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A rapid accurate and precise method for simultaneous determination of β-glucan and protein content in naked oat samples, based on the coupling of near-infrared spectroscopy and chemometrics, is presented. In particular, three different spectroscopic approaches [near infrared reflectance (NIR) and transmittance (NIT) on flour and NIT on whole grains] and various spectral pretreatments were considered. To account for the possibility of outlying samples, a robust version of the PLS algorithm (namely partial robust M-regression) was used. All the models resulted as accurate as the reference methods, reflectance spectroscopy being the technique providing the best outcomes. Variable reduction by inclusion of the most relevant predictors only (as evaluated by VIP scores) resulted in simpler and, in one case, more parsimonious models, without loss in accuracy.