The level of reducing sugars and asparagine in raw potatoes is critical for potato breeders and the food industry for production of commonly consumed food products including potato chips and French fries. Our objective was to evaluate the use of a portable infrared instrument for the rapid quantitation of major sugars and amino acids in raw potato tubers using single-bounce attenuated total reflectance (ATR) and dial path accessories as an alternative to time-consuming chromatographic techniques. Samples representing a total of 84 experimental and commercial potato varieties harvested in two consecutive growing seasons (2012 and 2013) were used in this study. Samples had wide ranges of sugars determined by HPLC-RID (non-detectable (ND)-7.7 mg glucose, ND-9.4 mg fructose and 0.4–5.4 mg sucrose per 1 g fresh weight), and asparagine and glutamine levels determined by GC-FID (0.7–2.9 mg and 0.3–1.7 mg per 1 g fresh weight). Infrared spectra collected from 64 varieties were used to create partial least squares regression (PLSR) calibration models that predicted the sugar and amino acid levels in an independent set of 16 validation potato varieties. Excellent linear correlations between infrared predicted and reference values were obtained. PLSR models had a high correlation coefficient of prediction (rPred >0.95) and residual predictive deviation (RPD) values ranging between 3.1 and 5.5. Overall, the results indicated that the models could be used to simultaneously predict sugars, free asparagine and glutamine levels in the raw tubers, significantly benefiting potato breeding, certain aspects of crop management, crop production and research.