Determinação de carboidratos estruturais em forrageiras tropicais por cromatografia líquida de alta eficiência (HPLC-UV)

Abstract

The nutritional quality of forage can be identified through structural analysis of carbohydrates present in the species. The structural carbohydrates in plants are distributed between the cellulose, composed of glucose, hemicellulose that has in its structure arabinose and xylose, and pectin which is composed of arabinose and others compounds. These fractions of the cell wall is that wounds the intestines of ruminant animals and are absorbed in its fullness. It is extreme importance the knowledge of this concentration to improve the performance of cattle on a pasture. This paper proposes a method for extraction with ultrasound and the heating plate, HPLC separation and quantification of glucose, arabinose and xylose in tropical forages. The method of analysis was developed in a high performance liquid chromatography with UV detection and reverse-phase C-18. The composition and pH of the mobile phase, wavelength in the detection parameters were studied in the optimization of the method. The best condition was achieved with isocratic elution using as mobile phase acetonitrile / aqueous H3PO4 pH 2.15 (1:99). In order to reduce the time and cost of the treatment and extraction of samples, this process was studied using the heating plate, which showed a more rapid, reproducible and efficient compared than extraction using the autoclave. The parameters of precolumn derivatization using p-aminobenzoic acid were studied. The best condition seen with the concentration of acid of 0.12 mol.L-1, a reaction time of 10 minutes at a temperature of 80 ° C. Tests were performed to evaluate the method, using external standardization, including evaluation of linear response, detection and quantification limits, repeatability, selectivity and recovery. The methodology implemented was applied to analysis of 68 samples (3 species of forage), leaf and stem: Pennisetum purpureum Schum cv. anão e Pennisetum purpureum Schum cv. pioneiro, Brachiaria brizantha, Brachiaria decumbens, Panicum maximum cv. Tanzania and Panicum maximum cv. Masai. The concentration ranges found expressed in mg.g-1 dry weight were: 31.98 to 188.38 for glucose, 6.28 to 30.95 for arabinose and 42.66 to 176.04 for xylose.