The objectives of this study were to determine the reflectance properties of volunteer potato and sugar beet and to assess the potential of separating sugar beet and volunteer potato at different fields and in different years, using spectral reflectance characteristics. With the ImspectorMobile, vegetation reflection spectra were successfully repeatedly gathered in two fields, on seven days in 2 years that resulted in 11 datasets. Both in the visible and in the near-infrared reflection region, combinations of wavelengths were responsible for discrimination between sugar beet and volunteer potato plants. Two feature selection methods, discriminant analysis (DA) and neural network (NN), succeeded in selecting sets of discriminative wavebands, both for the range of 450–900 and 900–1650 nm. First, 10 optimal wavebands were selected for each of the 11 available datasets individually. Second, by calculating the discriminative power of each selected waveband, 10 fixed wavebands were selected for all 11 datasets analyses. Third, 3 fixed wavebands were determined for all 11 datasets. These three wavebands were chosen because these had been selected by both DA and NN and were for sensor 1: 450, 765, and 855 nm and for sensor 2: 900, 1440, and 1530 nm. With the resulting three sets of wavebands, classifications were performed with a DA, a neural network with 1 hidden neuron (NN1) and a neural network with two hidden neurons (NN2). The maximum classification performance was obtained with the near-infrared sensor coupled to the NN2 method with an optimal adapted set of 10 wavebands, where the percentages were 100 ± 0.1 and 1 ± 1.3% for true negative (TN) classified volunteer potato plants and false negative (FN) classified sugar beet plants respectively. In general the NN2 method gave the best classification results, followed by DA and finally the NN1 method. When the optimal adapted waveband sets were generalized to a set of 10 fixed wavebands, the classification results were still at a reasonable level of a performance at 87% TN and 1% FN for the NN2 classification method. However, when a further reduction and generalization was made to 3 fixed wavebands, the classification results were poor with a minimum performance of 69% TN and 3% FN for the NN2 classification method. So, these results indicate that for the best classification results it is required that the sensor and classification system adapt to the specific field situation, to optimally discriminate between volunteer potato and sugar beet pixel spectra.