generates the analytical validation results (slope and intercept with their respective confidence limits, and expanded uncertainty of a concentration determined by the candidate method), hiding the intermediate functions and calculations. The Excel interface eases uncertainty calculations for undergraduate students, although the background mathematics can be quickly unveiled to students for didactic purposes. This tool has been applied to a laboratory exercise focused on validating experimental results obtained in the measurement of ozone levels in ambient air by passive sampling and spectrophotometric detection. The uncertainty calculation software has proved valuable by providing the student a resource to check the analytical quality of the data generated in the laboratory, while assimilating the fundamentals behind the calculations.