The amino acid composition of P. citrinopileatus powder was performed using previously methods (Wang et al. 2021 (link)). 10 mg P. citrinopileatus powder was hydrolyzed by incubation in 6 N HCl (10 mL) for 24 h at 110 ℃ in tubes sealed under nitrogen. After digestion, the samples were diluted to 50 mL with ultrapure water and ltered through two layers of lter paper. Subsequently, the supernatant was ltered using a 0.22 μm membrane lter, and the ltrate (20 μL) was determined using a Hitachi L-8900 automated amino acid analyzer (Hitachi, Tokyo, Japan).
For the analysis of free amino acids, the method was applied as described previously (Song et al. 2019 (link)). In general, the sample (20 mg) was dissolved thoroughly by adding 3.5% sulfo salicylic acid, further diluted to 50 mL, and centrifuged for 20 min at 10,000 rpm. The supernatant was taken and ltered through a membrane lter (0.22 μm). Afterwards, the ltrate (20 μL) was detected using Amino Acids Analyzer (L-8900, Hitachi, Tokyo, Japan).
For the analysis of free amino acids, the method was applied as described previously (Song et al. 2019 (link)). In general, the sample (20 mg) was dissolved thoroughly by adding 3.5% sulfo salicylic acid, further diluted to 50 mL, and centrifuged for 20 min at 10,000 rpm. The supernatant was taken and ltered through a membrane lter (0.22 μm). Afterwards, the ltrate (20 μL) was detected using Amino Acids Analyzer (L-8900, Hitachi, Tokyo, Japan).