A quantitative method was developed to characterize the short-range order in non-crystalline starch by Raman spectroscopy. The Raman spectra of three forms of non-crystalline starches (just-gelatinized starch, which was heated to the point of having just lost its long-range order but still retaining essentially all of its short-range order, gelatinized starch and amorphous starch) were resolved into subspectra to calculate the short-range ordered phases. By deducting the spectra of amorphous starch using a subtraction technique, the areas of subspectra for short-range ordered phases in just-gelatinized and gelatinized starches were obtained. The ratio of the area for short-range ordered phases in gelatinized starch relative to that in just-gelatinized starch was negatively correlated with water content for gelatinization. Based on this, we propose that this ratio of areas provides a quantitative measure for assessing the short-range order in non-crystalline starch. This study provides an alternative and simpler method to an X-ray diffraction protocol proposed previously.
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