USA), consisting of a degassing device, quaternary pump, column thermocompartment, and a refractive-index detector (Waters 410). An Aminex HPX-87C column ion exchange (7.8mm d.i. �� 300mm, Bio-Rad Hercules, CA, USA) Bicalutamide side effects was used as stationary phase. HPLC grade water with a flow of 0.6mL/min was used as the mobile phase. The volume of the injected sample was 50��l (with the injector completely full). The column temperature was kept at 75��C. Environmental temperature was kept constant at 20��C. The samples were filtered through nylon-membrane filters (0.45��m) coupled to 5mL polypropylene syringes, both from Waters (Milford, CT), and analyzed immediately.The Quick Start Empower 5.0 was used for system control and data analysis.

References sugars (arabinose, fructose, galactose, glucose, lactose, maltose, mannose, ribose, sucrose, and xylose) as well as the chicory inulin (reference compound) were purchased from Sigma-Aldrich. The chromatographic separation time was 20min and the carbohydrates present in samples were identified by comparing the retention times with those of references sugars. Quantification of the carbohydrates was performed as a functions of the calibration curves, derived for sugars between 0.1�C3.2% w/v (r = 0.9996) and for fructans between 0.2�C8.0% w/v (r = 0.9999). The minimum detection level was 0.068mg/mL. Correlation coefficients (r) were calculated between refraction values in samples and standards for each carbohydrate to estimate the detector consistency in terms of concentration amplitude.

All the determinations were carried out in duplicate, and a mixture of standards sugars was injected daily on order to identify any calibration variations.2.4. Solid-Phase Microextraction (SPME) of Volatiles CompoundsFor the identification of volatile compounds present in samples, SPME technique was used (SPME device from Supelco Inc. Bellefonte, PA, USA). The fiber used was 60��m diameter and 1cm length, polyethylene glycol (PEG) coated. For sampling, the SPME fiber was inserted directly into a 20mL vial with a valve cap with silicone septa containing 100mg of fructan sample. After that, the sample was heated to 90��C during 5min before the extraction. Then, the fiber was maintained 10min at 90��C. The desorption of analytes was performed by heating the fiber in the injection port of the GC-MS equipment at 250��C for 5seconds (split mode, split ratio 10:1).2.5. Analysis of Volatile Compounds by GC/MSGC-MS analysis was performed using an Agilent HP series 6890N gas chromatograph (Waldbronn, Germany), coupled to a mass selective detector 5973. A Stabilwax capillary column (30m �� 0.25mm i.d., 0.25��m film thickness; GSK-3 Restek) was used for separation.