Optimization of extraction with salicylic acid, rheological behavior and antiproliferative activity of pectin from Citrus sinensis peels

Abstract

A Box-Behnken design was used to optimize extraction temperature, extraction time and concentration of the salicylic acid to obtain a maximum polysaccharide yield from Citrus sinensis peels. The optimal settings were: extraction time 3 h, extraction temperature 80 °C and concentration of the salicylic acid 1.5%. Under these conditions, the experimental yield and uronic acid content were 11.74% and 66.9% respectively. Preliminary characterization was performed via FT-IR, SEC/MALS/VD/DRI and GC-MS after hydrolysis. SEC analysis showed that the extracted polysaccharide had a weight average molar mass of 350 kDa and an intrinsic viscosity of 640 mL/g. The GC-MS results revealed that the extracted polysaccharide was composed of arabinose 56.7%, galactose 17.8%, xylose 13.8%, rhamnose 5.1%, mannose 2.5% and glucose 1.5% suggested a rhamnogalacturonan pectin type I with a degree of esterification of 50.9% (IRTF). The flow curve and the dynamic frequency sweep were obtained at 10, 20, 30 and 40 g/L in water and at 30 g/L in presence of CaCl₂ or NaCl at 1 mol/L. The solutions showed shear-thinning behavior fitted with Ostwald-De Waele model, except 10 g/L with a Newtonian behavior. The apparent viscosity and, the G' and G" moduli increase with PACO concentration in agreement with a slow-down of the dynamic chain. In the presence of CaCl₂ or NaCl the reduction of electrostatic repulsions between pectin chains decreases the rheological parameters. The effect is less sensitive with CaCl₂ due to intermolecular interactions. The antiproliferative activity of the extracted pectin on human Caco-2 and Hep-2 cells was very interesting with an IC₅₀ 1.4 and 1.8 μg/mL respectively.

Keywords: Antiproliferative activity; Box-Behnken design; Pectin; Rheological behavior; Salicylic acid.