This study was conducted to develop calibration models for determining quality parameters of whole kernel barley using a rapid and nondestructive near-infrared (NIR) spectroscopic method. Two hundred and five samples of whole barley grains of three winter-habit types (hulled, malt, and hull-less) produced over three growing seasons and from various locations in the United States were used in this study. Among these samples, 137 were used for calibration and 68 for validation. Three NIR instruments with different resolutions, one Fourier transform instrument (4 cm<sup>−1</sup> resolution), and two dispersive instruments (8 nm and 10 nm bandpass) were utilized to develop calibration models for six components (moisture, starch, β-glucan, protein, oil, and ash) and the results were compared. Partial least squares regression was used to build models, and various methods for preprocessing of spectral data were used to find the best model. Our results reveal that the coefficient of determination for calibration models (NIR predicted versus reference values) ranged from 0.96 for moisture to 0.79 for β-glucan. The level of precision of the model developed for each component was sufficient for screening or classification of whole kernel barley, except for β-glucan. The higher resolution Fourier transform instrument gave better results than the lower resolution instrument for starch and β-glucan analysis. The starch model was most improved by the increased resolution. There was no advantage of using a higher resolution instrument over a lower resolution instrument for other components. Most of the components were best predicted using first-derivative processing, except for β-glucan, where second-derivative processing was more informative and precise.

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