New Bioactive Molecules with Potential Antioxidant Activity from Various Extracts of Wild Edible Gelam Mushroom (Boletus spp.)

Boletus spp. is a wild edible Gelam mushroom, which is only available at coastal area of Bachok, Kelantan. Locals have used this mushroom as a food source and as a traditional medicine. In this study, antioxidants were extracted using Mili-Q water and methanol. Extraction of 1.0 g of Boletus spp. mushroom pileus and stipe by water produced 0.22 ± 0.09 g and 0.3 ± 0.07 g of total aqueous soluble extracts, respectively. Methanol extraction of 1.0 g Boletus spp. mushroom pileus and stipe produced 0.31 ± 0.08 mg and 0.37 ± 0.05 mg of total methanol soluble extracts, respectively. DPPH assay has been used to determine the value of antioxidant efficiency of each extracts. DPPH assay revealed that the EC50 value of pileus aqueous soluble extract and both pileus and stipe methanol soluble extracts were 1.2 mg/ml. The EC50 value of stipe aqueous soluble extracts was 1.4 mg/ml. The EC50 value of ascorbic acid, an antioxidant control was 0.5 mg/ml. Liquid chromatography- DPPH (LC-DPPH) assay combined with Quadrupole Time-of-flight Mass Spectrometry (QTOF-MS) analysis of 3:2 ratio methanol and water extract has detected 43 antioxidant compounds that were involved in the reduction of DPPH. To identify these compounds, SIMCA 3.0 software that consists of PCA, OPLS-DA and s-plot analyses had been used. PCA analysis had shown the antioxidant activity from sample on DPPH as 73.6%. This antioxidant activity confirmed by OPLS-DA analysis had shown the variability between sample and control as 99.6% with 99.3% prediction. S-plot selection had shown ten antioxidant compounds significantly involved in the DPPH assay. Four of these compounds were identified by the METLIN and NIST databases. The antioxidant metabolites were 2,4,6-trimethylacetophenone imine, glutamyl tryptophan, azatadine and lithocholic acid glycine conjugate. In conclusion, this study revealed that Boletus spp. mushroom is rich with natural antioxidants, which are potentially useful for multiple nutritional and health applications.