Materials and Methods2.1. Obtention of the Agave Fructan PowderThe Agave salmiana plants used were from seven to nine years old, with approximately one year of castration, and collected at Ejido de Zaragoza de Sol��s, a municipality of Villa de Guadalupe, S Trichostatin A clinical trial L P. in the spring of 2010. The Agave juice was extracted using mill and expeller equipment from the local maguey processing plant located on the same Ejido land. This juice was filtered several times to eliminate the fibers and obtain samples with high-fructan content and convenient for spray drying. The first and second filtrations were carried out by means of a stainless steel press filter (Shriv, 405 Type), using bleached cellulose filter paper with a pore diameter of 22��m and 4��m and mixed with 1% of diatomaceous material (Celite) as a filtering aid.

The third filtration was performed under vacuum conditions, passing the juice through a Whatman no. 42 filter paper with a pore diameter of 2.5��m. To inactivate the saponins present in the juice, these were treated with heat at 80��C for 30min in a water bath with continuous agitation [38]. One batch of homogenized juice was obtained which was immediately dried with a B��chi mini spray dryer (Model B-290, Flawil, Switzerland). The atomizer pressure, the feed rate and the inlet air temperature were kept at 1.5 bar, 6.0mL/min and 170��C, respectively. Carrier agent was not used. The dried samples were subjected to quantitative and qualitative determination of sugars and used to measure the average-degree polymerization (DP) of agave fructan as described later.

These powders were also used as substrate during the hydrolysis experiments.2.2. DP Characterization of Agave FructanThe DP of the Agave fructan was determinated by the technique of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS), which has been reported as the best choice Cilengitide to establish the DP distribution of these types of carbohydrates. Experiments were performed in positive ion mode using an AutoFlex I mass spectrometer (Bruker Daltonics). The instrument was operated at an accelerating voltage of 20kV in linear mode and 19kV in reflectron mode. The pressure was 1.5 �� 10?7mbar. The sample was ionized with a nitrogen laser (�� = 337nm). The sample was dissolved in water, and the matrix was 2,5-Dihydroxybenzoic acid (10mg/mL), prepared in 1:1 methanol:water. Samples (0.5��L) and matrix solution were spotted and air dried on a stainless-steel plate. Spectra were acquired in linear mode (200�C300 laser shots) in the m/z range from 800 to 10,000 and reflectron mode (150�C200 laser shots) in the m/z range from 300 to 2,000. A pepmix calibration kit (Bruker Daltonics) and apomyoglobin were used for calibration.2.3.