TL;DR: A refined and shortened method suitable for the rapid measurement of total sugar and starch contents in woody plant tissues is developed.
Abstract: Several extraction and measurement methods currently employed in the determination of total sugar and starch contents in plant tissues were investigated with the view to streamline the process of total sugar and starch determination. Depending on the type and source of tissue, total sugar and starch contents estimated from samples extracted with 80% hot ethanol were significantly greater than from samples extracted with a methanol:chloroform:water solution. The residual ethanol did not interfere with the sugar and starch determination, rendering the removal of ethanol from samples unnecessary. The use of phenol-sulfuric acid with a phenol concentration of 2% provided a relatively simple and reliable colorimetric method to quantify the total soluble-sugar concentration. Performing parallel sugar assays with and without phenol was more useful for accounting for the interfering effects of other substances present in plant tissue than using chloroform. For starch determination, an enzyme mixture of 1000 U alpha-amylase and 5 U amyloglucosidase digested starch in plant tissue samples more rapidly and completely than previously recommended enzyme doses. Dilute sulfuric acid (0.005 N) was less suitable for starch digestion than enzymatic hydrolysis because the acid also broke down structural carbohydrates, resulting in overestimates of starch content. After the enzymatic digestion of starch, the glucose hydrolyzate obtained was measured with a peroxidase-glucose oxidase/o-dianisidine reagent; absorbance being read at 525 nm after the addition of sulfuric acid. With the help of this series of studies, we developed a refined and shortened method suitable for the rapid measurement of total sugar and starch contents in woody plant tissues.
TL;DR: In this paper, a rapid enzymic method for starch analysis, especially in cereal products, was presented, which includes a 15 min gelatinization step in a boiling water bath in the presence of a thermostable α-amylase, a 30 min amyloglucosidase incubation of a subsample, and subsequent determination of glucose with a glucose oxidase/peroxidase reagent.
Abstract: A rapid enzymic method for starch analysis, especially in cereal products, is presented. One person can analyze 30 samples per day. The method includes a 15 min gelatinization step in a boiling water bath in the presence of a thermostable α-amylase, a 30 min amyloglucosidase incubation of a subsample, and subsequent determination of glucose with a glucose oxidase/peroxidase reagent. The method was evaluated by analysis of the starch content in various raw and industrially processed wheat samples. The method showed high precision (CV=0.6–1.0%) and accuracy. Some factors which might affect the enzymic availability of starch and thus its recovery in the analysis are evaluated and discussed.
TL;DR: The anthrone method was applied for starch determination and the influence by cellulose on the determination of starch, as well as, the influence of perchloric acid on the anthrone reaction was investigated.
TL;DR: The acetate buffer-only method was performed in sealable vessels with dilution by weight; it gave greater starch values in the analysis of feed/food substrates than did the other methods and is a viable candidate for a collaborative study.
Abstract: Starch is a nutritionally important carbohydrate in feeds that is increasingly measured and used for formulation of animal diets. Discontinued production of the enzyme Rhozyme-S required for AOAC Method 920.40 invalidated this method for starch in animal feeds. The objective of this study was to compare methods for the determination of starch as potential candidates as a replacement method and for an AOAC collaborative study. Many starch methods are available, but they vary in accuracy, replicability, and ease of use. After assays were evaluated that differed in gelatinization method, number of reagents, and sample handling, and after assays with known methodological defects were excluded, 3 enzymatic-colorimetric assays were selected for comparison. The assays all used 2-stage, heat-stable, a-amylase and amyloglucosidase hydrolyses, but they differed in the gelatinization solution (heating in water, 3-(N-morpholino) propanesulfonic acid buffer, or acetate buffer). The measured values included both starch and maltooligosaccharides. The acetate buffer-only method was performed in sealable vessels with dilution by weight; it gave greater starch values (2-6 percentage units of sample dry matter) in the analysis of feed/food substrates than did the other methods. This method is a viable candidate for a collaborative study.
TL;DR: The earliest direct evidence for human processing of grass seeds, including barley and possibly wheat, in the form of starch grains recovered from a ground stone artefact from the Upper Palaeolithic site of Ohalo II in Israel is reported.
Abstract: Barley (Hordeum vulgare L.) and wheat (Triticum monococcum L. and Triticum turgidum L.) were among the principal 'founder crops' of southwest Asian agriculture. Two issues that were central to the cultural transition from foraging to food production are poorly understood. They are the dates at which human groups began to routinely exploit wild varieties of wheat and barley, and when foragers first utilized technologies to pound and grind the hard, fibrous seeds of these and other plants to turn them into easily digestible foodstuffs. Here we report the earliest direct evidence for human processing of grass seeds, including barley and possibly wheat, in the form of starch grains recovered from a ground stone artefact from the Upper Palaeolithic site of Ohalo II in Israel. Associated evidence for an oven-like hearth was also found at this site, suggesting that dough made from grain flour was baked. Our data indicate that routine processing of a selected group of wild cereals, combined with effective methods of cooking ground seeds, were practiced at least 12,000 years before their domestication in southwest Asia.