Asymmetrical-flow field-flow fractionation combined with multiangle light scattering and refractive index detection has been revealed to be a powerful tool for starch characterization. It is based on size separation according to the hydrodynamic diameter of the starch components. Starch from a wide range of different botanical sources were studied, including normal starch and high-amylose and high-amylopectin starch. The starch was dissolved by heat treatment at elevated pressure in a laboratory autoclave. This gave clear solutions with no granular residues. Amylose retrogradation was prevented by using freshly dissolved samples. Programmed cross flow starting at 1.0 mL min(-1) and decreasing exponentially with a half-life of 4 min was utilised. The starches showed two size populations representing mainly amylose and mainly amylopectin with an overlapping region where amylose and amylopectin were possibly co-eluted. Most of the first population had molar masses below 10(6) g mol(-1), and most of the second size population had molar masses above 10(7) g mol(-1). Large differences were found in the relative amounts of the two populations, the molar mass, and hydrodynamic diameters, depending on the plant source and its varieties.
The starch accumulation-degradation process as well as the structure of leaf starch are not completely understood. To study this, starch was isolated from potato leaves collected in the early morning and late afternoon in July and August, representing different starch accumulation rates. The starch content of potato leaves varied between 2.9 and 12.9% (dry matter basis) over the night and day in the middle of July and between 0.6 and 1.5% in August. Scanning electron microscopy analyses of the four isolated starch samples showed that the granules had either an oval or a round shape and did not exceed 5 microm in size. Starch was extracted by successive washing steps with dimethyl sulfoxide and precipitated with ethanol. An elution profile on Sepharose CL-6B of debranched starch showed the presence of a material with a chain length distribution between that generally found for amylose and amylopectin. Amylopectin unit chains of low molecular size were present in a higher amount in the afternoon than in the morning samples. What remains at the end of the night is depleted in specific chain lengths, mainly between DP 15 and 24 and above DP 35, relative to the end of the day.
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