Nitrogen (N) is one of the most important limiting nutrients for sugarcane production. Conventionally, sugarcane N concentration is examined using direct methods such as collecting leaf samples from the field followed by analytical assays in the laboratory. These methods do not offer real-time, quick, and non-destructive strategies for estimating sugarcane N concentration. Methods that take advantage of remote sensing, particularly hyperspectral data, can present reliable techniques for predicting sugarcane leaf N concentration. Hyperspectral data are extremely large and of high dimensionality. Many hyperspectral features are redundant due to the strong correlation between wavebands that are adjacent. Hence, the analysis of hyperspectral data is complex and needs to be simplified by selecting the most relevant spectral features. The aim of this study was to explore the potential of a random forest (RF) regression algorithm for selecting spectral features in hyperspectral data necessary for predicting sugarcane leaf N concentration. To achieve this, two Hyperion images were captured from fields of 6–7 month-old sugarcane, variety N19. The machine-learning RF algorithm was used as a feature-selection and regression method to analyse the spectral data. Stepwise multiple linear (SML) regression was also examined to predict the concentration of sugarcane leaf N after the reduction of the redundancy in hyperspectral data. The results showed that sugarcane leaf N concentration can be predicted using both non-linear RF regression (coefficient of determination, R 2 = 0.67; root mean square error of validation (RMSEV) = 0.15%; 8.44% of the mean) and SML regression models (R 2 = 0.71; RMSEV = 0.19%; 10.39% of the mean) derived from the first-order derivative of reflectance. It was concluded that the RF regression algorithm has potential for predicting sugarcane leaf N concentration using hyperspectral data.