Establishment of Acid Hydrolysis by Box–Behnken Methodology as Pretreatment to Obtain Reducing Sugars from Tiger Nut Byproducts
María del Carmen Razola-Díaz | Vito Verardo | Beatriz Martín-García | Elixabet Díaz-de-Cerio | Belén García-Villanova | Eduardo Jesús Guerra-Hernández
Carbohydrate-rich byproducts may be used as a growth substrate for microalgae. The tiger nut, compared to other tubers, has higher carbohydrate content, which mostly remain in the byproduct after the production of tiger nut milk (“Horchata”). The aim of this study is the optimization of a hydrolysis pre-treatment in tiger nut byproduct to obtain reducing sugars, objective framed in the European SUSPUFA (Sustainable production of health-promoting n-3 LC-PUFA using agro-food industry by-products through microalgae) Project. Two experimental 4-factor Box–Behnken designs (acid concentration (N), temperature (°C), time (min) and ratio acid/sample) were carried out with 27 experiments to optimize chemical hydrolysis conditions with H<sub>2</sub>SO<sub>4</sub> and H<sub>3</sub>PO<sub>4</sub> in duplicate. The sugars were determined by the Schoorl method and the extracts obtained by the optimal conditions were analyzed by high-performance liquid chromatography (HPLC). Protected designation of origin (PDO) and non-protected designation of origin (n-PDO) tiger nut byproducts were characterized by physicochemical methods. The validity of the experimental designs was confirmed by ANOVA. The optimum sugar values were obtained with H<sub>2</sub>SO<sub>4</sub> 2 N at 85 °C, 60 min and ratio 60 and H<sub>3</sub>PO<sub>4</sub> 21 N at 90 °C, 60 min and ratio 60. Glucose and xylose were the sugars detected in the hydrolyzed samples. In conclusion, almost 50% of the total carbohydrates present in the tiger nut byproduct were hydrolyzed to reducing sugars.
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