Amido resistente: metodologias de quantificação e resposta biológica em ratos

Abstract

The understanding of the role played by carbohydrates in human nutrition and health has made great strides in the last two decades, essentially regarding their differentiated physiological effects, ascribed especially to resistant starch. This fraction was recently discovered, and has been defined as the sum of starch and products of starch degradation not absorbed in the small intestine of healthy individuals. Due to its importance, several researches have been developed in order to quantify resistant starch in different foods, to better relate its levels with the beneficial effects. However, the existing data are very heterogeneous. Therefore, the present research was conducted aiming the optimization of in vitro resistant starch determination, through the modification of the protocol of an existing methodology, as well as relating increasing levels of this fraction in the diet with biological effects on rats. From the AOAC method 996.11 for the quantification of digestible and resistant starch, we evaluated the inclusion of protease in the hydrolysis process, the increase in sample amount and the use of phosphate buffer pH 6.8 instead of MOPS buffer pH 7.0, using as samples cornstarch, white rice, green banana and corn flakes. To evaluate the biological effects of increasing levels of resistant starch in the diet, male Wistar rats were fed diets supplemented with 0%, 3%, 9% and 18% of resistant starch. The animals were submitted to an adaptation period of 5 days. During the experimental period (15 days), data and samples were obtained to determine feed intake, body weight gain, apparent dry matter digestibility, apparent starch digestibility, wet and dry fecal production, fecal water content and pH, and fecal nitrogen excretion. The results obtained showed significant difference in the quantification of digestible and resistant starch in the same food, influenced by the presence or absence of proteolytic hydrolysis in the method. The utilization of increased amount of sample reduced the variation between replicates, and the use of phosphate buffer instead of MOPS buffer did not alter the obtained results, but reduced the cost of the analysis. So the recommendation resulting from this research is to use, associated with the increased amount of sample and phosphate buffer, the inclusion of protease in the digestion process. The addition of increasing levels of resistant starch in the diet of rats did not affect feed intake, but caused a reduction in body weight gain. Likewise, it was observed a reduction in apparent dry matter digestibility, apparent starch digestibility and fecal pH, allied to increased fecal production and water content, and fecal nitrogen excretion. Thus, we can conclude that resistant starch is capable of exerting biological effects, specially related to body weight reduction, which tend to be enhanced with increasing levels of this fraction, up to the tested 18%.