Optimization of Extraction Conditions to Improve Phenolic Content and In Vitro Antioxidant Activity in Craft Brewers’ Spent Grain Using Response Surface Methodology (RSM)

Abstract

Extraction temperature, extraction time and liquid (water) to solid ratio were optimized in order to extract antioxidant phenolic compounds from brewers’ spent grain (BSG). The extracts were analysed for their total phenol content (TPC) and antioxidant activity was measured using three different methods: 2,2-diphenyl-2-picrylhydrazyl (DPPH) free radical, 2,2′-azino-bis(3-ethylbenothiazoline-6-sulphonic acid) (ABTS), and reducing power (RP) assays. All the parameters except extraction time promoted different efficiencies for the extraction of antioxidant phenolic compounds. TPC extraction was higher at lower temperatures and lower liquid/solid ratios up to a certain point. In this sense, a decrease in TPC with increasing liquid/solid ratios took place until a 16:1 ratio and a plateau was reached beyond that ratio. The highest DPPH activity was reported for 30–35 °C and 60–90 min extraction and 60–90 min extraction with a 25 mL/g ratio. ABTS values increased as the liquid to solid ratio decreased, being positively correlated with TPC (R = 0.788; p < 0.01). The highest RP was achieved at 30–33 °C extraction temperature and 10–14 mg/mL v/w ratio and at 116–120 min extraction and 16–17 mg/mL ratio. Gallic acid accounted for the majority of the phenolic compounds found, followed by hydroxyphenylacetic acid, epicatechin, and protocatechuic acid. Sinapic, 4-hydroxy benzoic, and syringic acids were also found in lower quantities. Coumaric, vanillic, ferulic, and caffeic acids were present in very small amounts. All the extracts contained phenolics and showed in vitro antioxidant activity, but the extracts obtained by using 30 °C, 121.9 min, and 10 mL/g liquid/solid ratio exhibited the highest content in TPC and antioxidant potential. The aqueous extraction of a potentially bioactive extract from BSG was demonstrated to be an efficient and simple method to recover these value-added compounds.